scholarly journals High production of triterpenoids in Yarrowia lipolytica through manipulation of lipid components

2020 ◽  
Author(s):  
Jin Lai Zhang ◽  
Qiu Yan Bai ◽  
Yang Zi Peng ◽  
Jie Fan ◽  
Cong Cong Jin ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Cell Morphology ◽  
Extracellular Space ◽  
Unsaturated Fatty Acids ◽  
Microbial Cell ◽  
High Yield ◽  
Membrane Phospholipids ◽  
Microbial Cell Factories ◽  
Cell Factories ◽  
Lipid Components

Abstract BackgroundLupeol exhibits novel physiological and pharmacological activities, such as anti-cancer and immunity enhancement. However, cytotoxicity is still a challenge for triterpenoids overproduction in microbial cell factories. As lipophilic and relatively small-molecular compounds, triterpenes are generally secreted to the extracellular space. The effect of increasing triterpenes efflux on the synthesis capacity remains unknown.ResultsIn this study, we developed a strategy to enhance the triterpenes efflux through manipulation of lipid components in Y. lipolytica by overexpressing the enzyme of Δ9-fatty acid desaturase (OLE1) and disturbing the phosphatidic acid phosphatase (PAH1) and diacylglycerol kinase (DGK1). As a result, we obtained a high-yield lupeol strain with the highest lupeol production of 411.72 mg/L in shake flasks reported to date, reaching a 33.2-fold improvement over the initial strain. The lipid manipulation led to a two-fold increase of unsaturated fatty acids (UFAs) content, up to 61%~73%, and an exceptionally elongated cell morphology, which might be caused by enhanced membrane phospholipids biosynthesis flux. Both of the phenotypes accelerated the export of toxic products to the extracellular space and ultimately stimulated the capacity of triterpenoids synthesis, which were proved by 5.11-fold higher ratio of extra-/intra-cellular lupeol concentrations, 2.7-fold higher biomass accumulation and 2.60-fold higher lupeol productivity per unit OD in modified strains. This strategy was also highly efficient for biosynthesis of other triterpenes and sesquiterpenes, including α-, β-arymin, longifolene, longipinene and longicyclene.ConclusionsTo conclude, we successfully created a high-yield lupeol strain via lipid manipulation. And we demonstrated that the enhancement of lupeol efflux and synthesis capacity was induced by the increased unsaturated fatty acids content and elongated cell morphology. Our study provides a novel strategy to promote the biosynthesis of valuable but toxic products in microbial cell factories.

scholarly journals High production of triterpenoids in Yarrowia lipolytica through manipulation of lipid components

2020 ◽  
Author(s):  
Jin Lai Zhang ◽  
Qiu Yan Bai ◽  
Yang Zi Peng ◽  
Jie Fan ◽  
Cong Cong Jin ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Cell Morphology ◽  
Extracellular Space ◽  
Microbial Cell ◽  
High Yield ◽  
Total Production ◽  
Two Phase ◽  
Microbial Cell Factories ◽  
Cell Factories ◽  
Lipid Components

Abstract Background Lupeol exhibits novel physiological and pharmacological activities, such as anticancer and immunity-enhancing activities. However, cytotoxicity remains a challenge for triterpenoid overproduction in microbial cell factories. As lipophilic and relatively small-molecular compounds, triterpenes are generally secreted into the extracellular space. The effect of increasing triterpene efflux on the synthesis capacity remains unknown.Results In this study, we developed a strategy to enhance triterpene efflux through manipulation of lipid components in Y. lipolytica by overexpressing the enzyme Δ9-fatty acid desaturase (OLE1) and disturbing phosphatidic acid phosphatase (PAH1) and diacylglycerol kinase (DGK1). By this strategy combined with two-phase fermentation, the highest lupeol production reported to date was achieved, where the titer in the organic phase reached 381.67 mg/L and the total production was 411.72 mg/L in shake flasks, exhibiting a 33.20-fold improvement over the initial strain. Lipid manipulation led to a two-fold increase in the unsaturated fatty acid (UFA) content, up to 61%~73%, and an exceptionally elongated cell morphology, which might have been caused by enhanced membrane phospholipid biosynthesis flux. Both phenotypes accelerated the export of toxic products to the extracellular space and ultimately stimulated the capacity for triterpenoid synthesis, which was proven by the 5.11-fold higher ratio of extra/intracellular lupeol concentrations, 2.79-fold higher biomass accumulation and 2.56-fold higher lupeol productivity per unit OD in the modified strains. This strategy was also highly efficient for the biosynthesis of other triterpenes and sesquiterpenes, including α-amyrin, β-amyrin, longifolene, longipinene and longicyclene.Conclusions In conclusion, we successfully created a high-yield lupeol-producing strain via lipid manipulation. We demonstrated that the enhancement of lupeol efflux and synthesis capacity was induced by the increased UFA content and elongated cell morphology. Our study provides a novel strategy to promote the biosynthesis of valuable but toxic products in microbial cell factories.

