Screening of genes involved in chitinase production in Aeromonas caviae CB101 via transposon mutagenesis

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Abstract

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.

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