Letters in Applied Microbiology · 2001 · 10 citations · 14 references
Life SciencesBiochemistryMicrobiologyEnzymologyMetabolic EngineeringMicrobial MetabolismBacterial FermentationBacterial MetabolismMicrobial Metabolic EngineeringMicrobial Metabolite Production2, 3-Butanedione BiosynthesisEscherichia ColiStereochemical ModificationStereochemical Requirements
The L-2,3-butanediol dehydrogenase (L-BDH) gene of Brevibacterium saccharolyticum was strongly expressed in Escherichia coli using the tac promoter. However, the stereospecificity of the resulting L-BDH was reduced. The region upstream of the meso-BDH gene of Klebsiella pneumoniae was also involved in the expression of the B. saccharolyticum gene. However, in this case, the resulting L-BDH exhibited more stable stereospecificity. A stereospecificity recognition region was located within the rear sequence (Hpa I site, carboxy terminal) of the BDH open reading frame. Using a transformed strain of E. coli, the conversion of L-acetoin (L-AC), in the commercially available racemic mixture of AC, to L-2,3-butanediol (L-BD) was attempted. As a result, 0.37% L-BD was formed from 1% AC added to the culture.
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Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension
Robert M. Horton, Henry D. Hunt, Steffan N. Ho et al. · Gene · 1989 · 3.1K citations