Optimization Of Xylanase Production Research Articles

Optimization of xylanase production by Bacillus sp. MCC2212 under solid-state fermentation using response surface methodology

Xylanases have become increasingly significant due to their versatile applications in the food, paper, and pharmaceutical industries. Nevertheless, the current production of these enzymes relies on costly substrates, with estimates indicating that over 30% of the production expenses are attributed to these substrates. The objective of this study is to optimize the physico-chemical parameters for obtaining the maximum production of xylanase enzyme from Bacillus sp. MCC2212. The production conditions were statistically optimized using Plackett-Burman design (PBD) and Central Composite design (CCD). The significant variables identified through PB design including temperature, substrate to moisture ratio, MgSO4, peptone, tween 20, inoculum size, inoculum age, incubation time, and extracted xylan, were further optimized using the CCD approach. This optimization process revealed the most influential factors affecting xylanase production, with optimal conditions observed at a temperature, 35 °C; a substrate to moisture ratio, 1:1.75 g:mL; MgSO4, 1.1%; peptone, 1.5%; tween 20, 1.25%; inoculum size, 10 % (v/w); inoculum age, 20 h; an incubation time, 50 h; and crude xylan extracted from wheat bran at a concentration of 6% v/w. The R2 value of the model was found to be 0.9901, indicating its effectiveness. The optimized CCD model displayed a 1.84-fold greater xylanase production than the PB design approach. These findings suggest that wheat bran, a readily available agro-waste, could be a feasible alternative to the conventional substrate, beechwood xylan, for the production of xylanase enzyme with the possibility of achieving higher production levels optimized by using statistical design approach.

Optimization of xylanase production by Pichia kudriavzevii and Candida tropicalis isolated from the wood product workshop.

Enzymatic compounds can be found abundantly and provide numerous advantages in microbial organisms. Xylanases are used in various pharmaceutical, food, livestock, poultry, and paper industries. This study aimed to investigate xylanase-producing yeasts, xylose concentration curve and their enzymatic activity under various factors including carbon and nitrogen sources, temperature, and pH. Enzyme activity was evaluated under different conditions before, during, and after purification. The yeast strains were obtained from the wood product workshop and were subsequently cultivated on YPD (yeast extract peptone dextrose) medium. Additionally, the growth curve of the yeast and its molecular identification were conducted. The optimization and design process of xylan isolated from corn wood involved the use of Taguchi software to test different parameters like carbon and nitrogen sources, temperature, and pH, with the goal of determining the most optimal conditions for enzyme production. In addition, the Taguchi method was utilized to conduct a multifactorial optimization of xylanase enzyme activity. The isolated species were partially purified using ammonium sulfate precipitation and dialysis bag techniques. The results indicated that 3 species (8S, 18S, and 16W) after molecular identification based on 18S rRNA gene sequencing were identified as Candida tropicalis SBN-IAUF-1, Candida tropicalis SBN-IAUF-3, and Pichia kudriavzevii SBN-IAUF-2, respectively. The optimal parameters for wheat carbon source and peptone nitrogen source were found at 50°C and pH 9.0 through single-factor optimization. By using the Taguchi approach, the best combination for highest activity was rice-derived carbon source and peptone nitrogen source at 50°C and pH 6.0. The best conditions for xylanase enzyme production in single-factor optimization of wheat bran were 2135.6 U/mL, peptone 4475.25 U/mL, temperature 50°C 1868 U/mL, and pH 9.0 2002.4 U/mL. Among the tested yeast, Candida tropicalis strain SBN-IAUF-1 to the access number MZ816946.1 in NCBI was found to be the best xylanase product. The highest ratio of enzyme production at the end of the delayed phase and the beginning of the logarithmic phase was concluded by comparing the growth ratio of 8S, 16W, and 18S yeasts with the level of enzymatic activity. This is the first report on the production of xylan polymer with a relative purity of 80% in Iran. The extracellular xylanases purified from the yeast species of C. tropicalis were introduced as a desirable biocatalyst due to their high enzymatic activity for the degradation of xylan polymers.

