Engineering natural and noncanonical nicotinamide cofactor-dependent enzymes: design principles and technology development.

Engineering natural and noncanonical nicotinamide cofactor-dependent enzymes: design principles and technology development.
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工程天然和非规范烟酰胺辅助因素依赖性酶:设计原理和技术开发。

DOI:
10.1016/j.copbio.2020.08.005
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发表时间:
2020-12
影响因子:
7.7
通讯作者:
Li H
Li H
中科院分区:
工程技术1区
文献类型:
--
作者:
King E;Maxel S;Li H

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烟酰胺辅因子使氧化还原酶能够催化生物制造中的无数重要反应。数十年的研究集中在优化利用天然烟酰胺辅因子的酶,即烟酰胺腺嘌呤二核苷酸(磷酸)(NAD(P)+)。最近的研究结果重新点燃了工程酶的兴趣,利用非典型的辅因子,NAD+(mNADs)的模拟物,它表现出上级工业性能在体外,并使特定的电子传递在体内。我们比较了工程天然与非经典的辅因子利用酶的最新进展,讨论了发现的设计原则,并调查了新兴的高通量平台,超越了传统的96孔板为基础的方法。获得mNAD依赖性酶仍然具有挑战性,工具包有限。为此,我们强调的设计原则和技术,可以潜在地从工程天然翻译到非典型的辅因子依赖酶。
Nicotinamide cofactors enable oxidoreductases to catalyze a myriad of important reactions in biomanufacturing. Decades of research has focused on optimizing enzymes which utilize natural nicotinamide cofactors, namely nicotinamide adenine dinucleotide (phosphate) (NAD(P)+). Recent findings reignite the interest in engineering enzymes to utilize noncanonical cofactors, the mimetics of NAD+ (mNADs), which exhibit superior industrial properties in vitro and enable specific electron delivery in vivo. We compare recent advances in engineering natural versus noncanonical cofactor-utilizing enzymes, discuss design principles discovered, and survey emerging high-throughput platforms beyond the traditional 96-well plate-based methods. Obtaining mNAD-dependent enzymes remains challenging with a limited toolkit. To this end, we highlight design principles and technologies which can potentially be translated from engineering natural to noncanonical cofactor-dependent enzymes.
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