Production and Characterization of Bacterial Cellulose from Food waste: A sustainable approach

doi:10.21203/rs.3.rs-5207708/v1

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Production and Characterization of Bacterial Cellulose from Food waste: A sustainable approach

https://doi.org/10.21203/rs.3.rs-5207708/v1

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Cellulose is a natural polymer found in plant cell walls and has been used in many industries for ages. The increasing demand for cellulose is estimated to be 100–125 Giga tons per year globally, contributing to deforestation and exacerbated global warming. However, bacterial cellulose has emerged as a promising alternative to plant based cellulose due to its unique properties including biodegradability, high crystallinity, high mechanical strength, porosity and great water absorption and retention capabilities. This study aims to optimize the production of bacterial cellulose economically using readily available resources like food waste addressing environmental concerns and production costs. Utilized tea as nitrogen source and Bagasse and glucose as carbon sources were used for the production of Bacterial cellulose. SCOBY culture is used as an inoculum for production. Characterization was done by tests like FTIR, XRD, and FE-SEM. The results of bacterial cellulose formed from these substrates were then compared with the result of the standard HS media. The finding demonstrates that characterization of Bacterial cellulose layer produced from the HS media was much similar with the results of utilized tea substrate, and it has higher crystallinity. It was observed that the FTIR spectrum of BC layers formed by utilized tea media and bagasse were similar to the Standard HS media produced BC, indicating that they contain the same chemical functional bonds. In FE-SEM analysis of sugarcane bagasse bacterial cellulose are densely packed, discontinuously arranged, irregular shaped nanofibers of different diameters were observed.

Bacterial Cellulose

Utilized tea extract

Bagasse

Biomaterial Optimization

Food Waste valorization

Nanocellulose

1.1 Cellulose

Cellulose is a linear homopolysaccharide is composed of β-D-glucopyranose units that are linked by β-1, 4 glycosidic bonds (Raj et al., 2021). It is a polysaccharide consisting of 15,000 D- glucose monomers. Plants and bacteria are the primary sources of cellulose extraction (Rajeshkumar et al., 2021). Extraction of cellulose from plants remains challenging. To extract cellulose efficiently, the removal of other components such as lignin and hemicelluloses must be properly managed. Delignification and hemicellulose removal are required during the extraction of cellulose from any plant or agro waste. Bleaching is done to obtain bleached cellulose material (Gallegos et al., 2016).

1.2 Bacterial cellulose

In addition to plant cellulose, certain gram-negative bacteria possess the ability to produce cellulose (Iqbal et al., 2015). Bacterial cellulose (BC) is considered pure cellulose and is gaining significance in various industries such as paper, textiles, and leather (Iqbal et al., 2015). Various bacteria, including Gluconacetobacter, Aerobacter, Rhizobium, Sarcina, Azotobacter, Agrobacterium, Pseudomonas, and Alcaligenes, can synthesize BC. Among these, Gluconacetobacter xylinus, a Gram-negative, rod-shaped bacterium, has been the earliest and most extensively studied microorganism responsible for BC production, initially discovered by Brown in 1886 (Donini et al., 2010).

BC is composed of ribbon-like cellulose nanofibers (Fig. 1.1), that weave together to form a 3D reticulated network (Ruka et al., 2010). The structure of BC has both crystalline and non- crystalline areas, similar to that of plant cellulose, having up to 90% crystallinity (Wang et al., 2019; Sijabat et al., 2020). Additionally, it exhibits elevated levels of polymerization, mechanical strength, water retention capacity, chemical stability, and adaptability to biological environments (Watanabe et al., 1998). Bacterial cellulose demonstrates environmental friendliness and sustainability. It is biodegradable and has no deleterious or allergic side effects (Yim et al., 2017).

1.3 SCOBY symbiotic culture of bacteria and yeast

The symbiotic culture of bacteria and yeast, known as SCOBY, is composed of a diverse community of bacteria and yeast strains (Teoh et al., 2004). The most crucial bacteria in SCOBY are Gram-negative species such as Acetobacter and Gluconacetobacter, which play a vital role in the fermentation process responsible for producing kombucha and bacterial cellulose (Esa et al., 2014).

