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

页码：1428 / 1432

页数：5

共 41 条

[41] Taxonomic characterization of Ketogulonigenium vulgare gen. nov., sp nov and Ketogulonigenium robustum sp nov., which oxidize L-sorbose to 2-keto-L-gulonic acid
Urbance, JW
Bratina, BJ
Stoddard, SF
Schmidt, TM
INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 2001, 51 : 1059 - 1070

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