Increased Accumulation of Medium-Chain Fatty Acids by Dynamic Degradation of Long-Chain Fatty Acids in <i>Mucor circinelloides</i>
Concerns about global warming, fossil-fuel depletion, food security, and human health have promoted metabolic engineers to develop tools/strategies to overproduce microbial functional oils directly from renewable resources. Medium-chain fatty acids (MCFAs, C8–C12) have been shown to be important sou...
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MDPI AG
2020-08-01
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Online Access: | https://www.mdpi.com/2073-4425/11/8/890 |
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doaj-635bf2af051042feb39cca1667ed06e9 |
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record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Syed Ammar Hussain Alexis Garcia Md. Ahsanul Kabir Khan Shaista Nosheen Yao Zhang Mattheos A. G. Koffas Victoriano Garre Soo Chan Lee Yuanda Song |
spellingShingle |
Syed Ammar Hussain Alexis Garcia Md. Ahsanul Kabir Khan Shaista Nosheen Yao Zhang Mattheos A. G. Koffas Victoriano Garre Soo Chan Lee Yuanda Song Increased Accumulation of Medium-Chain Fatty Acids by Dynamic Degradation of Long-Chain Fatty Acids in <i>Mucor circinelloides</i> Genes <i>Mucor circinelloides</i> metabolic engineering acyl-ACP-Thioesterase acyl-CoA-oxidase acyl-CoA-thioesterase β-oxidation pathway |
author_facet |
Syed Ammar Hussain Alexis Garcia Md. Ahsanul Kabir Khan Shaista Nosheen Yao Zhang Mattheos A. G. Koffas Victoriano Garre Soo Chan Lee Yuanda Song |
author_sort |
Syed Ammar Hussain |
title |
Increased Accumulation of Medium-Chain Fatty Acids by Dynamic Degradation of Long-Chain Fatty Acids in <i>Mucor circinelloides</i> |
title_short |
Increased Accumulation of Medium-Chain Fatty Acids by Dynamic Degradation of Long-Chain Fatty Acids in <i>Mucor circinelloides</i> |
title_full |
Increased Accumulation of Medium-Chain Fatty Acids by Dynamic Degradation of Long-Chain Fatty Acids in <i>Mucor circinelloides</i> |
title_fullStr |
Increased Accumulation of Medium-Chain Fatty Acids by Dynamic Degradation of Long-Chain Fatty Acids in <i>Mucor circinelloides</i> |
title_full_unstemmed |
Increased Accumulation of Medium-Chain Fatty Acids by Dynamic Degradation of Long-Chain Fatty Acids in <i>Mucor circinelloides</i> |
title_sort |
increased accumulation of medium-chain fatty acids by dynamic degradation of long-chain fatty acids in <i>mucor circinelloides</i> |
publisher |
MDPI AG |
series |
Genes |
issn |
2073-4425 |
publishDate |
2020-08-01 |
description |
Concerns about global warming, fossil-fuel depletion, food security, and human health have promoted metabolic engineers to develop tools/strategies to overproduce microbial functional oils directly from renewable resources. Medium-chain fatty acids (MCFAs, C8–C12) have been shown to be important sources due to their diverse biotechnological importance, providing benefits ranging from functional lipids to uses in bio-fuel production. However, oleaginous microbes do not carry native pathways for the production of MCFAs, and therefore, diverse approaches have been adapted to compensate for the requirements of industrial demand. <i>Mucor circinelloides</i> is a promising organism for lipid production (15–36% cell dry weight; CDW) and the investigation of mechanisms of lipid accumulation; however, it mostly produces long-chain fatty acids (LCFAs). To address this challenge, we genetically modified strain <i>M. circinelloides</i> MU758, first by integrating heterologous acyl-ACP thioesterase (<i>TE</i>) into fatty acid synthase (FAS) complex and subsequently by modifying the β-oxidation pathway by disrupting the acyl-CoA oxidase (<i>ACOX</i>) and/or acyl-CoA thioesterase (<i>ACOT</i>) genes with a preference for medium-chain acyl-CoAs, to elevate the yield of MCFAs. The resultant mutant strains (M-1, M-2, and M-3, respectively) showed a significant increase in lipid production in comparison to the wild-type strain (WT). MCFAs in M-1 (47.45%) was sharply increased compared to the wild type strain (2.25%), and it was further increased in M-2 (60.09%) suggesting a negative role of <i>ACOX</i> in MCFAs production. However, MCFAs in M-3 were much decreased compared to M-1,suggesting a positive role of <i>ACOT</i> in MCFAs production. The M-2 strain showed maximum lipid productivity (~1800 milligram per liter per day or mg/L.d) and MCFAs productivity (~1100 mg/L.d). Taken together, this study elaborates on how the combination of two multidimensional approaches, <i>TE</i> gene over-expression and modification of the β-oxidation pathway via substantial knockout of specific <i>ACOX</i> gene, significantly increased the production of MCFAs. This synergistic approach ultimately offers a novel opportunity for synthetic/industrial biologists to increase the content of MCFAs. |
