Preserving Quality of Button Mushrooms (Agaricus bisporus) Using Nano-chitosan-Aloe Vera Coating with Tomato Seed Protein Hydrolyzate

doi:10.21203/rs.3.rs-4967669/v1

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Research Article

Preserving Quality of Button Mushrooms (Agaricus bisporus) Using Nano-chitosan-Aloe Vera Coating with Tomato Seed Protein Hydrolyzate

https://doi.org/10.21203/rs.3.rs-4967669/v1

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Button mushroom (Agaricus bisporus) is a widely consumed edible mushroom, but its quality deteriorates rapidly after harvest. Therefore, the use of edible coatings with natural preservative compounds is essential to delay and reduce microbial growth and maintain mushroom quality. This study examined the effects of a nano-chitosan (NC)-aloe vera (AV) edible coating combined with tomato seed protein hydrolyzate (TPH) as a natural preservative on the chemical, microbial, and organoleptic properties of button mushrooms. TPH was initially prepared using the enzyme Alcalase. Five edible films containing NC, NC-AV, and varying concentrations of TPH (0%, 0.5%, 1%, 1.5%) were then produced. The shelf lives of coated mushrooms were evaluated during 16 days of refrigerated storage (4 ± 1°C). Results showed that the TPH had high levels of protein (90.16%), hydrophobic amino acids (31.78%), and aromatic amino acids (11.74%). The produced films exhibited high antioxidant and antimicrobial activities, with improved results observed at increased concentrations of TPH. Compared to uncoated mushrooms, the nanocomposite coatings significantly reduced physicochemical changes, quality degradation, and microbial spoilage. Increased concentrations of TPH further enhanced browning inhibition, free radical scavenging, and microbial spoilage reduction (p < 0.05). Sensory evaluation indicated that the sample containing 1.5% TPH had the highest overall acceptance. The use of a NC-AV composite coating containing TPH can be an effective method for extending the shelf life of white button mushrooms.

button mushroom

nanocomposite coating

plant protein

preservation

shelf life

Edible mushrooms are important dietary protein sources and have been consumed by humans for centuries due to their sensory and nutritional properties (Pleșoianu and Nour, 2022). The button mushroom (Agaricus bisporus) is the most widely eaten edible mushroom globally, accounting for approximately 40% of total worldwide mushroom production, owing to its delectable flavor and nutritional value (Mohebbi et al., 2011; Gao et al., 2014). Button mushrooms are an excellent source of certain amino acids, vitamins (such as B2, folate), and minerals (including iron, calcium, phosphorus, potassium, selenium, zinc, and copper). Additionally, mushrooms contain triterpenoids, glycoproteins, polyphenols, and flavonoids, which possess various properties such as natural antioxidant and antibiotic activities (Vaziri et al., 2019; Pleșoianu and Nour, 2022). Due to the lack of a sufficient protective cuticle layer on their caps, mushrooms are susceptible to shrinkage and decay. The cuticle shields them from physical damage, water evaporation, and microbial attack; consequently, the loss of color and moisture ultimately leads to browning, taste alteration, and substantial quality degradation (Castellanos-Reyes et al., 2021; Khadem et al., 2020).

Fresh mushrooms have a high moisture content of approximately 85–95%, which gradually declines during the postharvest period, resulting in continuous weight loss. Moreover, the high respiration rate and moisture content contribute to the rapid deterioration of button mushrooms, leading to microbial attacks and enzymatic browning. Consequently, mushrooms are susceptible to various forms of physical and microbial spoilage (Khadem et al., 2020; Samadpour, 2020). The limited shelf life of this product (about 1 to 3 days at 25°C and 4–7 days at 4°C) presents a significant obstacle to its distribution and marketing. To overcome these challenges, the development and implementation of specific preservation methods and packaging techniques are essential (Pleșoianu and Nour, 2022).

Among these methods, coating is a promising technique for preserving fruits and vegetables. Edible coatings can enhance the postharvest quality of mushrooms, maintain their phytochemical content and physicochemical properties for extended durations, control moisture loss and respiration rates, delay deterioration, and ultimately prolong the shelf life of mushrooms (Vaziri et al., 2019; Pleșoianu and Nour, 2022). Nowadays, various antimicrobial compounds are also employed to further improve the preservation of mushrooms. Chitosan, a nitrogen-containing heteropolymer with a positive charge, is derived from the deacetylation of the natural polymer chitin. Due to its antimicrobial, biodegradable, and non-toxic properties, chitosan is used alone or in combination with other compounds as a coating to preserve various fruits (Rejane et al., 2016; Shahi et al., 2022). Nano-chitosan, a natural substance with excellent physicochemical properties, is environmentally friendly and bioactive. The antibacterial and antioxidant activities of chitosan nanoparticles have also been reported (Salimiraad et al., 2022; Shakour et al., 2021). The use of chitosan nanoparticles in food packaging is highly promising due to their compatibility with food materials (Shakour et al., 2021).

Aloe vera gel is a polysaccharide-based coating extracted from the inner parts of the Aloe vera plant leaves. It acts as a protective layer, reducing water loss and gas exchange in fruit skins, and decreases ethylene production in unripe fruits (Farajpour and Sheykhlouei, 2021; Amiri et al., 2022). Various studies have investigated the impact of Aloe vera gel in edible coatings to extend the shelf life of fresh fruits and vegetables, including mushrooms, bell peppers, kiwifruit, and strawberries (Farajpour and Sheykhlouei, 2021; Amiri et al., 2022; Mohammadi and Saidi, 2021; Mohebbi et al., 2011).

In recent years, hydrolyzed protein containing bioactive peptides has been utilized in various studies through enzymatic hydrolysis from different plant sources such as clover, wheat germ, sesame meal, canola, and watermelon seeds (Mirsadeghi Darabi, 2022; Ghelich et al., 2022; Ghanbarnia et al., 2022; Mirzapour et al., 2022; Mighan et al., 2024). Hydrolyzed proteins are produced through enzymatic, chemical, or fermentation methods. Among these, enzymatic hydrolysis of protein sources is favored due to more favorable conditions, better production outcomes, and fewer by-products (Nemati et al., 2024). Protein hydrolysis involves breaking down proteins into smaller peptides and free amino acids. The enzymes used for protein hydrolysis can originate from plants, animals, or microbes. Compared to plant and animal-derived enzymes, microbial enzymes offer more advantages, such as a broader range of proteolytic properties and greater stability under various pH and temperature conditions (Shahosseini et al., 2022). Among these enzymes, Alcalase has gained significant attention for producing hydrolyzed protein due to its activity in alkaline pH, higher degree of hydrolysis, and shorter peptide chain length (Nemati et al., 2012; Nemati et al., 2019).

In Iran, approximately 8,100 tons of wet tomato pomace is generated annually by processing plants, often discarded without considering their potential applications (Shariat et al., 2019). Tomato seeds contain 33.22–9.2% protein and 20.29–5.6% oil, making them an excellent source for edible oils and plant proteins (Sogi, 2001; Baker et al., 2022). Tomato seed protein includes amino acids such as arginine, lysine, histidine, phenylalanine, tryptophan, leucine, isoleucine, methionine, valine, cysteine, alanine, proline, glutamine, serine, asparagine, and threonine (Kumar et al., 2022; Persia et al., 2003). Therefore, considering the suitable protein content and high nutritional value, tomato waste can be used to produce hydrolyzed protein with desirable biological properties (Baker et al., 2022; Kumar et al., 2022).

The aim of this research is to investigate the properties of a nanocomposite film containing nano-chitosan and aloe vera along with hydrolyzed tomato seed protein and to evaluate the application of this edible nanocoating in improving the quality and extending the postharvest shelf life of button mushrooms. This innovative packaging method aims to maintain the highest quality and prolong the shelf life of button mushrooms. Previous studies in this area have been limited, and this investigation can provide greater insight into the potential of this nanocomposite coating to preserve the quality and extend the shelf life of button mushrooms.

Preparation of Raw Materials

Button mushrooms (Agaricus bisporus) were obtained from an agro-industrial company in Iran and transported to the laboratory at Ayatollah Amoli Islamic Azad University within 2 hours in plastic coolers at 1 ± 4°C. The mushrooms were then stored in a dark environment at 1 ± 4°C. Tomato pomace, a byproduct of tomato processing industries, was sourced from Ata Tomato Paste Factory in Iran. Fresh aloe vera leaves were purchased from a greenhouse in Amol, Iran, for gel extraction. Nano-chitosan was supplied by Nano Novin Polymer Company, Iran. Folin-Ciocalteu reagent and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals were obtained from Sigma-Aldrich, USA. Other chemicals used were of analytical grade, procured from Merck, Germany, and Sigma-Aldrich, USA.

