Discovery of Novel Bacterial Chalcone Isomerases by a Sequence-Structure-Function-Evolution Strategy for Enzymatic Synthesis of (S)-Flavanones.
Discovery of Novel Bacterial Chalcone Isomerases by a Sequence-Structure-Function-Evolution Strategy for Enzymatic Synthesis of (S)-Flavanones.
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DOI：
10.1002/anie.202107182
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发表时间：
2021-07-26
期刊：
影响因子：
--
通讯作者：
Bornscheuer UT
中科院分区：
文献类型：
--
作者：
Meinert H;Yi D;Zirpel B;Schuiten E;Geißler T;Gross E;Brückner SI;Hartmann B;Röttger C;Ley JP;Bornscheuer UT
查尔酮异构酶是植物黄酮类化合物生物合成的关键酶。最早从分枝真杆菌中鉴定出CHI (CHIera)，但其分布、进化来源、底物范围和立体选择性尚不清楚。在这里，我们描述了使用新的序列-结构-功能-进化（SSFE）策略从Genbank中鉴定出66种新型细菌CHIs。这些新型细菌CHIs对各种羟基化和甲氧基化查尔酮具有不同的底物特异性。根据这些新型细菌CHIs的底物结合模型，CHIera的诱变导致了对这些查尔酮的活性大大提高的几个变体。此外，对5种查尔酮进行了细菌CHIs催化的制备规模转化，结果表明(S)‐选择性高达96%，为高收率合成(S)‐黄酮提供了另一种生物催化途径。通过序列-结构-功能-进化（SSFE）策略鉴定的新型细菌查尔酮异构酶及其变体能够将查尔酮转化为具有不同底物范围和严格的(S) -立体偏好的黄酮，从而实现了(S) -黄酮的生物催化途径。
Chalcone isomerase (CHI) is a key enzyme in the biosynthesis of flavonoids in plants. The first bacterial CHI (CHIera) was identified from Eubacterium ramulus, but its distribution, evolutionary source, substrate scope, and stereoselectivity are still unclear. Here, we describe the identification of 66 novel bacterial CHIs from Genbank using a novel Sequence‐Structure‐Function‐Evolution (SSFE) strategy. These novel bacterial CHIs show diversity in substrate specificity towards various hydroxylated and methoxylated chalcones. The mutagenesis of CHIera according to the substrate binding models of these novel bacterial CHIs resulted in several variants with greatly improved activity towards these chalcones. Furthermore, the preparative scale conversion catalyzed by bacterial CHIs has been performed for five chalcones and revealed (S)‐selectivity with up to 96 % ee, which provides an alternative biocatalytic route for the synthesis of (S)‐flavanones in high yields. Novel bacterial chalcone isomerases and their variants identified by a Sequence‐Structure‐Function‐Evolution (SSFE) strategy are able to convert chalcones to flavanones with diverse substrate scopes and strict (S)‐stereopreference enabling a biocatalytic route to (S)‐flavanones.
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影响因子：
4.8
作者：
Jez, JM;Noel, JP
通讯作者：
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DOI：
10.1024/0300-9831.73.2.79
发表时间：
2003-03-01
影响因子：
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