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A two-step screening platform based on trans-aconitic acid assimilation unlocks novel bacterial resources for trans-aconitic acid production.

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Title: A two-step screening platform based on trans-aconitic acid assimilation unlocks novel bacterial resources for trans-aconitic acid production.
Authors: Zheng, Cao1, Hua, Jingyi1, Yang, Chengru1, Sun, Bowen1, Wu, Jingru1, Li, Anming1, Dai, Yujun1, Du, Cuiying1 ducuiying.123@163.com
Source: Applied & Environmental Microbiology. May2026, Vol. 92 Issue 5, p1-13. 13p.
Subjects: Microbial diversity, Bacteria classification, Organic acids, Biosynthesis, Microbiological assay, Isomerases, Biotechnology
Abstract: trans-Aconitic acid (TAA) possesses significant advantages in bioactivity, biosafety, and chemical modifiability, leading to its wide application in biopesticides, pharmaceuticals, and biobased synthesis. However, the scarcity of microbial resources for TAA production limits its large-scale commercial production, making it difficult to replace currently costly and low-yield chemical synthesis and plant extraction methods. In bacteria, aconitate isomerases (AIs) catalyze the biosynthesis of TAA. AIs are bifunctional enzymes that can also facilitate the reverse reaction, using TAA as the sole carbon source for bacterial growth. This provides a readily observable marker for the selection of TAA-producing bacteria. Using this dual functionality, we developed a novel two-step screening strategy comprising: (i) high-throughput screening of TAA-assimilating strains using a minimal medium with TAA as the sole carbon source and (ii) secondary screening of these assimilating strains to detect their ability to produce TAA in citrate medium. A total of 219 TAA-assimilating bacteria were isolated from a variety of environmental samples (n = 135). Subsequent LC-Q-TOF-MS analysis identified 19 strains (8.68%) capable of producing TAA at concentrations ranging from 34.5 to 621.9 µM. Taxonomic analysis revealed two new orders, five new families, and seven new genera of TAA-producing bacteria. This study established an efficient AI-directed strategy for discovering novel TAA-producing bacteria and significantly expanded taxonomic diversity, providing a sustainable resource for microbial technology development and strain innovation, thereby promoting the industrial biomanufacturing of TAA and its applications in multiple fields. [ABSTRACT FROM AUTHOR]
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Database: Engineering Source
Description
Abstract:trans-Aconitic acid (TAA) possesses significant advantages in bioactivity, biosafety, and chemical modifiability, leading to its wide application in biopesticides, pharmaceuticals, and biobased synthesis. However, the scarcity of microbial resources for TAA production limits its large-scale commercial production, making it difficult to replace currently costly and low-yield chemical synthesis and plant extraction methods. In bacteria, aconitate isomerases (AIs) catalyze the biosynthesis of TAA. AIs are bifunctional enzymes that can also facilitate the reverse reaction, using TAA as the sole carbon source for bacterial growth. This provides a readily observable marker for the selection of TAA-producing bacteria. Using this dual functionality, we developed a novel two-step screening strategy comprising: (i) high-throughput screening of TAA-assimilating strains using a minimal medium with TAA as the sole carbon source and (ii) secondary screening of these assimilating strains to detect their ability to produce TAA in citrate medium. A total of 219 TAA-assimilating bacteria were isolated from a variety of environmental samples (n = 135). Subsequent LC-Q-TOF-MS analysis identified 19 strains (8.68%) capable of producing TAA at concentrations ranging from 34.5 to 621.9 µM. Taxonomic analysis revealed two new orders, five new families, and seven new genera of TAA-producing bacteria. This study established an efficient AI-directed strategy for discovering novel TAA-producing bacteria and significantly expanded taxonomic diversity, providing a sustainable resource for microbial technology development and strain innovation, thereby promoting the industrial biomanufacturing of TAA and its applications in multiple fields. [ABSTRACT FROM AUTHOR]
ISSN:00992240
DOI:10.1128/aem.01831-25