Mutated L-amino acid ligase and process for preparing L-glutamic acid-L-trp-trp by adopting enzyme catalysis method

Abstract

The invention relates to the technical field of biochemistry, and discloses mutated L-amino acid ligase and a process for preparing L-glutamic acid-L-trp-trp by adopting an enzyme catalysis method. Based on wild type L-amino acid ligase, the mutated L-amino acid ligase has site mutation of H276G / S, W332K / N / Q / S, M334D / S, L12A / S, Y75S / G and W76L / V. According to the mutated L-amino acid ligase and the process for preparing the L-glutamic acid-L-trp-trp by adopting the enzyme catalysis method disclosed by the invention, based on a known-reported research on non-specific L-amino acid ligase, systematical amino acid replacement on binding sites of the activity of a substrate of the mutated L-amino acid ligase is carried out, and finally, quick connection of L-glutamic acid and L-tryptophan is realkzed; and meanwhile, through combination of cyclic regeneration of coenzyme ATP and catalysis of immobilized enzyme, on one hand, the production cost is reduced; and on the other hand, the quality of trp-trp products and the stability of production of the trp-trp products are improved preferably, and therefore, an application requirement is met.

Description

technical field [0001] The invention relates to the field of biochemical technology, in particular to a mutated L-amino acid ligase and a process for preparing L-glutamic acid-L-tryptophan dipeptide by an enzyme-catalyzed method. Background technique [0002] L-glutamic acid-L-tryptophan (Oglufanide) is a dipeptide formed by connecting two basic amino acids L-glutamic acid and L-tryptophan through an amide bond at the Alpha position. Its molecular formula is C16H19N3O5 and its molecular weight is 333.3, CAS No. 122933-59-9. The dipeptide is an immunomodulator (immunomodulator) used in the treatment of chronic hepatitis C (Hepatitis C) infection. It is also used to regulate the human immune response in the clinical treatment of cancer. It is also a Drugs that inhibit angiogenesis. [0003] L-glutamic acid-L-tryptophan is mainly prepared by chemical synthesis, and there are also a small amount of enzymatic and fermentation methods on the market. Most of the chemical synthes...

AI Technical Summary

Problems solved by technology

Although many literatures have reported that some LaIs are active on L-glutamic acid and L-tryptophan, so far no ligase that can directly link this amino acid to form a dipeptide has been found.

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