| Title: | Production of indole by Corynebacterium glutamicum microbial cell factories for flavor and fragrance applications |
| Author(s): | Mindt M; Beyraghdar Kashkooli A; Suarez-Diez M; Ferrer L; Jilg T; Bosch D; Martins Dos Santos V; Wendisch VF; Cankar K; |
| Address: | "Business Unit Bioscience, Wageningen Plant Research, Wageningen University & Research, Wageningen, The Netherlands. Axxence Aromatic GmbH, Emmerich am Rhein, Germany. Laboratory of Systems and Synthetic Biology, Wageningen University & Research, Wageningen, The Netherlands. Genetics of Prokaryotes, Faculty of Biology & CeBiTec, Bielefeld University, Bielefeld, Germany. Laboratory of Bioprocess Engineering, Wageningen University & Research, Wageningen, The Netherlands. Business Unit Bioscience, Wageningen Plant Research, Wageningen University & Research, Wageningen, The Netherlands. katarina.cankar@wur.nl" |
| DOI: | 10.1186/s12934-022-01771-y |
| ISSN/ISBN: | 1475-2859 (Electronic) 1475-2859 (Linking) |
| Abstract: | "BACKGROUND: The nitrogen containing aromatic compound indole is known for its floral odor typical of jasmine blossoms. Due to its characteristic scent, it is frequently used in dairy products, tea drinks and fine fragrances. The demand for natural indole by the flavor and fragrance industry is high, yet, its abundance in essential oils isolated from plants such as jasmine and narcissus is low. Thus, there is a strong demand for a sustainable method to produce food-grade indole. RESULTS: Here, we established the biotechnological production of indole upon L-tryptophan supplementation in the bacterial host Corynebacterium glutamicum. Heterologous expression of the tryptophanase gene from E. coli enabled the conversion of supplemented L-tryptophan to indole. Engineering of the substrate import by co-expression of the native aromatic amino acid permease gene aroP increased whole-cell biotransformation of L-tryptophan to indole by two-fold. Indole production to 0.2 g L(-1) was achieved upon feeding of 1 g L(-1) L-tryptophan in a bioreactor cultivation, while neither accumulation of side-products nor loss of indole were observed. To establish an efficient and robust production process, new tryptophanases were recruited by mining of bacterial sequence databases. This search retrieved more than 400 candidates and, upon screening of tryptophanase activity, nine new enzymes were identified as most promising. The highest production of indole in vivo in C. glutamicum was achieved based on the tryptophanase from Providencia rettgeri. Evaluation of several biological aspects identified the product toxicity as major bottleneck of this conversion. In situ product recovery was applied to sequester indole in a food-grade organic phase during the fermentation to avoid inhibition due to product accumulation. This process enabled complete conversion of L-tryptophan and an indole product titer of 5.7 g L(-1) was reached. Indole partitioned to the organic phase which contained 28 g L(-1) indole while no other products were observed indicating high indole purity. CONCLUSIONS: The bioconversion production process established in this study provides an attractive route for sustainable indole production from tryptophan in C. glutamicum. Industrially relevant indole titers were achieved within 24 h and indole was concentrated in the organic layer as a pure product after the fermentation" |
| Keywords: | *Corynebacterium glutamicum/genetics/metabolism Escherichia coli/metabolism Indoles/metabolism Odorants Tryptophan/metabolism Bioconversion Bioprospecting Corynebacterium glutamicum Flavor and fragrance applications In situ product recovery Indole Tryptop; |
| Notes: | "MedlineMindt, Melanie Beyraghdar Kashkooli, Arman Suarez-Diez, Maria Ferrer, Lenny Jilg, Tatjana Bosch, Dirk Martins Dos Santos, Vitor Wendisch, Volker F Cankar, Katarina eng 722361/ERA CoBioTech project INDIE, European Union's Horizon 2020 research and innovation programme/ 053.80.732/Dutch research council (NWO)/ 22023517/German funding agency (FNR)/ England 2022/03/26 Microb Cell Fact. 2022 Mar 24; 21(1):45. doi: 10.1186/s12934-022-01771-y" |
