Study on energy metabolic control strategies based on non-natural coenzymes.

Recently, Zhao Zongbao, a researcher in the Biomass Efficient Conversion Research Group of the Biotechnology Department of Dalian Institute of Chemical Physics of the Chinese Academy of Sciences, made new progress in the research of microbial energy metabolism control, successfully transmitting inorganic energy to a biomass compatible energy carrier and realizing a selective drive for the conversion pathway of in-cell matter, the results of which were published in ACS Catalysis (DOI:10.1021/acscatal.6b3579).
metabolism is one of the basic characteristics of life activities and needs to be mediated by energy carriers (coenzymes).
Natural energy vectors such as Nicotinamide adenosine dinucleotide (NADH), which can participate in many metabolic reactions and other biological processes, make energy metabolic regulation less selective, low biological effect predictability and other outstanding problems.
, the team proposed energy metabolic control strategies based on non-natural coenzymes.
work, the design and synthesis of non-natural coenzyme nicosamine dinucleotide (Nicotinamide cytosine dinucleotide, NCD), modified the coenzyme binding pocket of redoxase, screening for non-natural coenzyme preference mutant enzymes; Experiments have shown that mutant enzymes maintain catalytic properties such as catalytic efficiency and substrate/product selectivity comparable to wild enzymes, thus creating a bio-orthosis redox catalytic system (J.Am.Chem.Soc., 2011,133,20857).
In this study, the research team successfully modified and obtained the NCD preference subphosphate dehydrogenase mutant Pdh, which is used to catalyz phosphates to efficiently produce and prototype non-natural coenzyme NCDH, and designed a series of in-body polyenzyme catalysis systems that found that NCDH-mediated reactions were independent of other reactions that relied on natural coenzymes, i.e. selective energy transfer properties.
With E. coli as the host, using genetically engineered means to over-express Pdh, NCD preferred magnase mutant Mae and nucleotide transporter protein Ndt, the resulting engineering bacteria incubated under the presence of phosphate, glucose and catalytic NCD, the yield and yield of apple acid than the system without adding NCD were significantly improved;
results show that engineering bacteria ingest NCD, using phosphate as an energy source, produce NCDH, selectively drive Mae-catalytic acetone acid reduction reaction to synthesize feline acid.
the results provide a scientific basis for chemical biology and synthetic biology based on non-natural coenzymes.
the above-mentioned work has been funded by the Ministry of Science and Technology's "973" program, the National Natural Science Foundation of China and the National Key Laboratory of Catalytic Foundations.
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