Phenotypic and Genetic Characterization of Polyhydroxyalkanoate Producing Bacterial Isolates from Hypersaline Water Body, Atlantic Ocean, Nigeria

Fayemi Scott O., Akiibinu Moses O., Fatokun Evelyn N., Orukotan Abimbola A.
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Abstract

Introduction: The importance of polyhydroxylalkanoate (PHA) producing bacteria is hinged on the basis that PHA itself possess environmentally advantages over synthetic plastics therefore, the proper identity of these bacteria is inevitable. Aim: To determine the bacteria identification methods of phenotypic and genetic identification for marine sourced effective polyhydroxylalkanoate (PHA) production. Study Design: Randomised design was employed for PHA producing bacterial isolates. Place and Duration of Study: Samples containing PHA-producing bacteria from the hypersaline water body of Nigerian Southern Atlantic Ocean (6.35ᴼ N 3.28ᴼ E -St1; 6.35ᴼ N 3.40ᴼ E -St2; and 6.36ᴼ N 3.47ᴼ E -St3); water depths (0.07m (top), 50m (middle) and 100m (bottom) were collected and investigated between August, 2016-September, 2019. Methodology: Bacterial isolates was characterized using standard microbiological and biochemical tests for the phenotypes, and 16S rDNA for the genotypes.  Isolates were also screened for PHA potentials using specific primers. Results: Genetically identified Bacillus cereus strains LB17 and Bacillus paramycoides SA27 were phenotypically identified as Bacillus badius. Similarly, Acinetobacter calcoaceticus JL11, five strains of Alcaligenes faecalis, seven strains of Bacillus spp., Enterobacter cloacae, Falsochrobactrum ovis, Ochrobactrum ciceri, Providencia stuartii including two species of Pseudomonas and Bordetella trematum were all phenotypically identified as Corynebacterium kutsceri. Invariably, Alcaligenes faecalis (strains PSD10 and DEP8), Pseudomonas aeruginosa (H47921) and Vagococcus fluvialis (AWW1) were identified as Staphylococcus species. All 34 bacterial isolates expressed phaC genes for PHA potential, while Alcaligenes faecalis strains and Bacillus humi (NBPP9) expressed phaC1 for short PHA chain. Conclusion: This study established variations in the genetical identity of Corynebacterium kutsceri, Enterobacter amnigenus, Micrococcus luteus, Micrococcus varians, Staphylococcus epidermidis and Staphylococcus saprophyticus when compared with the phenotypic identification methods. But, there was synchronization in the genotypic and phenotypic identity of bacillus species. Records of varietal differences of PHA potential were also obtained from same species in this research. It is therefore pertinent to rely on the genetic identification of bacteria for the effective determination of PHA-producing character as displayed in Bacillus spp.
尼日利亚大西洋低盐度水体中产生聚羟基烷酸的细菌分离物的表型和遗传特征
导言:生产聚羟基烷酸(PHA)细菌的重要性在于 PHA 本身比合成塑料具有环境优势,因此,这些细菌的正确识别是不可避免的。目的:确定表型鉴定和基因鉴定的细菌鉴定方法,以有效生产海洋来源的聚羟基烷酸(PHA)。研究设计:对生产 PHA 的细菌分离物采用随机设计。研究地点和时间:含 PHA 生产细菌的样本来自尼日利亚南大西洋的高盐度水体(6.35ᴼ N 3.28ᴼ E -St1;6.35ᴼ N 3.40ᴼ E -St2;6.35ᴼ N 3.40ᴼ E -St3)。36ᴼ N 3.47ᴼ E -St3);水深(0.07 米(上层)、50 米(中层)和 100 米(下层)。研究方法:细菌分离物的表型采用标准微生物学和生物化学测试,基因型采用 16S rDNA。 还使用特定引物对分离物进行 PHA 电位筛选。结果经基因鉴定的蜡样芽孢杆菌菌株 LB17 和副溶血芽孢杆菌 SA27 经表型鉴定为巴迪乌斯芽孢杆菌。同样,醋酸钙化杆菌 JL11、5 株粪钙化杆菌、7 株芽孢杆菌属、泄殖腔肠杆菌、Falsochrobactrum ovis、Ochrobactrum ciceri、Providencia stuartii(包括 2 种假单胞菌)和震颤博德特氏菌都被表型鉴定为库茨法氏棒状杆菌。粪钙杆菌(菌株 PSD10 和 DEP8)、铜绿假单胞菌(H47921)和河口阴道球菌(AWW1)无一例外地被鉴定为葡萄球菌。所有 34 个细菌分离物都表达了 PHA 潜能的 phaC 基因,而粪钙球菌菌株和腐生芽孢杆菌(NBPP9)则表达了短 PHA 链的 phaC1 基因。结论与表型鉴定方法相比,本研究确定了库氏棒状杆菌(Corynebacterium kutsceri)、安氏肠杆菌(Enterobacter amnigenus)、黄体微球菌(Micrococcus luteus)、变异微球菌(Micrococcus varians)、表皮葡萄球菌(Staphylococcus epidermidis)和溶血性葡萄球菌(Staphylococcus saprophyticus)在基因鉴定方面的差异。但是,芽孢杆菌的基因型和表型鉴定是同步的。本研究还从同一物种中获得了 PHA 潜力的品种差异记录。因此,有必要依靠细菌的基因鉴定来有效确定芽孢杆菌属的 PHA 生产特性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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