Developing fourth-generation biofuels secreting microbial cell factories for enhanced productivity and efficient product recovery; a review

Fuel ◽  
2021 ◽  
Vol 298 ◽  
pp. 120858
Author(s):  
Sana Malik ◽  
Ayesha Shahid ◽  
Chen-Guang Liu ◽  
Aqib Zafar Khan ◽  
Muhammad Zohaib Nawaz ◽  
...  
Keyword(s):  
Product Recovery ◽  
Microbial Cell ◽  
Fourth Generation ◽  
Microbial Cell Factories ◽  
Cell Factories

scholarly journals Establishing new microbial cell factories for sustainable bioprocesses

2012 ◽  
Vol 29 ◽  
pp. S75-S76
Author(s):  
Mhairi Workman ◽  
Philippe Holt ◽  
Xiaoying Liu
Keyword(s):  
Microbial Cell ◽  
Microbial Cell Factories ◽  
Cell Factories

scholarly journals Overexpression of Thermophilic α-amylase in Bacillus Licheniformis using a High Efficiency Chromosomal Integration and Amplification Strategy

2021 ◽  
Author(s):  
Peili Shen ◽  
Dandan Niu ◽  
Xuelian Liu ◽  
Kangming Tian ◽  
Permaul Kugenthiren ◽  
...  
Keyword(s):  
Bacillus Licheniformis ◽  
High Efficiency ◽  
High Yield ◽  
Chromosomal Integration ◽  
Bacillus Sp ◽  
Strain Development ◽  
Microbial Cell Factories ◽  
Cell Factories ◽  
New Strategy ◽  
Amplification Strategy

Abstract Highly efficient preparation of industrially important enzymes depends on development of the genetically stable and high-yield microbial cell factories, which is often a challengeable laboratory hard work. In aims to simplify strain development with high efficiency for enzyme overproduction, a new strategy based on chromosomal integration and amplification in Bacillus sp . was developed. A pair of plasmids, an integrated expression plasmid pUB'-Ex1 and a thermosensitive replicable plasmid pUB-MazF, were constructed. pUB'-Ex1 conditionally self-replicated in Bacillus sp . when the RepF in pUB-MazF expressed. pUB-MazF thermosensitively self-replicated in Bacillus sp . , which was easily cured from the host by inducing MazF expression with IPTG. Bacillus licheniformis BL-UBM that integrated with pUB-MazF was then transformed with pUB'-amyS derived from pUB'-Ex1 by in-frame cloning of amyS encoding a thermophilic α-amylase from Geobacillus stearothermophilus ATCC 31195. The transformant of B. licheniformis BL-UBM with pUB'-amyS was cultivated at 42 o C with the existence of 1 mmol/l IPTG and 500 μg/ml kanamycin and the recombinants with high α-amylase activities were selected. All tested recombinants were extremely high genetic stability. One of which, recombinant BLiS-002, carried five copies of amyS and produced the highest yield of α-amylase. It could yield 50,753 U/ml of α-amylase in a 50-l bioreactor. The strategy developed in this study is of application potential for convenient and quick strain development for industrially important enzyme overexpression.

scholarly journals Effect of atorvastatin and rosuvasta tin on the fatty acid spectrum of lymphocyte membranes in patients with unstable angina

2020 ◽  
pp. 92-95
Author(s):  
T.V. Bogdan ◽  
Keyword(s):  
Cardiovascular Disease ◽  
Fatty Acids ◽  
Fatty Acid ◽  
Unstable Angina ◽  
Unsaturated Fatty Acids ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Membrane Phospholipids ◽  
Stabilization Of Membranes ◽  
Dynamic Structures

Numerous studies have demonstrated the superiority of rosuvastatin over other statins in the treatment of cardiovascular disease. It has been proven that rosuvastatin is more effectively lowers low-density lipoprotein cholesterol in patients with cardiovascular disease than other members of this drug group. Despite the known mechanisms of action of statins on blood lipids, their effective use in patients with cardiovascular disease, as well as side effects, the influence of these drugs on the fatty acid spectrum of lymphocyte (LC) membrane phospholipids in patients with ischemic heart disease remains unexplored. The results of the studies cited in the article indicate that, in patients with unstable angina who received the therapy that included rosuvastatin, unlike patients receiving the basic treatment with atorvastatin, the relative phosphate lipid contents of palmitic, stearic, and stearin arachidonic polyunsaturated fatty acids and the amount of unsaturated fatty acids are normalized, which testifies to the stabilization of membranes as dynamic structures.

scholarly journals Redirection of lipid flux toward phospholipids in yeast increases fatty acid turnover and secretion

2018 ◽  
Vol 115 (6) ◽  
pp. 1262-1267 ◽  
Author(s):  
Raphael Ferreira ◽  
Paulo Gonçalves Teixeira ◽  
Verena Siewers ◽  
Jens Nielsen
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Lipid Metabolism ◽  
Metabolic Network ◽  
Feedback Regulation ◽  
Acid Activation ◽  
Phospholipid Hydrolysis ◽  
Microbial Cell Factories ◽  
Cell Factories ◽  
High Level

Bio-based production of fatty acids and fatty acid-derived products can enable sustainable substitution of petroleum-derived fuels and chemicals. However, developing new microbial cell factories for producing high levels of fatty acids requires extensive engineering of lipid metabolism, a complex and tightly regulated metabolic network. Here we generated a Saccharomyces cerevisiae platform strain with a simplified lipid metabolism network with high-level production of free fatty acids (FFAs) due to redirected fatty acid metabolism and reduced feedback regulation. Deletion of the main fatty acid activation genes (the first step in β-oxidation), main storage lipid formation genes, and phosphatidate phosphatase genes resulted in a constrained lipid metabolic network in which fatty acid flux was directed to a large extent toward phospholipids. This resulted in simultaneous increases of phospholipids by up to 2.8-fold and of FFAs by up to 40-fold compared with wild-type levels. Further deletion of phospholipase genes PLB1 and PLB2 resulted in a 46% decrease in FFA levels and 105% increase in phospholipid levels, suggesting that phospholipid hydrolysis plays an important role in FFA production when phospholipid levels are increased. The multiple deletion mutant generated allowed for a study of fatty acid dynamics in lipid metabolism and represents a platform strain with interesting properties that provide insight into the future development of lipid-related cell factories.

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