2015 ; 25(2):
|Author||Mingxia Qi, Fei Mei, Hui Wang, Ming Sun, Gejiao Wang, Ziniu Yu, Yeonho Je, Mingshun Li|
|Affiliation||State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Hongshan District, Wuhan 430070, P.R. China|
|Title||Function of Global Regulator CodY in Bacillus thuringiensis BMB171 by Comparative Proteomic Analysis|
J. Microbiol. Biotechnol.2015 ; 25(2):
|Abstract||CodY is a highly conserved protein in low G+C gram-positive bacteria that regulates genes
involved in sporulation and stationary-phase adaptation. Bacillus thuringiensis is a grampositive
bacterium that forms spores and parasporal crystals during the stationary phase. To
our knowledge, the regulatory mechanism of CodY in B. thuringiensis is unknown. To study
the function of CodY protein in B. thuringiensis, BMB171codY- was constructed in a BMB171
strain. A shuttle vector containing the ORF of cry1Ac10 was transformed into BMB171 and
BMB171codY-, named BMB171cry1Ac and BMB171codY-cry1Ac, respectively. Some morphological
and physiological changes of codY mutant BMB171codY-cry1Ac were observed. A comparative
proteomic analysis was conducted for both BMB171codY-cry1Ac and BMB171cry1Ac through
two-dimensional gel electrophoresis and MALDI-TOF-MS/MS analysis. The results showed
that the proteins regulated by CodY are involved in microbial metabolism, including
branched-chain amino acid metabolism, carbohydrate metabolism, fatty acid metabolism, and
energy metabolism. Furthermore, we found CodY to be involved in sporulation, biosynthesis
of poly-β-hydroxybutyrate, growth, genetic competence, and translation. According to the
analysis of differentially expressed proteins, and physiological characterization of the codY
mutant, we performed bacterial one-hybrid and electrophoretic mobility shift assay
experiments and confirmed the direct regulation of genes by CodY, specifically those involved
in metabolism of branched-chain amino acids, ribosomal recycling factor FRR, and the late
competence protein ComER. Our data establish the foundation for in-depth study of the
regulation of CodY in B. thuringiensis, and also offer a potential biocatalyst for functions of
CodY in other bacteria.|
|Keywords||CodY, Bacillus thuringiensis, proteomics, poly-β-hydroxybutyrate, branched chain amino acids, FRR|
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