藻酸盐介导的固定化菠萝蜜(Artocarpus heterophyllus)胶乳丝氨酸蛋白酶具有更好的催化特性

IF 16.4 1区 化学 Q1 CHEMISTRY, MULTIDISCIPLINARY
Suman Natta, Swathi Chintala, Deepak Kumar, Nandita Sahana, Somnath Mandal
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引用次数: 0

摘要

丝氨酸蛋白酶因其热稳定性和在较宽 pH 值范围内的活性而受到工业界的特别关注。本研究利用色谱技术从菠萝蜜中纯化了一种丝氨酸蛋白酶(49.3 kDa)。在最后的纯化步骤中,丝氨酸蛋白酶的纯化倍数为 92.41,产率为 24%。丝氨酸蛋白酶特异性抑制剂 PMSF 能以最低浓度(100 µM)完全抑制该蛋白酶。纯化的丝氨酸蛋白酶被海藻酸钠(2.5%)和氯化钙(0.3 M)溶液固定。对固定化酶的动力学研究表明,该酶在 pH 值为 7 的条件下具有稳定的活性,并能承受高达 45 °C 的温度。与纯化的可溶性酶相比,固定化过程提高了酶的催化效率(Kcat 提高了 1.4 倍,Kcat/Km 提高了 1.21 倍)。我们的研究结果表明,藻酸盐介导的丝氨酸蛋白酶固定化过程大大改善了酶的最适 pH 值和温度,从而进一步拓宽了酶的使用范围。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Alginate-mediated immobilization of jackfruit (Artocarpus heterophyllus) latex serine protease enzyme exhibits improved catalytic properties

Alginate-mediated immobilization of jackfruit (Artocarpus heterophyllus) latex serine protease enzyme exhibits improved catalytic properties

Serine proteases have been receiving special attention from the industrial point of view, due to their thermo-stable properties and activity over wide ranges of pH. In the present investigation, a serine protease (49.3 kDa) from jackfruit latex has been purified using chromatographic techniques. Fold purification of the serine protease in the final purification step was 92.41 with 24% yield. The protease is completely inhibited by PMSF, a serine protease-specific inhibitor at a minimal concentration (100 µM). The purified serine protease was immobilized with sodium alginate (2.5%) and calcium chloride (0.3 M) solution. The kinetic studies of immobilized enzyme showed stable activity up to pH 7, and can withstand temperatures up to 45 °C. The immobilization process improves the catalytic efficiency of the enzyme over purified soluble enzyme (for Kcat 1.4 times and for Kcat/Km 1.21 times). Our results depicted that alginate mediated immobilization of serine protease greatly improves the pH and temperature optima which broadens the scope of usage of the enzyme further.

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来源期刊
Accounts of Chemical Research
Accounts of Chemical Research 化学-化学综合
CiteScore
31.40
自引率
1.10%
发文量
312
审稿时长
2 months
期刊介绍: Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.
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