High-level expression of improved thermo-stable alkaline xylanase variant in Pichia Pastoris through codon optimization, multiple gene insertion and high-density fermentation.
High-level expression of improved thermo-stable alkaline xylanase variant in Pichia Pastoris through codon optimization, multiple gene insertion and high-density fermentation.
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DOI：
10.1038/srep37869
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发表时间：
2016-11-29
影响因子：
4.6
通讯作者：
Ma Y
中科院分区：
文献类型：
--
作者：
Lu Y;Fang C;Wang Q;Zhou Y;Zhang G;Ma Y
在造纸工业中，木聚糖酶因其环保特性而被用来提高漂白过程中的纸浆性能。造纸工业中木聚糖酶的活性受到高温和碱性条件的阻碍，酶生产成本较高。在此，XynHB（一种来自短小芽孢杆菌 HBP8 的碱性稳定木聚糖酶）在 N188A 处突变为 XynHBN188A。在大肠杆菌中表达的突变体在 60°C 下表现出比 XynHB 高 1.5 倍的木聚糖酶活性。在毕赤酵母中表达的突变体被糖基化，在60°C下保持稳定30分钟。基于巴斯德毕赤酵母 (xynHBN188As) 密码子使用偏好进行优化的 XynHBN188A 显示酶活性增加了 39.5%。在木聚糖平板中形成最大水解晕的菌株Y16用于摇瓶实验，产生的酶活性为6,403U/ml。 qPCR显示，Y16菌株的基因组中有9个拷贝的重组xynHBN188As基因。在 5 L 发酵罐中，酶活性增加至 48,241 U/ml。漂白实验中添加15 U/g木聚糖酶可使纸制品白度提高2%，从而使拉伸强度和耐破度分别提高13%和6.5%。 XynHBN188As在造纸工业中具有巨大的潜力。
In paper industry, xylanases are used to increase the pulp properties in bleaching process as its eco-friendly nature. The xylanases activity is hindered by high temperature and alkaline conditions with high enzyme production cost in the paper industry. Here, XynHB, an alkaline stable xylanase from Bacillus pumilus HBP8 was mutated at N188A to XynHBN188A. Expressed mutant in E. coli showed 1.5-fold higher xylanase activity than XynHB at 60 °C. The mutant expressed in Pichia pastoris was glycosylated, remained stable for 30 min at 60 °C. XynHBN188A optimized based on codon usage bias for P. pastoris (xynHBN188As) showed an increase of 39.5% enzyme activity. The strain Y16 forming the largest hydrolysis halo in the xylan plate was used in shake flask experiments produced an enzyme activity of 6,403 U/ml. The Y16 strain had 9 copies of the recombinant xynHBN188As gene in the genome revealed by qPCR. The enzymatic activity increased to 48,241 U/ml in a 5 L fermentor. Supplement of 15 U/g xylanase enhanced the brightness of paper products by 2% in bleaching experiment, and thereby improved the tensile strength and burst factor by 13% and 6.5%, respectively. XynHBN188As has a great potential in paper industries.
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DOI：
10.1093/protein/15.2.141
发表时间：
2002-02-01
期刊：
PROTEIN ENGINEERING
影响因子：
--
作者：
Turunen, O;Vuorio, M;Leisola, M
通讯作者：
Leisola, M
影响因子：
2.8
作者：
Blanco, A;Díaz, P;Pastor, FIJ
通讯作者：
Pastor, FIJ
Caldicellulosiruptor sp 的糖苷水解酶 10 (GH10) 和 GH11 木聚糖酶的碳水化合物结合模块的不同作用。
影响因子：
4.4
作者：
Meng, Dong-Dong;Ying, Yu;Li, Fu-Li
通讯作者：
Li, Fu-Li
影响因子：
2.9
作者：
Luo, Huiying;Li, Jiang;Yao, Bin
通讯作者：
Yao, Bin
影响因子：
4.1
作者：
Fenel, F;Leisola, M;Turunen, O
通讯作者：
Turunen, O