In the SCOBY, acetic acid bacteria and yeast work in symbiosis to enhance cellulose production (Dima et al., 2017). Yeasts in SCOBY possess invertase enzymes, which break down sucrose into reducing sugars, releasing monosaccharides into the medium. These monosaccharides serve as a carbon source for the bacteria. Additionally, the bacteria utilize glucose as a carbon source to produce bacterial cellulose. Simultaneously, the yeast produces ethanol, which promotes the bacterial cellulose synthase process, ultimately leading to the production of a bacterial cellulose film (Hamed et al., 2023). This intricate interplay between bacteria and yeast is essential for successful fermentation and cellulose production in SCOBY cultures.

1.4 Kombucha

Kombucha, a popular fermented tea drink enjoyed globally, originated in China and later spread to Russia and beyond. Initially, microbiologists categorized Kombucha as a type of lichen because it involves a symbiotic relationship between fungi and acetic acid bacteria (AAB). However, unlike typical lichens, which are symbiotic associations of algae and fungi and rely on light for photosynthesis, Kombucha thrives in darkness (Brown et al., 1976). The Kombucha culture initially resembles a white, rubbery pancake and undergoes changes in appearance throughout the fermentation process. Initially, it has a creamy white color, but when brewed with black tea, the culture darkens due to the presence of tannins in the tea.

During Kombucha fermentation, microorganisms utilize sucrose as the primary carbon source, while nitrogen is derived from the tea extract. In the presence of oxygen, the SCOBY produces organic acids, carbon dioxide, and a dense cellulosic biofilm composed of cellulose. This biofilm thickens as fermentation progresses, often forming multiple layers resembling pancakes (McNamara et al., 2015).

1.5 Parameters affecting the growth of Bacterial Cellulose

1.5.1 pH

SCOBY typically thrives in an acidic environment with a pH range of about 2.5–4.5. This low pH is primarily due to the production of acetic acid and lactic acid during the fermentation process. The acidity of the environment helps create a suitable and protective habitat for the bacteria and yeast within the SCOBY community (Lee et al., 2014).

1.5.2 Temperature

Temperature is another critical parameter that influences the adaptation patterns of microorganisms for survival. A temperature range of 25 to 30°C is optimal to produce BC by certain Komagataeibacter species, while the optimum temperature for BC production by Acetobacter xylinum is 28°C (Wang et al., 2018). Temperature regulation is crucial for achieving efficient BC production in these bacterial cultures.

During the fermentation process, a biofilm often thickens and forms a matrix known as a pellicle. This pellicle's primary role is to protect the SCOBY (Symbiotic Culture of Bacteria and Yeast) from potential competitors or contaminants. Typically, after the Kombucha manufacturing process is completed, this biofilm is considered a byproduct and is commonly discarded (Borzani et al., 1995).

However, due to its composition primarily consisting of cellulose, this discarded pellicle has proven to be a valuable resource for various innovative applications. Despite being typically discarded as a byproduct of Kombucha production, the cellulose-rich biofilm pellicle offers a valuable resource for various industries seeking innovative and sustainable materials for a range of applications (Vandamme et al., 1998).

1.6 Raw material for Bacterial Cellulose production

Waste materials pose significant economic and environmental threats to the world. Food waste is a major concern, as food manufacturers and processors generate millions of tons annually, much of which could be prevented or recycled. Approximately one-third of all food produced worldwide is lost or wasted, leading to direct product loss and the discarding of valuable byproducts rich in sugars and other food ingredients by various industries. This not only impacts economies but also creates conditions favorable for microbial growth, posing environmental and health risks. In addition to reducing waste, it’s crucial to explore opportunities for recycling and converting waste into valuable products (Millati et al., 2019).

