为探究大型食药用真菌胞外蛋白酶的相关特性,从蜜环菌发酵的玉米醇溶蛋白产物中分离、纯化蛋白酶,通过饱和硫酸铵沉淀粗酶液、DEAE Sepharose Fast Flow阴离子层析柱及Sephadex G-50层析柱纯化,得到电泳纯的蛋白酶.结果表明,该蛋白酶的分子质量为18.4 ku;最适温度为60℃,在40～60 ℃具有良好稳定性;pH值为8时酶活力最高,pH值稳定性范围为6～8;Mn2+对该酶具有明显的促进作用,乙二胺四乙酸(EDTA)对中性蛋白酶有明显的抑制作用;米氏常数Km为3.786 mg/mL.研究结果为大型食药用真菌来源蛋白酶的研究与应用提供理论依据,为大型真菌源酶制剂的研发提出新思路.
Studies on the Enzymatic Properties of Zein from Armillaria mellea Fermentation
In order to explore the characteristics of extracellular proteases from large edible and medicinal fungi,proteas-es were isolated and purified from corn gliadin fermented by Armillaria mellea and purified by saturated ammonium sul-fate precipitation crude enzyme solution,DEAE Sepharose Fast Flow anion chromatography column and Sephadex G-50 chromatography column.The results showed that the molecular weight of the protease was 18.4 ku,the optimum tempera-ture of the protease was 60 ℃,it had good stability at 40-60 ℃,the enzyme activity was the highest when the pH val-ue was 8,the stability range of pH was 6-8;Mn2+could obviously promote the enzyme,ethylenediamine tetraacetic acid(EDTA)could obviously inhibit the neutral protease,and the Michaelis constant Km was 3.786 mg/mL.The results pro-vided a theoretical basis for the research and application of proteases derived from large edible and medicinal fungi,and put forward new ideas for the research and development of enzyme preparations from large fungi.
Armillaria melleazeinneutral proteaseisolation and purificationenzymatic properties
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