Optimization of xylanase production from Aspergillus Spp. hau-f-6 using plackett burman screening design and path of steepest ascent in submerged fermentation

Optimization of xylanase production from Aspergillus Spp. hau-f-6 using plackett burman screening design and path of steepest ascent in submerged fermentation

Safe production of Aspergillus terreus xylanase from Ricinus communis: gene identification, molecular docking, characterization, production of xylooligosaccharides, and its biological activities

BackgroundThe production of industrial enzymes such as xylanase using sufficient cost-effective substrates from potent microorganisms is considered economically feasible. Studies have reported castor cake (Ricinus communis) as the most potent and inexpensive alternative carbon source for production of xylanase C by using Aspergillus terreus (A. terreus). ResultsA. terreus strain RGS Eg-NRC, a local isolate from agro-wastes, was first identified by sequencing the internal transcribed spacer region of a nuclear DNA encoding gene cluster deposited in GenBank (accession number MW282328). Before optimization of xylanase production, A. terreus produced 20.23 U/g of xylanase after 7 days using castor cake as a substrate in a solid-state fermentation (SSF) system that was employed to achieve ricin detoxification and stimulate xylanase production. Physicochemical parameters for the production of xylanase were optimized by using a one-variable-at-a-time approach and two statistical methods (two-level Plackett–Burman design and central composite design, CCD). The maximum xylanase yield after optimization was increased by 12.1-fold (245 U/g). A 60–70% saturation of ammonium sulfate resulted in partially purified xylanase with a specific activity of 3.9 IU/mg protein. At 60 °C and pH 6, the partially purified xylanase had the highest activity, and the activation energy (Ea) was 23.919 kJmol. Subsequently, antioxidant capacity and cytotoxicity tests in normal Ehrlich ascites carcinoma human cells demonstrated xylooligosaccharides produced by the xylanase degradation of xylan as a potent antioxidant and moderate antitumor agent. Further investigations with sodium dodecyl sulfate polyacrylamide gel electrophoresis then determined the molecular weight of partially purified xylanase C to be 36 kDa. Based on the conserved regions, observations revealed that xylanase C belonged to the glycosyl hydrolase family 10. Next, the xylanase-encoding gene (xynC), which has an open reading frame of 981 bp and encodes a protein with 326 amino acids, was isolated, sequenced, and submitted to the NCBI GenBank database (accession number LC595779.1). Molecular docking analysis finally revealed that Glu156, Glu262, and Lys75 residues were involved in the substrate-binding and protein-ligand interaction site of modeled xylanase, with a binding affinity of −8.7 kcal. mol−1. ConclusionThe high production of safe and efficient xylanase could be achieved using economical materials such as Ricinus communis.

Optimization of xylanase production from Aspergillus tamarii SCBH2 using response surface methodology

Optimization of xylanase production from Aspergillus tamarii SCBH2 using response surface methodology

Optimization of Xylanase Production from Aspergillus spp. under Solid-state Fermentation Using Lemon Peel as Substrate

Xylanase is an enzyme that acts on xylan and degrades it into xylose. Xylanase may be produced by several different microbes such as bacteria and fungi. Xylanase has significant applications in food industries for improving dough handling and improving the quality of baked products in the extraction of starch, coffee, and plant oils. Xylanase, along with pectinase and cellulose, has enormous application in the clarification of fruit juices. In the present study, xylanase was produced by solid-state fermentation using lemon peel as substrates. Aspergillus Niger was induced to produce xylanase by frequent sub-culturing on a medium containing 2% xylan. OFAT was analyzed by using several fermentation parameters such as moisture content, particle size, incubation period, incubation temperature, peptone concentration, and pH of the extraction buffer. Under optimized conditions, the maximum xylanase activity was found for particle size of 1.7mm, moisture content of 80%, peptone concentration in nutrient solution at 0.3%, and extraction pH of 7.0. Hence, xylanase can be further subjected to down streaming processes and therefore for various applications.

Characterization of recombinant Bacillus halodurans CM1 xylanase produced by Pichia pastoris KM71 and its potential application in bleaching process of bagasse pulp

Thermoalkalophilic xylanases promise potential application in pulp biobleaching to reduce the use of toxic chlorinated chemical agents, which are harmful to the environment. In this study, a thermoalkalophilic endoxylanase gene (bhxyn3) originating from Indonesian indigenous Bacillus halodurans CM1 was cloned into yeast expression vector pPICZα A and expressed in Pichia pastoris KM71 under the control of AOX1 promoter. Recombinant P. pastoris expressed the highest final level of xylanase (146 U/mL) on BMGY medium after five days of cultivation. Optimization of xylanase production on a small scale was carried out by varying the methanol concentrations and the optimal xylanase production by the recombinant P. pastoris was observed in the culture with 2% (v/v) methanol after four days of the induction phase. The recombinant xylanase (BHxyn3E) was thermotolerant and alkalophilic, with an optimal temperature at around 55‐65 °C and under pH 8.0. The enzyme activity was slightly induced by K+, Fe2+, and MoO42‐. Enzymatic bleaching of bagasse pulp with no prior pH adjustment (pH 9) using BHxyn3E at 200 U/g oven dried pulp increased the lightness index (L*) and changed substantially the color a index (a*); however, the treatments did not change the whiteness index in a significant way. Therefore, further optimization and assessment such as adjustment of incubation temperature and pH in biobleaching were needed to reduce the use of harmful chemical agents in industrial applications.