Preparation of Tomato Seed Flour

The sedimentation method was used to separate seeds from tomato pomace. The pomace was immersed in large plastic containers filled with water, allowing the lighter peel to float while the heavier seeds settled. After separation, the seeds were sun-dried, ground using a household mill, and sieved through an 80-mesh screen. The resulting flour was defatted using hexane at a 1:10 ratio in four stages. The defatted flour was left to air-dry for one day and then stored in polyethylene bags in a refrigerator (Yemisi and Kayode, 2007).

Production of Tomato Seed Protein Concentrate

To extract protein from tomato seeds, the obtained flour was mixed with deionized water at a 1:10 ratio, and the pH was adjusted to 11.5 using 1N sodium hydroxide. The mixture was stirred for 30 min, then centrifuged at 2600 g for 20 min. The pH of the supernatant was adjusted to the isoelectric point of tomato seed proteins to precipitate the proteins, followed by centrifugation at 2600 g for 25 min. The protein concentrate was then freeze-dried and stored at -20°C until further analysis (Shariat et al., 2019).

Enzymatic Hydrolysis of Protein Concentrate

For the enzymatic hydrolysis, the protein concentrate was dissolved at a 5% (w/v) concentration in phosphate buffer at the optimal pH (8.5) for the Alcalase enzyme in 100 mL Erlenmeyer flasks. The mixture was stirred continuously for 30 min at room temperature (25°C) to ensure complete hydration. After reaching the desired incubation temperature, the enzyme was added to the solutions. The hydrolysis reaction was carried out for specific time intervals (30, 60, 90, and 120 min). To deactivate the enzyme, the flasks were placed in a 90°C water bath for 10 min, and then cooled to room temperature using an ice bath. The hydrolysate was centrifuged at 8000 g for 20 min, and the supernatant was freeze-dried and stored at -20°C until analysis (Mirzapour et al., 2022).

Degree of Hydrolysis

The degree of hydrolysis was calculated based on the α-amino acid content in the protein sample (Ovissipour et al., 2013).

Amino Acid Composition

To determine the amino acid composition of the hydrolyzed protein powder, the protein sample was completely hydrolyzed using 6 M hydrochloric acid at 110°C for 24 hours. The amino acids were then derivatized using phenyl isothiocyanate (PITC). The total amino acid content was measured using an HPLC system (Smart line, Germany) equipped with a C18 column and a fluorescence detector (RF-530) (Shahosseini et al., 2022).

Preparation of Nano-Chitosan-Aloe Vera Gel Film

The harvested Aloe vera leaves were washed with sterile distilled water. The tips and edges of the leaves were trimmed, and the middle part of the leaf was longitudinally sliced using a hand-held knife to separate the gel from the leaf skin. The gel was then extracted, blended for 5 min, and homogenized. The resulting mixture was passed through a cloth sieve to obtain pure gel, which was collected and diluted with sterile distilled water to achieve a 7.5% weight/volume concentration (Amiri et al., 2022). To prepare the composite gel, 1% chitosan nanoparticles, Aloe vera gel, and various concentrations of hydrolyzed protein (0.5%, 1%, and 1.5%) were added. These components were dissolved in 200 mL of distilled water at 70°C using a mixer for 45 min until fully homogenized (Shakour et al., 2021; Amiri et al., 2022). The casting method was employed to produce the films by pouring 50 mL of each prepared solution into a 12 cm diameter Petri dish and allowing them to dry at room temperature for 48 hours. Subsequently, the films were removed and stored in stomacher bags.

Antioxidant Activity of the Film

The antioxidant activity of the film was measured by its ability to scavenge DPPH free radicals (Siripatrawan and Harte, 2010). A 25 mg sample of the film was gently mixed with 3 mL of distilled water. Then, 8.2 mL of this solution was added to test tubes containing 0.2 mL of 1 mM DPPH solution in methanol and left at room temperature for 30 min. The absorbance of the samples and control was measured at 517 nm using a spectrophotometer. The decrease in absorbance compared to the control indicated the film's ability to scavenge DPPH radicals. The percentage of DPPH radical scavenging activity was calculated using Eq. 1:

Equation 1:

DPPH Radical Scavenging Activity Percentage = 1 - (Absorption rate of the sample / Absorption rate of the control) × 100

Measurement of Antibacterial Activity of the Film

Bacterial cultures of Staphylococcus aureus (PTCC 1189) and Escherichia coli (PTCC 1399) were obtained from the microbial collection of the University of Tehran. Using a sterile loop, a sample of each bacterium was taken from sterile ampoules and added to 10 mL of BHI Broth medium. The inoculated medium was incubated at 37°C for 24 hours. After incubation, the bacteria were cultured on nutrient agar plates using a sterile loop. The cultured plates were incubated again at 37°C for 24 hours. Three to five isolated colonies were homogenized and transferred using a sterile swab to tubes containing 5 mL of physiological serum. The turbidity (optical density) of the bacterial suspensions was measured at a wavelength of 625 nm using a spectrophotometer. The suspensions were diluted to a concentration equivalent to 0.5 McFarland, approximately 10⁸ CFU/mL. Using a sterile swab, the bacterial suspensions were uniformly spread over the surface of the culture media. Circular discs of the film, 6 mm in diameter, were cut using a circular knife. The film discs were placed at appropriate intervals on the bacteria-infused plates, which were then incubated at 37°C for 24 hours. After incubation, the diameter of the inhibition zones around the film discs was measured in mm and recorded. This method, known as the agar diffusion method, was employed to evaluate the antimicrobial activity of the edible films (Emiroğlu et al., 2010).

Preparation of Treatments

The mushrooms intended for coating were divided into six groups: one control group without coating and five groups with different coatings. Each group was coded accordingly. Except for the control sample, the mushrooms were immersed in the coating solutions for 5 min. After immersion, they were removed from the solution and placed on a mesh basket at room temperature for 10 min to allow excess coating material to drip off. The mushrooms were individually weighed and transferred into lidless disposable packaging containers. Each container was then placed inside a polyethylene zip-lock bag measuring 18×20 cm and finally stored in a refrigerator at 4 ± 1°C. The control sample was immersed in distilled water for 5 min (Vaziri et al., 2019).

Weight Loss Percentage

The weight of the mushrooms was measured after coating and before being transferred to the refrigerator, as well as on days 4, 8, 12, and 16 of storage using a scale with an accuracy of two decimal places. The weight loss percentage on different days was calculated using the following formula (Eq. 2) (Mohebbi et al., 2011).

Equation 2:

Weight Loss Percentage = Wo-W1/ Wo × 100

Where:

Wo = is the initial weight of the mushrooms on the first day after coating.

W1 = is the weight of the mushrooms on days 4, 8, 12, and 16.

This formula allows for the determination of weight loss over the storage period by comparing the initial weight to the subsequent weights measured at specified intervals.

Total Soluble Solids (TSS) Measurement

The mushroom extract was used to measure the total soluble solids (TSS). The TSS was determined using a refractometer, which was calibrated with distilled water prior to the measurements. The percentage of total soluble solids (expressed in degrees Brix) in the mushroom extract was obtained at a temperature of 25°C (Nasiri et al., 2019).

Color Index and Browning Index

The color of mushroom caps was measured on days 0, 4, 8, 12, and 16 using a portable digital colorimeter calibrated with a white standard. Randomly selected samples from each treatment were analyzed in triplicate. The color changes were evaluated by analyzing the L (lightness/darkness), a (red/green), and b (yellow/blue) values, where L ranges from 0 to 100, and a and b values range from − 120 to 120 (Pleșoianu and Nour, 2022). The browning index (BI) was calculated using Eq. 3 (Cavusoglu et al., 2021).

Equation 3:

BI = 100 (X- 0.31 )/ 0.17

X=(a + 1.75 L)/ (5.645 L + a-3.012 b)

Texture Firmness

The firmness of the mushroom texture was measured using a hand-held texture analyzer model (OSK 1618) with a 2 mm probe. Measurements were taken from two opposite sides of the mushroom cap (Mohebbi et al., 2011).

Total Phenol Content

The total phenol content was determined using the Folin-Ciocalteu reagent. A mixture of 0.1 mL of the extract, 5 mL of water, and 0.5 mL of 50% Folin-Ciocalteu reagent was prepared. Then, 1.5 mL of 2% sodium bicarbonate (NaHCO3) was added, and the volume was adjusted to 10 mL with distilled water. The reaction mixture was incubated at room temperature in the dark for 30 min. The absorbance was measured at 765 nm using a spectrophotometer. Gallic acid was used to construct a calibration curve by measuring the absorbance of different concentrations of gallic acid standard solutions at 765 nm (Pleșoianu and Nour, 2022).

