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一种新型血红素过氧化氢酶在枯草芽孢杆菌168中的异源表达与特性分析

Heterologous expression and characterization of a new heme-catalase in Bacillus subtilis 168.

作者信息

Philibert Tuyishime, Rao Zhiming, Yang Taowei, Zhou Junping, Huang Genshu, Irene Komera, Samuel Niyomukiza

机构信息

The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, 214122, Wuxi, Jiangsu, People's Republic of China.

出版信息

J Ind Microbiol Biotechnol. 2016 Jun;43(6):729-40. doi: 10.1007/s10295-016-1758-2. Epub 2016 Mar 26.

Abstract

Reactive oxygen species (ROS) is an inherent consequence to all aerobically living organisms that might lead to the cells being lethal and susceptible to oxidative stress. Bacillus pumilus is characterized by high-resistance oxidative stress that stimulated our interest to investigate the heterologous expression and characterization of heme-catalase as potential biocatalyst. Results indicated that recombinant enzyme significantly exhibited the high catalytic activity of 55,784 U/mg expressed in Bacillus subtilis 168 and 98.097 µmol/min/mg peroxidatic activity, the apparent K m of catalytic activity was 59.6 ± 13 mM with higher turnover rate (K cat = 322.651 × 10(3) s(-1)). The pH dependence of catalatic and peroxidatic activity was pH 7.0 and pH 4.5 respectively with temperature dependence of 40 °C and the recombinant heme-catalase exhibited a strong Fe(2+) preference. It was further revealed that catalase KatX2 improved the resistance oxidative stress of B. subtilis. These findings suggest that this B. pumilus heme-catalase can be considered among the industrially relevant biocatalysts due to its exceptional catalytic rate and high stability and it can be a potential candidate for the improvement of oxidative resistance of industrially produced strains.

摘要

活性氧(ROS)是所有需氧生物的固有产物,可能导致细胞死亡并易受氧化应激影响。短小芽孢杆菌具有高抗氧化应激能力,这激发了我们研究血红素过氧化氢酶作为潜在生物催化剂的异源表达和特性的兴趣。结果表明,重组酶在枯草芽孢杆菌168中显著表现出55,784 U/mg的高催化活性和98.097 μmol/min/mg过氧化物酶活性,催化活性的表观K m为59.6±​13 mM,周转速率较高(K cat = 322.651×​10(3) s(-1))。催化活性和过氧化物酶活性的pH依赖性分别为pH 7.0和pH 4.5,温度依赖性为40°C,重组血红素过氧化氢酶表现出强烈的Fe(2+)偏好性。进一步研究表明,过氧化氢酶KatX2提高了枯草芽孢杆菌的抗氧化应激能力。这些发现表明,这种短小芽孢杆菌血红素过氧化氢酶因其卓越的催化速率和高稳定性,可被视为具有工业相关性的生物催化剂之一,并且它可能是提高工业生产菌株抗氧化能力的潜在候选者。

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