Production of 1,3-propanediol from Glycerol by Recombinant E. coli Using Incompatible Plasmids System

Abstract

1,3-Propanediol (1,3-PD) has numerous applications in polymers, cosmetics, foods, lubricants, and medicines as a bifunctional organic compound. The genes for the production of 1,3-PD in Klebsiella pneumoniae, dhaB, which encodes glycerol dehydratase, and dhaT, which encodes 1,3-PD oxidoreductase, and gdrAB, which encodes glycerol dehydratase reactivating factor, are naturally under the control of different promoters and are transcribed in different directions. These genes were coexpressed in E. coli using two incompatible plasmids (pET28a and pET22b) in the presence of selective pressure. The recombinant E. coli coexpressed the glycerol dehydratase, 1,3-propanediol oxidoreductase and reactivating factor for the glycerol dehydratase at high levels. In a fed-batch fermentation of glycerol and glucose, the recombinant E. coli containing these two incompatible plasmids consumed 14.3 g/l glycerol and produced 8.6 g/l 1,3-propanediol. In the substitution case of yqhD (encoding alcohol dehydrogenase from E. coli) for dhaT, the final 1,3-propanediol concentration of the recombinant E. coli could reach 13.2 g/l.

Comparative Profiling and Discovery of Novel Glycosylated Mycosporine-Like Amino Acids in Two Strains of the Cyanobacterium Scytonema cf. crispum

“…E. coli BL21(DE3) cells (Novagen) cotransformed with the pRARE plasmid were used for heterologous expression. The possible expression of incompatible plasmids over short times has been reported in the past (27,28). Plasmid compatibility for the duration of culture expression was confirmed by the identification of soluble protein products expressed from both coexpressed plasmids (see the supplemental material).…”

Production of mesaconate in Escherichia coli by engineered glutamate mutase pathway

“…Co-overexpresion of MutL with glutamate mutase and MAL could enhance the mesaconate titer by 162.4%. Similar strategy has been used for producing 1,3-propanediol via glycerol dehydratase by recombinant E. coli (Ma et al, 2010;Przystalowska et al, 2015;Wang et al, 2007).…”

YqhD: a broad-substrate range aldehyde reductase with various applications in production of biorenewable fuels and chemicals

“…Encouraged by these positive results, Wang and coworkers replaced K. pneumonia's dhaT with yqhD in their recombinant E. coli strain, increasing 1,3-PD titer from 8.6 g/L to 13.2 g/L (Wang et al 2007). Others have used YqhD in combination with the Clostridium butyricum DHA pathway in E. coli: co-expression of yqhD with dhaB1 and dhaB2 resulted in a strain that produced 104.4 g/L 1,3-PD from glycerol with a productivity of 2.61 g/L/hr (Tang et al 2009).…”