Background and Objectives: Research on Xylanase has markedly increased due to its potential applications in pulping and bleaching processes using cellulose free preparations, textile processes, the enzymatic saccharification of lignocellulosic materials and waste treatment. The present study was aimed at isolation and characterization of xylan degrading strain of Bacillus cereus from soil for production of xylanase. Materials and Methods: Twelve isolates were obtained from soil samples of different areas in the Rajshahi University campus and studied for detection of xylanase activity. One of the strains was identified as Bacillus cereus on the basis of the nucleotide sequence of the 16S rRNA gene which produces xylanase extracellularly. We purified xylanase to homogeneity by a combination of ammonium-sulphate precipitation, DEAE-sepharose, Phenyl-5PW and Hydroxyapatite column chromatography using culture supernatant.Results: The SDS-PAGE gave a single band at 32 kDa. The optimum temperature and pH of the purified enzyme was 40°C and 6.0, respectively. The xylanase hydrolyzed oat spelt xylan, birch wood xylan and beech wood xylan efficiently but showed no activity towards cellulose, CM-cellulose and Avicell pH 101.Conclusion: Thus it was a true and neutral xylanase. The isolation of xylanase from Bacillus cereus is rare.

Crude fiber (CF) is a vital component in poultry nutrition with a notable phytonutrient effectively indicating the presence of indigestible biomass in food due to the absence of digestive enzymes for CF in broilers. This study aimed to analyze the properties of a multi-enzyme cocktail (MEC) Bacillus sonorensis BD92 (BsBD92) comprised of xylanase, β-glucosidase, exo-glucanase, and endo-glucanase enzymes. Also, this study intended to look at the growth performance and intestinal histology of broilers in the starter and finisher phases by the addition of MEC BsBD92 to their diet. To evaluate the efficacy of MEC BsBD92, 140 one-day-old unsexed Cobb500 broiler chicks were randomly divided into seven groups receiving different diets. The characterization of exo-glucanase, xylanase, β-glucosidase, and endo-glucanase showed that their peak activities were observed at a temperature of 50.00 ˚C and a pH of 5.50. The 6.00% CF and 2.00 X MEC BsBD92 improved the intestinal morphology and feed conversion ratio, demonstrating a synergistic effect on growth performance. Whereas, increasing meat percentages to 61.06 and 65.09 g per 100 g body weight during the starter and finisher phases was also observed, respectively. The lipid profiles revealed significant variations in triglyceride and cholesterol levels. This study provides an innovative approach, considering not only lowering the feed cost using inexpensive fibrous feedstuffs but also improving the feed efficiency through supplementation of MEC BsBD92.

The thermostable and alkaliphile xylanases are beneficial industrial enzymes in food industry (bakery and alcohol production or fermentation industries) pulp-kraft, and pharmaceutic industry. Xylanases hydrolyse xylan (the hemicellulose of plant cell wall) and produced by different kinds of microorganisms like bacteria, fungi and some algae. In this study, xylanase gene from thermoalkaline Anoxybacillus flavithermus was isolated from hot spring of Meshkinshahr (Iran) and was cloned in E. coli (BL21). At first, total DNA was extracted from bacteria. PCR was then accomplished and xylanase gene was isolated and amplified. Amplification of xylenase coding gene was confirmed by electrophoresis. Cloning was performed by digestion of xylanase gene and pET21a+ by restriction enzymes (NheI and XhoI), following by ligation and transformation to E. coli (BL21). Finally, the results were confirmed by colony-PCR and sequencing of selected colonies. PCR and sequencing results, demonstrated a high similarity between xylanases of Anoxybacillus species with xylanase of this study.

Background and Objective: Xylan the major portion of the hemicellulose of plant cell walls are heterogeneous polysaccharides. Xylanases (EC:3.2.1.8) are enzymes obtained from different species of microorganisms that degrade the xylosidic linkages of xylan’s backbone producing xylose with other monoresidues. In recent years, xylanases have many application in industry such as paper biobleaching and oil clearing.Materials and methods: In this study xylanase enzyme was purified from culture supernatant of Bacillus stearothermophilus (ATCC 12980) by ammonium sulfate precipitation, gel filtration on sephadex G- 100 followed by ion exchange chromatography on DEAE-cellulose. Also, the effect of pH and temperature on purified xylanase activity was studied.Results: In DEAE- cellulose column chromatography, three protein peaks F-1a, F-1b and F-1c were appeared. Among these peaks, only F-1b showed xylanase activity and the degree of purification attained 63.09 fold. The specific activity and purification fold of the purified xylanase was 87.7U/mg of protein and 17.45, respectively. The enzyme gave maximum activity against xylan as substrate at pH 7.0 and temperature at 60°C. In paper chromatography xylose was detected as the hydrolysis products of oat- spelt xylan by the xylanase at 16 h. These results indicate that xylanase of Bacillus stearothermophilus (ATCC 12980) was endoxylanase.Conclusion: These data includes, optimal pH and temprature suggested that purified xylanase can be suitable for industrial applications.

