Enhanced production of trans-cinnamic acid in Photorhabdus luminescens with homolog expression and deletion strategies

Gokduman F. U., YILMAZ S., Bode H. B.

Journal of Applied Microbiology, vol.135, no.7, 2024 (SCI-Expanded) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 135 Issue: 7
  • Publication Date: 2024
  • Doi Number: 10.1093/jambio/lxae149
  • Journal Name: Journal of Applied Microbiology
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Academic Search Premier, PASCAL, Agricultural & Environmental Science Database, Aqualine, Aquatic Science & Fisheries Abstracts (ASFA), BIOSIS, Biotechnology Research Abstracts, CAB Abstracts, Environment Index, Food Science & Technology Abstracts, Geobase, MEDLINE, Pollution Abstracts, Veterinary Science Database, DIALNET
  • Keywords: trans-cinnamic acid, phenylalanine ammonia-lyase, secondary metabolites, Photorhabdus luminescens, IRA402 resin extraction
  • Erciyes University Affiliated: Yes

Abstract

Aim: This study aimed to overproduce industrially relevant and safe bio-compound trans-cinnamic acid (tCA) from Photorhabdus luminescens with deletion strategies and homologous expression strategies that had not been applied before for tCA production. Methods and Results: The overproduction of the industrially relevant compound tCA was successfully performed in P. luminescens by deleting stlB (TTO1ΔstlB) encoding a cinnamic acid CoA ligase in the isopropylstilbene pathway and the hcaE insertion (knockout) mutation (hcaE::cat) in the phenylpropionate catabolic pathway, responsible for tCA degradation. A double mutant of both stlB deletion and hcaE insertion mutation (TTO1DM ΔstlB-hcaE::cat) was also generated. These deletion strategies and the phenylalanine ammonium lyase-producing (PI-PAL from Photorhabdus luminescens) plasmid, pBAD30C, carrying stlA (homologous expression mutants) are utilized together in the same strain using different media, a variety of cultivation conditions, and efficient anion exchange resin (Amberlite IRA402) for enhanced tCA synthesis. At the end of the 120-h shake flask cultivation, the maximum tCA production was recorded as 1281 mg l−1 in the TTO1pBAD30C mutant cultivated in TB medium, with the IRA402 resin keeping 793 mg l−1 and the remaining 488 mg l−1 found in the supernatant. Conclusion: TCA production was successfully achieved with homologous expression, coupled with deletion and insertion strategies. 1281 mg l−1is the highest tCA concentration that achieved by bacterial tCA production in flask cultivation, according to our knowledge.