New Invention Profile
| Publication No: | IN202311037138 [India] | Application No: | IN202311037138 |
| Title: | GENETICALLY ENGINEERED YEAST AND USE THEREOF FOR DE NOVO PRODUCTION OF ENANTIOPURE GERMACRENE D | ||
| Publication Date: | 06-12-2024 | File Date: | 29-05-2023 |
| Inventor(s): | SUMIT GHOSH; SHUBHA SHARMA; SEEMA CHAURASIA; SANDEEP; GAURAV SRIVASTAVA; ANAMIKA SINGH; CHANDAN SINGH CHANOTIYA | ||
| IPC Classification: | C12N 9/10, C12N 9/88, C12N 15/81, C12P 5/00, C12N 9/90 | ||
| Abstract: | Germacrene D, an industrially-important sesquiterpenoid, is found mainly in plant essential oils at a low level as (+) and/or (-) enantiomeric forms and the chemical process for enantiopure germacrene D is not yet established, which are the concerns for sustainable supply. The present invention provides a method of high-level de novo biosynthesis of (+) and (-)-germacrene D by expressing germacrene D synthases in engineered yeast (S. cerevisiae), which does not require an inducer of gene expression and thus, the biosynthetic platform is constitutive. To develop the yeast-producing enantiopure germacrene D in a constitutive manner, targeted genomic modifications were carried out in eight loci. These genomic modifications represented the knock-out of two yeast genes (Gal80 and LPP1), over-expression of eight ERG pathway genes (ERG10, ERG13, HMG1, ERG12, ERG8, ERG19, IDI, ERG20), replacement of ERG9 promoter with a regulated promoter (PCTR3) and multiple copy integration of germacrene D synthase expression cassette at gal80, lpp1 and rDNA loci. The present metabolic engineering strategy coupled with expression of a ATP citrate lyase provides high titers of (+)-germacrene D (290.28 g/ml) and (-)-germacrene D (2519.46 g/ml). |
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Industrial Applications
Medicinal and Aromatics Plants [Agriculture]
Synhtesis Procesess [Drugs and Pharmaceuticals]