Heterologous expression and characterization of an M4 family extracellular metalloprotease for detergent application.

IF 0.8 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Man Hao, Chaoshuo Shi, Weifeng Gong, Jia Liu, Xiangxin Meng, Fufeng Liu, Fuping Lu, Huitu Zhang
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引用次数: 0

Abstract

Proteolytic enzymes stand out as the most widely employed category utilized in manufacturing industry. A new protease was separated from Planococcus sp.11815 strain and named as nprS-15615 in this research. The gene of this protease has not been reported, and its enzymatic properties have been studied for the first time. To enhance enzyme production, the Planococcus sp. protease gene was expressed in Bacillus licheniformis 2709. The expression level of nprS-15615 was observed under the control of regulatory elements PaprE. nprS-15615 protease activity reached 1186.24±32.87 U/mL after 48 hours of cultivation in shake flasks which was nearly four times the output of the original bacteria (291.38±25.73U/mL). The optimum temperature and pH of the recombinant protease were 30 ℃ and 8.0, respectively.The enzyme exhibited the highest capacity for hydrolyzing casein and demonstrated resilience towards a NaCl concentration of 10.0% (wt/v). Furthermore, in the presence of 0.5% surfactants, the recombinant protease activity can maintain above 75%, and with the existence of 0.5% liquid detergents, there was basically no loss of enzyme activity which indicated that nprS-15615 had good compatibility with surfactants and liquid detergents. In addition, npS-15615 performed well in the washing experiment, and the washing effect at 20 ℃ can be significantly improved by adding crude enzyme solution in the washing process.

用于洗涤剂应用的M4家族胞外金属蛋白酶的异源表达和表征。
蛋白酶是制造业中应用最广泛的一类。本研究从Planococcus sp.11815菌株中分离得到一种新的蛋白酶,命名为nprS-15615。这种蛋白酶的基因尚未报道,其酶性质也首次得到研究。为了提高酶的产量,在地衣芽孢杆菌2709中表达了平球菌蛋白酶基因。在调节元件PaprE的控制下观察到npr-15615的表达水平。在摇瓶培养48小时后,npr-15615蛋白酶活性达到1186.24±32.87U/mL,几乎是原菌产量(291.38±25.73U/mL)的四倍。重组蛋白酶的最适温度和pH分别为30℃和8.0。该酶表现出最高的水解酪蛋白的能力,并对10.0%(wt/v)的NaCl浓度表现出弹性。此外,在0.5%的表面活性剂存在下,重组蛋白酶的活性可以保持在75%以上,在0.5%液体洗涤剂存在下,酶活性基本没有损失,这表明npr-15615与表面活性剂和液体洗涤剂具有良好的相容性。此外,npS-15615在洗涤实验中表现良好,在洗涤过程中加入粗酶溶液可以显著提高20℃的洗涤效果。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of General and Applied Microbiology
Journal of General and Applied Microbiology 生物-生物工程与应用微生物
CiteScore
2.40
自引率
0.00%
发文量
42
审稿时长
6-12 weeks
期刊介绍: JGAM is going to publish scientific reports containing novel and significant microbiological findings, which are mainly devoted to the following categories: Antibiotics and Secondary Metabolites; Biotechnology and Metabolic Engineering; Developmental Microbiology; Environmental Microbiology and Bioremediation; Enzymology; Eukaryotic Microbiology; Evolution and Phylogenetics; Genome Integrity and Plasticity; Microalgae and Photosynthesis; Microbiology for Food; Molecular Genetics; Physiology and Cell Surface; Synthetic and Systems Microbiology.
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