Filamentous fungi Trichoderma spp. are efficient hyper-producers of industrially important enzymes. Xylanases are essential hemicellulases having vast ranges of applications in the various industrial sectors. In this study, the protoplast fusion of two T. virens and T. harzianum strains resulted in 2. 5 Uml-1 xylanase activity in fusant X3 showing 4. 7-fold improvement in xylanase activity compared to that of parents with 0. 54 Uml-1. Moreover, to evaluate the influence of protoplast fusion on the xylanase activity, the expression patterns of the xylanase gene xyn3 was analyzed in the parental strains and the using qPCR. The results demonstrated that the relative expression of the xyn3 increased in X3 by 4. 9 fold compared to that of the parents. Finally, based on the results, it could be concluded that the protoplast fusion technology is a promising approach to generate new superior fungi with high production ability of the industrially important enzymes.

Xylanase enzymes produced by Trichoderma afroharzianum have significant industrial applications, including animal feed, food, biofuel, and textile industries. Creating novel sources of Trichoderma strains using induced gamma irradiation mutation can increase enzyme production. According to this, Co-60 gamma irradiation has been used to develop a mutant strain of T.afroharzianum. T.afroharzianum mutants were isolated, and qualitative and quantitative screening were used to evaluate the production of the extracellular enzymes with wheat bran waste as a substrate. The best T.afroharzianum mutant strain was identified using the DNA barcoding method. The highest xylanase activities were observed in the superior mutant of T.afroharzianum NAS107-M82, which is approximately 3.3 times higher than its parent strain. The electrophoretic pattern of proteins showed that synergistically, the exo-glucanase I, endo-glucanase III, and the xylanase I enzymes hydrolyzed the wheat bran. This study collectively highlights the diverse potential of gamma-radiated T. afroharzianum mutant-derived xylanases in various industrial and agricultural applications, showcasing their importance in enzyme production.

Xylan the major portion of the hemicellulose of plant cell walls are heterogeneous polysaccharides. Xylanases (EC:3.2.1.8) are enzymes obtained from different species of microorganisms that degrade the xylosidic linkages of xylan's backbone producing xylose with other monoresidues. In this study xylanase enzyme was purified from culture supernatant of Bacillus stearothermophilus (ATCC 12980) by ammonium sulfate precipitation, gel filtration on sephadex G-100 followed by ion exchange chromatography on DEAE-cellulose. In DEAE- cellulose column chromatography, three protein peaks F-1a, F-1b and F-1c were appeared. Among these peaks, only F-1b showed xylanase activity and the degree of purification attained 63.09 fold. The specific activity and purification fold of the purified xylanase was 87.7 U/mg of protein and 17.45, respectively. The enzyme gave maximum activity against xylan as substrate at pH 7.0 and temperature at 60oC. In paper chromatography xylose was detected as the hydrolysis products of oat-spelt xylan by the xylanase at 16 h. These results indicate that xylanase of Bacillus stearothermophilus (ATCC 12980) was endoxylanase. In conclusion, these data suggested that purified xylanase can be suitable for industrial applications.

Introduction: b-D-Xylosidases (EC 3.2.1.37), one of the most important hemicellulases with high potential industrial application such as bioethanol production, are exo-type glycosidases that catalyze the successive removal of b-xylosyl residues from the non-reducing termini of xylobiose and higher linear b- 1, 4-xylooligosaccharides and in conjunction with b-xylanases are essential in completely depolymerizing xylan. The aim of this study was secretive expression of bacterial b-xylosidase gene including hexahistidintag in Pichia pastoris in order to achieve high level expression and enzyme purification subsequently.Materials and Methods: In this study, after optimizing the sequence of protein encoding bacterial b- xylosidase based on the expression codon usage of Pichia pastoris and addition of 6xHis-tag sequence through suitable designed primers to gene, transformation of recombinant plasmids was performed through electroporation method into the expression yeast.Results: The results showed that the recombinant b-xylosidase production was 40 U/L with 0.6 U/mg specific activity in flask culture.Conclusion: Cloning and successful expression of bacterial b-xylosidase gene was carried out in order to obtain an active and stable enzyme with high level expression from yeast expression system.