一株新型甘露聚糖酶产生菌所产甘露聚糖酶的性质研究。

Characterization of mannanase from a novel mannanase-producing bacterium.

机构信息

Department of Sea Food Science, National Kaohsiung Marine University, No. 142 Hai-Chuan Road, Nan-Tzu, Kaohsiung 81143, Taiwan.

出版信息

J Agric Food Chem. 2012 Jun 27;60(25):6425-31. doi: 10.1021/jf301944e. Epub 2012 Jun 19.

DOI:10.1021/jf301944e

PMID:22694324

Abstract

Locust bean gum (LBG) was employed to screen mannanase-producing bacteria. The bacterium with highest mannanase ability was identified as Paenibacillus cookii. It revealed highest activity (6.67 U/mL) when cultivated in 0.1% LBG with 1.5% soytone and 0.5% tryptone after 4 days incubation at 27 °C. Its mannanase was purified to electrophoretical homogeneity after DEAE-Sepharose and Sephacryl S-100 separation. The purified mannanase, with an N-terminus of GLFGINAY, had pH and temperature optimum at 5.0 and 50 °C, respectively, and was stable at pH 5.0-7.0, ≤ 50 °C. It was strongly activated by β-mercaptoethanol, dithiothreitol, cysteine, and glutathione, but inhibited by Hg(2+), Cu(2+), Zn(2+), Fe(3+), PMSF, iodoacetic acid, and EDTA. According to substrate specificity study, the purified mannanase had high specificity to LBG and konjac.

摘要

槐豆胶（LBG）被用于筛选甘露聚糖酶产生菌。具有最高甘露聚糖酶活性的细菌被鉴定为蜡状芽孢杆菌。当在 0.1%的 LBG 中培养，添加 1.5%的大豆胨和 0.5%的胰蛋白胨，在 27°C 下培养 4 天后，其酶活最高（6.67 U/mL）。该甘露聚糖酶经 DEAE-Sepharose 和 Sephacryl S-100 分离后达到电泳纯。纯化的甘露聚糖酶的 N 端为 GLFGINAY，其最适 pH 和温度分别为 5.0 和 50°C，在 pH 5.0-7.0、≤50°C 条件下稳定。它被β-巯基乙醇、二硫苏糖醇、半胱氨酸和谷胱甘肽强烈激活，但被 Hg(2+)、Cu(2+)、Zn(2+)、Fe(3+)、PMSF、碘乙酸和 EDTA 抑制。根据底物特异性研究，纯化的甘露聚糖酶对 LBG 和魔芋具有很高的特异性。