Antioxidant Capacity

The antioxidant capacity was measured using the percentage inhibition of DPPH free radicals and expressed as the percentage of DPPH inhibition. First, a 0.1 mM solution of DPPH was prepared by dissolving 3.95 mg of DPPH in 100 mL of pure methanol. Then, 100 microliters of the samples were added to 900 µL of the DPPH solution, making the final volume 1 mL. The absorbance was read at 517 nm using a spectrophotometer after 20 min. The DPPH solution was used as the reference for comparison. The greater the antioxidant power, the greater the reduction in color. The radical scavenging activity (RSA) was calculated based on the percentage of DPPH radicals scavenged using Eq. 4:

Equation 4:

RSA% = 100 (Ac - As) / Ac

Where:

RSA is the radical scavenging activity (antioxidant capacity).

As is the absorbance of the sample containing the extract.

Ac is the absorbance of the control.

This formula allows for the determination of the antioxidant capacity of the samples by comparing the absorbance of the sample with the absorbance of the control.

Microbial tests

10 g of each treatment was removed under aseptic conditions and mixed well with 90 mL of physiological serum. In this way, the 10th dilution was obtained and then the required dilutions (10², 10³, etc.) were prepared from this dilution. Then 1 mL of each dilution was removed and poured into each plate. The culture medium that was prepared in advance and sterilized in the autoclave was removed and allowed to reach a temperature of 45°C. Then, it was poured onto the sample to fill approximately 1.3 of the plate space (about 15–20 mL). This process was repeated for 8 cycles. Finally, the culture medium was allowed to cool down and solidify. Then, for the total count of bacteria, the plates were placed upside down in the incubator at 37°C for 48 hours and for mold and yeast at 25°C for 3 days (Eissa, 2008). After 2 days, plates containing 60 to 300 colonies were selected and bacteria were counted. To calculate the number of bacteria, the number of colonies counted in the photo was multiplied and the results were obtained as log CFU/g (ISIRI, 2015). Mold and yeast counting was done according to Iranian national standard number 108991 (ISIRI, 2013). The culture medium used for total microbial count was Kant agar plate and Dichloran Rose Bengal Chloramphenicol Agar culture medium for mold and yeast.

Sensory Evaluation

Sensory evaluation for the coated samples was conducted using a 5-point hedonic scale to assess color, texture, and overall acceptability. Ten untrained panelists were employed for this evaluation. In this test, a score of 1 represented the lowest level of acceptance, and a score of 5 represented the highest level. Each panelist was provided with a form containing a table for scoring, along with a sufficient amount of the coded samples presented in standard mushroom packaging containers. The panelists evaluated the samples randomly to ensure unbiased results.

Statistical analysis

The obtained data were analyzed using SPSS software version 22 (SPSS Inc. Chicago, USA). All the studied parameters were compared between the control and treatment groups using One-way ANOVA. The data were processed in the form of mean and standard deviation. Also, the data were analyzed at the 5% probability level.

Degree of Hydrolysis

Degree of hydrolysis serves as a parameter monitoring the extent of protein hydrolysis. It is primarily used as an indicator for comparing various hydrolyzed proteins. Additionally, degree of hydrolysis is a crucial factor in studying the properties of hydrolyzed proteins as it reflects the extent of peptide bond cleavage and should be controlled (Dorvaj et al., 2013). The results regarding degree of hydrolysis (Table. 1) in this study demonstrate that as the hydrolysis process duration increases (P < 0.05), the degree of hydrolysis, indicating the extent of peptide bond cleavage, also increases. This occurs while the intensity and rate of hydrolysis, which refer to the degree of separation of soluble proteins from insoluble ones, decrease (Ghanbarnia et al., 2022; Liaset et al., 2002). No significant difference was observed between the 90 and 120-min durations (P > 0.05). With prolonged hydrolysis time, substrate depletion occurs, which affects the values of the hydrolysis degree (Golpaigani et al., 2023). Similar results were reported by Golpaigani et al. (2023) regarding the hydrolysis degree of hydrolyzed protein from rainbow trout eggs. They also stated that the hydrolysis degree increases with prolonged storage time, but after a certain period, the hydrolysis degree values become relatively constant.

Analysis of Initial Protein Content in Different Treatments:

The initial protein content of tomato seeds in this study was 19.54 ± 0.97%, whereas various studies have reported average initial protein contents of 17.71 ± 5.40% (ranging from 10.50–25.03%) for tomatoes (Ahmed et al., 2020; Ramos-Bueno et al., 2018; Elbadrawy and Sello, 2016; Ali et al., 2021; Oboulbiga et al., 2017). The isolated protein content of tomato seeds was 36.41 ± 0.91%. Furthermore, the protein content of hydrolyzed proteins ranged from 46.95–90.66% (Table. 1). These results align with the findings of Shariat alavi et al. (2019) concerning isolated and hydrolyzed protein contents, who also reported that the protein content after hydrolysis was higher than that of isolated tomato seed proteins. Additionally, a significant difference in protein content among the hydrolyzed samples, influenced by varying hydrolysis times, was observed. The protein content increased with extended hydrolysis time (P < 0.05), with no significant difference between the 90 and 120-min intervals (P > 0.05). With increased hydrolysis time, substrate reduction occurs, likely affecting the protein content.

Amino Acid Composition

Amino acids, often recognized as the building blocks of proteins, are compounds that play vital roles including maintaining cellular structure, repairing and rebuilding muscles and bones, and repairing damaged tissues (Elango et al., 2017). In the present study, the levels of 17 amino acids were identified (Table. 2). The highest amounts of amino acids were related to the non-essential amino acid glutamic acid at 16.99%, and the lowest amounts were related to the non-essential amino acid cysteine. Regarding essential amino acids, the highest amounts were related to leucine at 6.99%, and the lowest amounts were related to the essential amino acid methionine. These results are consistent with those of Ali et al. (2021) regarding tomato amino acids, where they also reported the highest amounts for the non-essential amino acid glutamic acid and the essential amino acid leucine. Additionally, similar results were reported by Elbadrawy and Sello (2016), who also stated that tomato skin contains 14.56% glutamic acid (the highest non-essential amino acid) and 5.07% leucine (the highest essential amino acid). The functionality of hydrolyzed protein is influenced by its amino acid composition. Hydrophobic amino acids (HAA) such as phenylalanine, proline, methionine, alanine, leucine, isoleucine, tyrosine, and valine, as well as aromatic amino acids (AAA) including phenylalanine, histidine, tryptophan, and tyrosine, contribute to the functional and biological properties of hydrolyzed proteins. These properties include antioxidant, anti-cancer, and antimicrobial activities. The present study found that the total amounts of HAA and AAA were 31.78% and 11.74% respectively, indicating their potentially significant bioactive effects on the host due to their high concentrations. The levels of valine, isoleucine, leucine, phenylalanine, lysine, and tyrosine in the hydrolyzed tomato seed protein were found to be higher than the recommended values for animal proteins by the FAO/WHO (1991), suggesting the high nutritional quality of this protein. Investigation of Antioxidant Properties of Films

The results (Fig. 1a) indicate that the antioxidant activity of nanochitosan films was 26.35%. The antioxidant property of chitosan has also been reported by other researchers (Muñoz-Tebar et al., 2023; Morachis et al., 2017). Furthermore, the addition of aloe vera gel was found to increase the DPPH free radical scavenging activity. This antioxidant effect of aloe vera gel has been associated with the presence of glutathione peroxidase, superoxide dismutase enzyme, and phenolic compounds (Langmead et al., 2004; Yoshida et al., 2021). Based on the findings, the antioxidant activity of the hydrolyzed protein films increased, and an increase in protein concentration had a positive impact on DPPH free radical inhibition (P < 0.05). The antioxidant properties of the hydrolyzed proteins have been attributed to their ability to scavenge free radicals, act as metal chelators, oxygen scavengers, or hydrogen donors, and prevent the penetration of lipid oxidation initiators by forming a protective layer around oil droplets (Yoshida et al., 2021). Other researchers have also reported that hydrolyzed plant proteins and peptides exhibit antioxidant properties under laboratory conditions (Mirsadeghi Darabi, 2022; Ghelich et al., 2022; Ghanbarnia et al., 2022; Mirzapour et al., 2022; Mighan et al., 2024). Pirveisi et al. (2023) also reported that nanocellulose films containing hydrolyzed pine nut protein have the ability to inhibit DPPH free radicals, and this ability increases with increasing protein concentration.

