Journal of Applied Microbiology, Vol.102, No.3, 640-649, 2007
Screening of genes involved in chitinase production in Aeromonas caviae CB101 via transposon mutagenesis
Aims: To find genes involved in chitinase production in chitinolytic bacterium Aeromonas caviae CB101. Methods and Results: By transposome mutagenesis, a high-quality mutant library containing around 20 000 insertion mutants was constructed in A. caviae CB101. Mutants with higher, lower and delayed chitinase-producing abilities were identified and analysed further. Genomic sequences flanking the insertion sites of these mutants were amplified by thermal asymmetric interlaced-PCR, cloned and sequenced. The mutated genes involved in chitinase production and/or secretion in CB101 include (i) nagA and nagB gene homologues that are related to the metabolism of the chitin digestion product GlcNAc; (ii) ftsX and exeL gene homologues that are related to transport or secretion systems; (iii) varA and rpoH gene homologues that are related to transcriptional regulator sequences; (iv) other genes with unknown functions. Conclusions: Transposome mutagenesis is an efficient method to identify genes involved in the chitinase production in CB101. Chitinase production in CB101 is a complex system, and genes with various functions were identified in this study. Significance and Impact of the study: Understanding regulation of chitinase production in CB101 would make molecular engineering of the bacterium for higher enzyme production possible.