Possible function and some properties of the CcpA protein of Bacillus subtilis Miwa, Yasuhiko and Saikawa, Mitsuyuki and Fujita, Yasutaro,, 140, 2567-2575 (1994), doi = https://doi.org/10.1099/00221287-140-10-2567, publicationName = Microbiology Society, issn = 1350-0872, abstract= The ccpAmutations alsA1(alsA1is allelic to ccpA) and ccpA::Tn917completely abolished catabolite repression of gluconate kinase and sorbitol dehydrogenase synthesis in Bacillus subtilis, whereas they only partially affected the catabolite repression of inositol dehydrogenase, histidase and xylose isomerase synthesis. The alsA1mutation also partially affected catabolite repression of sporulation. Analysis of revertants from the alsA1mutant by direct sequencing indicated that this mutation comprises a base substitution of guanine at nucleotide −14 to adenine within the Shine-Dalgarno sequence of the ccpAgene (ccpAtranslation starts at nucleotide +1). A 1.37 kb EcoRI fragment carrying the ccpAgene was cloned into Escherichia coliplasmid pUC19 and B. subtilisplasmid pUB110, resulting it plasmids pCCPA19 and pCCPA110, respectively. The ccpAgene carried in pCCPA110 complemented the alsA1mutation. Western blotting revealed that the level of the CcpA protein in B. subtiliscells, which seemed to be constitutively synthesized, was approximately 10 times lower for the alsA1mutant than for the wild-type. The CcpA protein synthesized by either E. colicells bearing pCCPA19 or B. subtiliscells bearing pCCPA110 was purified to over 90% homogeneity; the latter cells were grown in the presence of glucose The molecular mass of the protein purified from E. coliwas 74 kDa, suggesting that this protein exists as a dimer because its subunit molecular mass was 38 kDa as determined by SDS-PAGE. Gel retardation analysis indicated that the purified CcpA protein in both cases did not bind to the cissequence for catabolite repression of the gntoperon, but it bound non-specifically to DNA., language=, type=