Biosynthesis of 3-Hydroxy-3-Methylbutyrate from l-Leucine by Whole-Cell Catalysis.

Ruichen Gao, Zhimin Li

Journal of agricultural and food chemistry 2021 Mar 31

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3-Hydroxy-3-methylbutyrate (HMB) is an important compound that can be used for the synthesis of a variety of chemicals in the food and pharmaceutical fields. Here, a biocatalytic method using l-leucine as a substrate was designed and constructed by expressing l-amino acid deaminase (l-AAD) and 4-hydroxyphenylpyruvate dioxygenase (4-HPPD) in Escherichia coli. To reduce the influence of the rate-limiting step on the cascade reaction, two 4-HPPD mutants were screened by rational design and both showed improved catalytic activity. Under optimal reaction conditions, the maximum conversion rate and production rate were 80% and 0.257 g/L·h, respectively. HMB production could be realized with high efficiency without an additional supply of adenosine triphosphate (ATP), which successfully overcomes the shortcomings of chemical production and fermentation methods. This design-based strategy of constructing a whole-cell catalyst system from l-leucine might serve as an alternative route to HMB synthesis.

Citation

Ruichen Gao, Zhimin Li. Biosynthesis of 3-Hydroxy-3-Methylbutyrate from l-Leucine by Whole-Cell Catalysis. Journal of agricultural and food chemistry. 2021 Mar 31;69(12):3712-3719