Genetics and Molecular Biology of Industrial Organisms an Alkaline-active and Alkali-stable Pectate Lyase from Streptomyces Sp. S27 with Potential in Textile Industry
Genetics and Molecular Biology of Industrial Organisms an Alkaline-active and Alkali-stable Pectate Lyase from Streptomyces Sp. S27 with Potential in Textile Industry
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P. Yuan;bullet Kun;Meng bullet;P. Shi;bullet Huiying;Luo bullet;Huo-qing Huang;bullet Tao
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作者：
P. Yuan;bullet Kun;Meng bullet;P. Shi;bullet Huiying;Luo bullet;Huo-qing Huang;bullet Tao
从链霉菌S27中克隆了果胶酸裂解酶基因（pl-str），并在大肠杆菌Rosetta中表达。pl-str全长972 bp，编码一个323个氨基酸的蛋白质，不含信号肽，属于PF 00544家族。重组酶（r-PL-STR）纯化电泳均匀，使用Ni 2？-NTA层析，显示表观分子量为35 kDa。r-PL-STR的最适pH为10.0，在pH 12.0时，其最大活性为70%。在37°C下孵育1小时后，无底物，酶在pH 7.0-12.0下保持超过55%的活性。与Novozymes的商品化复合酶Scourzyme@301L相比，纯化的r-PL-STR显示出相似的降低聚半乳糖醛酸的特性粘度的功效（49.0vs.49.7%）。当与纤维素酶和α-淀粉酶组合时，r-PL-STR在黄麻织物的生物精练中具有相当的性能（22.39vs.22.99%）。因此，r-PL-STR可能代表用于碱性工业如纺织的良好候选者。
A pectate lyase gene (pl-str) was cloned from Streptomyces sp. S27 and expressed in Escherichia coli Rosetta. The full-length pl-str consists of 972 bp and encodes for a protein of 323 amino acids without signal peptide that belongs to family PF00544. The recombinant enzyme (r-PL-STR) was purified to electrophoretic homogeneity using Ni 2? –NTA chromatography and showed apparent molecular mass of *35 kDa. The pH optimum of r-PL-STR was found to be 10.0, and it exhibited [70% of the maximal activity at pH 12.0. After incubation at 37°C for 1 h without substrate, the enzyme retained more than 55% activity at pH 7.0–12.0. Compared with the commercial complex enzyme Scourzyme @ 301L from Novozymes, purified r-PL-STR showed similar efficacy in reducing the intrinsic viscosity of polygalacturonic acid (49.0 vs. 49.7%). When combined with cellulase and a-amylase, r-PL-STR had comparable performance in bi-oscouring of jute fabric (22.39 vs. 22.99%). Thus, r-PL-STR might represent a good candidate for use in alkaline industries such as textile.