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投稿时间：2021-02-15
投稿时间：2021-02-15
中文摘要: 在毕赤酵母中克隆表达黑曲霉CGMCC 3.7193中一个疑似金属蛋白酶基因MPase，分离纯化后对重组酶MPase进行理化特征分析。结果显示，纯化后的MPase：比酶活达到1 859.2 U/mg，最适反应温度35℃，最适反应pH7.0；在低于40℃，pH5.0～8.0之间稳定；偏好水解大豆分离蛋白，Km和Vmax分别为3.9 mg/mL、892.7 mg/（mL·min）；金属离子Co2+和Zn2+对其有激活作用，Cu2+和Fe2+对其有抑制作用；酶解大豆分离蛋白水解度可达到14.7%，并且产物分子量大小均匀；可耐受5 000 mmol/L NaCl，在50℃保持30 min即可完全失活。其酶学特征表明：MPase在食品工业尤其在发酵食品加工中具有较大应用潜力。
Abstract:A predicted metalloproteinase gene，MPase，from Aspergillus niger CGMCC 3.7193 was cloned and expressed in Pichia pastoris.The physicochemical characteristics of recombinant MPase were investigated after purification.The specific activity of the purified MPase reached 1 859.2 U/mg.Optimum temperature and pH for MPase activity were 35℃ and 7.0，respectively.MPase remained stable up to 40℃ and within the pH range of 5.0 to 8.0.MPase preferred soybean protein isolate as substrate，and Kmand Vmaxwere 3.9 mg/mL and 892.7 mg/（mL·min），respectively.MPase was activated by Co2+and Zn2+，and inhibited by Cu2+and Fe2+.The degree of hydrolysis of soybean protein isolate reached 14.7%，and hydrolysates were uniform in molecular weight.MPase could tolerate 5 000 mmol/L NaCl，and lost its activity completely after 30 min at 50 ℃.These enzymological characterizations indicated that MPase had great application potential in the food industry，especially in fermented food processing.
keywords: metalloproteinase Aspergillus niger cloning and expression enzymatic properties hydrolyzed protein
文章编号：202204028 中图分类号： 文献标志码：
基金项目:
| Author Name | Affiliation |
| FENG Wei，SONG Peng | School of Life Sciences，Liaocheng University，Liaocheng 252000，Shandong，China |
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