芽孢杆菌L_4菌株角蛋白酶的酶学性质研究  

Enzymatic Characteristics of Keratinase from Bacillus sp. L_4 Strains

作　　者：张奇[1] 孙丹[2] 杨文博[3] 刘茜[3] 白芳[1] 

机构地区：[1]南开大学药学院,天津300071 [2]南开大学学报编辑部,天津300071 [3]南开大学生命科学学院,天津300071

出　　处：《农业环境科学学报》2009年第10期2189-2193,共5页Journal of Agro-Environment Science

摘　　要：采用室内实验方法,对芽胞杆菌L4菌株产生的角蛋白酶进行了系列的酶学性质研究。结果表明,以角蛋白溶液为底物时该酶的最适酶促反应温度为40℃,最适酶促反应pH为7.0,其Km值为1.88mmol·L-1,Vmax为2.72×10-2mmol·L-·1s-1。利用不同的蛋白酶抑制剂进行酶活性抑制实验,发现该酶受邻啡罗啉和EDTA的抑制,推测该酶可能是一种含Zn2+的金属蛋白酶。微量元素对该酶活力的影响显著,Ca2+和Mg2+对酶活力有促进作用,高浓度的Fe2+和Cu2+明显抑制角蛋白活力。Keratin is one fibrous insoluble animal protein, do not dissolve in water and has a strong anti-decomposition. In this study, one keratinase was purified from Bacillus sp L4 strains, and its characterization was clarified. When keratin was regared as substrate, the optimum reaction temperature of keratinase was 40 ~C, the optimum reaction pH was 7.0, the Km and Vmax value were 1.88 mmol·L^-1 and 2.72 × 10^-2 mmol· L^-1· s^-1 respectively. The results indicated the keratinase had high properties and could be used in hydrolyzing feather to solve environmental pollution. At the same time, different protease inhibitors were selected to test the inhibition activity, and the results showed that the activity of keratinase could be inhibited by o-phenanthroline, eathylene diamine tetraacetic acid（ EDTA ）, but phenylmethanesulfonyl fluoride （PMSF） hardly. It suggested that this enzyme was a metal protease containing Zn2＋, and was difference from the reported keratinase that almost all were serine-type protease. In addition, ten kinds of trace element including Ba2＋, Sn2＋, Mg2＋, Li＋, Zn2＋, Fe2＋, Fe3＋, Ca2＋, Mn2＋ and Cu2＋ were used to analyze regulatory activity, the results showed that Ca2＋ and Mg2＋ could increased the enzyme activity and high concentration of Fe2＋ and Cu2＋ could obviously inhibit its activity.

关 键 词：芽孢杆菌L4 角蛋白酶 酶学性质 酶活力 

分 类 号：X172[环境科学与工程—环境科学]