Production Of Fungal Chitosan Research Articles

Optimization of fungal chitosan production from Cunninghamella echinulata using statistical designs.

The online version contains supplementary material available at 10.1007/s13205-024-03919-6.

Production of fungal chitosan and fabrication of fungal chitosan/polycaprolactone electrospun nanofibers for tissue engineering.

The present study investigated that chitosan production of Rhizopus oryzae NRRL 1526 and Aspergillus niger ATCC 16404. Fungal chitosans were characterized by scanning electron microscopy (SEM)-energy dispersive X-ray analysis, Fourier transform infrared spectroscopy (FTIR), differential scanning calorimeter and deacetylation degrees of fungal chitosans were determined. The percentage yield of Ro-chitosan and An-chitosan were determined as 18.6% and 12.5%, respectively. According to percentage of chitosan yield and the results of the characterization studies, chitosan that obtained from Rhizopus oryzae NRRL 1526 was selected for subsequent studies. Cytotoxicity of chitosan obtained from Rhizopus oryzae NRRL 1526 was determined by MTT assay on human dermal fibroblast cell line. Acording to results of the cytotoxicity test fungal chitosan was nontoxic on cells. The high cell viability was observed 375 μg/mL concentration at 24th, 48th h periods and at the 187.5μg/ml 72nd h periods on cells. The fungal chitosan obtained from Rhizopus oryzae NRRL 1526 was used to fabrication of electrospun nanofibers. Fungal chitosan based polymer solutions were prepared by adding different substances and different electrostatic spinning parameters were used to obtain most suitable nanofiber structure. Characterization studies of nanofibers were carried out by SEM, FTIR and X-ray diffraction. The most suitable nanofiber structure was determined as F4 formula. The nanofiber structure was evaluated to be thin, bead-free, uniform, flexible and easily remove from surface and taking the shape of the area. After the characterization analysis of fungal chitosan it was determined that the chitosan, which obtained from Rhizopus oryzae NRRL 1526 is actually chitosan polymer and this polymer is usable for pharmaceutical areas and biotechnological applications. The electrospun nanofiber that blends fungal chitosan and PCL polymers were fabricated successfully and that it can be used as fabrication wound dressing models. RESEARCH HIGHLIGHTS: Extraction of chitosan from Rhizopus oryzae NRRL 1526 and Aspergillus niger ATCC 16404 and characterization scanning electron microscopy-energy dispersive X-ray analysis, Fourier transform infrared spectroscopy, differential scanning calorimeter. Fabrication and characterization of the fungal chitosan/PCL electrospun nanofibers.

Banana peels as a cost effective substrate for fungal chitosan synthesis: optimisation and characterisation

ABSTRACT Massive accumulation of unprocessed banana peels enthralls sustainable issues as they are eventually dumped as landfills leading to emission of obnoxious gasses. To avoid these persisting challenges the present study shims lights on chitosan production from the characterised fungal strain using banana peel hydrolysate as an effective medium. Substantial amount of carbohydrate in banana peels serves as a potential solution for fungal chitosan production in a view to attain a circular bioeconomy and repurposed for synthesis of beneficial products in a cost effective manner. Presence of fermentable sugars in banana peels qualifies them as a feasible substrate which could be exploited for scaling up of fungal chitosan synthesis. Screened isolate was subjected to statistical optimisation using formulated medium to elucidate the influential factors that had significant effect on chitosan production. The harvested chitosan biomass was characterised through standardised techniques and evaluated for further studies. Statistical optimisation reveals that ammonium nitrate (5 g/L), pH (6) and incubation time (144 hrs) were the three PBD variables that had a greater influence on fungal chitosan yield. The validated developed model exhibited maximum yield of 200 mg/L, a 4.4 fold increase than unoptimised medium (45 mg/L). These findings emphasise the fermentative synthesis of chitosan through valorisation of banana peel prop up a complementary approach in concomitant with preserving renewable resources and bioproduct formation.

