Abstract
Increasing energy demands and health-related concerns worldwide have motivated researchers to adopt diverse strategies to improve medium-chain fatty acid (MCFA) biosynthesis for use in the functional food and aviation industries. The abundance of naturally produced MCFAs from botanical sources (i.e., coconut fruit/seeds and palm tree) has been observed to be insufficient compared with the various microorganisms used to cope with industrial demands. Mucor circinelloides is one of many promising microorganisms; it exhibits diverse biotechnological importance ranging from the production of functional lipids to applications in the manufacture of bio-fuel. Thus, research was conducted to acquire the desired elevated amounts of MCFAs (i.e., C8–C12) from metabolically engineered strains of M. circinelloides M65. To achieve this goal, four different acyl-acyl carrier protein (ACP) thioesterase (TE)-encoding genes exhibiting a substrate preference for medium-chain acyl-ACP molecules were expressed in M. circinelloides M65, resulting in the generation of C8–C12 fatty acids. Among all the engineered strains, M65-TE-03 and M65-TE-04 demonstrated the highest production of non-native C8–C10 and C12 fatty acids, respectively, in comparison to the control. These recombinant strains biosynthesized MCFAs de novo within the range from 28 to 46% (i.e., 1.14 to 2.77 g/L) of total cell lipids. Moreover, the reduction in chain length eventually resulted in a 1.5–1.75-fold increase in total lipid productivity in the engineered strains. The MCFAs were also found to be integrated into all lipid classes. This work illustrates how the integration of heterologous enzymes in M. circinelloides can offer a novel opportunity to edit the fatty acid synthases (FAS) complex, resulting in increased production of microbial MFCAs.
Highlights
Medium-chain fatty acids (MCFAs) and their derivates with a chain length of 8 to 12 [1] are involved in the production of green fuels
Despite the elevation in non-native fatty acid production, we noticed a significant increase in total lipid titers during the process of fermentation
We found that four different TEs from bacterial and plant sources led to significant increments in lipid accumulation and evident changes in the fatty acid profile when heterologously over-expressed in M circinelloides M65
Summary
Medium-chain fatty acids (MCFAs) and their derivates with a chain length of 8 to 12 [1] are involved in the production of green fuels. These MCFAs have gained attention as part of a healthy diet because they are absorbed and transported directly into the liver via portal veins and rapidly metabolized by the β-oxidation pathway, eventually augmenting diet-induced thermo-genesis. This process offers a platform for the prevention/treatment of diverse metabolic disorders, including atherosclerosis, hypertension, hyperlipidemia, obesity, type-II diabetes, and cardiovascular diseases [9]. These microorganisms represent a reliable option for the production of fatty-acid-derived products on an industrial scale [19,20,21,22,23]
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