BioWF: A Naturally-Fused, Di-Domain Biocatalyst from Biotin Biosynthesis Displays an Unexpectedly Broad Substrate Scope.
BioWF: A Naturally-Fused, Di-Domain Biocatalyst from Biotin Biosynthesis Displays an Unexpectedly Broad Substrate Scope.
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DOI：
10.1002/cbic.202200171
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发表时间：
2022-09-05
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生物素的碳链是由C7二酸的Pimelate组成的，它被依赖于ATP的Pimeloyl-CoA合成酶(PCAS，由Biow编码)转化为酰基-CoA硫酸酯。酰基硫酸酯与ʟ-丙氨酸在脱羧基、类克莱森反应中缩合，形成氨基酮(8-氨基-7-氧酮酸，AON)。这一步骤是由5‘-磷酸吡哆醛(PLP)依赖的酶(AON合成酶，AONS，由BioF编码)催化的。枯草杆菌Biow(BsBioW)和大肠杆菌BioF(EcBioF)的不同版本显示出严格的底物特异性。相比之下，来自淀粉棒状杆菌(CaBioWF)的Biow-BioF融合可以接受更广泛的单脂肪酸和双脂肪酸。对BsBioW：Pimeloyl-Adylate复合体的活性位点的分析表明，Phe(F192)残基在CaBioW结构域中起着关键作用；F192Y突变体恢复了底物对Pimelate的特异性。这种令人惊讶的底物灵活性也延伸到CaBioF结构域，它接受ʟ-丙氨酸、ʟ-丝氨酸和甘氨酸。CaBioWF融合的结构模型提供了对这两个结构域如何相互作用的洞察，并暗示了域内隧道的存在。CaBioWF融合体催化各种脂肪酸和氨基酸转化为一系列AON衍生物。如此意想不到的、天然的广泛底物范围表明，CaBioWF融合是一种多功能的生物催化剂，可用于制备许多氨基酮类似物。BioWF融合生物催化剂可以将各种脂肪酸底物转化为α-oxoamine(AON)产品。依赖于ATP的Biow结构域催化形成酰基辅酶A硫酯中间体。依赖于吡哆醛5‘-磷酸(PLP)的BioF结构域可以接受各种氨基酸，并催化一系列氨基酮类似物的形成。
The carbon backbone of biotin is constructed from the C7 di‐acid pimelate, which is converted to an acyl‐CoA thioester by an ATP‐dependent, pimeloyl‐CoA synthetase (PCAS, encoded by BioW). The acyl‐thioester is condensed with ʟ‐alanine in a decarboxylative, Claisen‐like reaction to form an aminoketone (8‐amino‐7‐oxononanoic acid, AON). This step is catalysed by the pyridoxal 5’‐phosphate (PLP)‐dependent enzyme (AON synthase, AONS, encoded by BioF). Distinct versions of Bacillus subtilis BioW (BsBioW) and E. coli BioF (EcBioF) display strict substrate specificity. In contrast, a BioW‐BioF fusion from Corynebacterium amycolatum (CaBioWF) accepts a wider range of mono‐ and di‐fatty acids. Analysis of the active site of the BsBioW : pimeloyl‐adenylate complex suggested a key role for a Phe (F192) residue in the CaBioW domain; a F192Y mutant restored the substrate specificity to pimelate. This surprising substrate flexibility also extends to the CaBioF domain, which accepts ʟ‐alanine, ʟ‐serine and glycine. Structural models of the CaBioWF fusion provide insight into how both domains interact with each other and suggest the presence of an intra‐domain tunnel. The CaBioWF fusion catalyses conversion of various fatty acids and amino acids to a range of AON derivatives. Such unexpected, natural broad substrate scope suggests that the CaBioWF fusion is a versatile biocatalyst that can be used to prepare a number of aminoketone analogues. A BioWF fusion biocatalyst can convert various fatty acid substrates to α‐oxoamine (AON) products. The ATP‐dependent BioW domain catalyses formation of an acyl‐CoA thioester intermediate. The pyridoxal 5’‐phosphate (PLP)‐dependent BioF domain then accepts various amino acids and catalyses the formation of a range of aminoketone analogues.
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影响因子：
16.1
作者：
Chen, Xiaoying;Zaro, Jennica L.;Shen, Wei-Chiang
通讯作者：
Shen, Wei-Chiang
影响因子：
64.8
作者：
Jumper J;Evans R;Pritzel A;Green T;Figurnov M;Ronneberger O;Tunyasuvunakool K;Bates R;Žídek A;Potapenko A;Bridgland A;Meyer C;Kohl SAA;Ballard AJ;Cowie A;Romera-Paredes B;Nikolov S;Jain R;Adler J;Back T;Petersen S;Reiman D;Clancy E;Zielinski M;Steinegger M;Pacholska M;Berghammer T;Bodenstein S;Silver D;Vinyals O;Senior AW;Kavukcuoglu K;Kohli P;Hassabis D
通讯作者：
Hassabis D
影响因子：
16.6
作者：
Pazmino, Daniel E. Torres;Snajdrova, Radka;Fraaije, Marco W.
通讯作者：
Fraaije, Marco W.
DOI：
10.1073/pnas.0805983105
发表时间：
2008-10-14
影响因子：
11.1
作者：
Cryle, Max J.;Schlichting, Ilme
通讯作者：
Schlichting, Ilme
影响因子：
11.6
作者：
Cobessi, David;Dumas, Renaud;Alban, Claude
通讯作者：
Alban, Claude