Standardization of fermentation process for mass production of fungal chitosan using agro-industrial waste

Standardization of fermentation process for mass production of fungal chitosan using agro-industrial waste

Exploitation of plant-derived food waste for valuable products using bioprocesses: a narrative review

Globally, a million tons of plant-derived food waste are generated annually from various cultivation activities and agro-industrial manufacturing. When food waste decomposes, it releases methane gas that causes global warming. The exploitation of food wastes for valuable products provides an alternative approach to environmental protection. This paper aimed to review the exploitation of plant-derived food waste for valuable products and to evaluate different bioprocesses for optimum yields of the valuable products. This review study was conducted by using PubMed, ScienceDirect, and NCBI databases. A total of 169 publications were screened and 43 papers of high relevance were selected. Cereal wastes such as wheat straw, rice bran, root and tuber waste, and fruits and vegetable pomace contribute the most amounts to the total food waste worldwide. These wastes serve as renewable sources for bioactive production. Various fermentation processes involving enzymes, yeast, or bacteria were used to convert food waste into valuable products such as phenolics, flavonoids, dietary fibre, carotenoids, lignocellulose, phytosterols, tocotrienols, coumaric acid, oryzanol, citrulline, vitamins, and essential amino acids. Several bioprocesses and the optimization conditions for the maximum yield of valuable products were highlighted. In the production of xylitol from corn cobs using yeast (Debaryomyces hansenii), a higher yield (0.216 g xylitol/g xylose) was obtained through enzyme hydrolysate fermentation compared to acid hydrolysate fermentation (0.100 g xylitol/g xylose). In the production of fungal chitosan using solid-state fermentation, the yield was 3 folds higher (34.4 g/kg substrate) for soybean meals substrate than potato peels substrate (10.8 g/kg substrate). Plant-derived food waste serves as a renewable substrate for the production of health-promoting bioactive compounds through the optimization of various bioprocesses. The exploitation of plant-derived food waste for high-value products has significant implications for environmental impact, public health, and economic gain.

Green synthesis of chitosan by Cunninghamella elegans UCP 1306 using sustainable substrates mediated morphological changes

Currently, the commercial production of chitosan takes place through the thermochemical deacetylation of crustacean shells, which process requires the use of chemical substances, such as strong alkaline solutions, which are important sources of environment pollution. We present a method for the production of chitosan by Cunninghamella elegans UCP 1306 using a green metabolic conversion of agro-industrial residues, based on a 22 Factorial Design and describe the main morphological changes observed in the specie. The highest yield of biomass (6.375 g/L) and chitosan (101.7 mg/g) were observed in the assay 2 (Corn steep liquor -CSL 4%, Cassava wastewater-CWW 4%), where the specie presented loose and branched hyphae with the presence of septations, pellets of 0.4 to 0.5 mm and clumps of up to 0.1 mm. Statistical analysis showed that higher concentration of CSL contributed significantly to the growth of the species. The bands determined in Fourier transform infrared spectroscopy confirmed the degree of deacetylation of 84.61% of the chitosan obtained. This research showed that the use of CSL and CWW in association was very promising, being able to be used as sustainable sources of nitrogen and carbon in the production of fungal chitosan with a high degree of deacetylation and can be applied to the industrial process.

Chitosan Production by Fungi: Current State of Knowledge, Future Opportunities and Constraints

Conventionally, the commercial supply of chitin and chitosan relies on shellfish wastes as the extraction sources. However, the fungal sources constitute a valuable option, especially for biomedical and pharmaceutical applications, due to the batch-to-batch unsteady properties of chitin and chitosan from conventional ones. Fungal production of these glycans is not affected by seasonality enables accurate process control and, consequently, more uniform properties of the obtained product. Moreover, liquid and solid production media often are derived from wastes, thus enabling the application of circular economy criteria and improving the process economics. The present review deals with fungal chitosan production processes focusing on waste-oriented and integrated production processes. In doing so, contrary to other reviews that used a genus-specific approach for organizing the available information, the present one bases the discussion on the bioprocess typology. Finally, the main process parameters affecting chitosan production and their interactions are critically discussed.

