Establishment of synthetic microbial consortia with Corynebacterium glutamicum and Pseudomonas putida: Design, construction, and application to production of γ‐glutamylisopropylamide and l‐theanine

• Published in: Microbial biotechnology 2024-01, Vol.17 (1), p.e14400-n/a
• Main Authors: Benninghaus, Leonie, Schwardmann, Lynn S., Jilg, Tatjana, Wendisch, Volker F.
• Format: Article
• Language: English
• Subjects: Amines; Auxotrophy; Biosynthesis; Biotechnology; Consortia; Cooperation; Corynebacterium glutamicum; Design; Division of labor; E coli; Energy utilization; Enzymes; Formamidase; Genes; Green tea; Metabolism; Microorganisms; Monoculture; Plasmids; Pseudomonas putida; Raw materials; Substrates; Syntrophism; Theanine
• ISSN: 1751-7915; 1751-7915
• DOI: 10.1111/1751-7915.14400

Microbial synthetic consortia are a promising alternative to classical monoculture for biotechnological applications and fermentative processes. Their versatile use offers advantages in the degradation of complex substrates, the allocation of the metabolic burden between individual partners, or the division of labour in energy utilisation, substrate supply or product formation. Here, stable synthetic consortia between the two industrially relevant production hosts, Pseudomonas putida KT2440 and Corynebacterium glutamicum ATCC13032, were established for the first time. By applying arginine auxotrophy/overproduction and/or formamidase‐based utilisation of the rare nitrogen source formamide, different types of interaction were realised, such as commensal relationships (+/0 and 0/+) and mutualistic cross‐feeding (+/+). These consortia did not only show stable growth but could also be used for fermentative production of the γ‐glutamylated amines theanine and γ‐glutamyl‐isopropylamide (GIPA). The consortia produced up to 2.8 g L−1 of GIPA and up to 2.6 g L−1 of theanine, a taste‐enhancing constituent of green tea leaves. Thus, the advantageous approach of using synthetic microbial consortia for fermentative production of value‐added compounds was successfully demonstrated. Stable synthetic consortia between the two industrially relevant production hosts, Pseudomonas putida KT2440 and Corynebacterium glutamicum, were established for the first time. By applying arginine auxotrophy/overproduction and formamidase‐based utilisation of the rare nitrogen source formamide, commensal (0/+ and +/0) and mutualistic (+/+) consortia were constructed, characterised and used for fermentative production of the γ‑glutamylated amines γ‑glutamyl‑isopropylamide (GIPA) and l‐theanine.