Molecular cloning and antimicrobial activity of enterolysin A and helveticin J of bacteriolysins from metagenome of Chinese traditional fermented foods

Zhang, Tiantian; Pan, Yingjie; Li, Bailin; Ou, Jie; Zhang, Jianmin; Chen, Yourong; Peng, Xu; Chen, Lanming

doi:10.1016/j.foodcont.2012.11.015

Cited by 17 publications

(8 citation statements)

References 31 publications

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“…Oligonucleotide primers were synthesized by Shanghai Sangon Biological Engineering Technology and Services Co., Ltd. (Shanghai, China). The PCR amplification was performed in a 20 μL reaction volume as described previously [ 39 ]. Amplification was performed in a MastercyclerW pro PCR thermal cycler (Eppendorf, Hamburg, Germany) with a touch down PCR protocol [ 38 ].…”

Section: Methodsmentioning

confidence: 99%

Identification of Bacterial Community Composition in Freshwater Aquaculture System Farming of Litopenaeus vannamei Reveals Distinct Temperature-Driven Patterns

Tang

Tao

Tan

et al. 2014

IJMS

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Change in temperature is often a major environmental factor in triggering waterborne disease outbreaks. Previous research has revealed temporal and spatial patterns of bacterial population in several aquatic ecosystems. To date, very little information is available on aquaculture environment. Here, we assessed environmental temperature effects on bacterial community composition in freshwater aquaculture system farming of Litopenaeus vannamei (FASFL). Water samples were collected over a one-year period, and aquatic bacteria were characterized by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and 16S rDNA pyrosequencing. Resulting DGGE fingerprints revealed a specific and dynamic bacterial population structure with considerable variation over the seasonal change, suggesting that environmental temperature was a key driver of bacterial population in the FASFL. Pyrosequencing data further demonstrated substantial difference in bacterial community composition between the water at higher (WHT) and at lower (WLT) temperatures in the FASFL. Actinobacteria, Proteobacteria and Bacteroidetes were the highest abundant phyla in the FASFL, however, a large number of unclassified bacteria contributed the most to the observed variation in phylogenetic diversity. The WHT harbored remarkably higher diversity and richness in bacterial composition at genus and species levels when compared to the WLT. Some potential pathogenenic species were identified in both WHT and WLT, providing data in support of aquatic animal health management in the aquaculture industry.

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Section: Methodsmentioning

confidence: 99%

Identification of Bacterial Community Composition in Freshwater Aquaculture System Farming of Litopenaeus vannamei Reveals Distinct Temperature-Driven Patterns

Tang

Tao

Tan

et al. 2014

IJMS

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“…Sediment samples were collected from a few aquaculture ponds of fishery farms located in Shanghai, China. Samples were homogenized in nine volumes of a sterilized PBS solution buffer (pH 7.4) [ 35 ], and microbial cells in supernatant were collected as previously described by Zhang et al [ 37 ]. Serial dilutions were made and spread onto the selective skim milk agar plates, which were incubated at 30°C for 48 h. Single colonies were scored positive for protease activity using a skim milk assay in which the isolates form clear zone around their colonies resulting from skim milk hydrolysis are scored as positive in protease activity.…”

Section: Methodsmentioning

confidence: 99%

A novel carboxyl-terminal protease derived from Paenibacillus lautusCHN26 exhibiting high activities at multiple sites of substrates

