极端耐碱甘露聚糖酶来源于地衣芽孢杆菌 subsp. inaquosorum CSB31，具有生物工业应用前景。

Prospects for Bio-Industrial Application of an Extremely Alkaline Mannanase From Bacillus subtilis subsp. inaquosorum CSB31.

机构信息

Department of Pharmacy, Chosun University, 309, Pilmun-daero, Dong-Gu, Gwangju 61452, Republic of Korea.

Department of Biotechnology, Sangmyung University, 20, Hongjimun 2-Gil, Jongno-Gu, Seoul 03016, Republic of Korea.

出版信息

Biotechnol J. 2017 Nov;12(11). doi: 10.1002/biot.201700113. Epub 2017 Sep 18.

DOI:10.1002/biot.201700113

PMID:28843040

Abstract

Mannan-degrading enzymes have been growing interest in bio-industrial applications, such as the pulp and paper, food, and pharmaceutical industries. In this study, an extremely alkaline mannanase (MnB31) is produced by Bacillus subtilis subsp. inaquosorum CSB31. MnB31 is purified to 17.92-fold with a 21.51% yield and specific activity of 1,796.13 U mg by anion-exchange and gel filtration column chromatography. The biochemical characterization of MnB31 is performed, and the results are as follows: molecular weight of ≈47 kDa with an optimum temperature of 60 °C and pH of 12.5. The enzyme is strongly activated by Co , Mn , Na , and K , and inhibited by Zn , Ni , and Mg . Halo-tolerance (10% NaCl), urea stability (3 M), and protease resistance are also observed. The kinetic parameters of MnB31 are found to be K of 0.043 mg ml , and V of 1,046 ± 3.605 U mg , respectively. In addition, the thermodynamical parameters are investigated; the activation energy (E ) is found to be 31.36 kJ mol with a K value of 156.9 × 10 s , ΔH (28.59 kJ mol ), ΔG (42.38 kJ mol ), ΔS (-41.39 J mol K ), Q (1.40), ΔG (-8.697 kJ mol ), and ΔG (-48.22 kJ mol ). These results suggest that MnB31 has potential bio-industrial application, due to its greater hydrolytic efficiency and feasibility of enzymatic reaction.

摘要

甘露聚糖降解酶在生物工业应用中越来越受到关注，例如纸浆和造纸、食品和制药等行业。在这项研究中，从地衣芽孢杆菌亚种 CSB31 中生产出一种极其碱性的甘露聚糖酶（MnB31）。MnB31 通过阴离子交换和凝胶过滤柱层析纯化至 17.92 倍，产率为 21.51%，比活为 1796.13 U mg。对 MnB31 的生化特性进行了研究，结果如下：分子量约为 47 kDa，最适温度为 60°C，最适 pH 为 12.5。该酶被 Co、Mn、Na 和 K 强烈激活，被 Zn、Ni 和 Mg 抑制。还观察到耐卤（10%NaCl）、脲稳定性（3 M）和蛋白酶抗性。MnB31 的动力学参数为 K 为 0.043 mg ml，V 为 1046±3.605 U mg。此外，还研究了热力学参数；发现活化能（E）为 31.36 kJ mol，K 值为 156.9×10 s，ΔH（28.59 kJ mol），ΔG（42.38 kJ mol），ΔS（-41.39 J mol K），Q（1.40），ΔG（-8.697 kJ mol），和 ΔG（-48.22 kJ mol）。这些结果表明，由于 MnB31 具有更高的水解效率和酶反应的可行性，因此具有潜在的生物工业应用价值。