Investigation of Antimicrobial Properties of Films

According to the results, nanochitosan films exhibited antimicrobial activity against both bacteria (Fig. 1. b and c). There are two theories about the mechanism of chitosan antimicrobial activity. First, chitosan, with its poly cationic property, can chelate metals and essential elements and remove them from the reach of bacteria. Second, by forming bonds with anions in the bacterial cell wall, chitosan disrupts their cell walls (Ghafari et al., 2024; Barzegar et al., 2008). Adding gel to the film increased the antimicrobial properties of the films (P < 0.05) Anthraquinones and dihydroxy anthraquinones are effective substances in aloe vera gel that have antimicrobial properties (Emamifar, 2015). In another study, the antimicrobial activity of aloe vera gel against a number of positive and negative bacteria has been demonstrated by several different methods (Habeeb et al., 2007). Additionally, the addition of hydrolyzed protein further increased the antimicrobial activity of the films. The highest level of antimicrobial activity against both bacteria was observed in nanocomposite film + 1.5% hydrolyzed protein (P < 0.05). Antimicrobial peptides, by penetrating the membrane, can disrupt it, leading to imbalance in cellular contents, disrupting processes such as replication, transcription, and translation of DNA sequences by binding to specific intracellular targets (Tkaczewska et al., 2020; de Oliveira et al., 2019; Ghafari et al., 2024).

Weight Loss in Mushrooms during Storage

This factor indicates the trend of weight loss in mushrooms as a percentage. Water loss is the most important physiological process that affects the qualitative characteristics of mushrooms. In more severe amounts, this phenomenon leads to wrinkling. This factor directly affects the commercial value of the product and, if severe, reduces its value (Zhang et al., 2018). The aim of this research is to prevent severe weight loss in button mushrooms through their coating. Wrinkling, moisture loss, and intense respiration are among the main reasons for weight loss in button mushrooms (Gholami et al., 2017).

A comparison of the percentage of weight loss between coated samples and control samples during the storage period showed that the control sample had a higher percentage of weight loss compared to the coated samples. According to the results (Fig. 2a), coating has led to reduced weight loss compared to the control sample (P < 0.05). The role of chitosan in reducing the weight of mushrooms can be attributed to its poly-cationic property. This coating, by breaking into polymeric fragments and re-forming polymeric chains, forms a gel-like surface coating film (Nasiri et al., 2018). This coating leads to the formation of a hydrophilic layer around the fruit, which acts as a barrier to gas exchanges, thus reducing respiration and preventing surface moisture loss (Liu et al., 2019). Additionally, the use of composite nano films had a more effective impact on weight loss amounts. The hygroscopic property of aloe vera gel is capable of forming a barrier against water diffusion between the fruit and the environment. Furthermore, these coatings protect the fruit skin from mechanical damage by repairing small injuries, thus preserving moisture and delaying drying (Mohammadi and Saidi, 2021). The lowest weight loss amounts were observed in mushrooms coated with composite nano films along with hydrolyzed protein (P < 0.05). During the hydrolysis process, hydrophobic groups are converted to hydrophilic groups through the production of carbonyl and amine groups (Halim et al., 2016). The use of coatings containing hydrophilic compounds can affect the formation of pores and surface breakage in mushrooms and can cause a reduction in mushroom weight loss by affecting the vapor permeability values (da Costa et al., 2020). da Costa et al. (2020) reported that the use of edible coatings along with hydrolyzed protein from black pacu fish resulted in less weight loss in cherry tomatoes during the storage period compared to the control.

Total Solid Soluble of Mushrooms during Storage

When comparing the total solid soluble at the beginning and end of the storage period, all mushroom samples exhibited an increase in Brix, although the magnitude of this increase varied slightly among different treatments. The reason for this gradual increase is the gradual reduction in the moisture content of the mushrooms over time during the storage period, resulting in a lower amount of solid soluble substances in the water and consequently a higher Brix concentration (Hosseini and Moradinezhad, 2018). The increase in total solid soluble (Fig. 2b) in mushrooms treated with nanocamposite film with hydrolyzed protein was slower than in other samples (P < 0.05), as the coating provides a semi-permeable film layer around the mushrooms, preventing water evaporation from the fruit. Additionally, it modifies the internal atmosphere by reducing oxygen and increasing carbon dioxide, thus altering the internal atmosphere (Pleșoianu and Nour, 2022; Zhu et al., 2019; Nasiri et al., 2019). It appears that preventing further increases in solid soluble content depends on maintaining moisture inside the coating. The control, from which moisture easily escapes, showed a higher increase in solid soluble content. Similar results were obtained by Gholami et al. (2020), demonstrating a slower increase in solid soluble content in mushrooms coated with chitosan and nanorose. The effectiveness of polysaccharide coatings on the solid soluble content of mushrooms during storage has also been reported by Pleșoianu and Nour (2022).

Texture Firmness of Mushrooms during Storage

Texture firmness is an important parameter related to the mechanical and structural properties of food materials. The firmness of a food material depends on cell size, wall resistance, and internal cell adhesion. In some fruits and vegetables, the firmness of their flesh is an indicator of the optimal harvest time (Gholami et al., 2020). Over time, the firmness (Fig. 2c) of all samples significantly decreased (P < 0.05). Guillaume et al. (2010) noted that during mushroom storage, cell swelling decreases, leading to soft and spongy tissue. The decrease in tissue firmness may be due to a decrease in the moisture content of the mushrooms, bacterial growth, enzymatic activity, and degradation of the cell walls of mushroom tissue. During ripening and senescence, enzymatic activity affects the tissue, leading to pectin breakdown and, consequently, softening of the tissue (Sanchis et al., 2015). According to the results, the greatest changes in firmness were observed in the control (P < 0.05). Coating provides a lower oxygen surface and a higher carbon dioxide surface, thus limiting enzyme activity and preserving tissue firmness during storage. Furthermore, the results showed that coating with a composite nano film along with hydrolyzed protein can prevent bacterial softening of the tissue, as tissue softening in samples coated with hydrolyzed protein was much lower compared to the control samples. The inhibitory effect of these coatings is due to the presence of hydrolyzed protein (Khademi et al., 2019) and the covering of the cuticle surface and the pores present on the mushroom surface due to the use of aloe vera gel, thus reducing respiration and the ripening process during storage (Mohammadi et al., 2021). Rokayya et al. (2021) reported that packaged mushrooms with a combination of chitosan and nanotitanium have higher firmness compared to control mushrooms during the storage period.

Phenolic Compound Levels in Mushrooms during Storage

Phenols are responsible for reducing oxidative damage and scavenging free radicals. Oxidative damage and browning of tissues are initial processes resulting from the combination of phenols with oxygen due to the activity of enzymes such as polyphenol oxidase (PPO) (Bach et al., 2019; Liu et al., 2013). Phenolic compounds are secondary plant metabolites responsible for aroma, flavor, and color in horticultural products, and they are degraded after harvest due to the activity of polyphenol oxidase and peroxidase enzymes (Jiang, 2013). The levels of phenolic compounds (Fig. 3a) in mushrooms showed a declining trend in all treatments (P < 0.05). The reduction in phenolic compounds at the end of the storage period is likely due to the breakdown of cellular structure during senescence throughout the storage period. However, this trend was faster in the control. This can be attributed to the defensive response of mushrooms to external stress, due to the presence of the coating, which preserves the overall phenolic content and consequently enhances its antioxidant properties (Pleșoianu and Nour, 2022). Similar results were reported by Kumar et al. (2023), who also found that mushrooms packaged with aloe vera gel had higher phenolic content compared to untreated mushrooms during storage.

Antioxidant Properties in Mushrooms during Storage

The results of the present study showed that with the increase in storage time from day zero to day 16 in the all treatments, the antioxidant capacity (Fig. 3b) showed a declining trend (P < 0.05). The lowest levels were observed in the control (P < 0.05). During the storage of fruits and vegetables, the reduction of phenolic compounds and ascorbic acid leads to a decrease in antioxidant levels. Coating with nano-chitosan helps preserve the antioxidant capacity; chitosan maintains antioxidant enzyme activity at higher levels. Multiple studies have demonstrated that chitosan, as a biological elicitor, may possess the potential to eliminate free radicals (Chen et al., 2021). The findings of the current study were in line with those reported by Khojah et al. (2021), who found that mushrooms coated with chitosan exhibited higher levels of phenols during the storage period compared to untreated mushrooms. Additionally, chitosan maintains antioxidant enzyme activity at higher levels, as indicated by various researches. Additionally, the consumption of aloe vera coating was effective due to its antioxidant properties (Hassanpour, 2015). Edible coatings improve antioxidant properties in products by reducing respiration intensity and ethylene production. Moreover, the best results were observed in treatments containing hydrolyzed protein, and an increase in concentration also had a positive effect on this trend, attributed to the antioxidant activity of hydrolyzed protein (Mirsadeghi Darabi et al., 2022; Ghelich et al., 2022; Ghanbarnia et al., 2022; Mirzapour et al., 2022; Mighan et al., 2024). Phenolic compounds are known as scavengers of free radicals and the main substrate for enzymatic browning reactions (Lu et al., 2016), and the levels of phenolic compounds in the current study were consistent with the levels of antioxidant activity. Pleșoianu and Nour (2022) also stated that there is a positive correlation between antioxidant activity and total phenolic content in mushrooms.

