A high throughput method to screen companion bacterium for 2-keto-L-gulonic acid biosynthesis by co-culturing Ketogulonicigenium vulgare

被引：14
|
作者：
Zhu, Yibo [1 ,2 ,3 ,4 ]
Liu, Jidong [1 ,2 ,3 ]
Liu, Jie [5 ]
Du, Guocheng [1 ,2 ,3 ]
Zhou, Jingwen [1 ,2 ,3 ]
Chen, Jian [1 ,2 ,3 ]
机构：
[1] Jiangnan Univ, Sch Biotechnol, Wuxi 214122, Jiangsu, Peoples R China
[2] Jiangnan Univ, Key Lab Ind Biotechnol, Minist Educ, Wuxi 214122, Jiangsu, Peoples R China
[3] Jiangnan Univ, State Key Lab Food Sci & Technol, Wuxi 214122, Jiangsu, Peoples R China
[4] Changshu Inst Technol, Sch Biotechnol & Food Engn, Changshu 215500, Jiangsu, Peoples R China
[5] Jiangsu Jiangshan Pharmaceut Co Ltd, Jingjiang 214500, Peoples R China
来源：
PROCESS BIOCHEMISTRY | 2012年 / 47卷 / 09期
基金：
国家高技术研究发展计划(863计划);
关键词：
Microtiter plates; High throughput; Co-culture process; Companion bacterium; 2-keto-L-gulonic acid biosynthesis; BACILLUS-MEGATERIUM; GROWTH; THURINGIENSIS; FERMENTATION; BIOREACTORS; MUTAGENESIS;
D O I：
10.1016/j.procbio.2012.05.010
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
Bacillus megaterium is widely used as companion bacterium in the two-step biosynthesis of 2-keto-L-gulonic acid (2-KLG) by Ketogulonicigenium vulgare. To screen efficiently target companion strains from large numbers of random mutants, a screen method based on spectrophotometry and 24-well microtiter plates was developed and validated on an integrated library of 450 transposon random insertional mutants and two sporulation-defective mutants. The co-culture processes were classified into three groups (low, intermediate and high performance) by K-mean clustering analysis. In addition, mutant m71 was successfully screened out from the library. The substrate conversion ratio of m71 and K. vulgare co-culture process after 72 h was decreased by about 38% compared with that of the wild-type co-culture process in 750 ml flasks. These results indicated that the proposed high throughput method is feasible for screening target companions for the co-culture process of 2-KLG biosynthesis. (C) 2012 Elsevier Ltd. All rights reserved.
引用
收藏
页码：1428 / 1432
页数：5
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