Biotransformation of 2-keto-4-hydroxybutyrate via aldol condensation using an efficient and thermostable carboligase from Deinococcus radiodurans  

作　　者：Yeon-Ju Jeong Min-Ju Seo Bong Hyun Sung Jeong-Sun Kim Soo-Jin Yeom 

机构地区：[1]School of Biological Sciences and Biotechnology,Graduate School,Chonnam National University,Gwangju,Republic of Korea [2]School of Biological Sciences and Technology,Chonnam National University,Gwangju,Republic of Korea [3]Institute of Synthetic Biology for Carbon Neutralization,Chonnam National University,Gwangju 61186,Republic of Korea [4]Synthetic Biology Research Center,Korea Research Institute of Bioscience and Biotechnology,Daejeon 34141,Republic of Korea [5]Department of Chemistry,Chonnam National University,Gwangju 61186,Republic of Korea.

出　　处：《Bioresources and Bioprocessing》2024年第1期111-123,共13页生物资源与生物加工(英文)

基　　金：supported by Grants from the C1 Gas Refinery Program,funded by the Ministry of Science and ICT(NRF-2018M3D3A1A01056181);the Enzyme engineering for next generation biorefinery(NRF-2022M3J5A1056169 and NRF-2022M3J5A1085239)from National Research Foundation(NRF);the Korea Research Institute of Bioscience and the Biotechnology(KRIBB)Research Initiative Program.

摘　　要：The bioconversion of 4-hydroxy-2-keto acid derivatives via aldol condensation of formaldehyde and pyruvate has received substantial attention as potential source of chemicals for production of amino acids,hydroxy carboxylic acids,and chiral aldehydes.We developed an environmentally friendly biocatalyst consisting of a novel thermostable class II pyruvate aldolase from Deinococcus radiodurans with maltose-binding protein(MBP-DrADL),which has specific activity of 46.3μmol min-1 mg-1.Surprisingly,MBP-DrADL maintained over 60%of enzyme activity for 4 days at 50 to 65°C,we used MBP-DrADL as the best candidate enzyme to produce 2-keto-4-hydroxybutyrate(2-KHB)from formaldehyde and pyruvate via aldol condensation.The optimum reaction conditions for 2-KHB production were 50°C,pH 8.0,5 mM Mg2+,100 mM formaldehyde,and 200 mM pyruvate.Under these optimized conditions,MBP-DrADL produced 76.5 mM(8.94 g L-1)2-KHB over 60 min with a volumetric productivity of 8.94 g L-1 h-1 and a specific productivity of 357.6 mg mg-enzyme-1 h-1.Furthermore,2-KHB production was improved by continuous addition of substrates,which produced approximately 124.8 mM(14.6 g L-1)of 2-KHB over 60 min with a volumetric productivity and specific productivity of 14.6 g L-1 h-1 and 583.4 mg mg-enzyme-1 h-1,respectively.MBP-DrADL showed the highest specific productivity for 2-KHB production yet reported.Our study provides a highly efficient biocatalyst for the synthesis of 2-KHB and lays the foundation for large-scale production and application of high-value compounds from formaldehyde.

关 键 词：Formaldehyde PYRUVATE Pyruvate aldolase 2-keto-4-hydroxybutyrate Deinococcus radiodurans 

分 类 号：O62[理学—有机化学]