Optimization of xylanase production by Bacillus tequilensis strain UD-3 using economical agricultural substrate and its application in rice straw pulp bleaching

Thermo-alkali-stable xylanase producing Bacillus tequilensis strain UD-3 was isolated from the hot spring, and the biobleaching effect on rice straw pulp was evaluated. Bacillus tequilensis strain UD-3 produced xylanase (8.54 IU mL−1) employing on wheat bran. The xylanase production was increased about 2 fold after optimization of media via central composite design. The maximum xylanase activity was achieved at 8.0 pH, 50 °C temperature, and boosted at 1.0 mM concentration of metal ions such as Mn+2, Fe+3, Cu+4, and Zn+4. Furthermore, Zn+4 concoction with xylanase was evaluated for pre-bleaching of rice straw pulp in which xylanase significantly reduce the Kappa number (17.24 ± 0.02) after Xyl + Zn bleaching. A significant reduction was also observed in the reducing sugars (50%), phenolic (29.19%), hydrophobic (33.20%), and lignin compounds (35.86 and 40.48%) during the Xyl + Zn treatment of pulp samples in comparison to the only xylanase.

Optimization of Xylanase Production by Streptomyces costaricanus 45I-3 Using Various Substrates through Submerged Fermentation

Xylanase is an important hydrolytic enzymes with many application in several industries, but to obtain enzyme derived products is not easy. Thus, the optimization of efficient xylanases production is a great interest for biotechnological application. This study aims to determine the type of substrate, medium composition, and optimum conditions of xylanase production by S. costaricanus 45I-3. Determination of substrate type was done by growing the tested bacteria on birchwood xylan, beechwood xylan, oat spelled xylan, corn cobs xylan, and tobacco xylan substrate, meanwhile the determination of medium composition and enzyme production were done by measuring xylanase activity at various substrate concentration and replacing the carbon, nitrogen, phosphate and surfactants source. The results showed that the highest enzymatic index (EI) produced from corn cob xylan substrate at 3.60 meanwhile the second highest was beechwood xylan substrate at 2.87 EI, however this substrate is purer, thus this substrate was selected and used as xylan sources for further optimization measurement. The best xylanase activity (2.29 U/mL) obtained on eighth day after inoculation on rotary incubator at 120 rpm in 28 ºC. Arabinose as the source of carbon generate the highest activity at 3.161 U/mL meanwhile the most preferred source of phosphate is Na2HPO4 (2.37 U/mL). Both source of nitrogen i.e. nitrogen ammonium sulphate (NH4)2SO4 and yeast extract were able to produce xylanase at 2.57 and 2.36 U/mL. The addition of surfactant in production medium showed addition of SDS surfactant (0.146 U/mL) and Tween 80 (0.438 U/mL) showed a negative response by decreasing the activity. The conclusion showed that the xylanase activity was increased after optimization at various C, N, and P sources, and the use of nitrogen source (NH4)2SO4), become a more economical alternative to replacing a nitrogen source yeast extract so it can lower the production costs of xylanase enzyme.

Optimization of xylanase production using ragi (Eleusine coracana) husk as a substrate by Aspergillus fumigatus JCM 10253 through response surface methodology

Hydrolytic enzymes play a significant role in hydrolyzing lignocellulosic polysaccharides into fermentable sugars. Due to the complex structure of lignocellulosic materials, developing robust processes for high titer production is a major challenge. The present study was investigated on xylanase production by filamentous fungi Aspergillus fumigatus strain JCM 10253 using agricultural residue ragi husk as a substrate for microbial fermentation. Central composite design (CCD) was employed to optimize the independent process variables and their effect on xylanase activity. Experimental results with optimized process variables showed 156.7 IU/mL activity under 1.24% substrate concentration and pH 2.0 at 49.9 °C after 120 h of the incubation period. The morphological study of isolate JCM 10253 was observed by scanning electron microscopy analysis and Fourier transform infrared spectroscopy was used to characterize the crude enzyme for secondary protein helical structures. Thus, this study demonstrates that ragi husk could be a potential substrate for xylanase production and can be further used as value-added industrial products.

