Characterization and high expression of recombinant Ustilago maydis xylanase in Pichia pastoris.
A recombinant xylanase gene (rxynUMB) from Ustilago maydis 521 was expressed in Pichia pastoris, and the enzyme was purified and characterized. Phylogenetic analysis demonstrated that rxynUMB belongs to glycosyl hydrolase family 11. The Trp84, Trp95, Glu93, and Glu189 residues are proposed to be present at the active site. The apparent molecular mass of the recombinant xylananse was approximately 24 kDa, and the optimum pH and temperature were 4.3 and 50 °C, respectively. Xylanase activity was enhanced by 166 and 115% with Fe(2+) and Mn(2+), respectively. The biochemical properties of this recombinant xylanase suggest that it may be a useful candidate for a variety of commercial applications.
Shanghai Key Laboratory of Agricultural Genetics and Breeding, Agro-Biotechnology Research Institute, Shanghai Academy of Agricultural Sciences, 2901 Beidi Rd, Shanghai, 201106, People's Republic of China, hhj2006@163.com.
Hongjuan
H
Han
Shuang
S
You
Bo
B
Zhu
Xiaoyan
X
Fu
Baihui
B
Sun
Jin
J
Qiu
Chengye
C
Yu
Lei
L
Chen
Rihe
R
Peng
Quanhong
Q
Yao
D000595:Amino Acid Sequence; D020134:Catalytic Domain; D002413:Cations, Divalent / Q000378:metabolism; D020536:Enzyme Activators / Q000378:metabolism; D004795:Enzyme Stability; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D007501:Iron / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010802:Phylogeny; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013696:Temperature; D014588:Ustilago / Q000201:enzymology* / Q000235:genetics; D014995:Xylosidases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D002413:Cations, Divalent; D020536:Enzyme Activators; D011994:Recombinant Proteins; D008345:Manganese; D007501:Iron; D014995:Xylosidases