Globally, there are about 2 billion tons of agricultural waste that accumulates every year (Pakistan Sugar Mills Association 2021). Pakistan is ranked fifth among all sugar-producing countries and in 2022 (Knoema 2023), it produced 88 million tons of sugar cane (Afghan et al., 2023). Sugarcane bagasse is highly valued by sugar producers as a primary feedstock source for bioenergy and biofuel production due to its rich carbon content (Jozala et al., 2015). Additionally, it has the potential to be used in the production of bacterial cellulose.

For over a century, there has been considerable interest in developing cost-effective methods for producing BC. The main challenges have been the expense of commonly used culture media and the slow production processes, hindering large-scale industrial production. Various approaches, including novel synthetic methods and the use of different bioreactors, have been explored to improve BC production. Despite some success, achieving economically viable BC production remains elusive. Efforts have focused on using inexpensive and waste sources such fruit juices, bagasse, molasses, and industrial wastes. While these alternatives can reduce production costs, challenges like purification expenses, environmental concerns, and subpar physiological qualities persist. Finding affordable materials with easy processing methods for high-quality BC production remains a crucial endeavor (Costa et al., 2017).

This research aimed to synthesize bacterial cellulose using waste materials such as carbon and nitrogen sources. Furthermore, bacterial cellulose production was qualitatively and quantitatively estimated. This work eventually will be helpful to produce a sustainable and cost-effective product and will additionally reduce food waste.

2.1 Study Location

The bacterial cellulose production and optimization was done at the Biochemistry lab, Kinnaird College for Women, Lahore. Characterization tests such as Fourier Transform Infrared (FTIR), X-Ray Diffraction (XRD), and Scanning Electron Microscopy (SEM) were done commercially at Comsats Research Centre, Lahore.

2.2 Materials Required

Kombucha SCOBY was purchased from Nature's store (https://thenaturesstore.com/). The sugarcane bagasse was obtained from commercial vendor. Glucose and utilized tea were obtained from the Biochemistry laboratory and Cafeteria of Kinnaird College for Women, Pakistan.

2.3 Production of Bacterial Cellulose (BC)

Bacterial cellulose production was done by using different carbon and nitrogen sources in medium size plastic containers. The following combinations were used as the medium for bacterial cellulose production by SCOBY.

HS medium as standard.
Bagasse and utilized tea.
Glucose and utilized tea.

2.3.1 Production of BC from Hestrin-Schramm (HS) media

The media was prepared by adding carbon and nitrogen sources in water. 800µl SCOBY culture was added in 200 ml HS media and then, pH was checked to be 4.5. The media components were gently stirred and mixed with bacterial culture. Then, the containers were covered with gauze to allow oxygen passage through the media. These containers were placed in an incubator at a temperature of 30°C for 7–10 days. The bacterial cellulose growth was examined and further processed by washing with distilled water and air dried.

2.3.2 Production of BC from Bagasse and utilized tea

2.3.2.1 Pre-Treatment of sugarcane bagasse

The pulpy fibrous material of sugarcane bagasse after the removal of pith and rind was subjected to a thorough washing process with distilled water. After drying, it was weighed to about 100g and boiled in a beaker using a Bunsen burner for 90 minutes. Initially, 1000ml distilled water was added to 100g of bagasse and its volume reduced to the desired concentration. The resulting mixture was then strained with a gauze cloth to obtain the bagasse water. This process ensured that the sugarcane bagasse was properly treated for further use.

2.3.2.2 Bagasse water with varying concentration

200ml bagasse water and 4g of utilized tea extract were used as carbon and nitrogen sources for 200ml setup. 800ul SCOBY was used as the inoculum for bacterial cellulose synthesis.

100ml bagasse water and 2g of utilized tea extract were used as the carbon and nitrogen sources for 100ml setup, and distilled water was added to make the volume up to 100 ml. 800ul SCOBY was used as the inoculum for bacterial cellulose synthesis.

300ml bagasse water and 6g of utilized tea extract were used as the carbon and nitrogen sources for 300ml setup. 1200ul SCOBY was used as the inoculum for bacterial cellulose synthesis.

1500ml bagasse water and 30g of utilized tea extract were used as the carbon and nitrogen sources for 1500ml setup. 6000ul SCOBY was used as the inoculum for bacterial cellulose synthesis.