Effect of optimization conditions on submerged fermentation of corn bran for the production of xylanase enzyme

UV- rays were used as mutagens on wild type of Aspergillus niger for enhanced production of Enzyme – Xylanase. The xylanase producing species were isolated from soil and exposed to UV rays at various timings of 20 mins, 40 mins, 60 mins, 80 mins, 100 mins and 120 mins. The mutants AN20 – AN100 showed 19.96, 21.22, 22.66, 27.88, and 37.77 % increase in activities of cellular xylanase respectively in comparison with the parental strain (0.556± 0.06). Mutant AN100 exposed to UV ray for 100 minutes was therefore selected as the best producer of xylanase after 7 days of fermentation using Corn bran as a substrate. As seen in the result, increase in time of exposure resulted to a decrease in cellular xylanase production. Optimization of xylanase production was carried out and the result obtained revealed that mutant AN100 showed best xylanase activity when cultivated at PH 5, 30 °C and without addition of any other Carbon or Nitrogen source. Aspergillus niger is a good producer of xylanase under the conditions determined in this assay

Optimization of xylanase production from penicillium funiculosum using agricultural (Corn cob) waste

Optimization of xylanase production from <i>penicillium funiculosum</i> using agricultural (Corn cob) waste

Assessment and optimization of xylanase production using co-cultures of Bacillus subtilis and Kluyveromyces marxianus.

Co-cultures of Bacillus subtilis and Kluyveromyces marxianus were investigated in submerged fermentation for xylanase production at shake-flask scale. Xylanase production markedly increased when arabinose, xylose, or hazelnut shells were used as the single carbon source. Maximal xylanase of 49.5IU/mL was achieved with 4% B. subtilis, 4% K. marxianus, 40% solid load of hazelnut shells, and pH 7.0. Overall, xylanase was enhanced by 4.4-fold compared to initial un-optimized monoculture production and 2.8-fold compared to initial un-optimized co-cultured production, after optimization by response surface method.

Optimization of Xylanase Production by Trichoderma orientalis Using Corn Cobs and Wheat Bran via Statistical Strategy

In order to obtain the best conditions for xylanase production, statistical experimental designs were applied for improvement of xylanase yield by Trichoderma orientalis EU7-22 using corn cobs and wheat bran as raw materials. Plackett–Burman design was applied to evaluate the effects of eight variables (concentration of peptone, tween-80, CaCl2, MgSO4, FeSO4, as well as initial pH, fermentation temperature, fermentation time). The key factors influencing on xylanase production were identified as the fermentation time, concentration of MgSO4, and fermentation temperature. And then the path of steepest ascent was carried out to approach the optimal region of the three significant factors. These variables were subsequently further investigated by Box–Behnken design of response surface methodology. The optimum conditions for xylanase production were obtained as fermentation time 72 h, concentration of 0.08% for MgSO4, fermentation temperature 37.3 °C, and the xylanase activity increased from 107.6 to 269.4 IU/mL, which was a 150% increase. It was found that the higher temperature (37.3 °C) was more suitable for xylanase production than low temperature (30.0 °C) in T. orientalis EU7-22. The statistical experimental design was proven as an effective method to obtain optimum parameters for xylanase production. The mixture of corn cobs and wheat bran as substrates could greatly improve the xylanase yield, in comparison with corn cobs alone, wheat bran alone, and Avicel supplement with wheat bran as inducer substrates, respectively.

Optimization of Xylanase Production from Aspergillus Flavus in Solid State Fermentation Using Agricultural Waste as a Substrate

Optimization of Xylanase Production from Aspergillus Flavus in Solid State Fermentation Using Agricultural Waste as a Substrate

Optimization of Xylanase Production from Some Agro-lignocellulosic Wastes by Aspergillus niger Via Solid State Fermentation

