Identification and Functional Characterization of Adenosine Deaminase in <i>Mucor circinelloides</i>: A Novel Potential Regulator of Nitrogen Utilization and Lipid Biosynthesis
Adenosine deaminase (ADA) is an enzyme distributed in a wide variety of organisms that cleaves adenosine into inosine. Since inosine plays an important role in nitrogen metabolism, ADA may have a critical function in the regulation of fatty acid synthesis. However, the role of ADA in oleaginous fung...
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MDPI AG
2022-07-01
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| Series: | Journal of Fungi |
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| Online Access: | https://www.mdpi.com/2309-608X/8/8/774 |
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| author | Shaoqi Li Junhuan Yang Hassan Mohamed Xiuwen Wang Shuxian Pang Chen Wu Sergio López-García Yuanda Song |
| author_facet | Shaoqi Li Junhuan Yang Hassan Mohamed Xiuwen Wang Shuxian Pang Chen Wu Sergio López-García Yuanda Song |
| author_sort | Shaoqi Li |
| collection | DOAJ |
| description | Adenosine deaminase (ADA) is an enzyme distributed in a wide variety of organisms that cleaves adenosine into inosine. Since inosine plays an important role in nitrogen metabolism, ADA may have a critical function in the regulation of fatty acid synthesis. However, the role of ADA in oleaginous fungi has not been reported so far. Therefore, in this study, we identified one <i>ada</i> gene encoding ADA (with ID scaffold0027.9) in the high lipid-producing fungus, <i>Mucor circinelloides</i> WJ11, and investigated its role in cell growth, lipid production, and nitrogen metabolism by overexpressing and knockout of this gene. The results showed that knockout of the <i>ada</i> altered the efficiency of nitrogen consumption, which led to a 20% increment in the lipid content (25% of cell dry weight) of the engineered strain, while overexpression of the <i>ada</i> showed no significant differences compared with the control strain at the final growth stage; however, interestingly, it increased lipid accumulation at the early growth stage. Additionally, transcriptional analysis was conducted by RT-qPCR and our findings indicated that the deletion of <i>ada</i> activated the committed steps of lipid biosynthesis involved in acetyl-CoA carboxylase (<i>acc1</i> gene), cytosolic malic acid enzyme (<i>cme1</i> gene), and fatty acid synthases (<i>fas1</i> gene), while it suppressed the expression of AMP-activated protein kinase (<i>ampk α1</i> and <i>ampk β</i> genes), which plays a role in lipolysis, whereas the <i>ada</i>-overexpressed strain displayed reverse trends. Conclusively, this work unraveled a novel role of ADA in governing lipid biosynthesis and nitrogen metabolism in the oleaginous fungus, <i>M. circinelloides</i>. |
| first_indexed | 2024-03-09T09:53:36Z |
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| language | English |
| last_indexed | 2024-03-09T09:53:36Z |
| publishDate | 2022-07-01 |
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| series | Journal of Fungi |
| spelling | doaj.art-29c61d1d6b9749588964dcc0e9e873822023-12-01T23:52:08ZengMDPI AGJournal of Fungi2309-608X2022-07-018877410.3390/jof8080774Identification and Functional Characterization of Adenosine Deaminase in <i>Mucor circinelloides</i>: A Novel Potential Regulator of Nitrogen Utilization and Lipid BiosynthesisShaoqi Li0Junhuan Yang1Hassan Mohamed2Xiuwen Wang3Shuxian Pang4Chen Wu5Sergio López-García6Yuanda Song7Colin Ratledge Center for Microbial Lipids, School of Agricultural Engineering and Food Science, Shandong University of Technology, Zibo 255000, ChinaDepartment of Food Sciences, College of Food Science and Engineering, Lingnan Normal University, Zhanjiang 524048, ChinaColin Ratledge Center for Microbial Lipids, School of Agricultural Engineering and Food Science, Shandong University of Technology, Zibo 255000, ChinaColin Ratledge Center for Microbial Lipids, School of Agricultural Engineering and Food Science, Shandong University of Technology, Zibo 255000, ChinaColin Ratledge Center for Microbial Lipids, School of