Identification and characterization of a protease produced by Vibrio parahaemolyticus in iron-limited medium.

H C Wong, J T Shyu
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Abstract

Two proteolytic proteins (about 43 and 90 kDa) were produced by clinical strains of Vibrio parahaemolyticus cultured in iron-limited medium. The 43 kDa-protease was partially purified by ammonium sulfate precipitation, ultrafiltration fractionation and DEAE-Sephacel chromatography. This protease had an optimum pH range of 7 to 8, and an optimum reaction temperature of about 40 degrees C. It was heat-labile, being partially inactivated by heat-treatment at 60 or 90 degrees C for 10 min. The protease hydrolyzed casein, gelatin, elastin, collagen and hemoglobin. As a chymotrypsin-like protease, it was inhibited only by the chymostatin among seven protease inhibitors tested. Activity of this protease was partially inhibited by 1 mM of Co2+, Cu2+, Zn2+ and Hg2+ and slightly enhanced by Ca2+ and Ba2+. It was completely inactivated by orthophenanthroline (OPA), and the OPA-inactivated sample was partially reactivated by Ca2+ and Fe2+. In conclusion, this 43-kDa protease of V. parahaemolyticus was an unstable neutral chymotrypsin-like metalloprotease; Ca2+ and/or Fe2+ was essential for its activity or stability.

副溶血性弧菌在限铁培养基中产生的一种蛋白酶的鉴定和特性。
在限铁培养基中培养的副溶血性弧菌临床菌株产生了两种蛋白水解蛋白(约43和90 kDa)。通过硫酸铵沉淀法、超滤分馏法和deae - sepacel层析法对43 kda蛋白酶进行了部分纯化。该蛋白酶的最适pH范围为7 ~ 8,最适反应温度为40℃左右,热不稳定,可在60 ~ 90℃下加热10 min使部分失活。该蛋白酶可水解酪蛋白、明胶、弹性蛋白、胶原蛋白和血红蛋白。作为一种凝乳胰蛋白酶样蛋白酶,在7种蛋白酶抑制剂中只有凝乳抑素对其有抑制作用。1 mM的Co2+、Cu2+、Zn2+和Hg2+对该蛋白酶的活性有部分抑制作用,Ca2+和Ba2+对其活性有轻微增强作用。正henanthroline (OPA)使其完全失活,OPA失活的样品被Ca2+和Fe2+部分复活。综上所述,副溶血性弧菌的43-kDa蛋白酶是一种不稳定的中性凝乳蛋白酶样金属蛋白酶;Ca2+和/或Fe2+对其活性或稳定性至关重要。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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