Scientists have developed unnatural coenzyme synthase

Recently, Zhao Zongbao’s team, a researcher at the Dalian Institute of Chemical Physics, Chinese Academy of Sciences, has made new progress in the field of unnatural coenzyme research. They have developed nicotinamide cytosine dinucleotide (NCD) synthetase and NCD self-sufficient microbial cells, which have been successfully used to construct high Selective material metabolism pathway. Relevant research results were published in “Nature-Communication”.

Nicotinamide adenine dinucleotide (NAD) is an indispensable coenzyme in cells and is widely involved in processes such as metabolism. Therefore, the biological effects of changing NAD levels are difficult to predict. In order to break through the limitations of regulating natural coenzymes, Zhao Zongbao’s team is committed to synthetic biology research based on unnatural coenzymes.

In this study, nicotinic acid mononucleotide adenylyltransferase was used as a template to semi-rationally design and modify the binding pockets of its substrates nicotinic acid mononucleotide and adenosine triphosphate to obtain preference for nicotinamide mononucleotide (NMN ) And cytidine triphosphate (CTP) mutants, namely NCD synthase (NcdS). The team expressed NcdS in Escherichia coli and realized the use of endogenous CTP and NMN to synthesize NCD; by strengthening the precursor synthesis step, the intracellular NCD content can exceed the natural coenzyme, reaching 5.0 millimoles/liter, obtaining the NCD self-sufficient platform strain. In addition, the team also expressed NCD-preferred malic enzyme and D-lactate dehydrogenase to achieve NCD-mediated conversion of L-malic acid into D-lactic acid with high selectivity.

This work lays the foundation for the synthesis of unnatural coenzymes in cells and provides a reference for subsequent research in synthetic biology and chemical biology.

Related paper information: https://doi.org/10.1038/s41467-021-22357-z