Genetic manipulation of acid and solvent formation inClostridium acetobutylicum ATCC 824

Autor: Edward M. Green, George N. Bennett
Rok vydání: 1998
Předmět:
Zdroj: Biotechnology and Bioengineering. 58:215-221
ISSN: 1097-0290
0006-3592
DOI: 10.1002/(sici)1097-0290(19980420)58:2/3<215::aid-bit14>3.0.co;2-b
Popis: The genes coding for enzymes involved in butanol or butyrate formation were subcloned into a novel Escherichia coli-Clostridium acetobutylicum shuttle vector constructed from pIMP1 and a chloramphenicol acetyl transferase gene. The resulting replicative plasmids, referred to as pTHAAD (aldehyde/alcohol dehydrogenase) and pTHBUT (butyrate operon), were used to complement C. acetobutylicum mutant strains, in which genes encoding aldehyde/alcohol dehydrogenase (aad) or butyrate kinase (buk) had been inactivated by recombination with Emr constructs. Complementation of strain PJC4BK (buk mutant) with pTHBUT restored butyrate kinase activity and butyrate production during exponential growth. Complementation of strain PJC4AAD (aad mutant) with pTHAAD restored NAD(H)-dependent butanol dehydrogenase activity, NAD(H)-dependent butyraldehyde dehydrogenase activity and butanol production during solventogenic growth. The development of an alternative selectable marker makes it is possible to overexpress genes, via replicative plasmids, in mutant strains that lack specific enzyme activities, thereby expanding the number of possible genetic manipulations that can be performed in C. acetobutylicum. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:215–221, 1998.
Databáze: OpenAIRE
Externí odkaz: