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设计 (S) -β-氨酸基化酶HitB的基质特异性

Dawei Wang¹, Akimasa Miyanaga^1,2,3, Taichi Chisuga^1,4

¹Department of Chemistry, Tokyo Institute of Technology, 2-12-1 Ookayama, Meguro-ku, Tokyo, 152-8551, Japan.

Chembiochem : a European journal of chemical biology

|May 28, 2024

概括

研究人员设计了像HitB这样的β-氨基酸腺化酶,以创建新的天然产品类似物. 突变改变了基质的特异性,扩大了天然产品生物合成中的潜在应用.

科学领域:

生物化学生化学
酵素工程是什么? 酶工程是什么
自然产品生物合成自然产品生物合成

背景情况:

基化酶对于将氨基酸单元纳入天然产品合成至关重要.
工程β-氨基酸腺化酶仍然没有得到充分的探索.
HitB 是 hitachimycin 生物合成中的 (S) -β-氨酸乙化酶.

研究的目的:

为了设计 (S) - β - 氨酸腺化酶的基质特异性,HitB.
了解β-氨基酸激活中的酶基质乱交的结构基础.
探索生产含有β-氨基酸的新型天然产品类似物.

主要方法:

在位点Phe328和Ser293.3的HitB的位点定向突变发生.
酶活性测定用甲基和正基替代 (S) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 的类似物.
对HitB突变体的结构分析与中间类型复合.

主要成果:

突变HitB F328V和F328L突变有效地激活了甲基替代 (S) - - 贝塔-氨类型.
HitB T293G和T293S突变有效地激活了正位置换的 (S) - 贝塔 - 氨酸类似物.
结构数据显示了一个扩大的基质结合口袋,解释了基质特异性的改变.

结论:

在Phe328和Ser293的突变成功地改变了HitB对 (S) - 贝塔 - 氨酸类似物的基质特异性.
酶工程为创造基质乱交提供了洞察力.
这些发现支持开发各种含有β-氨基酸的天然产品类似物.

关键词:

生物合成生物合成自然产品是天然产品.聚基基化物聚基基化物聚基基化物蛋白质工程是一种蛋白质工程.这是一种β-氨基酸.

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