RETRACTED: Purification and characterization of organic solvent and detergent stable protease isolated from marine Saccharopolyspora sp A9: Application of protease for wound healing (Retracted Article)

被引:4
作者
Raut, Ghanshyam [1 ,2 ]
Vetal, Sachin [2 ]
Biao, Ren [3 ]
Liu, Xiang-Yang [3 ]
Zhang, Lixin [3 ]
Kokare, Chandrakant [1 ,2 ]
机构
[1] Sinhgad Inst Pharm, STES, Dept Pharmaceut, Pune 411041, Maharashtra, India
[2] Bharati Vidyapeeth Deemed Univ, Dept Pharmaceut Biotechnol, Poona Coll Pharm, Pune 411038, Maharashtra, India
[3] Chinese Acad Sci, Inst Microbiol, Key Lab Pathogen Microbiol & Immunol, Beijing 100101, Peoples R China
关键词
Filamentous bacteria; Submerged culture; Protease; Purification; Wound healing; Topical gel; ALKALINE PROTEASE; SERINE-PROTEASE; OPTIMIZATION; STABILITY; GELS;
D O I
10.1016/j.bej.2012.09.002
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
An extracellular organic solvent stable protease producing strain was isolated from west coast of India and identified as Saccharopolyspora sp. A9. The protease was purified from the culture supernatant to homogeneity using ammonium sulfate precipitation, DEAE Cellulose and Sephadex G-200 gel filtration with a 23.28-fold increase in specific activity having approximately 32 kDa molecular weight. The enzyme stability was investigated over broad range of pH, temperature, salts, different organic solvents, commercial detergent and surfactants. In present study, carbopol 940 gel containing protease enzyme was developed for evaluation of its wound healing activity by excision wound model. The enzyme was stable in pH range of 8.0-12.0 with optimum of pH 10.0, whereas thermostable nature of enzyme was suggested by its stability over 70 and 80 degrees C. In addition, enzyme showed excellent stability with a wide range of organic solvents having log P-2.0 and below 2.0. In excision wound model wounds were found to contract to 80.40% (p<0.001) in first nine days at higher rate with formulation containing 300 U/g of protease enzyme. Therefore, topical protease formulation offers promise for its pharmaceutical use in wound healing and also proves its application in protein biosynthesis in non-aqueous media. (C) 2012 Elsevier B.V. All rights reserved.
引用
收藏
页码:196 / 203
页数:8
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