Rapid cloning, expression and purification of a novel high-activity alkaline phosphatase with detoxification of lipopolysaccharide
文献类型: 外文期刊
第一作者: Wu, Daichao
作者: Wu, Daichao;Teng, Da;Xi, Di;Wang, Xiumin;Wang, Xiaojie;Mao, Ruoyu;Zhang, Yong;Wang, Jianhua;Wu, Daichao;Dai, Hua;Wu, Daichao;Teng, Da;Xi, Di;Wang, Xiumin;Wang, Xiaojie;Mao, Ruoyu;Zhang, Yong;Dai, Hua;Wang, Jianhua
作者机构:
关键词: Heterologous expression;Alkaline phosphatase;Saccharomyces boulardii;Lipopolysaccharide;Dephosphorylation;TNF-alpha
期刊名称:PROCESS BIOCHEMISTRY ( 影响因子:3.757; 五年影响因子:3.665 )
ISSN: 1359-5113
年卷期: 2014 年 49 卷 3 期
页码:
收录情况: SCI
摘要: Lipopolysaccharide (LPS) is a bacterial endotoxin leading to endotoxemia. Its virulence factor 'diphosphoryl lipid A' can be abolished by alkaline phosphatase (AP). A novel AP gene (without introns) was cloned from Saccharomyces boulardii ATCC MYA-796 with a GenBank accession number KF471017, and the recombinant AP (rAP) was expressed as a soluble protein in Pichia pastoris X-33 with a yield of 43.66 mg/I at the end of 120 h of induction in a shaker flask. After purification by affinity-column chromatography, the purity of rAP was over 90%. The optimal reaction conditions of rAP were pH 9.6, temperature at 60 degrees C and 2 mM Mg2+ in diethanolamine buffer, and EDTA was a potent inhibitor of rAP activity. The specific activity of rAP was 9912.01 U/mg under the optimal conditions. Furthermore, rAP showed a broad dephosphorylation activity to LPS over a broad pH range (pH 2-10) in vitro and peaked at pH 4 in Tris-HCI buffer. After LPS dephosphorylated by rAP was injected intraperitoneally into mice, the serum level of tumor necrosis factor (TNF)-alpha was significantly reduced compared to that of the LPS group (p < 0.01). These findings suggest that rAP has great potential to cure diseases caused by LPS. (C) 2014 Elsevier Ltd. All rights reserved.
分类号:
- 相关文献
作者其他论文 更多>>
-
Versatile plant genome engineering using anti-CRISPR-Cas12a systems
作者:He, Yao;Xu, Tang;Zheng, Xuelian;Zhang, Yong;He, Yao;Liu, Shishi;Pu, Dongkai;Zhong, Zhaohui;Ren, Qiurong;Dong, Chuan;Wang, Yawei;Wang, Danning;Zhang, Yong;Liu, Shishi;Chen, Long;Zhang, Tao;Chen, Long;Zhang, Tao;Guo, Fengbiao;Guo, Fengbiao;Guo, Fengbiao;Qi, Yiping;Qi, Yiping
关键词:anti-CRISPR; Cas12a; AcrVA1; off-target effects; fine-tuning genome editing; inducible and tissue specific genome editing; CRISPRa; synthetic logic circuit
-
Effects of plant tissue permeability on invasion and population bottlenecks of a phytopathogen
作者:Jiang, Gaofei;Shi, Xiaojun;Chen, Xinping;Ding, Wei;Zhang, Yong;Jiang, Gaofei;Zhang, Yuling;Jousset, Alexandre;Zhao, Fang-Jie;Xu, Yangchun;Shen, Qirong;Wei, Zhong;Chen, Min;Zhang, Yong;Ramoneda, Josep;Han, Liangliang;Shi, Yu;Peyraud, Remi;Wang, Yikui;Hikichi, Yasufumi;Ohnishi, Kouhei;Dini-Andreote, Francisco;Dini-Andreote, Francisco;Dini-Andreote, Francisco
关键词:
-
Bacillus subtilis Simultaneously Detoxified Aflatoxin B1 and Zearalenone
作者:Wu, Jianwen;Li, Huanrong;Wu, Jianwen;Wang, Zhenlong;An, Wei;Gao, Boquan;Li, Chunxiao;Han, Bing;Tao, Hui;Wang, Jinquan;Wang, Xiumin;Wu, Jianwen;Wang, Zhenlong;An, Wei;Gao, Boquan;Li, Chunxiao;Han, Bing;Tao, Hui;Wang, Jinquan;Wang, Xiumin
关键词:mycotoxins; degradation; aflatoxin; zearalenone; Bacillus subtilis; active components; feed
-
Polylactic Glycolic Acid-Mediated Delivery of Plectasin Derivative NZ2114 in Staphylococcus epidermidis Biofilms
作者:Ma, Xuanxuan;Yang, Na;Mao, Ruoyu;Hao, Ya;Teng, Da;Wang, Jianhua;Ma, Xuanxuan;Huang, Yinhua;Ma, Xuanxuan;Yang, Na;Mao, Ruoyu;Hao, Ya;Teng, Da;Wang, Jianhua
关键词:antimicrobial peptide NZ2114; drug delivery system; nanoparticles; PLGA; Staphylococcus epidermidis; biofilms
-
Interaction between Dietary Lactoferrin and Gut Microbiota in Host Health
作者:Li, Bing;Zhang, Fuli;Liu, Xiaomeng;Zhang, Yunxia;Peng, Weifeng;Zhang, Bo;Teng, Da;Mao, Ruoyu;Yang, Na;Hao, Ya;Wang, Jianhua;Teng, Da;Mao, Ruoyu;Yang, Na;Hao, Ya;Wang, Jianhua
关键词:lactoferrin; gut microbiota; host health; interaction; noncommunicable chronic diseases
-
Efficiency of NZ2114 on Superficial Pyoderma Infected with Staphylococcus pseudintermedius
作者:Yang, Na;Huang, Yan;Li, Yuanyuan;Teng, Da;Mao, Ruoyu;Hao, Ya;Wang, Jianhua;Yang, Na;Huang, Yan;Li, Yuanyuan;Teng, Da;Mao, Ruoyu;Hao, Ya;Wang, Jianhua;Yang, Na;Huang, Yan;Li, Yuanyuan;Teng, Da;Mao, Ruoyu;Hao, Ya;Wang, Jianhua;Huang, Yan;Wei, Lingyun
关键词:antimicrobial peptides; NZ2114; Staphylococcus pseudintermedius; transdermal delivery; chemical permeation enhancers; superficial pyoderma
-
Salivary Protein Cyclin-Dependent Kinase-like from Grain Aphid Sitobion avenae Suppresses Wheat Defense Response and Enhances Aphid Adaptation
作者:Zhang, Yumeng;Ding, Tianbo;Zhang, Yumeng;Liu, Xiaobei;Sun, Yu;Zhang, Yong;Chen, Julian;Liu, Yong
关键词:Sitobion avenae; salivary protein; defense response; aphid adaptation; bacterial type III secretion system