High-level extracellular production of alkaline polygalacturonate lyase in Bacillus subtilis with optimized regulatory elements.

High-level extracellular production of alkaline polygalacturonate lyase in Bacillus subtilis with optimized regulatory elements.
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具有优化调节元件的枯草芽孢杆菌中碱性多聚半乳糖醛酸裂解酶的高水平胞外生产。

DOI:
10.1016/j.biortech.2013.07.129
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发表时间:
2013-10
期刊:
Bioresour Technol
影响因子:
--
通讯作者:
Chen, Jian
Chen, Jian
中科院分区:
其他
文献类型:
--
作者:
Liu, Long;Wang, Miao;Du, Guocheng;Chen, Jian

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本工作旨在构建一种功能强大的重组芽孢杆菌,以实现碱性多聚半乳糖醛酸裂解酶(PGL)的分泌生产。首先,用半理性方法筛选出6个信号肽(AMYX、BPR、VPR、YVGO、WAPA和nprE),比较它们对PGL产生的影响。通过优化和应用强启动子P43和Shine−Dalgarno序列,获得了较高滴度的−1PGL,滴度为446.3 U·mLPgL。最后,重组子B的容量。在3号L发酵罐中,采用分批补料策略对枯草杆菌WB43CB进行了发酵评价。PGL效价达632.6单位/毫升−-1,产生量为17.6U/毫升−-1h-−-1,是该菌最高的pGL分泌量。枯草杆菌系统。重组子B。本工作构建的枯草杆菌WB43CB菌株在生产碱性PGL方面具有很大的潜力。
The present work aims to construct a robust recombinantBacillus subtilisto achieve secretory production of alkaline polygalacturonate lyase (PGL). First, 6 signal peptides (amyX, bpr, vpr, yvgO, wapA and nprE) were screened with a semi-rational approach and comparatively investigated their effects on the production of PGL. The signal peptide bpr directed efficient PGL secretory expression and increased PGL titer to 313.7 U mL−1. By optimizing and applying strong promoter P43 and Shine−Dalgarno sequence, higher titer of 446.3 U mL−1PGL was achieved. Finally, the capacity of the recombinantB. subtilisWB43CB was evaluated with a fed-batch strategy in 3 L fermentor. The PGL titer reached 632.6 U mL−1with a productivity of 17.6 U mL−1h−1, which was the highest secretory production of PGL by theB. subtilissystem. The recombinantB. subtilisstrain WB43CB constructed in the present work has great potential in production of alkaline PGL.
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