Abstract
In Escherichia coli, disruption of ptsG, which encodes the glucose-specific permease of the phosphotransferase transport system (PTS) protein EIICBGlc, is crucial for high succinate production. This mutation can, however, cause very slow growth and low glucose consumption rates. The ptsG mutant (TUQ2), from wild type E. coli W1485, and E. coli galP (encoding galactose permease) and glk (encoding glucose kinase) gene expression plasmids were constructed. TUQ2 increased the generation time to approximately 4 h and gave a higher final cell density of 0.5 g/l by expression of galP. However, glk expression had no effect on the mutant. After expression of pyruvate carboxylase (PYC) and galactose permease, the ptsG mutant showed higher succinate yield (1.2 mol/mol glucose) and the specific rate of glucose consumption from 0.33 to 0.6 g/1 h.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
PK Bunch F Mat-Jan N Lee DP Clark (1997) ArticleTitleThe ldhA gene encoding the fermentative lactate dehydrogenase of Escherichia coli Microbiology 143 187–195 Occurrence Handle1:CAS:528:DyaK2sXptFOluw%3D%3D Occurrence Handle9025293 Occurrence Handle10.1099/00221287-143-1-187
R Chatterjee CS Millard K Champion DP Clark MI Donnelly (2001) ArticleTitleMutation of the ptsG gene results in increased production of succinate in fermentation of glucose by Escherichia coli Appl. Environ. Microbiol. 67 148–154 Occurrence Handle10.1128/AEM.67.1.148-154.2001 Occurrence Handle1:CAS:528:DC%2BD3MXjtVWgsw%3D%3D Occurrence Handle11133439
KA Datsenko BL Wanner (2000) ArticleTitleOne-step inactivation of chromosomal genes in Escherichia coli K−12 using PCR products Proc. Natl. Acad. Sci. 97 6640–6645 Occurrence Handle10.1073/pnas.120163297 Occurrence Handle1:CAS:528:DC%2BD3cXktFais7c%3D Occurrence Handle10829079
B Erni (2002) Glucose transport by the bacterial phosphotransferase system (PTS): an interface between energy and signal transduction G Winkelmann (Eds) Microbial Transport System Wiley-VCH Press Weinheim 115–138
B Erni B Zanolari HP Kocher (1987) ArticleTitleThe mannose permease of Escherichia coli consists of three different proteins: amino acid sequence and function in sugar transport, sugar phosphorylation, and penetration of phage lambda DNA J. Biol. Chem. 262 5238–5247 Occurrence Handle1:CAS:528:DyaL2sXkslehu7c%3D Occurrence Handle2951378
RR Gokarn MA Eiteman E Altman (1998) ArticleTitleExpression of pyruvate carboxylase enhances succinate production in Escherichia coli without affecting glucose uptake Biotechnol. Lett. 20 795–798 Occurrence Handle10.1023/B:BILE.0000015925.52287.1f Occurrence Handle1:CAS:528:DyaK1cXmvVSqsLc%3D
RR Gokarn MA Eiteman E Altman (2000) ArticleTitleMetabolic analysis of Escherichia coli in the presence and absence of the carboxylating enzymes phosphoenolpyruvate carboxylase and pyruvate carboxylase Appl. Environ. Microbiol. 66 1844–1850 Occurrence Handle10.1128/AEM.66.5.1844-1850.2000 Occurrence Handle1:CAS:528:DC%2BD3cXjtV2lt78%3D Occurrence Handle10788348
R Gutlnecht K Flukiger R Lanz B Erni (1999) ArticleTitleMechanism of phosphoryl transfer in the dimeric IIABMan subunit of the Escherichia coli mannose transporter J. Biol. Chem. 274 6091–6096
TG Lessie JC Wyk (1972) ArticleTitleMultiple forms of pseudomonas multivoransglucose−6-phosphate and 6-phosphogluconate dehydrogenases: differences in size, pyridine nucleotide specificity, and susceptibility to inhibition by adenosine 5-triphosphate J. Bacteriol. 110 1107–1117 Occurrence Handle1:CAS:528:DyaE38XktlSrsrc%3D Occurrence Handle4402279
P Maeba BD Sanwal (1969) ArticleTitlePhosphoenolpyruvate carboxylase from Salmonella typhimurium strain LT2 Methods Enzymol. 13 283–288 Occurrence Handle1:CAS:528:DyaE3cXktFSrsLo%3D
Postma PW, Lengeler JW, Jacobson GR (1996) Phosphoenolpyruvate: carbohydrate phosphotransferase systems. In: Neidhardt FC, Curtis R III, Ingraham JL, Lin ECC, Brooks K, Magasanik B, Rezniko WS, Riley M, Schaechter M & Umbarger HE, eds. In Escherichia coli and Salmonella: Cellular and Molecular Biology, 1Washington, DC: ASM Press, pp. 1149–1174.
AR Poteete AC Fenton KC Murphy (1999) ArticleTitleRoles of ruvC and recG in phage λ red-mediated recombination J. Bacteriol. 181 5402–5408 Occurrence Handle1:CAS:528:DyaK1MXlvVSltr4%3D Occurrence Handle10464213
AM Sanchez GN Bennett KY San (2005) ArticleTitleEfficient succinic acid production from glucose through overexpression of pyruvate carboxylase in an Escherichia coli alcohol dehydrogenase and lactate dehydrogenase mutant Biotechnol. Prog. 21 358–365 Occurrence Handle10.1021/bp049676e Occurrence Handle1:CAS:528:DC%2BD2MXptlSitg%3D%3D Occurrence Handle15801771
JH Tchieu V Norris JS Edwards MH Saier SuffixJr (2001) ArticleTitleThe complete phosphotransferase system in Escherichia coli J. Mol. Microbiol. Biotechnol. 3 329–346 Occurrence Handle1:CAS:528:DC%2BD3MXjsVyntLw%3D Occurrence Handle11361063
GN Vemuri MA Eiteman E Altman (2002) ArticleTitleEffects of growth mode and pyruvate carboxylase on succinic acid production by metabolically engineered strains of Escherichia coli Appl. Environ. Microbiol. 68 1715–1727 Occurrence Handle10.1128/AEM.68.4.1715-1727.2002 Occurrence Handle1:CAS:528:DC%2BD38XivFGltLo%3D Occurrence Handle11916689
Author information
Authors and Affiliations
Corresponding author
Additional information
Received 31 August 2005; Revisions requested 27 September 2005; Revisions received 1 November 2005; Accepted 2 November 2005
An erratum to this article is available at http://dx.doi.org/10.1007/s10529-006-6907-7.
Rights and permissions
About this article
Cite this article
Wang, Q., Wu, C., Chen, T. et al. Expression of Galactose Permease and Pyruvate Carboxylase in Escherichia coli ptsG Mutant Increases the Growth Rate and Succinate Yield under Anaerobic Conditions. Biotechnol Lett 28, 89–93 (2006). https://doi.org/10.1007/s10529-005-4952-2
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s10529-005-4952-2