| Summary: | The effects of various carbon sources and cultivation conditions on the growth kinetics, lipid accumulation, and medium-chain fatty acid (MCFA) production of <i>Mucor circinelloides</i> (MC) was investigated for 72 h in shake flask cultivation. Our previous investigation reported increments of 28 to 46% MCFAs among total cell lipids when the MC genome was genetically modified, in comparison to the wild-type. However, the growth of the engineered strain M65-TE-04 was adversely affected. Therefore, the current study was designed to enhance the growth, lipid production, and MCFA productivity of engineered <i>M. circinelloides</i> by optimizing the pH, agitation speed, temperature, and carbon sources. The findings for individual variables disclosed that the highest biomass (17.0 g/L) was obtained when coconut oil mixed with glucose was used as a carbon source under normal culture conditions. Additionally, the maximum lipid contents (67.5% cell dry weight (CDW)), MCFA contents (53% total fatty acid (TFA)), and overall lipid productivity (3.53 g/L·d) were attained at 26 °C, pH 6.0, and 150 rpm, respectively. The maximum biomass (19.4 g/L), TFA (14.3g/L), and MCFA (4.71 g/L) contents were achieved with integration of a temperature of 26 °C, pH 6.0, agitation speed 300 rpm, and coconut oil mixed medium as the carbon source. This work illustrates that biomass, TFA, and MCFA contents were increased 1.70–2.0-fold by optimizing the initial pH, agitation speed, temperature, and carbon sources in the <i>M. circinelloides</i> engineered strain (M65-TE-04) in comparison to initial cultivation conditions.
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