Simultaneous overexpression of ∆6-, ∆12- and ∆9-desaturases enhanced the production of γ-linolenic acid in Mucor circinelloides WJ11

Wang, Xiuwen; Yang, Junhuan; Mohamed, Hassan; Shah, Aabid Manzoor; Li, Shaoqi; Pang, Shuxian; Wu, Changfan; Xue, Futing; Shi, Wenyue; Sadaqat, Beenish; Song, Yuanda

doi:10.3389/fmicb.2022.1078157

Cited by 3 publications

(3 citation statements)

References 64 publications

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“…Plasmid pMAT2076 (Supplementary Figure S1), which was used to overexpress the target genes in M. circinelloides, was constructed previously by our laboratory [37,38]. It contained a selectable marker (LeuA) and a strong promoter (Pzrt1) flanked by up-and down-stream sodit-a sequences.…”

Section: Plasmids Constructionmentioning

confidence: 99%

Construction of Eicosatetraenoic Acid Producing Cell Factory by Genetic Engineering of Mucor circinelloides

Yang

et al. 2023

Fermentation

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Eicosatetraenoic acid (ETA, 20:4, ω-3) is the desaturation product of dihomo-gamma linolenic acid (DGLA, 20:3, ω-6) catalyzed by delta-17 desaturase, which is considered as a healthy product that helps to lower risk of heart diseases. The oleaginous filamentous fungus, Mucor circinelloides, has been used for a long time as a model micro-organism for GLA production at industrial scales. However, M. circinelloides lacks the key enzymes to synthesize C20 polyunsaturated fatty acids (PUFAs). M. circinelloides could produce DGLA by overexpressing the D6E(GLELO) gene, which could be a useful tool to produce ETA due to the availability of established genetic manipulation tools. Therefore, in this study, delta-17 desaturase (PpD17 and PaD17) genes from Phytophthora parasitica and Pythium aphanidermatum, respectively, were introduced into M. circinelloides to construct an ETA-producing cell factory. Our results showed that the PaD17 and PpD17 overexpression strains’ biomass increased by 25.98 and 23.34 g/L (39.98 and 25.75%), respectively, compared with the control strain. Meanwhile, the lipid contents of the recombinant strains also increased and reached up to 28.88% in Mc-PaD17 and 30.95% in Mc-PpD17, respectively, compared with the control strain (23.38% in Mc-2076). The RT-qPCR results showed that overexpression of delta-17 desaturase genes promoted the expression of cme2, fas2, and D6E, thereby contributing to lipid biosynthesis in M. circinelloides. Meanwhile, the content of ETA reached up to 1.95%, and the yield of ETA was up to 114.69 mg/L in PpD17 overexpression mutants at 96 h. This study provided the first report on the construction of an ETA-producing cell factory by heterologous overexpression of the PpD17 gene in M. circinelloides, which established a new scope for further research in the production of ETA in oleaginous fungi.

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Section: Plasmids Constructionmentioning

confidence: 99%

Construction of Eicosatetraenoic Acid Producing Cell Factory by Genetic Engineering of Mucor circinelloides

Yang

et al. 2023

Fermentation

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“…Fatty acid methyl esters were separated by n-hexane and analyzed by gas Chromatography (GC) equipped with a 30 m ×0.32 mm DB-waxetr column with a film thickness of 0.25 µm. The program was set as follows: 120°C for 3 min, rising to 200°C at a rate of 5°C per min, and then rising to 220°C at a rate of 4°C per min and holding for 2 min (Wang et al, 2022;Zhang et al, 2017;.…”

Section: Determination Of Lipid Content and Fatty Acid Compositionmentioning

confidence: 99%

“…Generally, GLA biosynthesis requires a series of fatty acid desaturases to introduce double bonds at the 9, 12, and 6 carbon atoms of the stearic acid chain (Nykiforuk et al, 2012;Ratledge, 2005;Wang et al, 2022;Zhang et al, 2017). The final step in GLA formation is catalyzed by delta-6 fatty acid desaturase, which is the rate-limiting enzyme and a crucial regulatory point of the GLA biosynthetic pathway (Wang et al, 2023;Zhang et al, 2017;.…”

Section: Gla Accumulation Of Engineered M Circinelloides Strains From...mentioning

confidence: 99%

Effect of Adding Cellulose on Lipid Accumulation in Cellobiohydrolase and Delta-6 Desaturase Engineered Mucor circinelloides Strains

Zhang,

Wang

et al. 2023

American Journal of Biochemistry and Biotechnology

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Despite microbial production of multifunctional gamma-linolenic acid (GLA) being an important and promising way, the high cost is still a major factor limiting industrial applications. Previously, we proposed a novel approach to promote the production of GLA from cellulose, a widely available and cheapest resource, by overexpression of the cellobiohydrolase (CBH2) and delta-6 desaturase (D6) in Mucor circinelloides. In the present study, an indepth analysis of growth and lipid accumulation in engineered M. circinelloides strains using cellulosic substrate was to be explored. When cultivated in modified Kendrick and Ratledge (K and R) medium supplemented with the ratio of 1: 1 for glucose and carboxymethyl cellulose (CMC), the cellobiohydrolase and delta-6 desaturase overexpressing strains led to increases in the biomass (up to 10.4 g/L) and lipid yield (up to 2.6 g/L) of 68% and 2.2-fold, respectively, compared with that of the control strain. The yield of GLA in the cellobiohydrolase and delta-6 desaturase co-expression strains reached 415 mg/L, which was a remarkable increase of 3.1-fold compared to that of the control strain. These results proved that the introduction of CBH2 and D6 two key enzymes could significantly enhance the utilization of CMC and effectively convert cellulose into GLA by engineered M. circinelloides. This study provided a reference for the further potential application of the engineered M. circinelloides strains in the conversion of functional GLA from cellulose.

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Oleaginous fungi: a promising source of biofuels and nutraceuticals with enhanced lipid production strategies

Hassane,

Eldiehy,

Saha

et al. 2024

Arch Microbiol

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Simultaneous overexpression of ∆6-, ∆12- and ∆9-desaturases enhanced the production of γ-linolenic acid in Mucor circinelloides WJ11

Cited by 3 publications

References 64 publications

Construction of Eicosatetraenoic Acid Producing Cell Factory by Genetic Engineering of Mucor circinelloides

Construction of Eicosatetraenoic Acid Producing Cell Factory by Genetic Engineering of Mucor circinelloides

Effect of Adding Cellulose on Lipid Accumulation in Cellobiohydrolase and Delta-6 Desaturase Engineered Mucor circinelloides Strains

Oleaginous fungi: a promising source of biofuels and nutraceuticals with enhanced lipid production strategies

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