MOLECULAR CHARACTERIZATION OF 2 CLOSTRIDIUM-ACETOBUTYLICUM ATCC 824 BUTANOL DEHYDROGENASE ISOZYME GENES

被引:110
作者
WALTER, KA
BENNETT, GN
PAPOUTSAKIS, ET
机构
[1] NORTHWESTERN UNIV,DEPT CHEM ENGN,EVANSTON,IL 60208
[2] RICE UNIV,DEPT BIOCHEM & CELL BIOL,HOUSTON,TX 77251
关键词
D O I
10.1128/jb.174.22.7149-7158.1992
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
A 4-kb segment of DNA containing two previously cloned butanol dehydrogenase (BDH) isozyme genes (D. Petersen, R. Welch, F. Rudolph, and G. Bennett, J. Bacteriol. 173:1831-1834, 1991) was sequenced. Two complete open reading frames (ORFs) were identified (bdhA and bdhB), along with a third truncated ORF (ORF1). The translation products of bdh.4 and bdhB corresponded to the N-terminal sequences of the purified BDH I and BDH II proteins, respectively. The two isozymes had a high amino acid identity (73%) and showed homology to a newly described class of alcohol dehydrogenases. Northern blots revealed that bdhA and bdhB did not form an operon. Primer extension experiments located single transcriptional start sites 37 and 58 bp upstream of the start codons of bdhA and bdhB, respectively. The -10 and -35 promoter regions for these genes were almost identical. bdhA and bdhB were found to be induced or derepressed immediately prior to significant butanol production in controlled pH 5.0 batch fermentations.
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页码:7149 / 7158
页数:10
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