Agricultural Engineering and Food Science, Shandong University of Technology, Zibo 255000, ChinaColin Ratledge Center for Microbial Lipids, School of Agricultural Engineering and Food Science, Shandong University of Technology, Zibo 255000, ChinaDepartment of Genetics and Microbiology (Associated Unit to IQFR-CSIC), Faculty of Biology, University of Murcia, 3100 Murcia, SpainColin Ratledge Center for Microbial Lipids, School of Agricultural Engineering and Food Science, Shandong University of Technology, Zibo 255000, ChinaAdenosine deaminase (ADA) is an enzyme distributed in a wide variety of organisms that cleaves adenosine into inosine. Since inosine plays an important role in nitrogen metabolism, ADA may have a critical function in the regulation of fatty acid synthesis. However, the role of ADA in oleaginous fungi has not been reported so far. Therefore, in this study, we identified one <i>ada</i> gene encoding ADA (with ID scaffold0027.9) in the high lipid-producing fungus, <i>Mucor circinelloides</i> WJ11, and investigated its role in cell growth, lipid production, and nitrogen metabolism by overexpressing and knockout of this gene. The results showed that knockout of the <i>ada</i> altered the efficiency of nitrogen consumption, which led to a 20% increment in the lipid content (25% of cell dry weight) of the engineered strain, while overexpression of the <i>ada</i> showed no significant differences compared with the control strain at the final growth stage; however, interestingly, it increased lipid accumulation at the early growth stage. Additionally, transcriptional analysis was conducted by RT-qPCR and our findings indicated that the deletion of <i>ada</i> activated the committed steps of lipid biosynthesis involved in acetyl-CoA carboxylase (<i>acc1</i> gene), cytosolic malic acid enzyme (<i>cme1</i> gene), and fatty acid synthases (<i>fas1</i> gene), while it suppressed the expression of AMP-activated protein kinase (<i>ampk α1</i> and <i>ampk β</i> genes), which plays a role in lipolysis, whereas the <i>ada</i>-overexpressed strain displayed reverse trends. Conclusively, this work unraveled a novel role of ADA in governing lipid biosynthesis and nitrogen metabolism in the oleaginous fungus, <i>M. circinelloides</i>.https://www.mdpi.com/2309-608X/8/8/774adenosine deaminase<i>Mucor circinelloides</i>lipid accumulationnitrogen metabolism |
| spellingShingle | Shaoqi Li Junhuan Yang Hassan Mohamed Xiuwen Wang Shuxian Pang Chen Wu Sergio López-García Yuanda Song Identification and Functional Characterization of Adenosine Deaminase in <i>Mucor circinelloides</i>: A Novel Potential Regulator of Nitrogen Utilization and Lipid Biosynthesis Journal of Fungi adenosine deaminase <i>Mucor circinelloides</i> lipid accumulation nitrogen metabolism |
| title | Identification and Functional Characterization of Adenosine Deaminase in <i>Mucor circinelloides</i>: A Novel Potential Regulator of Nitrogen Utilization and Lipid Biosynthesis |
| title_full | Identification and Functional Characterization of Adenosine Deaminase in <i>Mucor circinelloides</i>: A Novel Potential Regulator of Nitrogen Utilization and Lipid Biosynthesis |
| title_fullStr | Identification and Functional Characterization of Adenosine Deaminase in <i>Mucor circinelloides</i>: A Novel Potential Regulator of Nitrogen Utilization and Lipid Biosynthesis |
| title_full_unstemmed | Identification and Functional Characterization of Adenosine Deaminase in <i>Mucor circinelloides</i>: A Novel Potential Regulator of Nitrogen Utilization and Lipid Biosynthesis |
| title_short | Identification and Functional Characterization of Adenosine Deaminase in <i>Mucor circinelloides</i>: A Novel Potential Regulator of Nitrogen Utilization and Lipid Biosynthesis |
| title_sort | identification and functional characterization of adenosine deaminase in i mucor circinelloides i a novel potential regulator of nitrogen utilization and lipid biosynthesis |
| topic | adenosine deaminase <i>Mucor circinelloides</i> lipid accumulation nitrogen metabolism |
| url | https://www.mdpi.com/2309-608X/8/8/774 |
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