通过能够进行吡咯并喹啉醌（PQQ）生物合成的代谢工程大肠杆菌菌株提高重组吡咯并喹啉醌葡萄糖脱氢酶的产量。

Increased production of recombinant pyrroloquinoline quinone (PQQ) glucose dehydrogenase by metabolically engineered Escherichia coli strain capable of PQQ biosynthesis.

作者信息

Sode K, Ito K, Witarto A B, Watanabe K, Yoshida H, Postma P

机构信息

Department of Biotechnology, Faculty of Technology, Tokyo University of Agriculture and Technology, Japan.

出版信息

J Biotechnol. 1996 Aug 20;49(1-3):239-43. doi: 10.1016/0168-1656(96)01540-4.

DOI:10.1016/0168-1656(96)01540-4

PMID:8879174

Abstract

We have previously shown that the production of recombinant Escherichia coli PQQGDH was greatly improved by using a medium supplemented with the cofactor PQQ, which is not synthesized in E. coli. We show here that the increase in the accumulated PQQGDH is due to the increased stability of the holo-enzyme over apo-enzyme, using recombinant Acinetobacter calcoaceticus PQQGDH. In order to achieve cost-effective PQQGDH production, we incorporated the genes for PQQ biosynthetic pathway from Klebsiella pneumoniae into E. coli, which as a result allowed E. coli to produce PQQ. Using this metabolically engineered E. coli strain as a host, a 10-fold increase in the production of recombinant A. calcoaceticus PQQGDH was achieved, compared to the condition without PQQ and MgCl2.

摘要

我们之前已经表明，通过使用添加了大肠杆菌无法合成的辅因子吡咯喹啉醌（PQQ）的培养基，重组大肠杆菌PQQGDH的产量得到了极大提高。我们在此表明，使用重组乙酸钙不动杆菌PQQGDH时，积累的PQQGDH增加是由于全酶相对于脱辅基酶的稳定性增加。为了实现具有成本效益的PQQGDH生产，我们将肺炎克雷伯菌PQQ生物合成途径的基因整合到大肠杆菌中，结果使大肠杆菌能够产生PQQ。与没有PQQ和MgCl2的条件相比，使用这种经过代谢工程改造的大肠杆菌菌株作为宿主，重组乙酸钙不动杆菌PQQGDH的产量提高了10倍。