Effects of mutations in acetate metabolism on high-cell-density growth of Escherichia coli

被引:67
作者
Contiero, J
Beatty, C
Kumari, S
DeSanti, CL
Strohl, WR
Wolfe, A
机构
[1] Merck & Co Inc, Merck Sharp & Dohme Res Labs, Nat Prod Drug Discovery Microbiol, Rahway, NJ 07065 USA
[2] Ohio State Univ, Dept Microbiol, Columbus, OH 43210 USA
[3] Loyola Univ, Dept Microbiol & Immunol, Maywood, IL 60153 USA
[4] UNESP, Inst Quim, BR-14800900 Araraquara, SP, Brazil
关键词
acetate metabolism; acetate mutants; glucose-controlled high cell density fermentation; fed-batch fermentation; rpoS;
D O I
10.1038/sj.jim.7000014
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
To study the role played by acetate metabolism during high-cell-density growth of Escherichia coli cells, we constructed isogenic null mutants of strain W3100 deficient for several genes involved either in acetate metabolism or the transition to stationary phase. We grew these strains under identical fed-batch conditions to the highest cell densities achievable in 8 h using a predictive-plus-feedback-controlled computer algorithm that maintained glucose at a set-point of 0.5 g/l, as previously described. Wild-type strains, as well as mutants lacking the sigma(s) subunit of RNA polymerase (rpoS), grew reproducibly to high cell densities (44-50 g/l dry cell weights, DCWs). In contrast, a strain lacking acetate kinase (ackA) failed to reach densities greater than 8 g/l. Strains lacking other acetate metabolism genes (pta, acs, poxB, iciR, and fadR) achieved only medium cell densities (15-21 g/l DCWs). Complementation of either the acs or the ackA mutant restored wild-type high-cell-density growth, On a dry weight basis, poxB and fadR strains produced approximately threefold more acetate than did the wild-type strain. In contrast, the pta, acs, or rpoS strains produced significantly less acetate per cell dry weight than did the wild-type strain. Our results show that acetate metabolism plays a critical role during growth of E. coli cultures to high cell densities. They also demonstrate that cells do not require the sigma(s) regulon to grow to high cell densities, at least not under the conditions tested.
引用
收藏
页码:421 / 430
页数:10
相关论文
共 34 条
[1]   MODIFICATION OF CENTRAL METABOLIC PATHWAY IN ESCHERICHIA-COLI TO REDUCE ACETATE ACCUMULATION BY HETEROLOGOUS EXPRESSION OF THE BACILLUS-SUBTILIS ACETOLACTATE SYNTHASE GENE [J].
ARISTIDOU, AA ;
SAN, KY ;
BENNETT, GN .
BIOTECHNOLOGY AND BIOENGINEERING, 1994, 44 (08) :944-951
[2]   IMPROVED EXPRESSION OF HUMAN INTERLEUKIN-2 IN HIGH-CELL-DENSITY FERMENTER CULTURES OF ESCHERICHIA-COLI K-12 BY A PHOSPHOTRANSACETYLASE MUTANT [J].
BAUER, KA ;
BENBASSAT, A ;
DAWSON, M ;
DELAPUENTE, VT ;
NEWAY, JO .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1990, 56 (05) :1296-1302
[3]   Regulation of RssB-dependent proteolysis in Escherichia coli:: a role for acetyl phosphate in a response regulator-controlled process [J].
Bouché, S ;
Klauck, E ;
Fischer, D ;
Lucassen, M ;
Jung, K ;
Hengge-Aronis, R .
MOLECULAR MICROBIOLOGY, 1998, 27 (04) :787-795
[4]   ENZYMIC INTERCONVERSION OF ACETATE AND ACETYL-COENZYME-A IN ESCHERICHIA-COLI [J].
BROWN, TDK ;
JONESMORTIMER, MC ;
KORNBERG, HL .
JOURNAL OF GENERAL MICROBIOLOGY, 1977, 102 (OCT) :327-336
[5]   EXPRESSION OF ESCHERICHIA-COLI PYRUVATE OXIDASE (POXB) DEPENDS ON THE SIGMA-FACTOR ENCODED BY THE RPOS(KATF) GENE [J].
CHANG, YY ;
WANG, AY ;
CRONAN, JE .
MOLECULAR MICROBIOLOGY, 1994, 11 (06) :1019-1028
[6]   EFFECT OF MODIFIED GLUCOSE-UPTAKE USING GENETIC-ENGINEERING TECHNIQUES ON HIGH-LEVEL RECOMBINANT PROTEIN-PRODUCTION IN ESCHERICHIA-COLI DENSE CULTURES [J].
CHOU, CH ;
BENNETT, GN ;
SAN, KY .
BIOTECHNOLOGY AND BIOENGINEERING, 1994, 44 (08) :952-960
[7]   Regulation of acetate metabolism by protein phosphorylation in enteric bacteria [J].
Cozzone, AJ .
ANNUAL REVIEW OF MICROBIOLOGY, 1998, 52 :127-164
[8]   EFFECT OF ALTERATION OF THE ACETIC-ACID SYNTHESIS PATHWAY ON THE FERMENTATION PATTERN OF ESCHERICHIA-COLI [J].
DIAZRICCI, JC ;
REGAN, L ;
BAILEY, JE .
BIOTECHNOLOGY AND BIOENGINEERING, 1991, 38 (11) :1318-1324
[9]  
ELMANSI EMT, 1989, J GEN MICROBIOL, V135, P2875
[10]   Reduction of aerobic acetate production by Escherichia coli [J].
Farmer, WR ;
Liao, JC .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1997, 63 (08) :3205-3210