Waghmare Shailesh R, Gurav Aparna A, Mali Sonal A, Nadaf Naiem H, Jadhav Deepak B, Sonawane Kailas D
Department of Microbiology, Shivaji University, Kolhapur, Maharashtra, India.
Department of Microbiology, Shivaji University, Kolhapur, Maharashtra, India; Department of Biochemistry, Shivaji University, Kolhapur, Maharashtra, India.
Protein Expr Purif. 2015 Mar;107:1-6. doi: 10.1016/j.pep.2014.11.002. Epub 2014 Nov 21.
Ability of microorganisms to grow at alkaline pH makes them an attractive target for several industrial applications. Thus, search for new extremozyme producing microorganisms must be a continuous exercise. Hence, we isolated a potent alkaline protease producing bacteria from slaughter house soil. The morphological, biochemical and 16S rDNA gene sequencing studies revealed that the isolated bacteria is Stenotrophomonas maltophilia strain SK. Alkaline protease from S. maltophilia strain SK was purified by using ammonium sulphate precipitation and DEAE-cellulose ion exchange column chromatography. The purified enzyme was optimally active at pH 9.0 and temperature 40°C with broad substrate specificity. It was observed that the metal ions such as Ca(++), Mg(++) and Fe(+++) completely repressed the enzyme activity. The enzyme was stable in presence of various water miscible solvents like ethanol, methanol, isopropanol at 25% (v/v) concentration and less stable at 37.5% (v/v) concentration. These robust properties of enzyme might be applicable for various applications in detergent and pharmaceutical industries.
微生物在碱性pH条件下生长的能力使其成为多种工业应用的理想目标。因此，寻找新的产极端酶微生物必须是一项持续的工作。为此，我们从屠宰场土壤中分离出了一种高效产碱性蛋白酶的细菌。形态学、生物化学和16S rDNA基因测序研究表明，分离出的细菌是嗜麦芽窄食单胞菌菌株SK。通过硫酸铵沉淀和DEAE-纤维素离子交换柱色谱法对嗜麦芽窄食单胞菌菌株SK产生的碱性蛋白酶进行了纯化。纯化后的酶在pH 9.0和温度40°C时具有最佳活性，底物特异性广泛。观察到Ca(++)、Mg(++)和Fe(+++)等金属离子完全抑制了酶的活性。该酶在25%（v/v）浓度的乙醇、甲醇、异丙醇等各种与水混溶的溶剂中稳定，在37.5%（v/v）浓度时稳定性较差。酶的这些强大特性可能适用于洗涤剂和制药行业的各种应用。
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