混合碳源流加对重组毕赤酵母生产碱性果胶酶的影响

Effects of mixed carbon sources in cultivation of recombinant Pichia pastoris for polygalacturonate lyase production

摘要:

为提高重组毕赤酵母生产碱性果胶酶(PGL)的产量和生产强度, 在诱导期采用多种碳源与甲醇混合添加的模式。实验结果发现: 甘油、山梨醇、乳酸与甲醇的混合添加均可以提高PGL的产量, 其中山梨醇与甲醇的混合流加效果最为显著。研究表明, 通过双碳源混合流加可以提高细胞活力, 增强醇氧化酶活力, 提高毕赤酵母表达外源蛋白效率。当山梨醇的流速为3.6 g/(h·L)时, PGL酶活可达1593 U/mL, 生产强度为16.7 U/(mL·h), 比对照分别提高了84.6%和45.2%, 实现了碱性果胶酶的高效生产。

Abstract:

In order to increase the production and productivity of alkaline polygalacturonate lyase (PGL), we studied the mixed carbon sources feeding strategies during the induction phase by recombinant Pichia pastoris GS115. Glycerol, sorbitol or lactic acid co-feeding with methanol all enhanced the PGL production. Among all the feeding strategies, the sorbitol co-feeding strategy was most significant. By using this strategy, the PGL activity and productivity reached 1593 U/mL and 16.7 U/(mL·h). Compared to the control, the enhancements of PGL activity and productivity were 84.6% and 45.2 % respectively, when we set the sorbitol feeding rate at 3.6 g/(h·L).