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Crosslinking Mechanism on a Novel Bacillus cereus Transglutaminase-Mediated Conjugation of Food Proteins.

ABSTRACT: Until now, Streptoverticillium mobaraense transglutaminase (TG) is the only commercialized TG, but limited information is known about its selection tendency on crosslinking sites at the protein level, restricting its application in the food industry. Here, four recombinant Bacillus TGs were stable in a broad range of pH (5.0−9.0) and temperatures (<50 °C), exhibiting their maximum activity at 50−60 °C and pH 6.0−7.0. Among them, TG of B. cereus (BCETG) demonstrated the maximal specific activity of 177 U/mg. A structural analysis indicated that the Ala147-Ala156 region in the substrate tunnel of BCETG played a vital role in catalytic activity. Furthermore, bovine serum albumin, as well as nearly all protein ingredients in soy protein isolate and whey protein, could be cross-linked by BCETG, and the internal crosslinking paths of three protein substrates were elucidated. This study demonstrated Bacillus TGs are a candidate for protein crosslinking and provided their crosslinking mechanism at the protein level for applications in food processing.

SUBMITTER: Wang H

PROVIDER: S-EPMC9689123 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature

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Crosslinking Mechanism on a Novel <i>Bacillus cereus</i> Transglutaminase-Mediated Conjugation of Food Proteins.

Wang Hongbin H Zhang Yuanfu Y Yuan Zhaoting Z Zou Xiaotong X Ji Yuan Y Hou Jiayi J Zhang Jinfang J Lu Fuping F Liu Yihan Y

Foods (Basel, Switzerland) 20221119 22

Until now, Streptoverticillium mobaraense transglutaminase (TG) is the only commercialized TG, but limited information is known about its selection tendency on crosslinking sites at the protein level, restricting its application in the food industry. Here, four recombinant Bacillus TGs were stable in a broad range of pH (5.0−9.0) and temperatures (<50 °C), exhibiting their maximum activity at 50−60 °C and pH 6.0−7.0. Among them, TG of B. cereus (BCETG) demonstrated the maximal specific activity ...[more]

PMID: 36429314

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Project description:IntroductionBacillus cereus is a ubiquitous opportunistic human pathogen that causes food intoxications worldwide. However, the genomic characteristics and pathogenic mechanisms of B. cereus are still unclear.MethodsHere, we isolated and purified nine strains of B. cereus (LY01-LY09) that caused vomiting, diarrhea and other symptoms from four foodborne outbreaks happened in Guizhou Province in southwest China from June to September 2021. After colony observation, Gram staining, microscopic examination and biochemical test, they were identified as B. cereus. The genomic characteristics, phylogenetic relationships and virulence factors of the isolated strains were analyzed at the genome level. Genome sequencing, comparative genomic analysis, secondary metabolite analysis and quantitative PCR were utilized to give a thorough exploration of the strains.ResultsWe obtained the genome maps of LY01-LY09 and found that LY01-LY09 had a complex interspecific relationship with B. anthracis and B. thuringiensis. We also observed a contraction of gene families in LY01-LY09, and the contracted families were mainly associated with prophage, which contributed to the species diversity of B. cereus. The Hsp20 gene family underwent a rapid evolution in LY01-LY09, which facilitated the adaptation of the strains to adverse environmental conditions. Moreover, the LY01-LY09 strains exhibited a higher copy number in the non-ribosomal polypeptide synthetase (NRPS) genes and carried the complete cereulide synthetase (ces) gene cluster sequences. Considering that the NRPS system is a classical regulatory mechanism for emetic toxin synthesis, we hypothesized that LY01-LY09 could synthesize emetic toxins through the regulation of ces gene clusters by the NRPS system.DiscussionThese findings are important for further investigation into the evolutionary relationship between B. cereus and their related species, as well as the underlying mechanisms governing the synthesis and secretion of bacterial toxins.

Dataset Information

Crosslinking Mechanism on a Novel Bacillus cereus Transglutaminase-Mediated Conjugation of Food Proteins.

Publications

Crosslinking Mechanism on a Novel <i>Bacillus cereus</i> Transglutaminase-Mediated Conjugation of Food Proteins.

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