Homologous Overexpression of Acyl-CoA Thioesterase 8 Enhanced Free Fatty Acid Accumulation in Oleaginous Fungus <i>Mucor circinelloides</i> WJ11
Thioesterases play an essential role in the metabolism of fatty acids since they are considered one of the key enzymes to change the total amount and composition of fatty acid in an organism’s cells. Acyl-coenzyme A thioesterase 8 (ACOT8) exhibits substrate specificity mainly for short- to long-chai...
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MDPI AG
2023-06-01
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| Series: | Fermentation |
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| Online Access: | https://www.mdpi.com/2311-5637/9/6/545 |
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| author | Feifei Xin Meng Gao Yufei Chang Wenrui Dang Ruixue Wang Hongjuan Yuan Zhike Xie Yanlei Zhao Yuanda Song Chunchi Rong Huaiyuan Zhang |
| author_facet | Feifei Xin Meng Gao Yufei Chang Wenrui Dang Ruixue Wang Hongjuan Yuan Zhike Xie Yanlei Zhao Yuanda Song Chunchi Rong Huaiyuan Zhang |
| author_sort | Feifei Xin |
| collection | DOAJ |
| description | Thioesterases play an essential role in the metabolism of fatty acids since they are considered one of the key enzymes to change the total amount and composition of fatty acid in an organism’s cells. Acyl-coenzyme A thioesterase 8 (ACOT8) exhibits substrate specificity mainly for short- to long-chain acyl-CoA. To identify and characterize the ACOT8 enzyme’s superfamily in <i>Mucor circinelloides</i>, three genes were characterized and homologously expressed in <i>M. circinelloides</i> WJ11 which has been used as a model organism to investigate the mechanism of lipid accumulation. Multiple sequence alignment showed that McACOT8s had significant conserved motifs in the ACOT8 family. Experimental data indicated that the biomass of the three recombinant strains (McACOT8a, McACOT8b and McACOT8c) was slightly lower than that of the control strain Mc2075, but the total fatty acid (TFA) content was significantly increased by 30.3, 21.5 and 23.9%, respectively, and the free fatty acid production increased from 12.9% (control strain) to 19.2, 25.2 and 26.2% (recombinant types), respectively, with a maximum increase of 103.1% in McACOT8c. The three strains did not show any significant improvement in fatty acid composition compared to the control strain. However, in terms of fatty acid composition of free fatty acid, McACOT8b and McACOT8c contained γ-linolenic acid (C18:3), which was not detected in the control strain or in McACOT8a, indicating that ACOT8b and ACOT8c had substrate specificity for C18:3. These results displayed that ACOT8 can increase TFA accumulation and that it may be an important target of genetic manipulation for microbial oil production. |
| first_indexed | 2024-03-11T02:29:27Z |
| format | Article |
| id | doaj.art-1d50d6644ab14feca659354e35de8d25 |
| institution | Directory Open Access Journal |
| issn | 2311-5637 |
| language | English |
| last_indexed | 2024-03-11T02:29:27Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Fermentation |
| spelling | doaj.art-1d50d6644ab14feca659354e35de8d252023-11-18T10:21:13ZengMDPI AGFermentation2311-56372023-06-019654510.3390/fermentation9060545Homologous Overexpression of Acyl-CoA Thioesterase 8 Enhanced Free Fatty Acid Accumulation in Oleaginous Fungus <i>Mucor circinelloides</i> WJ11Feifei Xin0Meng Gao1Yufei Chang2Wenrui Dang3Ruixue Wang4Hongjuan Yuan5Zhike Xie6Yanlei Zhao7Yuanda Song8Chunchi Rong9Huaiyuan Zhang10Colin Ratledge Center for Microbial Lipids, School of Agriculture Engineering and Food Science, Shandong University of Technology, 266 Xincun West Road, Zibo 255000, ChinaColin Ratledge Center for Microbial Lipids, School of Agriculture Engineering and Food Science, Shandong University of Technology, 266 Xincun West Road, Zibo 255000, ChinaColin Ratledge Center for Microbial Lipids, School of Agriculture Engineering and Food Science, Shandong University of Technology, 266 Xincun West Road, Zibo 255000, ChinaColin Ratledge Center for Microbial Lipids, School of Agriculture Engineering and Food Science, Shandong University of Technology, 266 Xincun West Road, Zibo 255000, ChinaColin Ratledge Center for Microbial Lipids, School of Agriculture Engineering and Food Science, Shandong University of Technology, 266 Xincun West Road, Zibo 255000, ChinaColin