topic |
<i>Mucor circinelloides</i> metabolic engineering acyl-ACP-Thioesterase acyl-CoA-oxidase acyl-CoA-thioesterase β-oxidation pathway |
url |
https://www.mdpi.com/2073-4425/11/8/890 |
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doaj-635bf2af051042feb39cca1667ed06e92020-11-25T01:58:45ZengMDPI AGGenes2073-44252020-08-011189089010.3390/genes11080890Increased Accumulation of Medium-Chain Fatty Acids by Dynamic Degradation of Long-Chain Fatty Acids in <i>Mucor circinelloides</i>Syed Ammar Hussain0Alexis Garcia1Md. Ahsanul Kabir Khan2Shaista Nosheen3Yao Zhang4Mattheos A. G. Koffas5Victoriano Garre6Soo Chan Lee7Yuanda Song8Colin Ratledge Center for Microbial Lipids, School of Agriculture Engineering and Food Science, Shandong University of Technology, Zibo 255049, ChinaDepartment of Biology, South Texas Center of Emerging Infectious Diseases (STCEID), University of Texas, San Antonio, TX 78249, USAColin Ratledge Center for Microbial Lipids, School of Agriculture Engineering and Food Science, Shandong University of Technology, Zibo 255049, ChinaColin Ratledge Center for Microbial Lipids, School of Agriculture Engineering and Food Science, Shandong University of Technology, Zibo 255049, ChinaColin Ratledge Center for Microbial Lipids, School of Agriculture Engineering and Food Science, Shandong University of Technology, Zibo 255049, ChinaDepartment of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USADepartamento de Genética y Microbiología (Unidadasociada al IQFR-CSIC), Facultad de Biología, Universidad de Murcia, 30071 Murcia, SpainDepartment of Biology, South Texas Center of Emerging Infectious Diseases (STCEID), University of Texas, San Antonio, TX 78249, USAColin Ratledge Center for Microbial Lipids, School of Agriculture Engineering and Food Science, Shandong University of Technology, Zibo 255049, ChinaConcerns about global warming, fossil-fuel depletion, food security, and human health have promoted metabolic engineers to develop tools/strategies to overproduce microbial functional oils directly from renewable resources. Medium-chain fatty acids (MCFAs, C8–C12) have been shown to be important sources due to their diverse biotechnological importance, providing benefits ranging from functional lipids to uses in bio-fuel production. However, oleaginous microbes do not carry native pathways for the production of MCFAs, and therefore, diverse approaches have been adapted to compensate for the requirements of industrial demand. <i>Mucor circinelloides</i> is a promising organism for lipid production (15–36% cell dry weight; CDW) and the investigation of mechanisms of lipid accumulation; however, it mostly produces long-chain fatty acids (LCFAs). To address this challenge, we genetically modified strain <i>M. circinelloides</i> MU758, first by integrating heterologous acyl-ACP thioesterase (<i>TE</i>) into fatty acid synthase (FAS) complex and subsequently by modifying the β-oxidation pathway by disrupting the acyl-CoA oxidase (<i>ACOX</i>) and/or acyl-CoA thioesterase (<i>ACOT</i>) genes with a preference for medium-chain acyl-CoAs, to elevate the yield of MCFAs. The resultant mutant strains (M-1, M-2, and M-3, respectively) showed a significant increase in lipid production in comparison to the wild-type strain (WT). MCFAs in M-1 (47.45%) was sharply increased compared to the wild type strain (2.25%), and it was further increased in M-2 (60.09%) suggesting a negative role of <i>ACOX</i> in MCFAs production. However, MCFAs in M-3 were much decreased compared to M-1,suggesting a positive role of <i>ACOT</i> in MCFAs production. The M-2 strain showed maximum lipid productivity (~1800 milligram per liter per day or mg/L.d) and MCFAs productivity (~1100 mg/L.d). Taken together, this study elaborates on how the combination of two multidimensional approaches, <i>TE</i> gene over-expression and modification of the β-oxidation pathway via substantial knockout of specific <i>ACOX</i> gene, significantly increased the production of MCFAs. This synergistic approach ultimately offers a novel opportunity for synthetic/industrial biologists to increase the content of MCFAs.https://www.mdpi.com/2073-4425/11/8/890<i>Mucor circinelloides</i>metabolic engineeringacyl-ACP-Thioesteraseacyl-CoA-oxidaseacyl-CoA-thioesteraseβ-oxidation pathway |