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Identification, Characterization, and Virulence Gene Expression of Marine Enterobacteria in the Upper Gulf of Thailand.

ABSTRACT: Enterobacteria that commonly inhabit marine environments have a great impact on human health. In this study, enterobacteria isolated from seawater in the Upper Gulf of Thailand were identified and characterized. Seawater from nine sampling sites along the Upper Gulf of Thailand contained presumptive enterobacteria that ranged from 0.22 ± 0.44 to 17.00 ± 3.97 CFU/mL. The 101 strains belonged to seven species in which Klebsiella pneumoniae was the majority (47.5% of strains). The highest prevalence was resistant to ampicillin (76.2%) and ticarcillin (72.3%), respectively, whereas none was resistant to imipenem. Forty-five antibiotic resistance patterns were observed and 33.7% exhibited multidrug resistance, emphasizing the concern about public health. Three β-lactamase genes, including ampC, blaSHV, and blaTEM, were detected at the frequencies of 47.5%, 21.8%, and 11.9%, respectively. Six virulence genes, including csgD, uge, kfu, eaeA, magA, and LTI, were detected at the frequencies of 37.6%, 31.7%, 19.8%, 16.8%, 12.9%, and 5.9%, respectively. The condition of 4% NaCl downregulated the expression of the kfu and uge genes. The 67.3% and 63.4% of strains synthesized silver nanoparticles ranging between 3.04 ± 0.64 and 20.64 ± 0.95 μg/mL and gold nanoparticles ranging between 7.77 ± 0.45 and 57.57 ± 8.00 μg/mL, respectively.

SUBMITTER: Nimnoi P

PROVIDER: S-EPMC8952428 | biostudies-literature | 2022 Feb

REPOSITORIES: biostudies-literature

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Identification, Characterization, and Virulence Gene Expression of Marine Enterobacteria in the Upper Gulf of Thailand.

Nimnoi Pongrawee P Pongsilp Neelawan N

Microorganisms 20220226 3

Enterobacteria that commonly inhabit marine environments have a great impact on human health. In this study, enterobacteria isolated from seawater in the Upper Gulf of Thailand were identified and characterized. Seawater from nine sampling sites along the Upper Gulf of Thailand contained presumptive enterobacteria that ranged from 0.22 ± 0.44 to 17.00 ± 3.97 CFU/mL. The 101 strains belonged to seven species in which <i>Klebsiella pneumoniae</i> was the majority (47.5% of strains). The highest pr ...[more]

PMID: 35336087

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Project description:BackgroundThe total bacterial community plays an important role in aquatic ecosystems. In this study, bacterial communities and diversity along the shores of the Upper Gulf of Thailand were first characterized. The association between bacterial communities and types of land use was also evaluated.ResultsThe bacterial communities and diversity of seawater in the Upper Gulf of Thailand, with regard to types of land use, were first revealed by using Illumina next-generation sequencing. A total of 4953 OTUs were observed from all samples in which 554 OTUs were common. The bacterial communities in sampling sites were significantly different from each other. The run-off water from three types of land use significantly affected the community richness and diversity of marine bacteria. Aquaculture sites contained the highest levels of community richness and diversity, followed by mangrove forests and tourist sites. Seawater physicochemical parameters including salinity, turbidity, TSS, total N, and BOD5, were significantly different when grouped by land use. The bacterial communities were mainly determined by salinity, total N, and total P. The species richness estimators and OTUs were positively correlated with turbidity. The top ten most abundant phyla and genera as well as the distribution of bacterial classes were characterized. The Proteobacteria constituted the largest proportions in all sampling sites, ranging between 67.31 and 78.80%. The numbers of the Marinobacterium, Neptuniibacter, Synechococcus, Candidatus Thiobios, hgcI clade (Actinobacteria), and Candidatus Pelagibacter were significantly different when grouped by land use.ConclusionsType of land use significantly affected bacterial communities and diversity along the Upper Gulf of Thailand. Turbidity was the most influential parameter affecting the variation in bacterial community composition. Salinity, total N, and P were the ones of the important factors that shaped the bacterial communities. In addition, the variations of bacterial communities from site-to-site were greater than within-site. The Proteobacteria, Bacteroidetes, Actinobacteria, Cyanobacteria, Verrucomicrobia, Euryarchaeota, Planctomycetes, Firmicutes, Deep Sea DHVEG-6, and Marinimicrobia were the most and common phyla distributed across the Upper Gulf of Thailand.

Project description:BACKGROUND: Fumarase catalyzes the reversible hydration of fumarate to L-malate and is a key enzyme in the tricarboxylic acid (TCA) cycle and in amino acid metabolism. Fumarase is also used for the industrial production of L-malate from the substrate fumarate. Thermostable and high-activity fumarases from organisms that inhabit extreme environments may have great potential in industry, biotechnology, and basic research. The marine environment is highly complex and considered one of the main reservoirs of microbial diversity on the planet. However, most of the microorganisms are inaccessible in nature and are not easily cultivated in the laboratory. Metagenomic approaches provide a powerful tool to isolate and identify enzymes with novel biocatalytic activities for various biotechnological applications. RESULTS: A plasmid metagenomic library was constructed from uncultivated marine microorganisms within marine water samples. Through sequence-based screening of the DNA library, a gene encoding a novel fumarase (named FumF) was isolated. Amino acid sequence analysis revealed that the FumF protein shared the greatest homology with Class II fumarate hydratases from Bacteroides sp. 2_1_33B and Parabacteroides distasonis ATCC 8503 (26% identical and 43% similar). The putative fumarase gene was subcloned into pETBlue-2 vector and expressed in E. coli BL21(DE3)pLysS. The recombinant protein was purified to homogeneity. Functional characterization by high performance liquid chromatography confirmed that the recombinant FumF protein catalyzed the hydration of fumarate to form L-malate. The maximum activity for FumF protein occurred at pH 8.5 and 55°C in 5 mM Mg(2+). The enzyme showed higher affinity and catalytic efficiency under optimal reaction conditions: K(m) = 0.48 mM, V(max) = 827 ?M/min/mg, and k(cat)/K(m) = 1900 mM/s. CONCLUSIONS: We isolated a novel fumarase gene, fumF, from a sequence-based screen of a plasmid metagenomic library from uncultivated marine microorganisms. The properties of FumF protein may be ideal for the industrial production of L-malate under higher temperature conditions. The identification of FumF underscores the potential of marine metagenome screening for novel biomolecules.

Dataset Information

Identification, Characterization, and Virulence Gene Expression of Marine Enterobacteria in the Upper Gulf of Thailand.

Publications

Identification, Characterization, and Virulence Gene Expression of Marine Enterobacteria in the Upper Gulf of Thailand.

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