Enhancement of fatty acid biosynthesis by exogenous acetyl-CoA carboxylase and pantothenate kinase in Escherichia coli

Satoh, Shusaku; Ozaki, Miho; Matsumoto, Saki; Nabatame, Takumi; Kaku, Moena; Shudo, Takashi; Asayama, Munehiko; Chohnan, Shigeru

doi:10.1007/s10529-020-02996-w

Cited by 27 publications

(13 citation statements)

References 36 publications

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“…Acetyl‐CoA is a very important intermediate metabolite, which can connect the metabolic processes of carbohydrates, amino acids, and fats. The up‐regulation of Acetyl‐CoA‐related genes in the treatment group promotes the de novo synthesis of fatty acids in the cell membrane of the bacterial cell and extends the carbon chain, thereby promoting the production of unsaturated fatty acids (Satoh et al, 2020). This allows the fluidity and integrity of the bacterial cell membrane to be better maintained, and effectively reduces the damage to the bacterial cell membrane caused by freeze–drying.…”

Section: Resultsmentioning

confidence: 99%

Improving the viability of powdered Lactobacillus fermentum Lf01 with complex lyoprotectants by maintaining cell membrane integrity and regulating related genes

Cheng

et al. 2022

Journal of Food Biochemistry

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In this study, Lactobacillus fermentum Lf01, which was screened out in the early stage of the experiment, had better fermentation performance as the research objectives, and was prepared into powder by vacuum freeze–drying technology. We used response surface methodology to optimize the composition of the mixture used to protect powdered L. fermentum. Our data demonstrated that 10% skim milk, 12% sucrose, 0.767% tyrosine, and 2.033% sorbitol ensured the highest survival rate (92.7%) of L. fermentum. We have initially explored the potential mechanism of the complex protectants through the protection effect under the electron microscope, and the analysis methods of Fourier transform infrared spectroscopy and transcriptomics. The complex protectants could effectively maintain the permeability barrier and structural integrity of cell membrane and avoid the leakage of cell contents. Transcriptomic data have also indicated that the protective effect of the complex protectants on bacteria during freeze–drying was most likely achieved through the regulation of related genes. We identified 240 differential genes in the treatment group, including 231 up‐regulated genes and 9 down‐regulated genes. Gene ontology (GO) and Kyoto encyclopaedia of genes and genomes (KEGG) analyses of differential expression genes (DEGs) indicated that genes involved in amino acid metabolism, carbohydrate metabolism, membrane transport, fatty acid biosynthesis and cell growth were significantly up‐regulated. These new results provided novel insights into the potential mechanism of lyoprotectants at the cellular level, morphological level, and gene level of the bacteria. Practical applications In our study, a strain of Lactobacillus fermentum Lf01 with good fermentation performance was selected to be prepared into powder by freeze–drying technique. Bacterial cells were unavoidably damaged during the freeze–drying process. As a result, we investigated the protective effects on L. fermentum of ten distinct freeze–dried protectants and their mixtures. We were also attempting to explain the mechanism of action of the complex protectants at the cellular level, morphological level, and gene level of the bacteria. This presents very important theoretical and practical significance for the preservation of strains and the production of commercial direct‐investment starter.

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Section: Resultsmentioning

confidence: 99%

Improving the viability of powdered Lactobacillus fermentum Lf01 with complex lyoprotectants by maintaining cell membrane integrity and regulating related genes

Cheng

et al. 2022

Journal of Food Biochemistry

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“…Previous studies have shown that the expression of acetyl-CoA carboxylase (encoded by accA, accB, accC and accD ) and thioesterase I (encoded by tesA ) in Escherichia coli can speed up the FA synthesis by six times. This suggests that the catalytic reaction of acetyl-CoA carbohydrase is a rate-limiting step for FA synthesis ( 150 – 153 ). Overexpression of both heterologous Δ-15 desaturase (Δ- 15D ) sourced from flax and endogenous genes ( SCD, ACC1, DGA1 and Δ- 12D ) along with the deletion of endogenous MFE1 and PEX10 can yield a superior lipid producer ( 140 ).…”

Section: Metabolic Engineering Of Oleaginous Fungimentioning

confidence: 99%

Production, Biosynthesis, and Commercial Applications of Fatty Acids From Oleaginous Fungi

Zhang

Huang

et al. 2022

Front. Nutr.

