1. Cloning and expression of the Zymomonas mobilis 'production of ethanol' genes in Lactobacillus casei
- Author
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Michael M. Meagher, Tyrrell Conway, Robert W. Hutkins, Suxiang Tong, and R. Shane Gold
- Subjects
Lactobacillus casei ,Transcription, Genetic ,Operon ,Blotting, Western ,Genetic Vectors ,Molecular Sequence Data ,Molecular cloning ,Applied Microbiology and Biotechnology ,Microbiology ,Zymomonas mobilis ,Open Reading Frames ,Transformation, Genetic ,Cloning, Molecular ,Promoter Regions, Genetic ,Recombination, Genetic ,Zymomonas ,Binding Sites ,biology ,Base Sequence ,Lactococcus lactis ,Alcohol Dehydrogenase ,General Medicine ,Gene Expression Regulation, Bacterial ,biology.organism_classification ,Blotting, Northern ,Molecular biology ,Ribosomal binding site ,Open reading frame ,Lacticaseibacillus casei ,Biochemistry ,Pyruvate Decarboxylase ,Pyruvate decarboxylase - Abstract
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.
- Published
- 1996