Proteome-based physiological analysis of the metabolically engineered succinic acid producer Mannheimia succiniciproducens LPK7.
Using two-dimensional gel electrophoresis (2-DE) and mass spectrometry, the proteome of a metabolically engineered succinic acid-overproducing bacterium, Mannheimia succiniciproducens LPK7, was examined and compared with that of its wild type strain, MBEL55E, to elucidate the physiological and metabolic changes responsible for succinic acid overproduction and cell growth. Comparative proteomic studies clearly showed that the expression levels of enzymes involved in the ATP formation and consumption (AtpD, Ppa, SerS, ProS, Pnp, PotD, MalK, RbsB, and TbpA), pyruvate metabolism (AceF and Lpd), glycolysis (GapA, Pgk, Fba, and TpiA), and amino acid biosynthesis (Asd, DapA, DapD, Gdh, ArgD, and ArgG) varied significantly in the LPK7 strain compared with those in the MBEL55E strain. Based on the comparative proteome profiling, the formation of pyruvic acid, a newly formed byproduct in the engineered LPK7 strain, could be reduced by adding into the culture medium pantothenate and L: -cysteine, which serve as precursors of CoA biosynthesis.
Lee
Jeong Wook
JW
Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 Program), BioProcess Engineering Research Center, KAIST, 335 Gwahangno, Yuseong-gu, Daejeon 305-701, Republic of Korea.
Lee
Sang Yup
SY
D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D003545:Cysteine / Q000494:pharmacology; D004798:Enzymes / Q000096:biosynthesis* / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods; D031121:Mannheimia / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D010205:Pantothenic Acid / Q000494:pharmacology; D020543:Proteome / Q000096:biosynthesis* / Q000235:genetics; D019289:Pyruvic Acid / Q000378:metabolism; D013045:Species Specificity; D019802:Succinic Acid / Q000378:metabolism*; D014803:Vitamin B Complex / Q000494:pharmacology
D001426:Bacterial Proteins; D004798:Enzymes; D020543:Proteome; D014803:Vitamin B Complex; D010205:Pantothenic Acid; D019289:Pyruvic Acid; D019802:Succinic Acid; D003545:Cysteine