Reconstructing the clostridial n-butanol metabolic pathway in Lactobacillus brevis.
A Lactobacillus brevis strain with the ability to synthesize butanol from glucose was constructed by metabolic engineering. The genes crt, bcd, etfB, etfA, and hbd, composing the bcs-operon, and the thl gene encode the enzymes of the lower part of the clostridial butanol pathway (crotonase, butyryl-CoA-dehydrogenase, two subunits of the electron transfer flavoprotein, 3-hydroxybutyryl-CoA dehydrogenase, and thiolase) of Clostridium acetobutylicum. They were cloned into the Gram-positive/Gram-negative shuttle plasmid vector pHYc. The two resulting plasmids pHYc-thl-bcs and pHYc-bcs (respectively, with and without the clostridial thl gene) were transferred to Escherichia coli and L. brevis. The recombinant L. brevis strains were able to synthesize up to 300 mg l(-1) or 4.1 mM of butanol on a glucose-containing medium. A L. brevis strain carrying the clostridial bcs-operon has the ability to synthesize butanol with participation of its own thiolase, aldehyde dehydrogenase, and alcohol dehydrogenase. The particular role of the enzymes involved in butanol production and the suitability of L. brevis as an n-butanol producer are discussed.
Berezina
Oksana V
OV
State Research Institute of Genetics and Selection of Industrial Microorganisms, 1st Dorojniy pr. 1, 117545, Moscow, Russian Federation. mashchenko@yandex.ru
Zakharova
Natalia V
NV
Brandt
Agnieszka
A
Yarotsky
Sergey V
SV
Schwarz
Wolfgang H
WH
Zverlov
Vladimir V
VV
D020001:1-Butanol / Q000378:metabolism*; D015094:3-Hydroxyacyl CoA Dehydrogenases / Q000235:genetics / Q000378:metabolism; D000214:Acyl Coenzyme A / Q000235:genetics / Q000378:metabolism; D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D053898:Biosynthetic Pathways; D003001:Cloning, Molecular; D046969:Clostridium acetobutylicum / Q000201:enzymology* / Q000235:genetics; D005818:Genetic Engineering; D052196:Levilactobacillus brevis / Q000235:genetics* / Q000378:metabolism*; D053858:Metabolic Networks and Pathways
D000214:Acyl Coenzyme A; D001426:Bacterial Proteins; C024343:butyryl-coenzyme A; D020001:1-Butanol; D015094:3-Hydroxyacyl CoA Dehydrogenases; D000426:Alcohol Dehydrogenase