Extracellular xylanase production by A. niger was studied under solid-state fermentation using six agricultural wastes individually (wheat bran, rice husk, corn cobs, rice straw, clover straw and wheat straw) as well as fifteen treatment represent different combinations of the above agro-lignocellulosic wastes. The best substrate was the combination of wheat bran and clover straw. The initial moisture content, additive carbon source, additive nitrogen source, initial pH of the culture, incubation temperature and inoculum size affected the production of xylanase. The optimal fermentation medium was at 80% of initial moisture and composed of 5 g WB+CS (1:1 w/w) supplemented with 1% (w/w) of xylan and 1% (w/w) of (NH4)3PO4 and inoculated by adding 1 ml of spore suspension containing 2x105spores/ml. Xylanase production reached a peak of 1294.15 IU/gdw after 7 days of incubation at 35 °C under SSF. The produced enzyme has maximum activity at 70 oC and pH 5.5. Addition of 10 mM FeSO4 and 5mM of KCl, CaCl2 and MnCl2 to the reaction mixture enhanced xylanase activity to 127.46, 118.86, 115.76 and 111.15%, respectively. A. niger xylanase showed superior heat and pH tolerance, it remained 86.47 and 76% of its activity after 60 min of incubation at 70 oC with and without 10 mM CaCl2.The enzyme showed more stability in a pH ranged from 4 to 5 retaining more than 75% of its original activity, therefore it may have significant applications in biofuel and paper industries.

Batch Submerged Fermentation in Shake Flask Culture and Bioreactor: Influence of Different Agricultural Residuals as the Substrate on the Optimization of Xylanase Production by Bacillus subtilis and Aspergillus brasiliensis

Batch Submerged Fermentation in Shake Flask Culture and Bioreactor: Influence of Different Agricultural Residuals as the Substrate on the Optimization of Xylanase Production by Bacillus subtilis and Aspergillus brasiliensis

Optimization of Xylanase Production through Response Surface Methodology by Fusarium sp. BVKT R2 Isolated from Forest Soil and Its Application in Saccharification.

Xylanses are hydrolytic enzymes with wide applications in several industries like biofuels, paper and pulp, deinking, food, and feed. The present study was aimed at hitting at high yield xylanase producing fungi from natural resources. Two highest xylanase producing fungal isolates—Q12 and L1 were picked from collection of 450 fungal cultures for the utilization of xylan. These fungal isolates—Q12 and L1 were identified basing on ITS gene sequencing analysis as Fusarium sp. BVKT R2 (KT119615) and Fusarium strain BRR R6 (KT119619), respectively with construction of phylogenetic trees. Fusarium sp. BVKT R2 was further optimized for maximum xylanase production and the interaction effects between variables on production of xylanase were studied through response surface methodology. The optimal conditions for maximal production of xylanase were sorbitol 1.5%, yeast extract 1.5%, pH of 5.0, Temperature of 32.5°C, and agitation of 175 rpm. Under optimal conditions, the yields of xylanase production by Fusarium sp. BVKT R2 was as high as 4560 U/ml in SmF. Incubation of different lignocellulosic biomasses with crude enzyme of Fusarium sp. BVKT R2 at 37°C for 72 h could achieve about 45% saccharification. The results suggest that Fusarium sp. BVKT R2 has potential applications in saccharification process of biomass.

Characterization and valuable applications of xylanase from endophytic fungus Aspergillus terreus KP900973 isolated from Corchorus olitorius

Abstract Isolated fungal strains from molokhia stalks (Corchorus olitorius) were screened for the production of xylanase enzyme. Aspergillus terreus KP900973 was selected as potential xylanase producer and was identified on the basis of 18S rDNA gene homology. Screening was done by Congo red test for xylanase, based on the light coloured zone around the sample in xylan agar plates. Optimization of xylanase production was performed using low cost substrates through submerged fermentation (SMF). Using a mixture of untreated molokhia stalks and pea peel with no nutrients added enhanced enzyme production by 100.73%. Optimal assay temperature and pH were 50 °C and 5.5, respectively. At pH 4.0 the activity decreased to 91.23%, moreover, at pH 9.0 the enzyme had 67.25% of its activity. The enzyme retained about 97% of its activity after being stored for 46 days at −18 °C. A. terreus is free from tested mycotoxins and considered as a non-toxigenic strain, so, some industrial applications of crude xylanase were tested. Wheat bran was found to be the most susceptible substrate for hydrolysis with the highest level of fermentable sugars and saccharification yield (16%) after 48 h. The greatest clarification degree of orange juice (64%) was observed after 72 h using crude enzyme preparation. Xylanase can even substitute the addition of emulsifiers and additives used in bread production enhancing dough-raising as compared to the control about 2-fold.

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