来自嗜热脂肪芽孢杆菌的高耐热性中性蛋白酶

Motoki Kubo , Keiichi Murayama , Koji Seto , Tadayuki Imanaka
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引用次数: 30

摘要

从自然环境中分离出嗜热脂肪芽孢杆菌MK232,该芽孢杆菌产生高度热稳定性的中性蛋白酶。经过几个步骤的诱变,获得了高产突变株YG185。酶产量是原菌株的两倍。这种细胞外中性蛋白酶被纯化和结晶。经sds -聚丙烯酰胺凝胶电泳和凝胶过滤,酶的分子量为34000。酶活性的最适pH和温度分别为7.5℃和70℃,酶在pH 5 ~ 10℃和70℃以下稳定。新蛋白酶的热稳定性和比活性分别比热溶酶(来自热水解芽孢杆菌的中性蛋白酶)高10%和40%左右。EDTA灭活了该酶,但苯甲基磺酰氟灭活了该酶。这些结果表明该酶是一种高度耐热的中性(金属)蛋白酶。
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Highly thermostable neutral protease from Bacillus stearothermophilus

Bacillus stearothermophilus MK232, which produced a highly thermostable neutral protease, was isolated from a natural environment. By several steps of mutagenesis, a hyper-producing mutant strain, YG185, was obtained. The enzyme productivity was twice as much as that of the original strain. This extracellular neutral protease was purified and crystallized. The molecular weight of the enzyme was 34,000 by SDS-polyacrylamide gel electrophoresis and gel filtration. The optimum pH and temperature for the enzyme activity were 7.5 and 70°C, respectively, and the enzyme was stable at pH 5–10 and below 70°C. The thermostability and specific activity of the new protease are around 10% and 40% higher than those of thermolysin (the neutral protease from Bacillus thermoproteolyticus), respectively. The enzyme was inactivated by EDTA, but not by phenylmethylsulfonyl fluoride. These results indicate that the enzyme is a highly thermostable neutral-(metallo)protease.

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