All setups were placed in an incubator at a temperature of 30°C under static conditions.

2.3.3 Production of Bacterial Cellulose from Glucose and Utilized tea

4g of utilized tea extract and 4g of glucose were added in the 200 ml water as nitrogen and carbon sources for 200ml reaction setup. 800µl SCOBY was added as an inoculum for bacterial cellulose synthesis. The entire fermentation took place under static conditions in an incubator at a temperature of 30°C. Following 7–14 days, bacterial cellulose growth was examined as per media conditions.

2.4 pH measurement

pH of the media was checked with the help of pH strips. pH was checked to allow adjustments in pH to a range of 4 − 5 for maximum bacterial cellulose yield.

2.5 Washing and Drying of Bacterial Cellulose

After the formation, the layer of bacterial cellulose was first washed with distilled water 3–4 times and then it was further washed with 0.1% NaOH to remove all the impurities as shown in Fig. 3. After the process of washing, the layer was placed on a plane object and dried in sunlight.

2.6 Weight Measurement

Weight of Bacterial cellulose produced from various culture media with different carbon and nitrogen sources was measured by the weighing machine.

The sequence in which each setup was completed is shown in Fig. 2.

2.7 Characterization of Bacterial Cellulose

2.7.1 X-Ray Diffraction

X-ray diffraction was performed by a diffractometer (Rigaku, MiniFlex II, Tokyo, Japan) operating at 220V. Bacterial cellulose was homogenized before analysis. The sample was placed in the sample tray where the surface was smoothed. The X-ray diffraction spectra were recorded using diffractometer in steps of 0.02̊ using Cu Kα radiation. Scans were performed at the range of 5–40 in 10 degree/min.

2.7.2 Fourier Transformed Infrared (FTIR) Spectroscopy

Fourier Transformed Infrared (FTIR) Spectroscopy associated with Attenuated Total Reflection (ATR) mode of bacterial cellulose was performed by using FTIR spectrometer (Nicolet 6700, Thermo Scientific, Waltham, USA). Spectra were recorded at wavelength 4000 − 650 cm-1, resolution 8cm-1, and number of scans 128 cm-1.

2.7.3 Field Emission Scanning Electron Microscope (FE-SEM)

Surface morphologies of the samples were performed using variable pressure Field Emission Scanning Electron Microscope (FE-SEM) Apreo S from ThermoFischer Scientific Eindhoven, The Netherlands. The sample was dried at 45 ̊C using hot oven before administering to the FE-SEM for analysis.

3.1 Production of Bacterial Cellulose

A fine layer of bacterial cellulose was produced by nitrogen and varied carbon sources as illustrated in Fig. 3. Results obtained from different culture media employed for bacterial cellulose production were given in Table 1. The layer was separated from the media and then washed and dried. After drying, bacterial cellulose layer can be used for different purposes in the pharmaceutical, textile, and paper industry.

3.1.2 Production of bacterial cellulose using different sources

Results obtained from various culture media employed for bacterial cellulose production are given in Table 1.

Table 1

Bacterial cellulose produced using different sources.
Media	Production of Bacterial cellulose
HS Media	A clear prominent layer of bacterial cellulose formed.
100ml Bagasse, 100ml Distilled water and Utilized Tea	A thin layer of Bacterial cellulose formed.
200ml Bagasse and Utilized Tea	A clear layer of Bacterial cellulose formed.
300ml Bagasse and Utilized Tea	Substantially thicker layer of Bacterial cellulose formed.
1500ml Bagasse and Utilized Tea	A significant clear and denser bacterial cellulose layer formed.
Glucose and Utilized Tea	The prominent layer of Bacterial cellulose layer formed.

3.2 Optimization of pH for production of Bacterial cellulose

The optimal pH for bacterial cellulose production by SCOBY is between 4 and 5. The pH adjustments were measured by pH strips to optimize the efficiency of SCOBY and increase Bacterial cellulose yield. Acidic pH increases the crystallinity and yield of bacterial cellulose, so the pH should not increase above 5 (Supplementary material).