Chitosan production by Penicillium citrinum using paper mill wastewater and rice straw hydrolysate as low-cost substrates in a continuous stirred tank reactor

ABSTRACT In this study, paper mill wastewater and hemicellulose hydrolysate were evaluated as low-cost substrates for fungal chitosan production using Penicillium citrinum. Submerged fermentation was first studied using a bioreactor operated under batch, fed-batch and continuous modes with paper mill wastewater as the substrate. Very high removal (91%) of organics as chemical oxygen demand (COD) in the wastewater with 160 mg L-1 chitosan production by P. citrinum was obtained using the bioreactor operated under fed-batch mode for 72 h. Moreover, 86% reduction of phenolics in the wastewater with 89% decolourization efficiency was achieved in the fed-batch experiments with the bioreactor. Under the continuous mode of operation with the bioreactor, maximum chitosan production of 170 mg L-1 was observed. The effect of acetic acid addition to the wastewater for enhancing chitosan production by the fungus was further studied in a batch system. Chitosan productivity of 2.33 mg L-1 h-1 was obtained with 50 mg/L acetic acid. Various models, viz. Monod, Haldane, Andrews, Webb and Yano, were fitted to the experimental data for understanding the kinetics involved in the process. Haldane model accurately fitted the experimental data on biomass specific growth rate, acetic acid consumption rate and chitosan production rate by P. citrinum with acetic acid addition to the wastewater. Fungal fermentation of another low-cost substrate, rice straw hydrolysate, was further studied using the batch-operated bioreactor; and a maximum chitosan titre of 911 mg L−1 was achieved using the detoxified rice straw hydrolysate. Highlights Low-cost substrates for chitosan production by Penicillium citrinum are reported Acetic acid addition to paper mill wastewater enhances chitosan production Biomass growth and chitosan production follow substrate inhibition kinetics Fed-batch -operated bioreactor resulted in 91% wastewater treatment efficiency Maximum chitosan titre of 911 mg L−1 was achieved with rice straw hydrolysate

Agroindustrial waste as ecofriendly and low-cost alternative to production of chitosan from Mucorales fungi and antagonist effect against Fusarium solani (Mart.) Sacco and Scytalidium lignicola Pesante

This study aimed to evaluate the production of fungal chitosan (FuChi) from Mucorales fungi cultivated in a cashew apple juice (CAJ) and cheese whey (CW) mixture, and to determine the growth-inhibitory effect of this biopolymer against Fusarium solani CFF109 and Scytalidium lignicola CMM1098, which cause root rot disease in cassava plants. Cunninghamella phaeospora UCP 1303 and Cunninghamella elegans UCP 1306 showed the highest FuChi production in screening assay, being selected to a CCRD 22 design to analyze the influence of different CAJ and CW concentrations in the increase of FuChi production. All nine Mucorales fungi cultivated in CAJ-CW medium, showing FuChi production in the range of 27.58 (Mucor hiemalis UCP 1309) to 65.40 mg/g (C. elegans UCP 1306). During CCRD 22 design, the highest FuChi production (64.09 mg/g) was achieved by C. elegans UCP 1306 cultivated in medium containing 40% (v/v) of CAJ and 30% (v/v) of CW, presenting 75% deacetylation degree and crystallinity indexes of 41.50%. FuChi at 16000 μg/mL showed a better inhibition against S. lignicola mycelial growth (81.70%) when compared with F. solani (22.13%) and induced alterations in hyphae morphology on both strains. CAJ and CW are promising substrates for FuChi production, and this biopolymer shows antimicrobial effect against F. solani and S. lignicola.

Chitosan Synthesis and Optimization of Root Endophytic Fungi

The cell wall component of root endophytic fungi is a main source of chitosan synthesis. This research aimed to optimize chitosan synthesis from three different isolated species of root endophytic fungi in Pulau Dua Nature Reserve, Banten Indonesia. Three species of root endophytic fungi, namely, Aspergillus niger, Aureobasidium sp., Basipetospora sp., were cultured to produce chitosan. To optimize the production of fungal chitosan, cultures were treated with different conditions such as growth medium (Oat Meal broth, Potato Dextrose Broth, CMMY Broth), pH value (4, 5, 6, 7), and temperature (25, 30, 35, 40°C). Chitosan was extracted from dried mycelium after treated on NaOH 10% at 121°C for 2 min. The degree of deacetylation (DD) of chitosan was then examined and compared with the control (shrimp chitosan). The optimum fungal chitosan condition was higher in CMMY medium, pH 5, and 30°C. The DD value of three root endophytic fungi species, Aspergillus niger, Aureobasidium sp., Basipetospora sp. were 88.5, 83.3, 80.8, respectively, which was lower than DD value resulted from control. This study indicates that Aspergillus niger was a potential endophytic fungi for chitosan synthesis.

