Open AccessCoexpression of pyruvate decarboxylase and alcohol dehydrogenase genes in Lactobacillus brevis
Author(s) -
Liu Siqing,
Dien Bruce S.,
Nichols Nancy N.,
Bischoff Kenneth M.,
Hughes Stephen R.,
Cotta Michael A.
Publication year - 2007
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2007.00849.x
Subject(s) - lactobacillus brevis , pyruvate decarboxylase , alcohol dehydrogenase , fermentation , biochemistry , biology , mutant , gene , zymomonas mobilis , ethanol fuel , ethanol , microbiology and biotechnology , chemistry , bacteria , genetics , lactic acid , lactobacillus plantarum
Lactobacillus brevis ATCC367 was engineered to express pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) genes in order to increase ethanol fermentation from biomass‐derived residues. First, a Gram‐positive Sarcina ventriculi PDC gene ( Svpdc ) was introduced into L. brevis ATCC 367 to obtain L. brevis bbc03. The Sv PDC was detected by immunoblot using an Sv PDC oligo peptide antiserum, but no increased ethanol was detected in L. brevis bbc03. Then, an ADH gene from L. brevis ( Bradh ) was cloned behind the Svpdc gene that generated a pdc/adh ‐coupled ethanol cassette pBBC04. The pBBC04 restored anaerobic growth and conferred ethanol production of Escheirichia coli NZN111 (a fermentative defective strain incapable of growing anaerobically). Approximately 58 kDa ( Sv PDC) and 28 kDa ( Br ADH) recombinant proteins were observed in L. brevis bbc04. These results indicated that the Gram‐positive ethanol production genes can be expressed in L. brevis using a Gram‐positive promoter and pTRKH2 shuttle vector. This work provides evidence that expressing Gram‐positive ethanol genes in pentose utilizing L. brevis will further aid manipulation of this microbe toward biomass to ethanol production.