Color Index Values of Mushrooms during Storage

As indicated in the results, with increasing storage time, the brightness index (L*) decreased in all treatments (P < 0.05) (Fig. 4a), and the rate of this decrease was higher for the control compared to the coated ones (P < 0.05). One of the reasons for this decrease is the increase in respiratory activity and enzymatic activity, including tyrosinase reactions, leading to browning of button mushrooms during storage, significantly reducing the brightness component of mushroom treatments. In treatments containing coatings, this may be attributed to the protection of mushroom surfaces against the atmosphere and the reduced penetration of oxygen into the tissue, consequently reducing the respiratory activity of mushrooms (Rokayya et al., 2021; Vargas et al., 2008). If their L* value is less than 80, button mushrooms may not be acceptable for sellers, and those with an L* value less than 70 may not even be acceptable for consumers (Vaziri et al., 2019; Gao et al., 2014). Therefore, treatments containing hydrolyzed protein had levels acceptable to consumers. With time, the a* (Fig. 4b), increased for all treatments (P < 0.05). This increase may be due to the increased respiratory rate and stimulation of enzymatic activities, including browning reactions and other quality-reducing reactions (Rad et al., 2020). The b* of button mushroom treatments showed significant differences among each other, as also observed in Fig. 4c, with the untreated button mushroom treatment showing the highest changes in yellow index (P < 0.05). The increase in yellow index during storage is also due to oxidation resulting from microbial activity and lipid oxidation. Rad et al. (2020), Rokayya et al. (2021), and Samadpour (2020) also stated that during the storage period, the color indices a*, b*, and L* of button mushrooms increase, and they further noted that the use of edible coatings slows down the rate of these changes.

Enzymatic Browning Index Values of Mushrooms during Storage

Enzymatic browning is the most significant physiological abnormality that severely affects the quality and marketability of button mushrooms after harvest and during storage. It is initiated by a group of enzymes called polyphenol oxidases or polyphenolases (Lagnika et al., 2014). According to the results, the enzymatic browning index (Fig. 4d) increased in all treatments with increasing storage time, and the rate of this increase was higher for the control compared to the coated ones (P < 0.05). In the control, the absence of a protective layer allows more oxygen to penetrate into the mushrooms, resulting in higher levels of enzymatic browning. Additionally, the addition of hydrolyzed protein reduces the surface microbial load of mushrooms due to its antimicrobial properties (Ghafari et al., 2024), thereby reducing the formation of brown spots caused by bacterial growth. Furthermore, based on the results of antioxidant compounds, substrates, and compounds involved in enzymatic browning reactions, they compete with and inhibit the formation of this reaction. Mechanisms for preventing enzymatic browning include: 1) controlling the entry and exit of gases, especially oxygen, which is one of the main factors in enzymatic browning; 2) eliminating or binding free radicals through coatings; 3) controlling enzymes involved in enzymatic browning reactions and preventing their negative effects (Sedaghat et al., 2012).

Total Bacterial Counts, Mold and Yeast of Mushrooms during Storage

The presence of bacterial populations in fresh mushrooms is an important factor that affects the quality of this product, leading to the occurrence of brown spots and a speckled appearance in mushrooms after harvest. These damages depend on the initial microbial contamination (Samadpour, 2020). The data obtained from the enumeration of total bacteria as well as molds and yeasts indicate an increasing trend over time. With time, the total bacterial counts increased in all treatments (P < 0.05). However, this increase was more pronounced in the control samples, and on all days, the bacterial load in the control samples was higher than that in the coated samples. Significant differences were observed between the control samples and the coated samples, as well as among the coated samples themselves, on all days (P < 0.05). The lower microbial load (total bacterial counts (Fig. 5a), mold and yeast (Fig. 5b) in the coated samples may be attributed to the effect of the nanofilm coating on mushrooms, which prevents microbial contamination in mushroom samples. Additionally, chitosan and aloe vera gel also have antimicrobial properties (Algarni et al., 2022; Duan et al., 2019; Emamifar, 2015). Similar results regarding the use of chitosan nanoparticles in reducing the microbial load of mushrooms (Agaricus bisporus) were reported by Karimirad et al. (2019). Furthermore, the use of hydrolyzed protein also led to a reduction in the microbial load of mushrooms, with the lowest microbial load observed on day 16 of storage in treatments containing nanocomposite film + 1.5% hydrolyzed protein (P < 0.05). The antimicrobial property of hydrolyzed protein has also been reported by other researchers (Pirveisi et al., 2023; Ghanbarnia et al., 2022; Shahosseini et al., 2021). The use of edible coatings with natural preservatives may reduce the respiration rate and prevent enzymatic browning and microbial spoilage (Mohebbi et al., 2012). The results regarding the microbial load of mushrooms are consistent with the findings reported by Kumar et al. (2023), where the use of edible coatings based on aloe vera gel along with orange peel essence resulted in a reduction in microbial growth (total bacterial counts, mold and yeast) during the storage period. Similar results were reported by Gull et al. (2021) regarding the use of nano-chitosan coatings with pomegranate peel extract on the microbial load of apricots during storage. These researchers attributed this to the antimicrobial property of nano-chitosan-aloe vera gel along with natural preservatives. Additionally, similar results were reported by da Costa et al. (2020) regarding the use of edible coatings along with hydrolyzed black paku fish protein on the microbial load of cherry tomatoes during storage. They also stated that the addition of hydrolyzed protein had a satisfactory effect on reducing the fungal and yeast load

Sensory Evaluation of Mushrooms during Storage

The color and appearance of edible mushrooms are qualitative characteristics that influence consumer acceptance and have significant economic implications. Therefore, this study evaluated sensory attributes based on color and appearance (Fig. 6). With time, enzymatic activity increases, causing the color of the mushrooms to gradually darken and become more brownish. These changes were more pronounced in the control. Additionally, during mushroom storage, the reduction in moisture content leads to a decrease in cellular turgor pressure and tissue firmness. Overall, these factors affect the overall acceptance of mushrooms by consumers. The differences in color and texture observed in button mushrooms are consistent with the findings of Samadpour et al. (2020). Accordingly, during the initial days, there was minimal difference in texture between the coated and control mushrooms, but over time, these differences became apparent, and the firmness and freshness of the tissue were preserved by coating. The sensory evaluation results were consistent with the findings of other tests, with the best results observed in mushrooms coated with hydrolyzed protein. The difference in color among mushrooms can be attributed to the effect of coating and hydrolyzed protein on bacteria. Additionally, the composite nanofilm with hydrolyzed protein, due to its potential enzymatic activity-inhibiting compounds, can delay enzymatic browning of mushrooms and postpone the spoilage of this sensitive product (Mohebbi et al., 2012).

The results indicated that compared to the untreated sample, this coating led to a reduction in physicochemical and microbial changes in the mushrooms and also improved their sensory properties. Addition of hydrolyzed protein to the coating resulted in decreased browning, increased antioxidant activity, and reduced microbial growth of the mushrooms. According to sensory evaluation, the sample nano composite coated with 1.5% hydrolyzed protein received the highest overall acceptance score. Therefore, the use of this composite coating containing hydrolyzed protein is suggested as an effective solution for enhancing the shelf life and maintaining the quality of button mushrooms.

DATA AVAI LABILITY STATEMENT

Data available on request from the authors.

CONFLICT OF INTEREST

The authors declare that they do not have any conflict of interest.

