Aromatic compound transporter mutant and application thereof

By performing site-directed mutagenesis on efflux proteins to create a one-way valve function, the reflux problem in aromatic compound production was solved, achieving high-efficiency production and improved host bacterial tolerance, thereby increasing the efflux efficiency and yield of aromatic compounds.

CN121991189AActive Publication Date: 2026-05-08INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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Patent Information

Application Number
CN202610458370.2
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2026-04-09
Publication Date
2026-05-08
Estimated Expiration
2046-04-09

AI Technical Summary

Technical Problem

In existing technologies, extracellular efflux proteins exhibit non-selective bidirectional diffusion during the production of aromatic compounds, leading to the reflux of aromatic compounds, weakening cellular detoxification efficiency, and causing toxicity accumulation, which makes it difficult to meet the needs of industrial production.

Method used

We developed an extracellular efflux protein mutant with a one-way valve function. By performing site-directed mutations on key amino acid residues, we created steric hindrance to block the backflow of aromatic compounds and optimized the channel structure to improve the efflux efficiency of small molecule aromatic compounds.

Benefits of technology

It achieved a synergistic improvement in the efficient production of aromatic compounds and the host bacteria's tolerance, significantly reduced the accumulation of intracellular toxicity, and increased the titer and production intensity of the target product.

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Abstract

The invention discloses an aromatic compound transporter mutant and application thereof, and belongs to the technical field of bioengineering. The mutant is obtained by carrying out site-specific substitution on a key residue of a gated ring on the basis of an outer membrane transporter derived from Pseudomonas putida, and specifically comprises one or a combination of more of substitution of 120th phenylalanine into glycine, substitution of 121st leucine into glycine and substitution of 122nd glutamine into glycine, and the mutant is obtained by carrying out site-specific substitution on a key residue of a gated ring of the outer membrane transporter, so that the mutant is obtained, and the mutant is obtained by carrying out site-specific substitution on the key residue of the gated ring of the Pseudomonas putida derived from Pseudomonas putida derived from Pseudomonas putida derived from Pseudomonas putida derived from Pseudomonas putida. Preferably, a single mutant F120G, a double mutant F120G / L121G or a three mutant F120G / L121G / Q122G can be included, and conservative substitution derivation can be carried out on the basis of the F120G / L121G / Q122G. The mutant can be used for microbial efficient production and cell tolerance regulation of aromatic compounds according to physicochemical characteristics of a target product, and has important application value in green biological manufacturing, fermentation engineering and bioremediation.
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