Comparative phenotypic analysis and genome sequence of Clostridium beijerinckii SA-1, an offspring of NCIMB 8052

被引:17
|
作者
Sandoval-Espinola, Walter J. [1 ]
Makwana, Satya T. [1 ]
Chinn, Mari S. [2 ]
Thon, Michael R. [3 ]
Andrea Azcarate-Peril, M. [4 ,5 ]
Bruno-Barcena, Jose M. [1 ]
机构
[1] N Carolina State Univ, Dept Microbiol, Raleigh, NC 27695 USA
[2] N Carolina State Univ, Dept Biol & Agr Engn, Raleigh, NC 27695 USA
[3] Univ Salamanca, Dept Microbiol & Genet, Ctr Hispano Luso Invest Agr CIALE, Villamayor 37185, Spain
[4] Univ N Carolina, Dept Cell Biol & Physiol, Sch Med, Chapel Hill, NC 27599 USA
[5] Univ N Carolina, Microbiome Core Facil, Ctr Gastrointestinal Biol & Dis, Chapel Hill, NC 27599 USA
来源
MICROBIOLOGY-SGM | 2013年 / 159卷
关键词
HYPERPRODUCING MUTANT BA101; SACCHAROBUTYLICUM DSM 13864; SHORT PALINDROMIC REPEATS; BUTANOL PRODUCTION; PHOSPHOTRANSFERASE SYSTEM; ACETOBUTYLICUM; GLUCOSE; STRAIN; FERMENTATION; METABOLISM;
D O I
10.1099/mic.0.069534-0
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Production of butanol by solventogenic clostridia is controlled through metabolic regulation of the carbon flow and limited by its toxic effects. To overcome cell sensitivity to solvents, stress-directed evolution methodology was used three decades ago on Clostridium beijerinckii NCIMB 8052 that spawned the SA-1 strain. Here, we evaluated SA-1 solventogenic capabilities when growing on a previously validated medium containing, as carbon- and energy-limiting substrates, sucrose and the products of its hydrolysis D-glucose and D-fructose and only D-fructose. Comparative small-scale batch fermentations with controlled pH (pH 6.5) showed that SA-1 is a solvent hyper-producing strain capable of generating up to 16.1 g l(-1) of butanol and 26.3 g l(-1) of total solvents, 62.3 % and 63% more than NCIMB 8052, respectively. This corresponds to butanol and solvent yields of 0.3 and 0.49 g g(-1), respectively (63% and 65 % increase compared with NCIMB 8052). SA-1 showed a deficiency in D-fructose transport as suggested by its 7 h generation time compared with 1 h for NCIMB 8052. To potentially correlate physiological behaviour with genetic mutations, the whole genome of SA-1 was sequenced using the IIlumina GA IIx platform. PCR and Sanger sequencing were performed to analyse the putative variations. As a result, four errors were confirmed and validated in the reference genome of NCIMB 8052 and a total of 10 genetic polymorphisms in SA-1. The genetic polymorphisms included eight single nucleotide variants, one small deletion and one large insertion that it is an additional copy of the insertion sequence ISCb1. Two of the genetic polymorphisms, the serine threonine phosphatase cbs_4400 and the solute binding protein cbs_0769, may possibly explain some of the observed physiological behaviour, such as rerouting of the metabolic carbon flow, deregulation of the D-fructose phosphotransferase transport system and delayed sporulation.
引用
收藏
页码:2558 / 2570
页数:13
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