title: High expression of recombinant Streptomyces sp. S38 xylanase in Pichia pastoris by codon optimization and analysis of its biochemical properties.
abstract: In recent years, the biotechnological use of xylanases has grown remarkably. To efficiently produce xylanase for food processing and other industry, a codon-optimized recombinant xylanase gene from Streptomyces sp. S38 was synthesized and extracellularly expressed in Pichia pastoris under the control of AOX1 promoter. SDS-PAGE and activity assay demonstrated that the molecular mass of the recombinant xylanase was estimated to be 25 kDa, the optimum pH and optimum temperature were 5.5 and 50°C, respectively. In shake flask culture, the specific activity of the xylanase activity was 5098.28 U/mg. The K ( m ) and V ( max ) values of recombinant xylanase were 11.0 mg/ml and 10000 μmol min(-1) mg(-1), respectively. In the presence of metal ions such as Ca(2+), Cu(2+), Cr(3+) and K(+), the activity of the enzyme increased. However, strong inhibition of the enzyme activity was observed in the presence of Hg(2+). This is the first report on the expression properties of a recombinant xylanase gene from the Streptomyces sp. S38 using Pichia pastoris. The attractive biochemical properties of the recombinant xylanase suggest that it may be a useful candidate for variety of commercial applications.
authors.affiliation: Agro-Biotechnology Research Institute, Shanghai Academy of Agricultural Sciences, 2901 Beidi Rd, Shanghai 201106, China.
authors.forename: Xiao-Yan
authors.initials: XY
authors.lastname: Fu
authors.forename: Wei
authors.initials: W
authors.lastname: Zhao
authors.forename: Ai-Sheng
authors.initials: AS
authors.lastname: Xiong
authors.forename: Yong-Sheng
authors.initials: YS
authors.lastname: Tian
authors.forename: Ri-He
authors.initials: RH
authors.lastname: Peng
mesh_terms: D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D043364:Endo-1,4-beta Xylanases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004795:Enzyme Stability / Q000187:drug effects; D015870:Gene Expression / Q000187:drug effects; D005822:Genetic Vectors / Q000235:genetics; D006031:Glycosylation / Q000187:drug effects; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D007477:Ions; D007700:Kinetics; D008670:Metals / Q000494:pharmacology; D008967:Molecular Biology / Q000379:methods*; D010843:Pichia / Q000187:drug effects / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*; D011995:Recombination, Genetic / Q000187:drug effects / Q000235:genetics; D013302:Streptomyces / Q000187:drug effects / Q000201:enzymology*; D013696:Temperature
chemical_list: D003062:Codon; D007477:Ions; D008670:Metals; D011994:Recombinant Proteins; D043364:Endo-1,4-beta Xylanases