3.3 Weight Measurement

Wet hydrated weight was measured to quantitatively determine the yield of Bacterial cellulose. It signifies the water retention capability of BC and the efficiency of the production method utilized. Under static conditions, the wet weight of different Bacterial cellulose film was given in Table 2.

Table 2

Wet weight measurements of Bacterial cellulose produced from different culture mediums.
Substrate	Weight measurement (g)
HS Media	15.7408
200ml (100ml Bagasse water, 100ml Distilled water) and Utilized Tea	2.8459
200ml Bagasse and Utilized Tea	9.2734
300ml Bagasse and Utilized Tea	18.5999
1500ml Bagasse and Utilized Tea	55.4854
Glucose and Utilized Tea	16.0142

3.4 Characterization of Bacterial Cellulose

To analyze the various properties of bacterial cellulose including its crystallinity, water holding capacity, and high degree of polymerization, different tests were performed. For the characterization of bacterial cellulose, tests of X-ray Diffraction, FTIR and Scanning Electron Microscopy were commercially performed.

3.4.1 X-Ray Diffraction

The XRD spectra of the bacterial cellulose membranes utilizing different carbon and nitrogen sources were generated in the form of graphs. The data obtained for bacterial cellulose using natural waste sources was compared with the bacterial cellulose produced using standard HS media (Supplementary material).

The graph shown above indicates the intensity on (y-axis) and 2 theta on (x-axis), which is typical for XRD analysis. In this graph, distinct peaks are shown around 2 theta values of approximately 15°, 22°, and 34°. The peaks around 2 theta = 15° and 22° are characteristic of cellulose I, indicating the presence of crystalline cellulose. The sharp peak at around 22° (approximately around 22.5°) is particularly indicative of the (200) plane of cellulose I. The sharpness and intensity of the peaks suggest a relatively high degree of crystallinity. The peak around 15° corresponds to the (110) plane, and its presence alongside the 22° peak confirms the typical pattern of cellulose I. The broad background signal and the less intense areas between the peaks represent amorphous regions in the cellulose.

The X-ray Diffraction (XRD) pattern shows the crystalline structure of bacterial cellulose produced from bagasse. The x-axis represents the 2-theta angle (degrees), and the y-axis represents the intensity (counts). The peak around 2-theta = 5 degrees is very prominent and sharp, indicating a high degree of crystallinity in the sample.

There is another significant broad peak around 2-theta = 20 to 35 degrees, which suggests the presence of amorphous regions within the cellulose. The sharp peak at a low 2-theta angle (around 5 degrees) corresponds to the crystalline regions of cellulose. This indicates well-ordered, periodic arrangements of cellulose chains. The broad feature between 20 and 35 degrees indicates the presence of amorphous regions where the cellulose chains are less ordered.

Three peaks at 2θ = 14.62° (1–10), 16.29° (110), and 22.48° (200) for bacterial cellulose derived from glucose, commercial glycerol, glycerol leftover from biodiesel production, grape bagasse, and sugarcane molasses indicate that only cellulose I was present in all BC samples [67]. Diffraction patterns obtained from the bacterial cellulose of HS media, glucose and bagasse are shown in Fig. 4.2–4.4. The peaks of cellulose produced by HS media around 2 theta = 15° and 22° are characteristic of cellulose I, which is the presence of crystalline cellulose. The sharp peak at around 22° (approximately around 22.5°) is particularly indicative of the (200) plane of cellulose I. The sharpness and intensity of the peaks suggest a relatively high degree of crystallinity. The peak around 15° corresponds to the (110) plane, and its presence alongside the 22° peak confirms the typical pattern of cellulose I [68].

3.3.2 Fourier Transform Infrared (FTIR) Spectroscopy

The spectra of the bacterial cellulose membranes were generated utilizing different carbon and nitrogen sources, and this data was compared with the standard HS media obtained bacterial cellulose (Supplementary material).

