产低温弹性蛋白酶菌株的筛选及酶学特性分析

doi:10.3969/j.issn.1673-1689.2012.10.016

首页 > 过刊浏览>2012年第31卷第10期 >1105-1111. DOI:10.3969/j.issn.1673-1689.2012.10.016

产低温弹性蛋白酶菌株的筛选及酶学特性分析
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                        10.3969/j.issn.1673-1689.2012.10.016
                    
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基金项目:徐州工程学院青年培育项目(XKY2009123)

Screening of Cold-Active Elastase Producing Strain and Characterization of Enzyme Activity

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筛选获得一株产生低温型弹性蛋白酶菌株并研究酶学性质,为该酶在肉类嫩化中应用奠定理论基础。在不溶性的弹性蛋白琼脂平板上,以弹性蛋白作为惟一氮源,根据水解圈直径和菌落直径的比值筛选到一株分泌低温型弹性蛋白酶的菌株,命名为XZE116,通过形态学、生理生化试验以及16s rDNA序列分析,该菌株鉴定为短芽孢杆菌(Bacillus brevis)。菌株所产弹性蛋白酶最适作用pH值8.0,在较广的pH值范围(7.0～12.0)内酶稳定性较好。酶最适作用温度25℃,在40℃以下保温2 h,酶稳定性好。浓度为5 mmol/L的Mg2+离子对酶有激活作用,但Hg2+、Pb2+强烈抑制酶活性。丝氨酸蛋白酶特异性抑制剂苯甲基磺酰氟(PMSF)强烈抑制酶活性,但E-64对酶活性没有影响,表明菌株XZE116弹性蛋白酶是丝氨酸蛋白酶。0.1%的阴离子表面活性剂SDS、阳离子表面活性剂十六烷基三甲基溴化铵(CTAB)以及非离子型表面活性剂Triton X-100对酶活性有轻微的抑制作用。这些酶的优良生化特性赋予了该酶在肉品工业中具有潜在的应用价值。

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The aim was to screen a cold-active elastase producing strain and study the enzymatic properties in order to establish the theoretical basis for its application of meat tenderization.One strain,named XZE116,which produced cold-active elastase was screened using elastin as sole nitrogen source on elastin agar plate according to the ratio of the diameter of the hydrolytic circle to the diameter of the colony.The strain was identified as Bacillus brevis by morphological,physiological,biochemical tests and 16S rDNA gene sequence analysis.The enzyme was highly active over a wide range of pH from 7.0 to 12.0,with an optimum at pH 8.0.It exhibited maximal activity at 25 ℃ and was stable within two hours below 40 ℃.The enzyme activity was activated by Mg2+,but was strongly inhibited by Hg2+ and Pb2+.Its activity was completely inhibited by phenylmethanesulfonyl fluoride(PMSF),but not by E-64,suggesting that the enzyme is a serine-protease.The elastase activity was slightly reduced by anionic(0.1% SDS),cationic(0.1% cetyltriethylammnonium bromide,CTAB),non-ionic surfactants(1% Tween-80 and 1% Triton X-100).Considering its promising properties,the elastase from Bacillus brevis XZE116 may find potential application in meat industry.

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高兆建,陈尚龙,巫有华,张莉.产低温弹性蛋白酶菌株的筛选及酶学特性分析[J].食品与生物技术学报,2012,31(10):1105-1111.

GAO Zhao-Jian, CHEN Shang-Long, WU You-hua, ZHANG Li. Screening of Cold-Active Elastase Producing Strain and Characterization of Enzyme Activity[J]. Journal of Food Science and Biotechnology,2012,31(10):1105-1111.

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