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首页> 外文期刊>Journal of Polymers and the Environment >Production and Purification of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Degrading Enzyme from Streptomyces sp. AF-111
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Production and Purification of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Degrading Enzyme from Streptomyces sp. AF-111

机译：链霉菌（Streptomyces sp。）的聚（3-羟基丁酸酯-co-3-羟基戊酸酯）降解酶的生产和纯化。 AF-111

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摘要

A poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) degrading bacterial strain designated as AF-111 was isolated from sewage sludge sample. The bacterium was identified by 16S rRNA gene sequencing. The results revealed that strain AF-111 showed 99 % similarity with Streptomyces althioticus strain NRRL B-3981 and designated as Streptomyces sp. strain AF-111. An extracellular PHBV depolymerase enzyme was produced under optimized conditions and purified through ammonium sulphate fractionation and column chromatography. The enzyme was purified to homogeneity, indicated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and molecular weight was found to be approximately 51 kDa. Effect of temperature, pH, metal ions and inhibitors on the PHBV depolymerase activity was determined. The enzyme was stable at wide range of temperature (35-55 ℃) and pH (6-8). PHBV depolymerase was stable in the presence of different metal ions except iron and zinc which had inhibitory effect on depolymerase activity. Both ethylene-diamine teteracetic acid and phenylmethyl sulphonyl fluoride strongly inhibited enzyme activity which indicates that this enzyme belongs to the serine hydrolase family like other polyhydroxyalkanoate depolymerases. The results show that a depolymerase from strain AF-111 can effectively degrade PHBV, therefore, it can be applied in the process of biochemical monomer recycling.
机译：从污水污泥样品中分离出降解为菌株AF-111的聚3-羟基丁酸酯-3-羟基戊酸共聚物（PHBV）。通过16S rRNA基因测序鉴定细菌。结果表明，菌株AF-111与硫链霉菌NRRL B-3981菌株具有99％的相似性，并命名为链霉菌sp。 AF-111株。在最佳条件下产生了细胞外PHBV解聚酶，并通过硫酸铵分级分离和柱色谱法纯化。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳表明该酶被纯化至均一，发现分子量约为51 kDa。确定了温度，pH，金属离子和抑制剂对PHBV解聚酶活性的影响。该酶在很宽的温度范围（35-55℃）和pH值（6-8）下均稳定。除铁和锌对解聚酶活性有抑制作用外，PHBV解聚酶在存在不同金属离子的情况下是稳定的。乙二胺四乙酸和苯甲基磺酰氟都强烈抑制酶活性，这表明该酶与其他聚羟基链烷酸酯解聚酶一样属于丝氨酸水解酶家族。结果表明，菌株AF-111的解聚酶可有效降解PHBV，因此可用于生化单体的回收利用。

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