Funding Declaration

No Funding

Ahmed, M. J., Iya, I. R., & Dogara, M. F. (2020). Proximate, mineral, and vitamin content of flesh, blanched and dried tomatoes (Lycopersicon esculentum). Asian Food Science Journal, 18, 11-18.
Ali, M. Y., Sina, A. A. I., Khandker, S. S., Neesa, L., Tanvir, E. M., Kabir, A., & Gan, S. H. (2021). Nutritional composition and bioactive compounds in tomatoes and their impact on human health and disease: A review. Foods, 10(1), 45.
Algarni, E. H. A., Elnaggar, I. A., Abd El-wahed, A. E. W. N., Taha, I. M., AL-Jumayi, H. A., Elhamamsy, S. M., ... & Fahmy, A. (2022). Effect of chitosan nanoparticles as edible coating on the storability and quality of apricot fruits. Polymers, 14(11), 2227.
Amiri, S., Rezazad Bari, L., & Ahmadi Partovi, G. (2022). The effect of edible coating based on Aloe vera gel containing copper oxide nanoparticles on freshly cut pieces of kiwi in refrigerator temperature. Journal of Packaging Science and Technology, 13(49), 19-34.
Baker, P. W., Preskett, D., & Krienke, D. (2022). Pre-processing waste tomatoes into separated streams with the intention of recovering protein: Towards an integrated fruit and vegetable biorefinery approach to waste minimization. Waste Biomass Valor, 13, 3463-3473.
Bach, F., Zielinski, A. A., Helm, C. V., Maciel, G. M., Pedro, A. C., Stafussa, A. P., ... & Haminiuk, C. W. (2019). Bio compounds of edible mushrooms: In vitro antioxidant and antimicrobial activities. LWT, 107, 214-220.
Barzegar, H., Carbasi, A., Jamalian, J., & Lari, M. A. (2008). The possibility of using chitosan as a natural preservative in mayonnaise. Journal of Agricultural Sciences and Natural Resources, 12, 43.
Castellanos-Reyes, K., Villalobos-Carvajal, R., & Beldarrain-Iznaga, T. (2021). Fresh mushroom preservation techniques. Foods, 10(9), 2126.
Cavusoglu, S., Uzun, Y., Yilmaz, N., Ercisli, S., Eren, E., Ekiert, H., ... & Szopa, A. (2021). Maintaining the quality and storage life of button mushrooms (Agaricus bisporus) with gum, agar, sodium alginate, egg white protein, and lecithin coating. Journal of Fungi, 7(8), 614.
Chen, C., Peng, X., Chen, J., Gan, Z., & Wan, C. (2021). Mitigating effects of chitosan coating on postharvest senescence and energy depletion of harvested pummelo fruit response to granulation stress. Food Chemistry, 348, 129113.
da Costa de Quadros, C., Lima, K. O., Bueno, C. H. L., dos Santos Fogaça, F. H., da Rocha, M., & Prentice, C. (2020). Effect of the edible coating with protein hydrolysate on cherry tomatoes shelf life. Journal of Food Processing and Preservation, 44(7), e14760.
de Oliveira Filho, J. G., Rodrigues, J. M., Valadares, A. C. F., de Almeida, M. A., & Dyszy, F. H. (2019). Active food packaging: Alginate films with cottonseed protein hydrolysates. Food Hydrocolloids, 92, 267-275.
Dorvaj, Z., Javadian, S. R., Oveissipour, M., & Nemati, M. (2013). Use of protein hydrolysates from Caspian Sea sprat (Clupeonella cultiventris) as a nitrogen source for bacteria growth media (Vibrio anguillarum, Bacillus licheniformis, Bacillus subtilis). Journal of Aquatic Animals & Fisheries, 4(15), 11-18.
Duan, C., Meng, X., Meng, J., Khan, M. I. H., Dai, L., Khan, A., ... & Ni, Y. (2019). Chitosan as a preservative for fruits and vegetables: A review on chemistry and antimicrobial properties. Journal of Bioresources and Bioproducts, 4(1), 11-21.
Eissa, H. A. (2008). Effect of chitosan coating on shelf-life and quality of fresh-cut mushroom. Polish Journal of Food and Nutrition Sciences, 58(2), 211-218.
Elbadrawy, E., & Sello, A. (2016). Evaluation of nutritional value and antioxidant activity of tomato peel extracts. Arabian Journal of Chemistry, 9, S1010-S1018.
Elango, R., & Laviano, A. (2017). Protein and amino acids: Key players in modulating health and disease. Current Opinion in Clinical Nutrition and Metabolic Care, 20(1), 69-70.
Emamifar, A. (2015). Evaluation of Aloe vera gel effect as an edible coating on microbial, physicochemical and sensorial characteristics of fresh strawberry during storage. New Food Technologies, 6, 15-29.
Emiroğlu, Z. K., Yemiş, G. P., Coşkun, B. K., & Candoğan, K. (2010). Antimicrobial activity of soy edible films incorporated with thyme and oregano essential oils on fresh ground beef patties. Meat Science, 86(2), 283-288.
Farajpour, P., & Sheykhlouei, H. (2021). Study on edible coating effect, based on Aloe vera gel and thymol on the postharvest quality and storage life of strawberry. Food Science and Technology, 18(112), 81-95.
Gao, M., Feng, L., & Jiang, T. (2014). Browning inhibition and quality preservation of button mushroom (Agaricus bisporus) by essential oils fumigation treatment. Food Chemistry, 149, 107-113.
Ghafari, E., Ariaii, P., Bagheri, R., Rezaei, M., & Soltanian, S. (2024). Investigating the effect of nanochitosan-Iranian tragacanth gum composite film along with Eryngium campestre essential oil on the shelf life of goat meat. Food Measurement, 18, 1543-1558.
Ghanbarinia, S. H., Ariaii, P., Safari, R., & Najafian, L. (2022). The effect of hydrolyzed sesame meal protein on the quality and shelf life of hamburgers during refrigerated storage. Animal Science Journal, 93(1), e13729.
Ghelich, S., Ariaii, P., & Ahmadi, M. (2022). Evaluation of Functional Properties of Wheat Germ Protein Hydrolysates and Its Effect on Physicochemical Properties of Frozen Yogurt. International Journal of Peptide Research and Therapeutics, 28, 69.
Gholami, R., Ahmadi, E., & Farris, S. (2017). Shelf life extension of white mushrooms (Agaricus bisporus) by low temperatures conditioning, modified atmosphere, and nanocomposite packaging material. Food Packaging and Shelf Life, 14, 88-95.
Guillaume, C., Schwab, I., Gastaldi, E., & Gontard, N. (2010). Biobased packaging for improving preservation of fresh common mushrooms (Agaricus bisporus L.). Innovative Food Science & Emerging Technologies, 11, 690-696.
Golpaigani, M. H., Ariaii, P., Ahmadi, M., Sharifi, A., & Rezaei, M. (2023). Preservation effect of protein hydrolysate of rainbow trout roe with a composite coating on the quality of fresh meat during storage at 4 ± 1 °C. Food Measurement, 17, 2416-2428.
Gull, A., Bhat, N., Wani, S. M., Masoodi, F. A., Amin, T., & Ganai, S. A. (2021). Shelf life extension of apricot fruit by application of nanochitosan emulsion coatings containing pomegranate peel extract. Food Chemistry, 349, 129149.
Halim, N. R. A., Yusof, H. M., & Sarbon, N. M. (2016). Functional and bioactive properties of fish protein hydolysates and peptides: A comprehensive review. Trends in Food Science & Technology, 51, 24-33.
Habeeb, F., Shakir, E., Bradbury, F., Cameron, P., Taravati, M. R., Drummond, A. J., ... & Ferro, V. A. (2007). Screening methods used to determine the anti-microbial properties of Aloe vera inner gel. Methods, 42(4), 315-320.
Hassanpour, H. (2015). Effect of Aloe vera gel coating on antioxidant capacity, antioxidant enzyme activities, and decay in raspberry fruit. LWT-Food Science and Technology, 60(1), 495-501.
Hasanzati Rostami, A., Motamedzadegan, A., Hosseini, S. E., Rezaei, M., & Kamali, A. (2017). Evaluation of plasticizing and antioxidant properties of silver carp protein hydrolysates in fish gelatin film. Journal of Aquatic Food Product Technology, 26(4), 457-467.
Hwang, J. Y., Shyu, Y. S., Wang, Y. T., & Hsu, C. K. (2010). Antioxidative properties of protein hydrolysate from defatted padnut kernels treated with esperase. Food Science and Technology, 43, 285-290.
Hosseini, A., & Moradinezhad, F. (2018). Effect of short-term high CO2 treatment on quality and shelf life of button mushroom (Agaricus bisporus) at refrigerated storage. Postharvest Biology and Technology, 1, 37-48.
Institute of Standards and Industrial Research of Iran. (2015). Microbiology of the food chain-Horizontal method for the enumeration of microorganism (ISIRI no 5272-2).
Institute of Standards and Industrial Research of Iran. (2013). Microbiology of food and animal feeding stuffs-Horizontal method for the enumeration of yeasts and molds (ISIRI no 10899-1.
Jiang, T., Feng, L., & Zheng, X. (2012). Effect of chitosan coating enriched with thyme oil on postharvest quality and shelf life of shiitake mushroom (Lentinus edodes). Journal of Agricultural and Food Chemistry, 60, 188-196.
Karimirad, R., Behnamian, M., & Dezhsetan, S. (2019). Application of chitosan nanoparticles containing Cuminum cyminum oil as a delivery system for shelf life extension of Agaricus bisporus. LWT, 106, 218-228.
Khadem, S., Khalili, M., Shahabi-Ghahfarrokhi, I., Dobakhti, F., & Almasi, H. (2020). Methods of Extending Shelf-Life and Use of New Packaging Systems in Button Mushroom. , 11(43), 50-63.
Khademi, M., & Nazarian-Firouzabadi, F. (2019). Expression and antimicrobial activity analysis of dermaseptin B1 recombinant peptides in tobacco transgenic plants. pgr, 6(1), 139-150.
Kumar, M., Chandran, D., Tomar, M., Bhuyan, D. J., Grasso, S., Sá, A. G. A., ... & Saleena, L. A. K. (2022). Valorization Potential of Tomato (Solanum lycopersicum L.) Seed: Nutraceutical Quality, Food Properties, Safety Aspects, and Application as a Health-Promoting Ingredient in Foods. Horticulturae, 8, 265.
Lagnika, C., Zhang, M., Atindana, J., & Bashari, M. (2014). Effect of ultrasound and chemical treatments on white mushroom (agaricus bisporus) prior to modified atmosphere packaging in extending shelf-life. Journal of food science and technology, 51(12), 3749-3757.
Saha, A., Kumar, N., & Rahul, K. (2023). Characterization of Aloe Vera Gel-Based Edible Coating with Orange Peel Essential Oil and Its Preservation Effects on Button Mushroom (Agaricus bisporus). Food and Bioprocess Technology.
Langmead, L., Makins, R. J., & Rampton, D. S. (2004). Anti-inflammatory effects of aloe vera gel in human colorectal mucosa in vitro. Alimentary Pharmacology & Therapeutics, 19(5), 521–527.
Liaset, B., Nortvedt, R., Lied, E., & Espe, M. (2002). Studies on the nitrogen recovery in enzymatic hydrolysis of Atlantic salmon (Salmo salar, L.) frames by Protamex™ protease. Process Biochemistry, 37(11), 1263–1269.
Liu, F., Chang, W., Chen, M., Xu, F., Ma, J., & Zhong, F. (2019). Tailoring physicochemical properties of chitosan films and their protective effects on meat by varying drying temperature. Carbohydrate Polymers, 212, 150–159.
