Immobilization of R-ω-transaminase on MnO2nanorods for catalyzing the conversion of (R)-1-phenylethylamine.
R-ɷ-transaminases transfer an amino group from an amino donor (e.g. (R)-1-phenylethylamine) onto an amino acceptor (e.g. pyruvate), resulting a co-product (e.g. d-alanine). This work intends to immobilize R-ɷ-Transaminase on MnO2nanorods to achieve multienzyme catalysis. R-ɷ-Transaminase (RTA) and d-amino acid oxidase (DAAO) have been fused to an elastin-like polypeptide (ELP) separately through genetic engineering of the enzymes. ELP-RTA and ELP-DAAO have been separately immobilized on polydopamine-coated MnO2nanorods. When the two immobilized enzymes were used together in one pot, the transformation of (R)-1-phenylethylamine was catalyzed by the immobilized ELP-RTA, and the co-product d-alanine was converted back to pyruvate under the catalysis of the immobilized ELP-DAAO, achieving the recycling of pyruvate in situ. Thus pyruvate was maintained at a low concentration in order to reduce its negative effect. On the other hand, the generated H2O2of ELP-DAAO was decomposed by the MnO2nanorods, and the evolved oxygen oxidized the reduced cofactors of ELP-DAAO. Forming the circles of hydrogen peroxide→oxygen→hydrogen peroxide accelerated the deamination reaction. The highly efficient conversion of the co-product d-alanine back to pyruvate accelerated the forming of the pyruvate→d-alanine→pyruvate cycle between the two immobilized enzymes. The coordination of the pyruvate→d-alanine→pyruvate and hydrogen peroxide→oxygen→hydrogen peroxide cycles accelerated the transformation of (R)-1-phenylethylamine. As a result, As a result, the immobilized enzymes achieved a conversion of 98±1.8% in comparison to 69.6±1.2% by free enzymes.
Department of Biochemical Engineering, Beijing University of Chemical Technology, Beijing 100029, China.
Jian
J
Sun
Department of Biochemical Engineering, Beijing University of Chemical Technology, Beijing 100029, China.
Wen-Hui
WH
Cui
Department of Biochemical Engineering, Beijing University of Chemical Technology, Beijing 100029, China.
Kun
K
Du
Department of Biochemical Engineering, Beijing University of Chemical Technology, Beijing 100029, China.
Qian
Q
Gao
Department of Chemical Engineering, Beijing University of Chemical Technology, Beijing 100029, China.
Mengmeng
M
Du
Department of Chemical Engineering, Beijing University of Chemical Technology, Beijing 100029, China. Electronic address: jipj@mail.buct.edu.cn.
Peijun
P
Ji
Department of Biochemical Engineering, Beijing University of Chemical Technology, Beijing 100029, China. Electronic address: fengwei@mail.buct.edu.cn.
Wei
W
Feng
D002384:Catalysis; D004800:Enzymes, Immobilized / Q000737:chemistry; D017895:Manganese Compounds / Q000737:chemistry*; D043942:Nanotubes / Q000737:chemistry*; D010087:Oxides / Q000737:chemistry*; D010627:Phenethylamines / Q000737:chemistry*; D000637:Transaminases / Q000737:chemistry*
D004800:Enzymes, Immobilized; D017895:Manganese Compounds; D010087:Oxides; D010627:Phenethylamines; C027424:manganese oxide; D000637:Transaminases
Elastin-like polypeptide; Immobilization; MnO(2) nanorods; Transaminase; d-amino acid oxidase