CLONING AND EXPRESSION OF THE ZYMOMONAS-MOBILIS PRODUCTION OF ETHANOLGENES IN LACTOBACILLUS-CASEI

This study describes the expression of the Zymomonas mobilis genes cod ing for pyruvate decarboxylase (pdc) and alcohol dehydrogenase (adh) i n Lactobacillus casei 686. To promote transcription, the promoter and ribosome binding site (RES) from the Lactococcus lactis subsp. lactis- derived vector, pMGE36e, were inserted upstream of the pde gene, The f ormer sequences were positioned such that translation of pdc was coupl ed to translation of an 81-base pair open reading frame terminating wi thin the pde initiation site. The recombinant plasmid (pRSG02) was ele ctroporated into L. casei, and transformants were obtained. Northern a nalysis confirmed the production of a 3.1-kb transcript corresponding to the predicted size of the PET operon. Western blot analyses reveale d that the recombinant strain expressed both enzymes. The recombinant produced more than twice the ethanol produced by the parental L. casei strain.