![]() ![]() Quantitation of the precursors provides information of the metabolic flux and intracellular metabolic state, giving guidance for precise pathway engineering. The biosynthesis of food components (protein, fats, carbohydrates or vitamins) in engineered microbial cells often involves cellular central metabolic pathways, where common precursors are processed into different proteins and products. Synthetic biology, using engineered microbial cell factories for production of various molecules, shows great advantages for generating food alternatives and additives, which not only relieve the pressure laid on tradition agriculture, but also create a new stage in healthy and sustainable food supplement. ![]() In addition, in the emerging of consumers’ awareness of health related issues bring a growing trend towards novel nature-based food additives. Nowadays, traditional agriculture faces challenges in balancing the need of sustainable environmental development and the rising food demand caused by an increasing population. The developed assay was used to quantitate ACoA and MCoA levels in various tissues of rat.įood is essential for human survival. ACoA and MCoA were stable in a battery of stability studies viz. The intra- and inter-day precision values for ACoA and MCoA met the acceptance as per FDA guidelines. A linear response function was established for the range of concentrations 1.09-2187 and 1.09-2193 ng/mL for ACoA and MCoA, respectively. ![]() The total run time was 3 min and the elution of both analytes (ACoA, MCoA) and IS occurred at 1.28 min this was achieved with a mobile phase consisting of 5 mM ammonium formate (pH 7.5)-acetonitrile (30:70, v/v) delivered at a flow rate of 1 mL/min on a monolithic RP-18e column. Simple acidification followed by dilution using an assay buffer process was used to extract ACoA, MCoA and IS from surrogate matrix and tissue samples. LC-MS/MS was operated under the multiple reaction-monitoring mode using the electrospray ionization technique. A highly sensitive and specific LC-MS/MS method was developed for simultaneous estimation of acetyl co-enzyme A (ACoA) and malonyl co-enzyme A (MCoA) in surrogate matrix using n-propionyl co-enzyme A as an internal standard (IS). ![]()
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