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Co-feeding enhances the yield of methyl ketones
Journal of Industrial Microbiology & Biotechnology ( IF 3.4 ) Pub Date : 2023-09-12 , DOI: 10.1093/jimb/kuad029 Anita L Ziegler 1 , Carolin Grütering 2, 3 , Leon Poduschnick 1, 2 , Alexander Mitsos 1, 4, 5 , Lars M Blank 2
Journal of Industrial Microbiology & Biotechnology ( IF 3.4 ) Pub Date : 2023-09-12 , DOI: 10.1093/jimb/kuad029 Anita L Ziegler 1 , Carolin Grütering 2, 3 , Leon Poduschnick 1, 2 , Alexander Mitsos 1, 4, 5 , Lars M Blank 2
Affiliation
The biotechnological production of methyl ketones is a sustainable alternative to the fossil-derived chemical production. To date, the best host for microbial production of methyl ketones is a genetically engineered Pseudomonas taiwanensis VLB120 ∆6 pProd strain, achieving yields of 101mgg−1 on glucose in batch cultivations. For competitiveness with the petrochemical production pathway, however, higher yields are necessary. Co-feeding can improve the yield by fitting the carbon-to-energy ratio to the organism and the target product. In this work, we developed co-feeding strategies for P. taiwanensis VLB120 ∆6 pProd by combined metabolic modeling and experimental work. In a first step, we conducted flux balance analysis with an expanded genome-scale metabolic model of iJN1463 and found ethanol as the most promising among five cosubstrates. Next, we performed cultivations with ethanol and found the highest reported yield in batch production of methyl ketones with P. taiwanensis VLB120 to date, namely 154 mg g−1 methyl ketones. However, ethanol is toxic to the cell, which reflects in a lower substrate consumption and lower product concentrations when compared to production on glucose. Hence, we propose cofeeding ethanol with glucose and find that, indeed, higher concentrations than in ethanol-fed cultivation (0.84 g Laq−1 with glucose and ethanol as opposed to 0.48 g Laq−1 with only ethanol) were achieved, with a yield of 85 mg g−1. In a last step, comparing experimental with computational results suggested the potential for improving the methyl ketone yield by fed-batch cultivation, in which cell growth and methyl ketone production are separated into two phases employing optimal ethanol to glucose ratios.
更新日期：2023-09-12
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