Construction and application of fusion proteins of d-amino acid oxidase and glutaryl-7-aminocephalosporanic acid acylase for direct bioconversion of cephalosporin C to 7-aminocephalosporanic acid |
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Authors: | Hui Luo Qiang Li Huimin Yu Zhongyao Shen |
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Institution: | Department of Chemical Engineering, Institute of Biochemical Engineering, Tsinghua University, Beijing 100084, PR China. |
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Abstract: | Two fusion proteins of D-amino acid oxidase (DAAO) and glutaryl-7-aminocephalosporanic acid acylase (GLA) were designed to simplify the bioconversion process of cephalosporin C to 7-aminocephalosporanic acid (7-ACA), which is conventionally produced in a two-step enzymatic process. Two recombinant plasmids, pET-DLA and pET-ALD, were constructed to express fusion proteins of DAAO-linker-GLA (DLA) and GLA-linker-DAAO (ALD), respectively. When the recombinant plasmids were expressed in E. coli, the fusion protein DLA was not correctly folded and only DAAO activity could be detected. ALD, however, possessed activities of both DAAO and GLA, which directly catalyze the conversion of cephalosporin C into 7-ACA. |
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Keywords: | d-amino acid oxidase 7-aminocephalosporanic acid cephalosporin C fusion protein glutaryl-7-aminocephalosporanic acid acylase |
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