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在模块化聚酸合成中通过转移酶域交换进行非自然聚酸基质选择

Elias Englund^1,2, Matthias Schmidt^1,3,4, Alberto A Nava^3,5

¹Joint BioEnergy Institute, Emeryville, California 94608, United States.

Journal of the American Chemical Society

|April 14, 2023

概括

通过交换乙基转移酶 (AT) 域来利用新型扩展基质来设计聚乙基合成酶 (PKS). 这种策略成功地在体外产生了多种以前未报告的多基基.

科学领域:

生物化学
合成生物学
酵素学

背景情况:

模块化聚化合成酶 (PKS) 对于合成各种聚化天然产品至关重要.
PKS模块通常包含马洛尼尔-CoA或甲基马洛尼尔-CoA,但自然多样性来自各种马洛尼尔-CoA类似物.
乙转移酶 (AT) 域决定PKS模块内的扩展基质的特异性.

研究的目的:

通过交换AT域来设计PKS模块以改变扩展器基板的特异性.
使用工程PKS和多种扩展基质探索新型聚基的生物合成.
开发用于预测AT基板范围的计算方法.

主要方法:

在模块化多基合成酶 (PKS) 中交换了转移酶 (AT) 域,以改变扩展剂基质的特异性.
使用野生型和人工PKS与14种不同的扩展基质进行了体外多基酸生物合成反应.
开发了一个计算工作流来预测基于活跃站点体积的AT基板范围.

主要成果:

从大约200个体外反应中产生了13种结构上不同的多基基,包括几种新型化合物.
在某些情况下,与野生类型的PKS相比,人工PKS产生的目标聚化物超过100倍.
不寻常的AT域优先结合稀有扩展基质,其尺寸与自然基质相似或略大,而不是马洛尼尔-CoA或甲基马洛尼尔-CoA.

结论:

乙转移酶 (AT) 域交换是一种可行的策略,用于工程PKS在体外产生新型多基体.
了解罕见的AT域的基质特异性扩大了聚基生物合成的工具箱.
开发的计算工作流程有助于选择适合PKS工程和新化学合成的AT领域.

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