title: Genetic engineering of Lactobacillus diolivorans.
abstract: In this study, we developed a toolbox for genetic manipulation of Lactobacillus diolivorans, a promising production organism for 1,3-propanediol from glycerol. Two major findings play a key role for successful transformation of this organism: (1) the absence of a native plasmid, because a native plasmid is a major obstacle for transformation of L. diolivorans, and (2) the absence of DNA methylation. A suitable expression plasmid, pSHM, for homologous and heterologous protein expression in L. diolivorans was constructed. This plasmid is based on the replication origin repA of L. diolivorans. The native glyceraldehyde-3-phosphate dehydrogenase promoter is used for constitutive expression of the genes of interest. Functional expression of genes in L. diolivorans was shown with two examples: production of green fluorescent protein resulted in a 40- to 60-fold higher fluorescence of the obtained clones compared with the wild-type strain. Finally, the homologous overexpression of a putatively NADPH-dependent 1,3-propanediol oxidoreductase improved 1,3-propanediol production by 20% in batch cultures.
authors: School of Bioengineering, FH Campus Wien - University of Applied Sciences, Vienna, Austria.
authors: Stefan
authors: S
authors: Pflügl
authors: Hans
authors: H
authors: Marx
authors: Diethard
authors: D
authors: Mattanovich
authors: Michael
authors: M
authors: Sauer
mesh_terms: D019175:DNA Methylation; D005818:Genetic Engineering / Q000379:methods*; D007778:Lactobacillus / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D011409:Propylene Glycols / Q000378:metabolism
chemical_list: D011409:Propylene Glycols; C041787:1,3-propanediol