Lu, Y., Zhang, J., Wang, X., Lin, Q., Liu, W., Xie, X., ... & Guan, W. (2016). Effects of UV-C irradiation on the physiological and antioxidant responses of button mushrooms (Agaricus bisporus) during storage. International Journal of Food Science & Technology, 51(6), 1502–1508.
Mighan, N. M., Ariaii, P., Soltani, M. S., Mohsenin, S. S., Jafari, S. M., & Nazari, B. (2024). Investigating the possibility of increasing the microbial and oxidative stability of silver carp burgers using hydrolyzed protein of watermelon seeds. Food Science and Biotechnology, 33, 375–388.
Mirsadeghi Darabi, D., Ariaii, P., Safari, R., & Ahmadi, M. (2022). Effect of clover sprouts protein hydrolysates as an egg substitute on physicochemical and sensory properties of mayonnaise. Food Science & Nutrition, 10, 253–263.
Mirzapour, Z., Ariaii, P., Safari, R., Jafari, S. M., & Khodaverdian, B. (2022). Evaluation the Effect Hydrolyzed Canola Meal Protein with Composite Coating on Physicochemical and Sensory Properties of Chicken Nugget. International Journal of Peptide Research and Therapeutics, 28, 97.
Mohammadi, M., & Saidi, M. (2021). Effect of Aloe vera gel and Arabic gum coating on quality characteristics of green bell peppers (Capsicum annuum L.) during storage. Journal of Food Processing and Preservation, 12(2), 39–52.
Mohebbi, M., Ansarifar, E., Hasanpour, N., & Amiryousefi, M. R. (2012). Suitability of Aloe vera and gum tragacanth as edible coatings for extending the shelf life of button mushroom. Food and Bioprocess Technology, 5(8), 3193–3202.
Morachis-Valdez, A. G., Gómez-Oliván, L. M., García-Argueta, I., Hernández-Navarro, M. D., Díaz-Bandera, D., & Dublán-García, O. (2017). Effect of chitosan edible coating on the biochemical and physical characteristics of carp fillet (Cyprinus carpio) stored at −18°C. International Journal of Food Science, 2017, 2812483.
Muñoz-Tebar, N., Pérez-Álvarez, J. A., Fernández-López, J., & Viuda-Martos, M. (2023). Chitosan edible films and coatings with added bioactive compounds: Antibacterial and antioxidant properties and their application to food products: A review. Polymers, 15, 396.
Nasiri, M., Barzegar, M., Sahari, M. A., & Niakousari, M. (2018). Application of tragacanth gum impregnated with Satureja khuzistanica essential oil as a natural coating for enhancement of postharvest quality and shelf life of button mushroom (Agaricus bisporus). International Journal of Biological Macromolecules, 106, 218–226.
Nasiri, M., Barzegar, M., Sahari, M., & Niakousari, M. (2019). Efficiency of tragacanth gum coating enriched with two different essential oils for deceleration of enzymatic browning and senescence of button mushroom (Agaricus bisporus). Food Science & Nutrition, 7, 1520–1528.
Najafian-Jazi, M., Jafarpour, M., Kahvaei, M., Raeisi-Dehkordi, B., & Alamarvdasht, N. (2022). Physicochemical properties and shelf life of button mushroom under essential oils with chitosan coating treatment. Research on Crop Ecophysiology, 15(2), 97–109.
Nemati, M., Javadian, S. R., & Keshavarz, M. (2019). Production of protein hydrolysates from Caspian shad (Alosa caspia) by-products using Alcalase enzyme. Journal of Marine Biology, 11(43), 87–95.
Nemati, M., Javadian, S. R., Ovissipour, M., & Keshavarz, M. (2012). A study on the properties of alosa (Alosa caspia) by-products protein hydrolysates using commercial enzymes. World Applied Sciences Journal, 18(7), 950–956.
Oboulbiga, E. B., Parkouda, C., Sawadogo-Lingani, H., Compaoré, E. W. R., Sakira, A. K., & Traoré, A. S. (2017). Nutritional composition, physical characteristics and sanitary quality of the tomato variety Mongol F1 from Burkina Faso. Food and Nutrition Sciences, 8, 444–455.
Ovissipour, M., Rasco, B., Shiroodi, S. G., Modanlow, M., Gholami, S., & Nemati, M. (2013). Antioxidant activity of protein hydrolysates from whole anchovy sprat (Clupeonella engrauliformis) prepared using endogenous enzymes and commercial proteases. Journal of the Science of Food and Agriculture, 93(7), 1718-1726.
Persia, M. E., Parsons, C. M., Schang, M., & Azcona, J. (2003). Nutritional evaluation of dried tomato seeds. Poultry Science, 82(1), 141-146.
Pirveisi, N., Ariaii, P., Esmaeili, M., Sharifi, M., & Afshari, A. (2023). Investigating active packaging based on cellulose nanofibers oxidized by TEMPO method containing hydrolyzed protein obtained from pine tree fruit on the quality of pacific white shrimp (Litopenaeus vannamei) during the storage period. Food Measurement, 17(3), 3323-3337.
Pleșoianu, A. M., & Nour, V. (2022). Effect of some polysaccharide-based edible coatings on fresh white button mushroom (Agaricus bisporus) quality during cold storage. Agriculture, 12(10), 1491.
Rad, M., Ghafouri, H., & Gholami, Z. (2020). Effect of edible coating containing carboxymethyl cellulose and sodium metabisulfite on the shelf life of the button mushroom. Iranian Food Science and Technology Research Journal, 16(5), 581-605.
Ramos-Bueno, R. P., Romero-González, R., González-Fernández, M. J., & Guil-Guerrero, J. L. (2017). Phytochemical composition and in vitro anti-tumour activities of selected tomato varieties. Journal of the Science of Food and Agriculture, 97(2), 488-496.
Rejane, C. G., Sinara, T. B. M., & Odilio, B. G. A. (2016). Evaluation of the antimicrobial activity of chitosan and its quaternized derivative on E. coli and S. aureus growth. Revista Brasileira de Farmacognosia, 26, 122-127.
Rokayya, S., Khojah, E., Elhakem, A., Benajiba, N., Chavali, M., Vivek, K., ... & Helal, M. (2021). Investigating the nano-films effect on physical, mechanical properties, chemical changes, and microbial load contamination of white button mushrooms during storage. Coatings, 11(1), 44.
Salimiraad, S., Safaeian, S., Akhondzadeh Basti, A., Khanjari, A., & Mousavi Nadoushan, R. (2022). Characterization of novel probiotic nanocomposite films based on nano chitosan/nano cellulose/gelatin for the preservation of fresh chicken fillets. LWT - Food Science and Technology, 162, 113429.
Samadpour, R. (2020). The effect of edible coating with combined Thymus Vulgaris extract and glycerol monoestearate on oyster mushroom's shelf life. Future of Food: Journal on Food, Agriculture and Society, 8 (2).
Sanchis, E., Mateos, M., & Perez-Gao, M. B. (2015). Effect of maturity stage at processing and antioxidant treatments on the physicochemical, sensory and nutritional quality of fresh-cut 'Rojo Brillante' persimmon. Postharvest Biology and Technology, 105, 34-44.
Sedaghat, N., & Zahedi, Y. (2012). Application of edible coating and acidic washing for extending the storage life of mushrooms (Agaricus bisporus). Food Science and Technology International, 18(6), 523-530.
Shahi, T., Ghorbani, M., Jafari, S. M., Sadeghi Mahoonak, A., Maghsoudlou, Y., & Beigbabaei, A. (2022). Effect of chitosan nano-coating loaded with pomegranate peel extract on physicochemical and microbial characteristics of pomegranate arils during storage. Food Science and Technology, 19(126), 71-85.
Shahosseini, S. R., Javadian, S. R., & Safari, R. (2021). Evaluation of antibacterial and antioxidant activities of Liza abu viscera protein hydrolysate. Journal of Innovation in Food Science and Technology, 30(2), 123-146.
Shahosseini, S. R., Javadian, S. R., & Safari, R. (2022). Effects of molecular weights-assisted enzymatic hydrolysis on antioxidant and anticancer activities of Liza abu muscle protein hydrolysates. International Journal for Peptide Research & Therapeutics, 28, 72.
Shakour, N., Khoshkhoo, Z., Akhondzadeh Basti, A., Khanjari, A., & Mahasti Shotorbani, P. (2021). Investigating the properties of PLA-nanochitosan composite films containing Ziziphora Clinopodioides essential oil and their impacts on oxidative spoilage of Oncorhynchus mykiss fillets. Food Science & Nutrition, 00, 1-13.
Shariat alavi, M., Sadeghi Mahoonak, A., Ghorbani, M., Alami, M., & Mohammadzadeh, J. (2019). Determination of optimum conditions for production of hydrolyzed protein with antioxidant capability and decrease of nitric oxide from tomato wastes by Alcalas. Food Science and Technology, 15(84), 137-151.
Siripatrawan, U., & Harte, B. R. (2010). Physical properties and antioxidant activity of an active film from chitosan incorporated with green tea extract. Food Hydrocolloids, 24(8), 770-775.
Sogi, D. S. (2001). Functional properties and characterization of tomato waste seed proteins (Doctoral dissertation). Guru Nanak Dev University, Amritsar, India.
Sun, T., Xu, Z., Wu, C. T., Janes, M., Prinyawiwatkul, W., & No, H. K. (2007). Antioxidant activities of different colored sweet bell peppers (Capsicum annuum L.). Journal of Food Science, 72, 98-102.
Tkaczewska, J. (2020). Peptides and protein hydrolysates as food preservatives and bioactive components of edible films and coatings - A review. Trends in Food Science & Technology, 106, 298-311.
Vaziri, A., Safekordi, A. A., Shekarabi, A., & Shamlou, S. (2019). Extending the shelf life of edible button mushroom (Agaricus bisporus) by edible coatings on the basis of natural polymers. Food Science and Technology, 16(91), 243-256.
Yathisha, U. G., Bhat, I., Karunasagar, I., & Mamatha, B. S. (2018). Antihypertensive activity of fish protein hydrolysates and its peptides. Critical Reviews in Food Science and Nutrition, 59(15), 2363-2374.
Yoshida, C. M. P., Pacheco, M. S., de Moraes, M. A., Lopes, P. S., Severino, P., Souto, E. B., & da Silva, C. F. (2021). Effect of Chitosan and Aloe Vera Extract Concentrations on the Physicochemical Properties of Chitosan Biofilms. Polymers, 13, 1187.
Yemisi, A., & Kayode, O. A. (2007). Evaluation of the gelation characteristics of mucuna bean flour and protein isolate. Journal of Agricultural and Food Chemistry, 6(8), 2243-2262.
Zhang, K., Pu, Y. Y., & Sun, D. W. (2018). Recent advances in quality preservation of postharvest mushrooms (Agaricus bisporus): A review. Trends in Food Science & Technology, 78, 72-82.
Zhu, D., Guo, R., Li, W., Song, J., & Cheng, F. (2019). Improved postharvest preservation effects of Pholiota nameko mushroom by sodium alginate–based edible composite coating. Food and Bioprocess Technology, 12, 587-598.