The FTIR ATR of BC samples were prepared from the culture media having Bagasse and Glucose as carbon source and utilized tea as nitrogen source and compared with the FTIR ATR spectrum of BC produced by the Standard HS media. It was observed that these spectra were similar to the HS media produced BC, indicating that they contain the same chemical functional bonds. The FTIR absorption ranges are alcohol and phenol O-H stretching bond 3000–3800 cm- 1 with high absorption peak is 3343.10 cm-1 confirming the presence of cellulose structure as shown in Fig. 3.6, C ≡ C bond isolated stretch present in alkynes 2100–2250 cm-1 with an absorption peak at 2162.42 cm-1. Carbonyl groups C = O stretching 1630–1690 cm-1 wavelength with absorption peak at 1638.30 cm-1 often present in cellulose derivatives. 1314.50 cm-1 absorption peak of O-H bond bending might be present in phenols. 1160.04 cm-1 peak as present in asymmetric stretching vibrations from C–O–C bonds reflecting β 1, 4- glycosidic linkages present in cellulose. 1107.64 cm-1, 1056.08 cm-1 and 1032.65 cm-1 vibration stretch of C-O bond as present in primary and secondary alcohol which signifies the production of ethanol by yeast that promotes the bacterial cellulose synthase process, ultimately leading to the production of a bacterial cellulose film.

The absence of an absorption peak around 1540 cm-1 signifies amine bonds present in proteins from culture media. Hence, their absence confirms that the BC film produced by HS media, Bagasse and utilized tea, Glucose and utilized media were washed correctly and are pure. However, an unexpected peak of C ≡ C bond stretch 2100–2250 cm-1, present in alkynes having an absorption peak at 2161.70 cm- 1 identify some kind of noise in the measurement process by spectroscopy or contamination in the layer. Moreover, there were no peaks around 3440 cm − 1 to 3495 cm − 1, which signifies OH bond stretching due to intramolecular hydrogen bonds confirming the absence of cellulose II. The peaks visible in the FTIR ATR spectrum of Standard HS media, Bagasse and utilized tea, Glucose and utilized tea produced BC are consistent with the structure of Cellulose I [69].

3.3.3 Field Emission Scanning Electron Microscopy (FE-SEM)

Surface morphological characteristics of the sample sugarcane bagasse and utilized tea produced bacterial cellulose was performed using the variable pressure Field Emission Scanning Electron Microscope (FE-SEM) with an accelerating voltage at 5.00 KV. An image magnification of approximately 5000X, 8000X, 10,000X, 15,000X, 20,000X, 50,000X, and 100,000X were used. The micrograph of the bacterial cellulose sample was collected and recorded.

The Fig. 6 (a-b) showed that sugarcane bagasse bacterial cellulose FESEM analysis at a magnification of 5000X and 8000X gave a diameter of 10µm, 4µm, fibrils size and densely packed, fibrous network of fibrils are present. The fibrils are irregular in shape and structure. The Fig (c-f) showed that with a Sugarcane bagasse bacterial cellulose at magnification of 10,000X, 20,000X, 50,000X, and 100,000X with a diameter ranging from 1µm, 4µm, and 5µm size fibrils. The fibrils were more fragile and fragmented structures containing the small pores that are connected with the fibrils with the neighboring cell walls in a packed form. Besides, this bagasse bacterial cellulose poses smoother surface, strong fibrous network of nanofibers with different diameters, irregular in shape and arranged in a discontinuous manner. The morphological characteristics and microstructure of BC were identified as they contain characteristics of nanostructure and microfibrillar arrangement of cellulose that is naturally present in plant-based cellulose. This suggests that sugarcane bagasse bacterial cellulose produced from inner pitch have nanofibers and nanostructure that would be used in many future applications for the product formation from sugarcane bagasse bacterial cellulose.

