Pliot 규모에서의 재조합 대장균을 이용한(R)-3-Hydroxybutyric acid 생산
Pilot Scale Production of (R)-3-Hydroxybutyric acidby Metabolically Engineered Escherichia coli
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Production of (R)-3-hydroxybutyric acid (R3HB) by fed-batch culture and continuous culture of metabolically engineered Escherichia coli harboring Ralstonia eutropha PHB biosynthesis and depolymerase genes was examined in a 30 l pilot-scale fermentor. A new stable two-plasmid system, pBRRed containing the R. eutropha PHB depolymerase gene and pMCS105 containing the R. eutropha PHB biosynthesis genes, was developed. Among a variety of E. coli strains harboring plasmids, recombinant E. coli XL-10 Gold (pBRRed, pMCS105) was able to produce R3HB with the highest efficiency in a batch culture. By the fed-batch culture of recombinant E. coli XL-10 Gold(pBRRed, pMCS105) in a 30 l fermentor, the final R3HB concentration was 22.4 g/l giving a productivity of 0.97 g/l-h. To produce R3HB to a high concentration with high productivity, a new strategy of fed-batch culture followed by a continuous culture was investigated. The maximum productivity and R3HB concentration were 5.06 g/l-h and 25.3 g/l, respectively. These results show that economical production of R3HB is possible by recombinant E. coli in large scale.
Abstract
Production of (R)-3-hydroxybutyric acid (R3HB) by fed-batch culture and continuous culture of metabolically engineered Escherichia coli harboring Ralstonia eutropha PHB biosynthesis and depolymerase genes was examined in a 30 l pilot-scale fermentor. A new stable two-plasmid system, pBRRed containing the R. eutropha PHB depolymerase gene and pMCS105 containing the R. eutropha PHB biosynthesis genes, was developed. Among a variety of E. coli strains harboring plasmids, recombinant E. coli XL-10 Gold (pBRRed, pMCS105) was able to produce R3HB with the highest efficiency in a batch culture. By the fed-batch culture of recombinant E. coli XL-10 Gold(pBRRed, pMCS105) in a 30 l fermentor, the final R3HB concentration was 22.4 g/l giving a productivity of 0.97 g/l-h. To produce R3HB to a high concentration with high productivity, a new strategy of fed-batch culture followed by a continuous culture was investigated. The maximum productivity and R3HB concentration were 5.06 g/l-h and 25.3 g/l, respectively. These results show that economical production of R3HB is possible by recombinant E. coli in large scale.
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