当前位置: X-MOL 学术Prep. Biochem. Biotechnol. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Purification and characterization of Stenotrophomonas maltophilia chitinase with antifungal and insecticidal properties
Preparative Biochemistry & Biotechnology ( IF 2.9 ) Pub Date : 2022-11-11 , DOI: 10.1080/10826068.2022.2142942
Cigdem Aktas 1 , Damla Ruzgar 1 , Sumeyra Gurkok 2 , Arzu Gormez 3
Affiliation  

Abstract

This study aimed to determine the ability of bacteria to produce the chitinase enzyme, purify, and characterize the enzyme from the isolate with the best activity, and determine the use of this purified enzyme as a biocontrol agent. The chitinolytic bacterium was identified as Stenotrophomonas maltophilia. The chitinase enzyme was purified 1.4 times at a 30% ammonium sulfate concentration with a yield of 40.7%. Following partial purification, the enzyme was purified by ion-exchange chromatography using HiPrep Q XL 16/10 column and HiPrep™ 26/10 desalting column with 25.34% and 18.12% yields, respectively. It was calculated that the purified enzyme had a molecular weight of 52 kDa by sodium dodecyl sulfate–polyacrylamide gel electrophoresis. The optimum activity of the enzyme was determined at 50 °C and pH 7.0. Enzyme activity was most induced by Fe2+, while it was most inhibited by Zn2+ at 5 mM concentration. Km and Vmax values of the enzyme for colloidal chitin were calculated as 1.6419 mg/mL and 16.129 U/mg, respectively. The purified chitinase was used as a biocontrol agent against the fungus Fusarium oxysporum and potato beetle Leptinotarsa decemlineata. The enzyme was shown to be effective in reducing the growth of fungus and causing disruption of the chitin structure of potato beetle.



中文翻译:

具有抗真菌和杀虫特性的嗜麦芽寡养单胞菌几丁质酶的纯化和表征

摘要

本研究旨在确定细菌产生几丁质酶的能力,从具有最佳活性的分离物中纯化和表征该酶,并确定这种纯化酶作为生物防治剂的用途。该几丁质分解细菌被鉴定为嗜麦芽寡养单胞菌。几丁质酶在30%硫酸铵浓度下纯化1.4倍,产率40.7%。部分纯化后,使用 HiPrep Q XL 16/10 柱和 HiPrep™ 26/10 脱盐柱通过离子交换色谱纯化酶,产率分别为 25.34% 和 18.12%。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳计算纯化酶的分子量为52 kDa。酶的最佳活性在 50 °C 和 pH 7.0 下确定。在5 mM浓度下,酶活性最受Fe 2+诱导,而最受Zn 2+抑制。胶体几丁质酶的K mV max值分别计算为 1.6419 mg/mL 和 16.129 U/mg。纯化的几丁质酶被用作针对真菌尖镰孢和马铃薯甲虫Leptinotarsa decemlineata 的生物防治剂。该酶被证明可以有效减少真菌的生长并破坏马铃薯甲虫的几丁质结构。

更新日期:2022-11-11
down
wechat
bug