Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China.
Biotechnol Lett. 2012 Mar;34(3):507-14. doi: 10.1007/s10529-011-0790-6. Epub 2011 Nov 4.
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.
葡萄糖氧化酶（GOD）基因来自青霉被表达在巴斯德毕赤酵母中。1815bp 的基因，god-w，编码 604 个氨基酸。重组 GOD-w 在 35-40°C 和 pH6.2 时具有最佳活性，并且在 50°C 孵育 1 小时后，在 pH3 到 7 的范围内保持超过 75%的最大活性。GOD-w 像商业 GOD 一样能够改善面包制作。为了在巴斯德毕赤酵母中实现重组 GOD 的高水平表达，对涉及 228 个残基的 272 个核苷酸进行了突变，与巴斯德毕赤酵母的密码子偏爱一致。优化后的重组 GOD-m 在 3L 发酵罐中产生 615Uml(-1)（2.5g 蛋白 l(-1)）-比 GOD-w（148Uml(-1)）高 410%，因此是面包烘焙行业的低成本替代品。