Sustainable and green approach of chitosan production from Penicillium citrinum biomass using industrial wastewater as a cheap substrate

Marine sources especially crustaceans have been extensively used worldwide for the production of chitosan. However, limited availability as well as variations in the properties of the derived chitosan is a serious drawback of utilizing marine sources for chitosan production. This study investigated sustainable and green approach of fungal chitosan production using paper mill wastewater as a cheap and easily available substrate. The fungus Penicillium citrinum IITG_KP1 used in this study was initially isolated from an infected bamboo shoot. Addition of acetic acid at low levels led to a 150% increase in the yield of chitosan from 95 g/kg to 138 g/kg of dry fungal biomass. This result correlated well with an increase in xylose uptake rate due to acetic acid addition that was confirmed by enhanced activity of xylose reductase (XR) and xylitol dehydrogenase (XDH) enzymes in the presence of acetic acid. Very high COD removal efficiency (75%) along with 70% phenolic reduction and 84% decolourization efficiency of the raw paper mill wastewater without any prior pre-treatment was further achieved by carrying out the fungal fermentation using a bioreactor under batch mode of operation. The fungal chitosan showed properties comparable with those of a commercially available standard.

Efficient production of fungal chitosan utilizing an advanced freeze-thawing method; quality and activity studies

Fungal chitosan was produced from brown Agaricus bisporus, white Agaricus bisporus, and Pleurotus ostreatus mushrooms (commercially known as brown portobello, white protobello, and oyster mushrooms, respectively) by utilizing a classic alkali deacetylation protocol (PI), as well as a new protocol (PII) that involves a freeze-thawing cycle. The new protocol (PII) has led up to a twofold increase in the obtained chitosan's yield, a more efficient deacetylation process that resulted in up to a threefold decrease in its acetylated units, and a whiter color. For instance, using the PII protocol, 1 g of dry Pleurotus ostreatus mushrooms yielded 44.3 mg of chitosan that contained only 14.8% acetylated units, and had a whiteness index of 80.3. For comparison, using the classic PI protocol, 1 g of dry Pleurotus ostreatus mushrooms yielded only 22.4 mg of chitosan with 44.1% acetylated units, and a whiteness index of 75.7. Gel permeation chromatography, FTIR, NMR, UV, elemental analysis, X-ray diffractometry, and thermogravimetric analysis were all used to characterize the prepared fungal chitosan samples and examine their physical properties (viscosity, color, hydrophobicity, and solubility). Microbiological studies have revealed the yielded chitosan samples have demonstrated biological activity against Gram-positive Bacillus subtilis, Gram-negative Escherichia coli bacteria, and Saccharomyces cerevisiae yeast. The prepared fungal-sourced chitosan samples were also compared with the crustacean-sourced commercial chitosan all throughout this paper studies. It can be concluded that the PII protocol allows for the formation of fungal chitosan with quality and properties that do not fall short of those found in the crustacean-sourced chitosan. This study therefore presents an efficient method for the production of non-animal sourced chitosan, which is extremely desired in the food industry because it overcomes the limitations of animal-sourced chitosan and allows to expand this material use. In addition, the presented freeze-thaw cycle technique can also be used to yield and modify other edible hydrocolloids.

Fungal chitosan production using xylose rich of corn stover prehydrolysate by Rhizopus oryzae

ABSTRACTXylose rich of lignocellulosic prehydrolysate could be used for fungal chitosan production effectively by Rhizopus oryzae. When 15 g/L xylose was used as carbon source for R. oryzae AS 3.819 fermentation, almost all of the xylose was consumed for cell growth (4.33 g/L of biomass) and the chitosan extraction yield was 0.11 g/g biomass. Corn stover prehydrolysate by dilute acid-assisted steam explosion pretreatment that contained 30 g/L xylose and 10 g/L glucose was fermented by R. oryzae AS 3.819 for 72 h, the biomass and the chitosan extraction yield were 10.96 g/L and 0.09 g/g biomass. The degree of deacetylation of the fungal chitosan derived from corn stover prehydrolysate (91.27%) was higher than that of the commercial chitosan extracted from natural shellfish exoskeleton (87.25%), and the viscosity of the fungal chitosan derived from corn stover prehydrolysate (2.67 mPa⋅s) had a large decrease compared to the commercial chitosan (22.25 mPa⋅s). The functional groups and thermostability of the fungal chitosan were proofed by Infrared (IR) spectra and Derivative thermogravimetric (DTG), respectively.

