title: High-level expression of the Penicillium notatum glucose oxidase gene in Pichia pastoris using codon optimization.
abstract: The glucose oxidase (GOD) gene from Penicillium notatum was expressed in Pichia pastoris. The 1,815 bp gene, god-w, encodes 604 amino acids. Recombinant GOD-w had optimal activity at 35-40°C and pH 6.2 and was stable, from pH 3 to 7 maintaining >75% maximum activity after incubation at 50°C for 1 h. GOD-w worked as well as commercial GODs to improve bread making. To achieve high-level expression of recombinant GOD in P. pastoris, 272 nucleotides involving 228 residues were mutated, consistent with the codon bias of P. pastoris. The optimized recombinant GOD-m yielded 615 U ml(-1) (2.5 g protein l(-1)) in a 3 l fermentor--410% higher than GOD-w (148 U ml(-1)), and thus is a low-cost alternative for the bread baking industry.
authors: Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China.
authors: Zhaowei
authors: Z
authors: Gao
authors: Zhuofu
authors: Z
authors: Li
authors: Yuhong
authors: Y
authors: Zhang
authors: Huoqing
authors: H
authors: Huang
authors: Mu
authors: M
authors: Li
authors: Liwei
authors: L
authors: Zhou
authors: Yunming
authors: Y
authors: Tang
authors: Bin
authors: B
authors: Yao
authors: Wei
authors: W
authors: Zhang
mesh_terms: D003062:Codon; D004795:Enzyme Stability; D015870:Gene Expression; D005949:Glucose Oxidase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010408:Penicillium chrysogenum / Q000201:enzymology* / Q000235:genetics; D010843:Pichia / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013696:Temperature
chemical_list: D003062:Codon; D011994:Recombinant Proteins; D005949:Glucose Oxidase