title: Extracellular expression of alkali tolerant xylanase from Bacillus subtilis Lucky9 in E. coli and application for xylooligosaccharides production from agro-industrial waste.
abstract: An alkali tolerant xylanase gene from Bacillus subtilis Lucky9 was cloned and extracellular expressed in E. coli BL21. Xylanase amino acid sequence showed 99% identity with xylanase sequence from Bacillus subtilis 168, and was belonged to glycoside hydrolase family 11. The recombinant E. coli (pET-pelB-xynLC9) containing pelB signal peptide produced extracellular xylanase of 436.5U/mL for 8h, which was used arabinose as extra carbon source and inducer for enhancing extracellular production. The extracellular xylanase was determined by SDS-PAGE with a relative molecular mass of 21kDa. The recombinant xylanase was optimally activity at pH 6.5 and 60°C. The xylanase exhibited 80% residual activity over a broad pH range of 6.0-9.0 for 24h. Thermostability studies showed that xylanase retained 60% residual activity after 2h at 60°C. The main end-products of hydrolysis of beech-wood xylan and corncob by the extracellular xylanase were xylobiose and xylotriose. This extracellular xylanase without purification is a suitable candidate for application in the industrial production of xylooligosaccharides from agro-industrial waste for use as prebiotics.
authors.affiliation: College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, 30 Puzhunan Road, Nanjing 211816, Jiangsu, China.
authors.forename: Siyuan
authors.initials: S
authors.lastname: Chang
authors.affiliation: College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, 30 Puzhunan Road, Nanjing 211816, Jiangsu, China.
authors.forename: Yalan
authors.initials: Y
authors.lastname: Guo
authors.affiliation: College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, 30 Puzhunan Road, Nanjing 211816, Jiangsu, China; Jiangsu National Synergetic Innovation Center for Advanced Materials, 30 Puzhunan road, Nanjing 211816, Jiangsu, China. Electronic address: wubin1977@njtech.edu.cn.
authors.forename: Bin
authors.initials: B
authors.lastname: Wu
authors.affiliation: Jiangsu National Synergetic Innovation Center for Advanced Materials, 30 Puzhunan road, Nanjing 211816, Jiangsu, China; School of Pharmaceutical Sciences, Nanjing Tech University, 30 Puzhunan Road, Nanjing 211816, Jiangsu, China.
authors.forename: Bingfang
authors.initials: B
authors.lastname: He
mesh_terms: D000383:Agriculture; D000468:Alkalies / Q000494:pharmacology*; D001089:Arabinose / Q000494:pharmacology; D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D043364:Endo-1,4-beta Xylanases / Q000235:genetics* / Q000378:metabolism; D004795:Enzyme Stability / Q000187:drug effects; D004926:Escherichia coli / Q000166:cytology / Q000187:drug effects / Q000235:genetics*; D005110:Extracellular Space / Q000187:drug effects / Q000235:genetics; D029964:Fagus / Q000737:chemistry; D015870:Gene Expression; D005965:Glucuronates / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007220:Industrial Waste; D008670:Metals / Q000494:pharmacology; D009844:Oligosaccharides / Q000737:chemistry*; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity; D013696:Temperature; D003313:Zea mays / Q000737:chemistry
chemical_list: D000468:Alkalies; D005965:Glucuronates; D007220:Industrial Waste; D008670:Metals; D009844:Oligosaccharides; C570991:xylooligosaccharide; D001089:Arabinose; D043364:Endo-1,4-beta Xylanases
keywords: Alkali stable; Arabinose; Corncob; Extracellular expression; Xylanase; Xylooligosaccharides