Table 1. Effect of Hydrolysis Duration on Protein Content and Degree of Hydrolysis of tomato seed protein hydrolyzate (TPH)

Hydrolysis time (min)	Protein content (%)	Degree of hydrolysis (%)
30	46.95±1.01^c	14.09±0.87^c
60	76.10±0.79^b	21.80±0.48^b
90	90.66±1.20^a	24.90±0.30^a
120	90.16±0.91^a	24.73±0.54^a

Averages with the different letters (in same columns) indicate that there is significant difference at the P< 0.05.

Table 2: Amino Acid Composition of tomato seed protein hydrolyzate (TPH)

Amino acid (g 100 g^-1)	TPH	FAO/ WHO, 1990
Histidine^a	2.48	3.4
Isoleucine ^a	3.44	3.5
Leucine^a	6.99
Lysine^a	3.97	2.8
Methionine^a	0.74	6.6
Phenyl alanine^a	5.97	6.3
Threonine^a	2.59	5.8
Arginine^a	6.92
Valine^a	4.15
Aspartic acid	8.95
Glycine	3.95
Proline	4.11
Serine	3.47
Alanine	3.09	1.1
Tyrosine	2.59	1.9
Cystein	0.37
Glutamic acid	16.99
Total amino acid	80.02
Hydrophobic amino acids	31.78
Aromatic amino acids	11.74

^aEssential amino acids

No competing interests reported.

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You are reading this latest preprint version

Preserving Quality of Button Mushrooms (Agaricus bisporus) Using Nano-chitosan-Aloe Vera Coating with Tomato Seed Protein Hydrolyzate

Status:

Version 1

Abstract

Figures

Introduction

Materials and Methods

Preparation of Raw Materials

Preparation of Tomato Seed Flour

Production of Tomato Seed Protein Concentrate

Enzymatic Hydrolysis of Protein Concentrate

Degree of Hydrolysis

Amino Acid Composition

Preparation of Nano-Chitosan-Aloe Vera Gel Film

Antioxidant Activity of the Film

Measurement of Antibacterial Activity of the Film

Preparation of Treatments

Weight Loss Percentage

Equation 2:

Total Soluble Solids (TSS) Measurement

Color Index and Browning Index

Equation 3:

BI = 100 (X- 0.31 )/ 0.17

Texture Firmness

Total Phenol Content

Antioxidant Capacity

Microbial tests

Sensory Evaluation

Statistical analysis

Results and Discussion

Degree of Hydrolysis

Analysis of Initial Protein Content in Different Treatments:

Amino Acid Composition

Investigation of Antimicrobial Properties of Films

Weight Loss in Mushrooms during Storage

Total Solid Soluble of Mushrooms during Storage

Texture Firmness of Mushrooms during Storage

Phenolic Compound Levels in Mushrooms during Storage

Antioxidant Properties in Mushrooms during Storage

Color Index Values of Mushrooms during Storage

Enzymatic Browning Index Values of Mushrooms during Storage

Total Bacterial Counts, Mold and Yeast of Mushrooms during Storage

Sensory Evaluation of Mushrooms during Storage

Conclusion

Declarations

References

Tables

Additional Declarations

Status:

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