Ratledge Center for Microbial Lipids, School of Agriculture Engineering and Food Science, Shandong University of Technology, 266 Xincun West Road, Zibo 255000, ChinaColin Ratledge Center for Microbial Lipids, School of Agriculture Engineering and Food Science, Shandong University of Technology, 266 Xincun West Road, Zibo 255000, ChinaColin Ratledge Center for Microbial Lipids, School of Agriculture Engineering and Food Science, Shandong University of Technology, 266 Xincun West Road, Zibo 255000, ChinaColin Ratledge Center for Microbial Lipids, School of Agriculture Engineering and Food Science, Shandong University of Technology, 266 Xincun West Road, Zibo 255000, ChinaSchool of Food Science and Pharmaceutical Engineering, Nanjing Normal University, Nanjing 210023, ChinaColin Ratledge Center for Microbial Lipids, School of Agriculture Engineering and Food Science, Shandong University of Technology, 266 Xincun West Road, Zibo 255000, ChinaThioesterases play an essential role in the metabolism of fatty acids since they are considered one of the key enzymes to change the total amount and composition of fatty acid in an organism’s cells. Acyl-coenzyme A thioesterase 8 (ACOT8) exhibits substrate specificity mainly for short- to long-chain acyl-CoA. To identify and characterize the ACOT8 enzyme’s superfamily in <i>Mucor circinelloides</i>, three genes were characterized and homologously expressed in <i>M. circinelloides</i> WJ11 which has been used as a model organism to investigate the mechanism of lipid accumulation. Multiple sequence alignment showed that McACOT8s had significant conserved motifs in the ACOT8 family. Experimental data indicated that the biomass of the three recombinant strains (McACOT8a, McACOT8b and McACOT8c) was slightly lower than that of the control strain Mc2075, but the total fatty acid (TFA) content was significantly increased by 30.3, 21.5 and 23.9%, respectively, and the free fatty acid production increased from 12.9% (control strain) to 19.2, 25.2 and 26.2% (recombinant types), respectively, with a maximum increase of 103.1% in McACOT8c. The three strains did not show any significant improvement in fatty acid composition compared to the control strain. However, in terms of fatty acid composition of free fatty acid, McACOT8b and McACOT8c contained γ-linolenic acid (C18:3), which was not detected in the control strain or in McACOT8a, indicating that ACOT8b and ACOT8c had substrate specificity for C18:3. These results displayed that ACOT8 can increase TFA accumulation and that it may be an important target of genetic manipulation for microbial oil production.https://www.mdpi.com/2311-5637/9/6/545Acyl-CoA thioesterase 8free fatty acidhomologous overexpression<i>Mucor circinelloides</i>total fatty acid |
| spellingShingle | Feifei Xin Meng Gao Yufei Chang Wenrui Dang Ruixue Wang Hongjuan Yuan Zhike Xie Yanlei Zhao Yuanda Song Chunchi Rong Huaiyuan Zhang Homologous Overexpression of Acyl-CoA Thioesterase 8 Enhanced Free Fatty Acid Accumulation in Oleaginous Fungus <i>Mucor circinelloides</i> WJ11 Fermentation Acyl-CoA thioesterase 8 free fatty acid homologous overexpression <i>Mucor circinelloides</i> total fatty acid |
| title | Homologous Overexpression of Acyl-CoA Thioesterase 8 Enhanced Free Fatty Acid Accumulation in Oleaginous Fungus <i>Mucor circinelloides</i> WJ11 |
| title_full | Homologous Overexpression of Acyl-CoA Thioesterase 8 Enhanced Free Fatty Acid Accumulation in Oleaginous Fungus <i>Mucor circinelloides</i> WJ11 |
| title_fullStr | Homologous Overexpression of Acyl-CoA Thioesterase 8 Enhanced Free Fatty Acid Accumulation in Oleaginous Fungus <i>Mucor circinelloides</i> WJ11 |
| title_full_unstemmed | Homologous Overexpression of Acyl-CoA Thioesterase 8 Enhanced Free Fatty Acid Accumulation in Oleaginous Fungus <i>Mucor circinelloides</i> WJ11 |
| title_short | Homologous Overexpression of Acyl-CoA Thioesterase 8 Enhanced Free Fatty Acid Accumulation in Oleaginous Fungus <i>Mucor circinelloides</i> WJ11 |
| title_sort | homologous overexpression of acyl coa thioesterase 8 enhanced free fatty acid accumulation in oleaginous fungus i mucor circinelloides i wj11 |
| topic | Acyl-CoA thioesterase 8 free fatty acid homologous overexpression <i>Mucor circinelloides</i> total fatty acid |
| url | https://www.mdpi.com/2311-5637/9/6/545 |
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