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Oleaginous fungi (including fungus-like protists) are attractive in lipid production due to their short growth cycle, large biomass and high yield of lipids. Some typical oleaginous fungi including Galactomyces geotrichum, Thraustochytrids, Mortierella isabellina, and Mucor circinelloides, have been well studied for the ability to accumulate fatty acids with commercial application. Here, we review recent progress toward fermentation, extraction, of fungal fatty acids. To reduce cost of the fatty acids, fatty acid productions from raw materials were also summarized. Then, the synthesis mechanism of fatty acids was introduced. We also review recent studies of the metabolic engineering strategies have been developed as efficient tools in oleaginous fungi to overcome the biochemical limit and to improve production efficiency of the special fatty acids. It also can be predictable that metabolic engineering can further enhance biosynthesis of fatty acids and change the storage mode of fatty acids.

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“…A cell engineered for fatty acid overproduction, accumulates overproduced fatty acids in the membrane, or excretes it across the membrane in the form of FFA [13,31]. Depending upon the pH of the medium, these fatty acids either exist in its acidic form (protonated) or in the form of its conjugate base (deprotonated) [32].…”

Section: Stresses Generated Due To Fatty Acid Overproduction Ph and Osmotic Stressmentioning

confidence: 99%

“…The basal ability of E. coli to produce fatty acids can be further increased by overexpressing/deleting few of the key genes or rate-limiting enzymes of fatty acid biosynthesis pathway [10][11][12][13]. Acetyl-CoA Carboxylase (ACC), a ratelimiting enzyme of fatty acid synthesis pathway, plays an important role in catalysing carboxylation of acetyl-CoA to form malonyl-CoA.…”

Section: Introductionmentioning

confidence: 99%

“…The conversion of acetyl-CoA to malonyl-CoA is the committing step towards fatty acid synthesis. Hence, fatty acid titres can be increased by overexpressing acc gene along with other genetic alterations [13]. According to their results, a fatty acid producing strain yielded six times more fatty acids (691 mg/L) than the control strain (100 mg/L) by overexpressing an exogenous acc, pantothenate kinase (coaA) and fasA to supplement high levels of malonyl-CoA, coenzyme-A and enhancement of fatty acid pathway, respectively.…”

Section: Introductionmentioning

confidence: 99%

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Overview of the Cellular Stress Responses Involved in Fatty Acid Overproduction in E. coli

2021

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Research on microbial fatty acid metabolism started in the late 1960s, and till date, various developments have aided in elucidating the fatty acid metabolism in great depth. Over the years, synthesis of microbial fatty acid has drawn industrial attention due to its diverse applications. However, fatty acid overproduction imparts various stresses on its metabolic pathways causing a bottleneck to further increase the fatty acid yields. Numerous strategies to increase fatty acid titres in Escherichia coli by pathway modulation have already been published, but the stress generated during fatty acid overproduction is relatively less studied. Stresses like pH, osmolarity and oxidative stress, not only lower fatty acid titres, but also alter the cell membrane composition, protein expression and membrane fluidity. This review discusses an overview of fatty acid synthesis pathway and presents a panoramic view of various stresses caused due to fatty acid overproduction in E. coli. It also addresses how certain stresses like high temperature and nitrogen limitation can boost fatty acid production. This review paper also highlights the interconnections that exist between these stresses.

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Enhancement of fatty acid biosynthesis by exogenous acetyl-CoA carboxylase and pantothenate kinase in Escherichia coli

Cited by 27 publications

References 36 publications

Improving the viability of powdered Lactobacillus fermentum Lf01 with complex lyoprotectants by maintaining cell membrane integrity and regulating related genes

Improving the viability of powdered Lactobacillus fermentum Lf01 with complex lyoprotectants by maintaining cell membrane integrity and regulating related genes

Production, Biosynthesis, and Commercial Applications of Fatty Acids From Oleaginous Fungi

Overview of the Cellular Stress Responses Involved in Fatty Acid Overproduction in E. coli

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