ORIGINAL RESEARCH article
Front. Bioeng. Biotechnol.
Sec. Industrial Biotechnology
Volume 13 - 2025 | doi: 10.3389/fbioe.2025.1631063
Gas Chromatography for Analysis and Estimation of 13 C at Natural Abundance Level in Fatty Acids produced from Aurantiochytrium limacinum, a sustainable source of Polyunsaturated Fatty Acid
Provisionally accepted
- 1khalifa University of Sciences and Technology, Abu Dhabi, United Arab Emirates
- 2khalifa University of Sciences and Technology, ABU FJH, United Arab Emirates
- 3united arab emirates university, Al Ain, United Arab Emirates
- 4khalifa University of Sciences and Technolog, Abu Dhabi, United Arab Emirates
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Aurantiochytrium limacinum (A. limacinum) is a promising microbial source of polyunsaturated fatty acids (PUFAs), particularly Docosahexaenoic Acid (DHA, C22:6n-3). In this study, we first optimized the culture conditions of A. limacinum ATCC MYA-1381 (strain SR21). Cell growth was monitored via optical density, cell counts, and glucose concentration. Cells were harvested at exponential and stationary phases, and lipids were extracted using a green method. Fatty Acid Methyl Esters (FAMEs) were prepared and analyzed using Gas Chromatography-Flame Ionisation Detection (GC-FID). At the exponential phase, DHA was the most abundant (65.6% of total fatty acids) followed by palmitic acid (C16:0) at 34.4%. At the stationary phase, Docosapentaenoic acid (DPA, C22:5n-3) and DHA were the most abundant at 45.4% and 33.9% respectively. Myristic acid (C14:0), myristoleic acid (C14:1n-9), palmitic acid (C16:0) were present at 4.6%, 6.2% and 9.9%, respectively. Compound-specific isotope analysis (CSIA) using Gas Chromatography-Combustion-Isotope Ratio Mass Spectrometry (GC-C-IRMS) revealed that all FAMEs had negative δ 13 C values, indicating depletion in 13 C. At the exponential phase, δ 13 C (‰) of C16:0 and DHA were -16.8 ±0.2 and -18.5 ±0.1 ‰, respectively. At the stationary phase, δ 13 C (‰) of C14:0, C14:1n-9, C16:0, C22:5n-3 and DHA were -10.6 ±1.1, -11.3 ±0.1, -11.1 ±0.2, -8.3 ±0.2 and -10.6 ±0.1‰, respectively. Overall, our findings emphasized the importance of A. limacinum as a viable microbial platform for environmentally friendly production of PUFA such as DHA. Also, the study reinforced the utility of CSIA in tracking PUFA metabolic fate, which has latent applications in biomedical research, particularly in neurodegenerative disease frameworks where DHA plays a vital role. Finally, these results may also contribute to understanding isotopic fractionation patterns and metabolic flux variations across different microalgal growth phases.
Keywords: Aurantiochytrium limacinum, PUFA, DHA, CSIA, GC-FID, GC-C-IRMS
Received: 19 May 2025; Accepted: 11 Jul 2025.
Copyright: © 2025 Dirir, Chokshi, Ali, Alhanawi, Rommala and Hachem. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Mayssa Hachem, khalifa University of Sciences and Technolog, Abu Dhabi, United Arab Emirates
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