The Isolation, Screening, and Characterization of Polyhydroxyalkanoate-Producing Bacteria from Hypersaline Lakes in Kenya

Martin N. Muigano, S. Anami, J. Onguso, Godfrey M. Omare
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Abstract

Extremophilic microorganisms such as those that thrive in high-salt and high-alkaline environments are promising candidates for the recovery of useful biomaterials including polyhydroxyalkanoates (PHAs). PHAs are ideal alternatives to synthetic plastics because they are biodegradable, biocompatible, and environmentally friendly. This work was aimed at conducting a bioprospection of bacteria isolated from hypersaline-alkaliphilic lakes in Kenya for the potential production of PHAs. In the present study, 218 isolates were screened by Sudan Black B and Nile Red A staining. Of these isolates, 31 were positive for PHA production and were characterized using morphological, biochemical, and molecular methods. Through 16S rRNA sequencing, we found that the isolates belonged to the genera Arthrobacter spp., Bacillus spp., Exiguobacterium spp., Halomonas spp., Paracoccus spp., and Rhodobaca spp. Preliminary experiments revealed that Bacillus sp. JSM-1684023 isolated from Lake Magadi had the highest PHA accumulation ability, with an initial biomass-to-PHA conversion rate of 19.14% on a 2% glucose substrate. Under optimized fermentation conditions, MO22 had a maximum PHA concentration of 0.516 g/L from 1.99 g/L of cell dry weight and 25.9% PHA conversion, equivalent to a PHA yield of 0.02 g/g of biomass. The optimal PHA production media had an initial pH of 9.0, temperature of 35 °C, salinity of 3%, and an incubation period of 48 h with 2.5% sucrose and 0.1% peptone as carbon and nitrogen sources, respectively. This study suggests that bacteria isolated from hypersaline and alkaliphilic tropical lakes are promising candidates for the production of polyhydroxyalkanoates.
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肯尼亚高盐湖聚羟基烷酸产菌的分离、筛选和鉴定
嗜极微生物,如那些在高盐和高碱性环境中茁壮成长的微生物,是回收有用生物材料(包括聚羟基烷酸酯(pha))的有希望的候选者。pha是合成塑料的理想替代品,因为它们具有可生物降解、生物相容性和环境友好性。这项工作的目的是对肯尼亚高盐嗜碱湖泊中分离的细菌进行生物展望,以用于生产pha的潜力。本研究用苏丹黑B和尼罗红A染色对218株分离株进行了筛选。在这些分离株中,31株PHA生产阳性,并通过形态学、生化和分子方法进行了鉴定。通过16S rRNA测序发现,分离菌株分别属于节杆菌属、芽孢杆菌属、出iguobacterium属、Halomonas属、副球菌属和Rhodobaca属。初步实验表明,从马加迪湖分离的芽孢杆菌sp. JSM-1684023具有最高的PHA积累能力,在2%葡萄糖底物下,其初始生物量转化为PHA的转化率为19.14%。在优化的发酵条件下,在细胞干重1.99 g/L的条件下,MO22的最大PHA浓度为0.516 g/L, PHA转化率为25.9%,相当于0.02 g/g生物质的PHA产量。最佳PHA生产培养基初始pH为9.0,温度为35℃,盐度为3%,碳源为2.5%蔗糖,氮源为0.1%蛋白胨,培养时间为48 h。这项研究表明,从高盐和亲碱的热带湖泊中分离出来的细菌是生产聚羟基烷酸盐的有希望的候选者。
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