Cloning, Secretory Expression, Partial Characterization, and Structural Modeling of an Alkaline Protease from Bacillus subtilis D-2
文献类型： 外文期刊
作者： Xu, Yayuan 1 ; Xu, Fazhi 3 ; Ding, Xiaoling 3 ; Qian, Kun 2 ; Li, Lvmu 1 ;
作者机构： 1.Anhui Agr Univ, Sch Life Sci, Hefei 230036, Anhui, Peoples R China
2.Anhui Acad Agr Sci, Inst Agr Food, Hefei 230031, Anhui, Peoples R China
3.Anhui Agr Univ, Sch Anim Sci & Technol, Hefei 230036, Anhui, Peoples R China
关键词： Bacillus subtilis strain D-2; Secretory expression; Escherichia coli; Alkaline protease; Structural model
期刊名称：BIORESOURCES （ 影响因子：1.614； 五年影响因子：1.923 ）
ISSN： 1930-2126
年卷期： 2019 年 14 卷 3 期
页码：
收录情况： SCI
摘要： To develop a large-scale production of the protease of Bacillus subtilis strain D-2, the full-length gene apr-D2 (1,149 bp) encoding the alkaline protease was cloned into plasmid pET-32a and expressed as a secretory protein in Escherichia coli. Sequence analysis of the deduced amino acid sequence revealed high homology with the catalytic domains of the subtilisin serine proteases. From SDS-PAGE analysis, the recombinant protein had a molecular mass of 60.4 kDa. The expressed protease was secreted into the culture medium in a functional active form. The purified recombinant protease showed a pH optimum of 10.5 and temperature optimum of 55 degrees C, and it was stable in the pH range from 5.0 to 13.0. The enzyme activity was slightly enhanced by Ca2+, Mg2+, Ba2+, and SBT1. However, it was highly inhibited by Ag+ and PMSF. A theoretical structural model of mature protein was constructed by comparative modeling, which showed a putative catalytic triad (Asp-32, His-64 and Ser-221) with high similarity to the template. The structural characteristics that confer enzymatic specificity of the protease were analyzed. Taken together, the data suggested that the secretory expression system with pET-32a in E. coli was successfully constructed. Additionally, enzymatic specificity analysis of the alkaline protease indicated that it was suitable for various processing industries.
- 相关文献
作者其他论文 更多>>
-
In silico and in vitro evaluation of antiviral activity of wogonin against main protease of porcine epidemic diarrhea virus
作者：Wang, Jieru;Zeng, Xiaoyu;Yin, Dongdong;Yin, Lei;Shen, Xuehuai;Dai, Yin;Pan, Xiaocheng;Xu, Fazhi
关键词：porcine epidemic diarrhea virus (PEDV); wogonin; main protease; antiviral activity; binding affinity
-
Tofu Whey Wastewater as a Beneficial Supplement to Poultry Farming: Improving Production Performance and Protecting against Salmonella Infection
作者：Shen, Xuehuai;Yin, Lei;Yin, Dongdong;Zhao, Ruihong;Dai, Yin;Hu, Xiaomiao;Hou, Hongyan;Pan, Xiaocheng;Shen, Xuehuai;Liu, Yongjie;Shen, Xuehuai;Yin, Lei;Yin, Dongdong;Zhao, Ruihong;Dai, Yin;Hu, Xiaomiao;Hou, Hongyan;Pan, Xiaocheng;Xu, Yayuan;Cheng, Jianghua;Qian, Kun
关键词：tofu whey wastewater; poultry; production performance; antioxidant capacity; immunity; gut microbiota; Salmonella enteritidis infection
-
The transcriptome sequencing analysis reveals immune mechanisms of soybean fermented powder on the loach (Misgurnus anguillicaudatus) in response to Lipopolysaccharide (LPS) infection
作者：Xu, Yayuan;Li, Dongqi;Qian, Kun;Cheng, Jianghua;Xu, Yayuan;Li, Dongqi;Qian, Kun;Cheng, Jianghua;Zhang, Xinxin;Liu, Yu;Dai, Lishang;Xu, Tingjuan
关键词：transcriptome sequencing; soybean fermentation powder; loach; LPS; immune function
-
Upregulated beta-arrestin1 predicts poor prognosis and promotes metastasis via AKT/ERK signaling pathway in gastric cancer
作者：Xu, Tingjuan;Shen, Guodong;Cheng, Min;Wu, Xinchun;Hu, Shilian;Xu, Tingjuan;Shen, Guodong;Cheng, Min;Wu, Xinchun;Hu, Shilian;Xu, Yayuan
关键词：Gastric cancer; beta-arrestin1; Prognosis; Metastasis; Epithelial-mesenchymal transition
-
Roles of Probiotic Lactobacilli Inclusion in Helping Piglets Establish Healthy Intestinal Inter-environment for Pathogen Defense
作者：Yang, Jiajun;Qian, Kun;Wang, Chonglong;Wu, Yijing
关键词：Lactobacillus; Microbiome; Piglets; Pathogen; Defense
-
Effects of chromium-enriched bacillus subtilis KT260179 supplementation on chicken growth performance, plasma lipid parameters, tissue chromium levels, cecal bacterial composition and breast meat quality
作者：Yang, Jiajun;Qian, Kun;Zhang, Wei;Xu, Yayuan;Wu, Yijing
关键词：Chromium-enriched bacillus subtilis;Chicken;Growth performance;Meat quality;Bacterial composition
-
Sequence analysis of the haemagglutinin genes of H9N2 avian influenza viruses isolated in China during 1996 to 2011
作者：Dai, Yin;Zhao, Ruihong;Hu, Xiaomiao;Zhang, Danjun;Wang, Junjun;Xu, Fazhi
关键词：Avian influenza virus;H9N2 subtype;hemagglutinin gene;sequence analysis