Pan

She

et al. 2013

BMC Biotechnol

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BackgroundCarboxyl-terminal protease (CtpA) plays essential functions in posttranslational protein processing in prokaryotic and eukaryotic cells. To date, only a few bacterial ctpA genes have been characterized. Here we cloned and characterized a novel CtpA. The encoding gene, ctpAp (ctpA of Paenibacillus lautus), was derived from P. lautus CHN26, a Gram-positive bacterium isolated by functional screening. Recombinant protein was obtained from protein over-expression in Escherichia coli and the biochemical properties of the enzyme were investigated.ResultsScreening of environmental sediment samples with a skim milk-containing medium led to the isolation of a P. lautus CHN26 strain that exhibited a high proteolytic activity. A gene encoding a carboxyl-terminal protease (ctpAp) was cloned from the isolate and characterized. The deduced mature protein contains 466 aa with a calculated molecular mass of 51.94 kDa, displaying 29-38% amino acid sequence identity to characterized bacterial CtpA enzymes. CtpAp contains an unusual catalytic dyad (Ser309-Lys334) and a PDZ substrate-binding motif, characteristic for carboxyl-terminal proteases. CtpAp was expressed as a recombinant protein and characterized. The purified enzyme showed an endopeptidase activity, which effectively cleaved α S1- and β- casein substrates at carboxyl-terminus as well as at multiple internal sites. Furthermore, CtpAp exhibited a high activity at room temperature and strong tolerance to conventional protease inhibitors, demonstrating that CtpAp is a novel endopeptidase.ConclusionsOur work on CtpA represents the first investigation of a member of Family II CtpA enzymes. The gene was derived from a newly isolated P. lautus CHN26 strain exhibiting a high protease activity in the skim milk assay. We have demonstrated that CtpAp is a novel endopeptidase with distinct cleavage specificities, showing a strong potential in biotechnology and industry applications.

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“…The strains from kefir grains were close to the root of the phylogenetic tree, and so the other strains may have originated from kefir grains, which may have lost some genes in the evolution process and evolved to be more adapted to the environment of a single milk source. The presence of the bacteriocin operon predicted suggests that the strain has antibacterial ability and many phenotypic and animal tests have shown that this is associated with an inhibitory ability against pathogenic bacteria [ 39 ]. The distribution of bacteriocins in different strains was also different.…”

Section: Discussionmentioning

confidence: 99%

Comparative Genomics Analysis of Habitat Adaptation by Lactobacillus kefiranofaciens

Luo

Liu

et al. 2023

Foods

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Lactobacillus kefiranofaciens is often found in fermented dairy products. Many strains of this species have probiotic properties, contributing to the regulation of immune metabolism and intestinal flora. This species was added to the list of lactic acid bacteria that can be added to food in China, in 2020. However, research on the genomics of this species is scarce. In this study we undertook whole genome sequencing analysis of 82 strains of L. kefiranofaciens from different habitats, of which 9 strains were downloaded from the NCBI RefSeq (National Center for Biotechnology Information RefSeq). The mean genome size of the 82 strains was 2.05 ± 0.25 Mbp, and the mean DNA G + C content was 37.47 ± 0.42%. The phylogenetic evolutionary tree for the core genes showed that all strains belonged to five clades with clear aggregation in relation to the isolation habitat; this indicated that the genetic evolution of L. kefiranofaciens was correlated to the isolation habitat. Analysis of the annotation results identified differences in the functional genes, carbohydrate active enzymes (CAZy) and bacteriocins amongst different isolated strains, which were related to the environment. Isolates from kefir grains had more enzymes for cellulose metabolism and a better ability to use vegetative substrates for fermentation, which could be used in feed production. Isolates from kefir grains also had fewer kinds of bacteriocin than isolates from sour milk and koumiss; helveticin J and lanthipeptide class I were not found in the isolates from kefir grains. The genomic characteristics and evolutionary process of L. kefiranofaciens were analyzed by comparative genomics and this paper explored the differences in the functional genes amongst the strains, aiming to provide a theoretical basis for the research and development of L. kefiranofaciens.

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Molecular cloning and antimicrobial activity of enterolysin A and helveticin J of bacteriolysins from metagenome of Chinese traditional fermented foods

Cited by 17 publications

References 31 publications

Identification of Bacterial Community Composition in Freshwater Aquaculture System Farming of Litopenaeus vannamei Reveals Distinct Temperature-Driven Patterns

Identification of Bacterial Community Composition in Freshwater Aquaculture System Farming of Litopenaeus vannamei Reveals Distinct Temperature-Driven Patterns

A novel carboxyl-terminal protease derived from Paenibacillus lautusCHN26 exhibiting high activities at multiple sites of substrates

Comparative Genomics Analysis of Habitat Adaptation by Lactobacillus kefiranofaciens

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