Effects of Plant Growth Hormones on Mucor indicus Growth and Chitosan and Ethanol Production.

The objective of this study was to investigate the effects of indole-3-acetic acid (IAA) and kinetin (KIN) on Mucor indicus growth, cell wall composition, and ethanol production. A semi-synthetic medium, supplemented with 0–5 mg/L hormones, was used for the cultivations (at 32 °C for 48 h). By addition of 1 mg/L of each hormone, the biomass and ethanol yields were increased and decreased, respectively. At higher levels, however, an inverse trend was observed. The glucosamine fraction of the cell wall, as a representative for chitosan, followed similar but sharper changes, compared to the biomass. The highest level was 221% higher than that obtained without hormones. The sum of glucosamine and N-acetyl glucosamine (chitin and chitosan) was noticeably enhanced in the presence of the hormones. Increase of chitosan was accompanied by a decrease in the phosphate content, with the lowest phosphate (0.01 g/g cell wall) being obtained when the chitosan was at the maximum (0.45 g/g cell wall). In conclusion, IAA and KIN significantly enhanced the M. indicus growth and chitosan production, while at the same time decreasing the ethanol yield to some extent. This study shows that plant growth hormones have a high potential for the improvement of fungal chitosan production by M. indicus.

Direct Solid-State Fermentation of Soybean Processing Residues for the Production of Fungal Chitosan by Mucor rouxii

The feasibility of utilizing soybean-processing residues such as soybean meal and hulls as substrates for chitosan production by the fungus Mucor rouxii ATCC 24905 via solid-state fermentation (SSF) was investigated. The effects of the type of soybean-based substrate, length of cultivation period, substrate moisture content, substrate pH, incubation temperature and extraction conditions on chitosan yield were determined. The results showed that a maximum fungal chitosan yield of up to 3.44% by dry substrate weight (34.4 g/kg) could be achieved using a pure soybean meal substrate with an initial moisture content of 50% (w/w) and pH of 5 - 6 incubated for six days at 25&degC. A more severe heat treatment (autoclaving vs. refluxing) resulted in higher chitosan extraction yields regardless of the strength of extraction reagents. Fourier transform infrared (FTIR) analysis of the fungal chitosan revealed its degree of deacetylation (DDA) to be between 55% and 60%.

Production of fungal chitosan from date wastes and its application as a biopreservative for minced meat

Raw and processed meat contaminated with pathogenic microorganisms is a continuing worldwide problem facing health and industry overseers.Fungal chitosan was extracted, purified and characterized from Aspergillus brasiliensis (niger) ATCC 16404 grown in date syrup (dips) and applied as a potential meat biopreservative. The main features of produced chitosan were a deacetylation degree of 81.3%, a molecular weight of 31,000Da, 96% solubility in 1% acetic acid solution and a harmonized IR-spectrum to standard commercial chitosan.The application of fungal chitosan, as a natural and safe biopreservative for minced meat, was conducted in comparison with potassium sorbate, as a commercial meat preservative.Treated meat samples with 0.02% chitosan was the least trials in microbial contents, i.e. total count, coliforms, β-glucuronidase-positive Escherichia coli, Enterobacteriaceae, yeasts and molds, Staphylococcus aureus and coagulase positive staphylococci. The antimicrobial activity of fungal chitosan was considerably greater than that of potassium sorbate or their combination at 0.01% from each.Sensory characteristics, e.g. color, odor and texture, of treated meat with chitosan, were higher than those of control and potassium sorbate treated samples.Fungal chitosan, however, could be recommended as a powerful, natural and eco-friendly alternative for meat preservation and overall quality maintenance.

