Tianjin Workers' Industry is making progress in using artificial fusion protein to increase methanol biotransformation rate
October 16, 2018 Source: Chinese Academy of Sciences
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Schematic diagram of improving the biotransformation efficiency of methanol by using artificial fusion protein
As an alternative carbon source, methanol has the advantages of abundant source, low price and high reduction compared with the existing fermentation raw materials. Therefore, it is of great social and economic significance to study methanol biotransformation technology and develop a methanol-based bio-manufacturing industry. Oxidation of methanol to formaldehyde is the first step in the bioconversion of methanol and is the rate limiting step. Increasing the oxidation rate of methanol while avoiding the accumulation of formaldehyde in highly toxic intermediates is the key to increasing the biotransformation rate of methanol.
Recently, Zheng Ping, a researcher at the Tianjin Institute of Industrial Biotechnology of the Chinese Academy of Sciences, led a team of systems and synthetic biotechnology research team and researcher Sun Jibin to lead the research team of the Center for Systems Biology to rationally design and construct key enzymes for the ribulose monophosphate methanol utilization pathway. The fusion protein increases the rate of conversion of methanol to the key metabolic intermediate fructose-6-phosphate (F6P). The researchers were active against NAD+-dependent methanol dehydrogenase (Mdh), 3-hexulose-6-phosphate synthase (Hps) and 6-phosphate-3-hexulose isomerase (Phi) from different sources. The bifunctional fusion proteins (Mdh-Hps and Hps-Phi) and the trifunctional fusion protein (Mdh-Hps-Phi) were screened and selected, and the effects of different lengths of flexible linker on the catalytic efficiency of the fusion protein were evaluated. The optimal fusion protein has a 4.8-fold increase in methanol oxidation activity and a 30% increase in F6P production rate. Dynamic light scattering and transmission electron microscopy analysis showed that protein fusion expression changed the state of the polymer and formed a larger assembly, which may affect the catalytic activity. This study provides an effective strategy for solving the toxicity of formaldehyde and increasing the biotransformation rate of methanol.
The research has been supported by the National Natural Science Foundation of China, the Chinese Academy of Sciences Key Deployment Project, the Chinese Academy of Sciences International Cooperation Bureau's Foreign Cooperation Key Project, the Tianjin Youth Talents Program and the Tianjin Special Support Project. The relevant research results have been published in the journal ChemBioChem. Tianjin Institute of Labor and the University of Science and Technology of China jointly cultivated Ph.D. student Fan Liwen, assistant researcher of Tianjin Institute of Labor and Social Welfare, the co-first author of the paper, and the engineer of the Technical Support Center of Tianjin Institute of Labor and Technology, Huan Huanhuan, provided assistance for TEM analysis.
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