Efficient Coproduction of Mannanase and Cellulase by the Transformation of a Codon-Optimized Endomannanase Gene from &ITAspergillus niger&IT into &ITTrichoderma reesei &IT
Efficient Coproduction of Mannanase and Cellulase by the Transformation of a Codon-Optimized Endomannanase Gene from &ITAspergillus niger&IT into &ITTrichoderma reesei &IT
复制标题
通过将来自 3 种黑曲霉的密码子优化的内甘露聚糖酶基因第二次转化到里氏木霉中，高效共生产甘露聚糖酶和纤维素酶
DOI：
10.1021/acs.jafc.7b05114
复制
发表时间：
2017-12-20
影响因子：
6.1
通讯作者：
Su, Xiaoyun
中科院分区：
文献类型：
--
作者：
Sun, Xianhua;Xue, Xianli;Su, Xiaoyun
纤维素酶和甘露聚糖酶都是动物饲料中重要的酶添加剂。在一个微生物宿主中同时表达这两种酶可能会潜在地降低动物饲养的成本。为此，我们对黑曲霉Man5A基因进行了密码子优化，以适应里氏木霉的密码子使用偏向。通过比较核苷酸序列的自由能和局部结构，最终筛选出一个优化的核苷酸序列，并转化里氏木霉吡啶营养缺陷型菌株TU-6。密码子优化后的基因表达水平高于原基因。通过补料分批培养，进一步将密码子优化的基因在里氏酵母突变株中表达，纤维素酶和甘露聚糖酶的产量分别达到1376和1204 umL(-1)。
Cellulase and mannanase are both important enzyme additives in animal feeds. Expressing the two enzymes simultaneously within one microbial host could potentially lead to cost reductions in the feeding of animals. For this purpose, we codon-optimized the Aspergillus niger Man5A gene to the codon-usage bias of Trichoderma reesei. By comparing the free energies and the local structures of the nucleotide sequences, one optimized sequence was finally selected and transformed into the T. reesei pyridine-auxotrophic strain TU-6. The codon-optimized gene was expressed to a higher level than the original one. Further expressing the codon-optimized gene in a mutated T. reesei strain through fed-batch cultivation resulted in co-production of cellulase and mannanase up to 1376 U.mL(-1) and 1204 U.mL(-1) , respectively.