Modulation of culture medium confers high-specificity production of isopentenol in Bacillus subtilis
| Autor: | Deepshi Chaurasia, Batul Diwan, Preeti Chaturvedi, Suresh Chandra Phulara, Pratima Gupta |
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| Rok vydání: | 2019 |
| Předmět: |
Microbiological Techniques
0106 biological sciences 0301 basic medicine Cost-Benefit Analysis Bioengineering Bacillus subtilis 01 natural sciences Applied Microbiology and Biotechnology Hydrolysate Prenol law.invention 03 medical and health sciences chemistry.chemical_compound Pentanols law 010608 biotechnology Pyruvic Acid Biomass chemistry.chemical_classification Organisms Genetically Modified biology Oryza Isoprenol biology.organism_classification Terpenoid Culture Media 030104 developmental biology Enzyme Metabolic Engineering chemistry Biochemistry Batch Cell Culture Techniques Recombinant DNA Sorbitol Metabolic Networks and Pathways Biotechnology |
| Zdroj: | Journal of Bioscience and Bioengineering. 127:458-464 |
| ISSN: | 1389-1723 |
| Popis: | Enthusiasm for mining isoprenoid-based flavors, pharmaceuticals, and nutraceuticals from GRAS (Generally Regarded as Safe) status microbial hosts has increased in the past few years due to the limitations associated with their plant-based extraction and chemical synthesis. Bacillus subtilis, a well-known GRAS microbe, is a promising alternative due to its fast growth rate and the ability to metabolize complex carbon sources. The study focused on the high-specificity production of isopentenol in B. subtilis by modulating the culture medium. Media modulation led to a 2.5 folds improvement in isopentenol titer in the wild-type strain. In the recombinant strain, optimization of physico-chemical factors, coupled with overexpression of the nudF enzyme resulted in a maximum isopentenol titer of ∼6 mg/L in a shake flask. The recombinant strain produced ∼5 mg/L isoprenol (∼80% of the total isopentenol production) and ∼1.8 mg/L prenol (∼65% of the total isopentenol production) by utilizing sorbitol and pyruvate as the carbon sources, respectively. Replacement of glucose with sorbitol and pyruvate reduced the production of the undesired metabolites and enhanced high-specificity production of isopentenol. Upon replacement of the carbon source with a low-cost substrate, a non-detoxified rice-straw hydrolysate, the engineered strain produced 2.19 mg/L isopentenol. This proof-of-concept study paves the path for the high-specificity production and cost-effective recovery of isopentenol from industrially competent microbial strains with engineered isoprenoid pathways. |
| Databáze: | OpenAIRE |
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