دورية أكاديمية

Fermentative production of enantiopure (S)-linalool using a metabolically engineered Pantoea ananatis

التفاصيل البيبلوغرافية
العنوان: Fermentative production of enantiopure (S)-linalool using a metabolically engineered Pantoea ananatis
المؤلفون: Nobuhisa Nitta, Yoshinori Tajima, Yoko Yamamoto, Mika Moriya, Akiko Matsudaira, Yasushi Hoshino, Yousuke Nishio, Yoshihiro Usuda
المصدر: Microbial Cell Factories, Vol 20, Iss 1, Pp 1-14 (2021)
بيانات النشر: BMC, 2021.
سنة النشر: 2021
المجموعة: LCC:Microbiology
مصطلحات موضوعية: Linalool, Biphasic fermentation, Mevalonate pathway, Solubility tag, Pantoea ananatis, Microbiology, QR1-502
الوصف: Abstract Background Linalool, an acyclic monoterpene alcohol, is extensively used in the flavor and fragrance industries and exists as two enantiomers, (S)- and (R)-linalool, which have different odors and biological properties. Linalool extraction from natural plant tissues suffers from low product yield. Although linalool can also be chemically synthesized, its enantioselective production is difficult. Microbial production of terpenes has recently emerged as a novel, environmental-friendly alternative. Stereoselective production can also be achieved using this approach via enzymatic reactions. We previously succeeded in producing enantiopure (S)-linalool using a metabolically engineered Pantoea ananatis, a member of the Enterobacteriaceae family of bacteria, via the heterologous mevalonate pathway with the highest linalool titer ever reported from engineered microbes. Results Here, we genetically modified a previously developed P. ananatis strain expressing the (S)-linalool synthase (AaLINS) from Actinidia arguta to further improve (S)-linalool production. AaLINS was mostly expressed as an insoluble form in P. ananatis; its soluble expression level was increased by N-terminal fusion of a halophilic β-lactamase from Chromohalobacter sp. 560 with hexahistidine. Furthermore, in combination with elevation of the precursor supply via the mevalonate pathway, the (S)-linalool titer was increased approximately 1.4-fold (4.7 ± 0.3 g/L) in comparison with the original strain (3.4 ± 0.2 g/L) in test-tube cultivation with an aqueous-organic biphasic fermentation system using isopropyl myristate as the organic solvent for in situ extraction of cytotoxic and semi-volatile (S)-linalool. The most productive strain, IP04S/pBLAAaLINS-ispA*, produced 10.9 g/L of (S)-linalool in “dual-phase” fed-batch fermentation, which was divided into a growth-phase and a subsequent production-phase. Thus far, this is the highest reported titer in the production of not only linalool but also all monoterpenes using microbes. Conclusions This study demonstrates the potential of our metabolically engineered P. ananatis strain as a platform for economically feasible (S)-linalool production and provides insights into the stereoselective production of terpenes with high efficiency. This system is an environmentally friendly and economically valuable (S)-linalool production alternative. Mass production of enantiopure (S)-linalool can also lead to accurate assessment of its biological properties by providing an enantiopure substrate for study.
نوع الوثيقة: article
وصف الملف: electronic resource
اللغة: English
تدمد: 1475-2859
Relation: https://doaj.org/toc/1475-2859
DOI: 10.1186/s12934-021-01543-0
URL الوصول: https://doaj.org/article/84b40a650d3e4ddd8672ca57cd4912b3
رقم الأكسشن: edsdoj.84b40a650d3e4ddd8672ca57cd4912b3
قاعدة البيانات: Directory of Open Access Journals
الوصف
تدمد:14752859
DOI:10.1186/s12934-021-01543-0