In this study, the comparison of HS media, Glucose and Sugarcane Bagasse medium was successfully done for bacterial cellulose production. The results concluded that sugarcane bagasse and utilized tea are cheaper alternative substrates for obtaining bacterial cellulose. This low-cost bacterial cellulose production has a characteristic feature of natural cellulose that can significantly contribute to production of sustainable products such as edible films, facemask, and cling sheets. Furthermore, this overall experimental research work highlights a new approach to provide an eco-friendly, low cost, biodegradable higher cellulose production from food waste. Sugarcane bacterial cellulose is ideal for the sustainable production of products that will reduce food waste and align with the mission of SDG 12 “Ensure sustainable consumption and production pattern'. There is a need to perform further analysis for characterizing the bacterial cellulose and its production at a higher scale.

FUTURE PERSPECTIVES

Bacterial Cellulose holds significant promise for future applications in several fields such as cosmetology, food industry and in biomedical industries.

Moreover, it can be utilized as an advanced wound healing dressing, biodegradable facial masks, biodegradable cling sheet and as a drug delivery vehicle.

Further research in enhancing the efficiency of BC production can lead to the development of more BC related products in society.

BC Bacterial Cellulose

FTIR-ATR Fourier Transform Infrared Spectroscopy-Attenuated Total Reflection

XRD X-Ray Diffraction

FE-SEM Field Emission Scanning Electron Microscopy

HS Media Hestrin-Schramm Media

Competing Interest

The authors have no relevant financial and non-financial interest to disclose.

Ethics approval

Not applicable. This research work does not contain any experimentation with human participants or animals.

Funding

The authors did not receive any funding from any organization for the submitted work.

Author Contribution

CRediT StatementConceptualization: S Marriam, S Ghazal, S Shahzadi, S FatimaMethodology: S M, Ghazal, S FatimaFormal analysis: S Marriam, S ShahzadiInvestigation: S Ghazal, S FatimaResources: S Marriam, S Ghazal, S Shahzadi, S Fatima Writing – Original Draft Preparation: S Marriam , S Ghazal, S FatimaWriting – Review & Editing: S Marriam, S Ghazal, Dr. Ruqyya KhalidSupervision: Dr. Amna Younus, Dr. Ruqyya Khalid, Dr. Razia Tassaduq

Acknowledgement

The authors like to extend their gratitude to Dr. Amna Younus, Dr. Ruqyya Khalid and Dr. Razia Tassaduq, Professors at Kinnaird College for Women.

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Production and Characterization of Bacterial Cellulose from Food waste: A sustainable approach

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Abstract

Figures

INTRODUCTION

1.1 Cellulose

1.2 Bacterial cellulose

1.3 SCOBY symbiotic culture of bacteria and yeast

1.4 Kombucha

1.5 Parameters affecting the growth of Bacterial Cellulose

1.5.1 pH

1.5.2 Temperature

1.6 Raw material for Bacterial Cellulose production

MATERIALS AND METHODS

2.1 Study Location

2.2 Materials Required

2.3 Production of Bacterial Cellulose (BC)

2.3.1 Production of BC from Hestrin-Schramm (HS) media

2.3.2 Production of BC from Bagasse and utilized tea

2.3.2.1 Pre-Treatment of sugarcane bagasse

2.3.2.2 Bagasse water with varying concentration

2.3.3 Production of Bacterial Cellulose from Glucose and Utilized tea

2.4 pH measurement

2.5 Washing and Drying of Bacterial Cellulose

2.6 Weight Measurement

2.7 Characterization of Bacterial Cellulose

2.7.1 X-Ray Diffraction

2.7.2 Fourier Transformed Infrared (FTIR) Spectroscopy

2.7.3 Field Emission Scanning Electron Microscope (FE-SEM)

RESULTS AND DISCUSSION

3.1 Production of Bacterial Cellulose

3.1.2 Production of bacterial cellulose using different sources

3.2 Optimization of pH for production of Bacterial cellulose

3.3 Weight Measurement

3.4 Characterization of Bacterial Cellulose

3.4.1 X-Ray Diffraction

3.3.2 Fourier Transform Infrared (FTIR) Spectroscopy

3.3.3 Field Emission Scanning Electron Microscopy (FE-SEM)

CONCLUSION

Abbreviations

Declarations

Competing Interest

Ethics approval

Funding

Author Contribution

Acknowledgement

References

Additional Declarations

Supplementary Files

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