TY - JOUR
T1 - A biocatalytic platform for asymmetric alkylation of α-keto acids by mining and engineering of methyltransferases
AU - Ju, Shuyun
AU - Kuzelka, Kaylee P.
AU - Guo, Rui
AU - Krohn-Hansen, Benjamin
AU - Wu, Jianping
AU - Nair, Satish K.
AU - Yang, Yang
N1 - We are grateful to Profs. Armen Zakarian (UCSB), Tom Pettus (UCSB), Liming Zhang (UCSB) and Yiming Wang (University of Pittsburgh) for helpful discussions and critical reading of this manuscript. We acknowledge the NSF CAREER award (CHE-2145749 to Y.Y.) and the NIH (R01GM131347 to S.K.N.) for financial support. Transaminase study is partially supported by the ACS Herman Frasch Foundation (947-HF22 to Y.Y.). We thank the NSF BioPACIFIC MIP (DMR-1933487) and NSF MRSEC at UCSB (DMR-2308708) for access to instrumentation. We acknowledge Drs. Hongcheng Shen and Dian Li for assistance with substrate synthesis, Dr. Lei Cheng for assistance with Marfey’s analysis of 4b , and Spencer Anderson and the staff at LS-CAT (Sector 21, APS, Argonne IL) for facilitating data collection.
We are grateful to Profs. Armen Zakarian (UCSB), Tom Pettus (UCSB), Liming Zhang (UCSB) and Yiming Wang (University of Pittsburgh) for helpful discussions and critical reading of this manuscript. We acknowledge the NSF CAREER award (CHE-2145749 to Y.Y.) and the NIH (R01GM131347 to S.K.N.) for financial support. Transaminase study is partially supported by the ACS Herman Frasch Foundation (947-HF22 to Y.Y.). We thank the NSF BioPACIFIC MIP (DMR-1933487) and NSF MRSEC at UCSB (DMR-2308708) for access to instrumentation. We acknowledge Drs. Hongcheng Shen and Dian Li for assistance with substrate synthesis, Dr. Lei Cheng for assistance with Marfey’s analysis of 4b , and Spencer Anderson and the staff at LS-CAT (Sector 21, APS, Argonne IL) for facilitating data collection.
PY - 2023/12
Y1 - 2023/12
N2 - Catalytic asymmetric α-alkylation of carbonyl compounds represents a long-standing challenge in synthetic organic chemistry. Herein, we advance a dual biocatalytic platform for the efficient asymmetric alkylation of α-keto acids. First, guided by our recently obtained crystal structures, we develop SgvMVAV as a general biocatalyst for the enantioselective methylation, ethylation, allylation and propargylation of a range of α-keto acids with total turnover numbers (TTNs) up to 4,600. Second, we mine a family of bacterial HMTs from Pseudomonas species sharing less than 50% sequence identities with known HMTs and evaluated their activities in SAM regeneration. Our best performing HMT from P. aeruginosa, PaHMT, displays the highest SAM regeneration efficiencies (TTN up to 7,700) among HMTs characterized to date. Together, the synergistic use of SgvMVAV and PaHMT affords a fully biocatalytic protocol for asymmetric methylation featuring a record turnover efficiency, providing a solution to the notorious problem of asymmetric alkylation.
AB - Catalytic asymmetric α-alkylation of carbonyl compounds represents a long-standing challenge in synthetic organic chemistry. Herein, we advance a dual biocatalytic platform for the efficient asymmetric alkylation of α-keto acids. First, guided by our recently obtained crystal structures, we develop SgvMVAV as a general biocatalyst for the enantioselective methylation, ethylation, allylation and propargylation of a range of α-keto acids with total turnover numbers (TTNs) up to 4,600. Second, we mine a family of bacterial HMTs from Pseudomonas species sharing less than 50% sequence identities with known HMTs and evaluated their activities in SAM regeneration. Our best performing HMT from P. aeruginosa, PaHMT, displays the highest SAM regeneration efficiencies (TTN up to 7,700) among HMTs characterized to date. Together, the synergistic use of SgvMVAV and PaHMT affords a fully biocatalytic protocol for asymmetric methylation featuring a record turnover efficiency, providing a solution to the notorious problem of asymmetric alkylation.
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U2 - 10.1038/s41467-023-40980-w
DO - 10.1038/s41467-023-40980-w
M3 - Article
C2 - 37709735
AN - SCOPUS:85171353302
SN - 2041-1723
VL - 14
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 5704
ER -