Cloning and expression of the Zymomonas mobilis "production of ethanol" genes in Lactobacillus casei.
This study describes the expression of the Zymomonas mobilis genes coding for pyruvate decarboxylase (pdc) and alcohol dehydrogenase (adh) in Lactobacillus casei 686. To promote transcription, the promoter and ribosome binding site (RBS) from the Lactococcus lactis subsp. lactis-derived vector, pMGE36e, were inserted upstream of the pdc gene. The former sequences were positioned such that translation of pdc was coupled to translation of an 81-base pair open reading frame terminating within the pdc initiation site. The recombinant plasmid (pRSG02) was electroporated into L. casei, and transformants were obtained. Northern analysis confirmed the production of a 3. 1-kb transcript corresponding to the predicted size of the PET operon. Western blot analyses revealed that the recombinant strain expressed both enzymes. The recombinant produced more than twice the ethanol produced by the parental L. casei strain.
Gold
R S
RS
Department of Food Science and Technology, University of Nebraska, Lincoln, NE 68583-0919, USA.
Meagher
M M
MM
Tong
S
S
Hutkins
R W
RW
Conway
T
T
D000426:Alcohol Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D015152:Blotting, Northern; D015153:Blotting, Western; D003001:Cloning, Molecular; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors; D007780:Lacticaseibacillus casei / Q000235:genetics; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D009876:Operon; D011401:Promoter Regions, Genetic; D011767:Pyruvate Decarboxylase / Q000235:genetics*; D011995:Recombination, Genetic; D014158:Transcription, Genetic; D014170:Transformation, Genetic; D017355:Zymomonas / Q000235:genetics*
D000426:Alcohol Dehydrogenase; D011767:Pyruvate Decarboxylase