酸性和碱性酶稳定性机制及其识别.doc

酸性和碱性酶稳定性机制及其识别华侨大学生物工程与技术系厦门 361021 : 了解酸性和碱性酶稳定性机制并对其进行识别具有重要理论和实践意义。通过分析105条酸性酶111条碱性酶序列酸性酶中Trp、Tyr、Thr和Ser的含量明显高于平均值而GluLys、Met和Arg的含量则明显低于平均值碱性酶中Trp、Ala和Cys的含量略高于平均值而Lys、Arg和Glu的含量则略低于平均值Ala、Glu、Leu、Asn、Arg、Ser和Thr86.1%, 5倍交叉验证的精度为83.3%。建立了一种基于序列识别和的新方法: 酸性酶碱性酶识别加权氨基酸组成and classification of acidic andalkaline enzymes Guangya Zhang, Jiaqiang Gao, and Baishan Fang Department of Bioengineering and Biotechnology, Huaqiao University, Xiamen 361021, China Abstract: It is of theoretical and practical significance to understand the mechanism of enzyme adaptation to acidic and alkaline environments and classification of them based on sequence information. In present work, the amino acid composition of 105 acidic and 111 alkaline enzyme sequences was systematically analyzed. Acidic enzymes contained significantly more Trp, Tyr, Thr and Ser, whereas less Glu, Lys, Met and Arg. On the other hand, alkaline enzymes have slightly more Trp, Ala and Cys, whereas less Lys, Arg and Glu. The amount of Ala, Glu, Leu, Asn, Arg, Ser and Thr in acidic and alkaline enzymes varied largely. Hence, a weighted amino acid composition method was developed for the discrimination of acidic and alkaline enzymes. Using the back-check and the 5-fold cross validation methods, the overall accuracy could reach 86.1% and 83.3%, respectively. A new method to classify acidic and alkaline enzymes based on their sequences was established. Keywords: acidic enzyme, alkaline enzyme, discrimination, weighted amino acid composition 近年来, 极端微生物引起了研究者极大兴趣, 对其研究日益升温, 嗜酸菌和嗜碱菌就是其中重要的极端微生物类群。对嗜酸和嗜碱菌的生理学进行了大量研究, 结果发现, 尽管嗜酸和嗜碱菌生长在酸性和碱性环境中, 但其细胞内部的pH却接近中性, 细胞内酶反应和生化代谢过程也与中性菌相似, 而只有它们产生的胞外酶由于处在酸性或碱性环境而成为酸性酶和碱性酶[1,2]。酸性酶和碱性酶在生物造纸、食品、医药、环境整治及其他化学工业中的应用潜力极大, 商业期望值很高, 前景诱人。因此, 寻找嗜酸和嗜碱菌并分离其酸性和碱性酶是当前研究的热点, 许多研究者致力于此[3,4]。此外, 探讨这些极端酶稳定性的机制也一直是关注的焦点, 有研究者[5]分析了Nocardiopsis alba中酸性蛋白酶A(NAPase)的结构以及去折叠(Unfolding)动力学特性并和其中性同源蛋白进行了比较, 结果表明二者包含的酸性氨基酸数量虽相近, 但NAPase对酸去折叠自