Open AccessamyP , a reporter gene to study strain degeneration in Clostridium acetobutylicum ATCC 824
Author(s) -
Sabathé Fabrice,
Croux Christian,
Cornillot Emmanuel,
Soucaille Philippe
Publication year - 2002
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.1574-6968.2002.tb11165.x
Subject(s) - clostridium acetobutylicum , gene , biology , extracellular , regulon , reporter gene , biochemistry , strain (injury) , gene expression , butanol , anatomy , ethanol
Clostridium acetobutylicum produces an extracellular α‐amylase when grown on glucose as the sole carbon source. This enzyme was previously characterized from a biochemical point of view but its encoding gene was never identified. The 2283‐bp amyP gene encodes a 83 013‐Da mature protein with an N‐terminal domain that exhibits strong identity to the family 13 glycosyl hydrolases such as the Bacillus α‐amylases. Transcriptional analysis revealed that amyP is transcribed in solventogenic but not in acidogenic chemostat cultures. These results are in agreement with the extracellular α‐amylase activities indicating that the expression of amyP is regulated at the transcriptional level. amyP is located on the pSOL1 megaplasmid that carries all the genes involved in the final steps of solvent formation. Degeneration of C. acetobutylicum has been associated to the loss of pSOL1. We demonstrate here that amyP can be used as a reporter system to quantitatively follow this phenomenon.