通过结构引导同源蛋白块的体内重组工程非特异性过氧酶。

4区生物学 Q3 Biochemistry, Genetics and Molecular Biology

Methods in enzymology Pub Date : 2025-01-01 Epub Date: 2025-02-06 DOI:10.1016/bs.mie.2025.01.008

Alejandro Beltran-Nogal, Ivan Mateljak, David Gonzalez-Perez, Miguel Alcalde

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引用次数: 0

摘要

真菌非特异性过氧酶（UPOs）是用于C-H氧官能化反应的高度通用的酶。多年来，为了提高异源功能表达、活性、稳定性和选择性，它们一直受到定向进化运动的影响。虽然可用于蛋白质工程的UPO基因数量正在稳步增加，但它们在酶嵌合实验中的应用却很少被探索。在本章中，我们描述了如何应用SCHEMA-RASPP计算算法结合体内DNA洗刷，从不同的同源物构建功能多样的UPO嵌合体。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Engineering unspecific peroxygenases by structure-guided in vivo recombination of homologous protein blocks.

Fungal unspecific peroxygenases (UPOs) are highly versatile enzymes for C-H oxyfunctionalization reactions. Over the years, they have been subjected to directed evolution campaigns in order to improve heterologous functional expression, activity, stability and selectivity. While the number of UPO genes available for protein engineering is steadily increasing, their use in enzyme chimeragenesis experiments has been little explored. In this chapter we describe how to construct functionally diverse UPO chimeras from different orthologs by applying the SCHEMA-RASPP computational algorithm in combination with in vivo DNA shuffling.

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来源期刊

Methods in enzymology 生物-生化研究方法

CiteScore

2.90

自引率

0.00%

发文量

308

审稿时长

3-6 weeks

期刊介绍： The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 500 volumes the series contains much material still relevant today and is truly an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research and genetics-just to name a few. Five of the 2013 Nobel Laureates have edited or contributed to volumes of MIE.