Apple pomace as a substrate for fungal chitosan production in an airlift bioreactor

Abstract The aim of this work was to evaluate apple pomace as carbon source for fungal chitosan production in an external-loop airlift bioreactor by Gongronella butleri. Apple pomace has high moisture content, insoluble carbohydrates such as cellulose, hemicellulose and lignin, reducing sugars like glucose and fructose and other sugars such as sucrose. Due to its high content of sugars, apple pomace can be a potential substrate for fungal chitosan production. To improve chitosan production from apple pomace, the effect of the nitrogen sources NaNO3 and (NH4)2SO4 and specific aeration rate of 0.1, 0.3 and 0.6 vvm was investigated. The ammonium sulfate and specific aeration rate of 0.6 vvm had significant effect (p

Production and isolation of chitosan from Aspergillus terreus and application in tin(II) adsorption

ABSTRACTFed‐batch fermentation was used for biomass and fungal chitosan production by Aspergillus terreus (BCRC 32068) grown in a potato dextrose agar medium. The polysaccharides were extracted by an alkali–acid treatment, and structural investigations by X‐ray diffraction, Fourier transform infrared analysis, and viscosity and thermal analysis were done. A high level of chitosan was extracted from A. terreus; this implied that it was feasible to produce chitosan from industrial waste mycelia. Fungal chitosan derived from A. terreus showed the highest adsorption capacity for Sn(II). The order of Sn(II) adsorption capacity for these chitosanaceous materials was Fungal chitosan > Chitin > Biomass. Fungal chitosan derived from A. terreus was well correlated with Langmuir's isotherm model. The maximum capacity for Sn(II) sorption deduced from the use of the Langmuir isotherm equation was 303 mg/g; this was significantly higher than that of A. terreus. Fungal chitosan is an easy and cost‐effective material for the abatement of pollution. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014, 131, 40436.

Antibacterial action of acetic acid soluble material isolated from Mucor rouxii and its application onto textile

Acetic acid soluble material (AcSM) is a chitosan-rich fraction isolated from the fungal cell wall materials. The final step in the traditional production of fungal chitosan is the separation of chitosan from the cell wall AcSM via raising the pH to 9–10 followed by centrifugation. This step results in further undesirable economic and environmental effects. The goal of this paper is to avoid that by investigating the antimicrobial effect of the whole AcSM from Mucor rouxii DSM-1191 cell wall and its application on cotton fabrics. The treated fabrics were characterized through monitoring the textile physical properties and for the antibacterial activity against Escherichia coli and Micrococcus luteus. Results showed that Mucor rouxii DSM-1191 has excellent potentials to be used for cell wall AcSM production on industrial scale with a maximum content of 40% in dry mycelia. The obtained results indicated that the physical properties of the treated fabrics, as well as the antibacterial activity, were improved after treatment with fungal AcSM.

Laboratory scale production of 13C labeled chitosan by fungi Absidia coerulea and Gongronella butleri grown in solid substrate and submerged fermentation

Nowadays, chitin and chitosan are applied in many medical and pharmaceutical products. However little is known about the metabolism of chitin and chitosan in vivo. In the human body, lysozyme will degrade chitin and chitosan into chito-oligosaccharides. 13C labeled chitosan is an essential prerequisite for the further study of the fate of chito-oligosaccharides in vivo. To fulfill this requirement, chitosans were extracted from mycelia of fungi, Absidia coerulea ATCC 14076 and Gongronella butleri USDB 0201 and ATCC 42618 grown in solid substrate fermentation (SSF) and submerged fermentation (SMF) to select the best fungus and fermentation method for the production of 13C labeled chitosan. Based on the production yield of chitosan, the SSF method is the best method for the production of fungal chitosan when compared with SMF methods (i.e., fed-batch fermentation and batch fermentation). However synthesis of 13C labeled chitosan in cell wall of G. butleri grown in SSF medium coated with 1- 13C-glucose was not observed. Alternatively, fungus A. coerulea was grown in SMF medium containing 2- 13C-glucose. The successful synthesis of 13C labeled glucosamine from 2- 13C-glucose was observed in mycelia of A. coerulea grown in SMF medium containing 2- 13C-glucose in a yield of about 13 g/100 g mycelia.