Genetic Modification of Mucor circinelloides to Construct Stearidonic Acid Producing Cell Factory
Abstract
:1. Introduction
2. Results
2.1. Generation of D15D-Overexpressing Strains of M. circinelloides by Genetic Engineering
2.2. Expression Levels of D15D Genes in the Recombinant Strains
2.3. Effect of D15D Over-Expression in Cell Growth and Lipid Accumulation
2.4. SDA Accumulation in D15D Overexpressing Strains
3. Discussion
4. Materials and Methods
4.1. Plasmids, Strains and Conditions of Culture Media
4.2. Plasmids Construction
4.3. Gene Expression and RT-qPCR Analysis
4.4. Calculation of Glucose and Nitrogen Concentration from Culture Medium
4.5. Determination of Cell Dry Weight, Total Fatty Acid and Fatty Acid Analysis
4.6. Statistical Analysis
4.7. Ethics Approval and Consent to Participate
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
SDA | Stearidonic acid |
LA | Linoleic acid |
ALA | Alpha linolenic acid |
GLA | Gamma linolenic acid |
ARA | Arachidonic acid |
EPA | Eicosapentaenoic acid |
DHA | Docosahexaenoic acid |
PUFA | Polyunsaturated fatty acid |
GC | Gas chromatography |
VLCPUFA | Very-long-chain polyunsaturated fatty acid |
CDW | Cell dry weight |
SAFA | Saturated fatty acids |
MUFA | Mono-unsaturated fatty acids |
TFA | Total fatty acids |
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Time | Fatty Acid Composition (relative %, w/w) | ||||||
---|---|---|---|---|---|---|---|
Hour | C(16:0) | C(18:0) | C(18:1) OA % | C(18:2) LA % | C(18:3) GLA % | C(18:3) ALA % | C(18:4) SDA % |
Mc-1552 | |||||||
12 h | - | - | 32.38 ± 1.33 | 30.87 ± 0.96 | 36.75 ± 0.88 | - | - |
24 h | 29.10 ± 0.75 | 7.84 ± 0.58 | 20.43 ± 0.44 | 16.50 ± 0.67 | 26.13 ± 0.37 | - | - |
36 h | 29.91 ± 0.52 | 7.60 ± 0.27 | 22.57 ± 0.11 | 14.39 ± 0.38 | 25.53 ± 0.23 | - | - |
48 h | 22.23 ± 0.64 | 4.32 ± 0.44 | 26.68 ± 0.32 | 16.56 ± 0.25 | 30.21 ± 0.10 | - | - |
60 h | 24.72 ± 0.07 | 3.70 ± 0.15 | 25.85 ± 0.08 | 15.78 ± 0.05 | 29.95 ± 0.05 | - | - |
72 h | 24.92 ± 0.25 | 2.71 ± 0.32 | 26.09 ± 0.22 | 15.68 ± 0.30 | 30.60 ± 0.20 | - | - |
Mc-D15D | |||||||
12 h | 37.23 ± 1.68 | 14.89 ± 1.93 | 16.24 ± 1.3 | 16.91 ± 1.13 | 14.73 ± 0.95 | - | - |
24 h | 28.47 ± 0.44 | 9.08 ± 1.96 | 22.15 ± 0.77 | 18.91 ± 0.99 | 15.05 ± 0.58 | 3.73 ± 0.44 | 2.62 ± 0.38 |
36 h | 17.79 ± 1.73 | 5.87 ± 0.68 | 26.99 ± 1.28 | 18.60 ± 0.97 | 21.41 ± 1.22 | 4.38 ± 1.1 | 4.97 ± 0.96 |
48 h | 13.74 ± 0.09 | 2.96 ± 0.87 | 30.08 ± 0.03 | 17.61 ± 0.29 | 27.50 ± 0.06 | 3.13 ± 0.05 | 5.09 ± 0.08 |
60 h | 12.60 ± 0.68 | 2.37 ± 0.28 | 32.76 ± 0.47 | 16.41 ± 0.49 | 28.55 ± 0.53 | 2.60 ± 0.47 | 4.70 ± 0.17 |
72 h | 12.65 ± 0.07 | 2.03 ± 0.55 | 34.26 ± 0.08 | 15.90 ± 0.59 | 28.84 ± 0.44 | 2.13 ± 0.37 | 4.20 ± 0.28 |
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Khan, M.A.K.; Yang, J.; Hussain, S.A.; Zhang, H.; Garre, V.; Song, Y. Genetic Modification of Mucor circinelloides to Construct Stearidonic Acid Producing Cell Factory. Int. J. Mol. Sci. 2019, 20, 1683. https://doi.org/10.3390/ijms20071683
Khan MAK, Yang J, Hussain SA, Zhang H, Garre V, Song Y. Genetic Modification of Mucor circinelloides to Construct Stearidonic Acid Producing Cell Factory. International Journal of Molecular Sciences. 2019; 20(7):1683. https://doi.org/10.3390/ijms20071683
Chicago/Turabian StyleKhan, Md. Ahsanul Kabir, Junhuan Yang, Syed Ammar Hussain, Huaiyuan Zhang, Victoriano Garre, and Yuanda Song. 2019. "Genetic Modification of Mucor circinelloides to Construct Stearidonic Acid Producing Cell Factory" International Journal of Molecular Sciences 20, no. 7: 1683. https://doi.org/10.3390/ijms20071683