Engineering lipid overproduction in the oleaginous yeast Yarrowia lipolytica. 2015

Kangjian Qiao, and Syed Hussain Imam Abidi, and Hongjuan Liu, and Haoran Zhang, and Sagar Chakraborty, and Nicki Watson, and Parayil Kumaran Ajikumar, and Gregory Stephanopoulos
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, United States.

Conversion of carbohydrates to lipids at high yield and productivity is essential for cost-effective production of renewable biodiesel. Although some microorganisms can convert sugars to oils, conversion yields and rates are typically low due primarily to allosteric inhibition of the lipid biosynthetic pathway by saturated fatty acids. By reverse engineering the mammalian cellular obese phenotypes, we identified the delta-9 stearoyl-CoA desaturase (SCD) as a rate limiting step and target for the metabolic engineering of the lipid synthesis pathway in Yarrowia lipolytica. Simultaneous overexpression of SCD, Acetyl-CoA carboxylase (ACC1), and Diacylglyceride acyl-transferase (DGA1) in Y. lipolytica yielded an engineered strain exhibiting highly desirable phenotypes of fast cell growth and lipid overproduction including high carbon to lipid conversion yield (84.7% of theoretical maximal yield), high lipid titers (~55g/L), enhanced tolerance to glucose and cellulose-derived sugars. Moreover, the engineered strain featured a three-fold growth advantage over the wild type strain. As a result, a maximal lipid productivity of ~1g/L/h is obtained during the stationary phase. Furthermore, we showed that the engineered yeast required cytoskeleton remodeling in eliciting the obesity phenotype. Altogether, our work describes the development of a microbial catalyst with the highest reported lipid yield, titer and productivity to date. This is an important step towards the development of an efficient and cost-effective process for biodiesel production from renewable resources.

UI MeSH Term Description Entries
D008055 Lipids A generic term for fats and lipoids, the alcohol-ether-soluble constituents of protoplasm, which are insoluble in water. They comprise the fats, fatty oils, essential oils, waxes, phospholipids, glycolipids, sulfolipids, aminolipids, chromolipids (lipochromes), and fatty acids. (Grant & Hackh's Chemical Dictionary, 5th ed) Lipid
D005656 Fungal Proteins Proteins found in any species of fungus. Fungal Gene Products,Fungal Gene Proteins,Fungal Peptides,Gene Products, Fungal,Yeast Proteins,Gene Proteins, Fungal,Peptides, Fungal,Proteins, Fungal
D000103 Acetyl-CoA Carboxylase A carboxylating enzyme that catalyzes the conversion of ATP, acetyl-CoA, and HCO3- to ADP, orthophosphate, and malonyl-CoA. It is a biotinyl-protein that also catalyzes transcarboxylation. The plant enzyme also carboxylates propanoyl-CoA and butanoyl-CoA (From Enzyme Nomenclature, 1992) EC 6.4.1.2. Acetyl Coenzyme A Carboxylase,Acetyl CoA Carboxylase,Carboxylase, Acetyl-CoA
D013230 Stearoyl-CoA Desaturase An enzyme that catalyzes the formation of oleoyl-CoA, A, and water from stearoyl-CoA, AH2, and oxygen where AH2 is an unspecified hydrogen donor. Stearyl-CoA Desaturase,Stearate Desaturase,delta-9 Desaturase,Desaturase, Stearate,Desaturase, Stearoyl-CoA,Desaturase, Stearyl-CoA,Desaturase, delta-9,Stearoyl CoA Desaturase,Stearyl CoA Desaturase,delta 9 Desaturase
D044943 Fatty Acid Desaturases A family of enzymes that catalyze the stereoselective, regioselective, or chemoselective syn-dehydrogenation reactions. They function by a mechanism that is linked directly to reduction of molecular OXYGEN. Acyl CoA Desaturase,Enoyl CoA Reductase,Fatty Acid Desaturase,Fatty Acid Desaturating Enzymes,Acyl CoA Desaturases,Enoyl CoA Reductases,Acid Desaturase, Fatty,CoA Desaturase, Acyl,CoA Reductase, Enoyl,Desaturase, Acyl CoA,Desaturase, Fatty Acid,Desaturases, Fatty Acid,Reductase, Enoyl CoA,Reductases, Enoyl CoA
D051048 Diacylglycerol O-Acyltransferase An enzyme that catalyses the last step of the TRIACYLGLYCEROL synthesis reaction in which diacylglycerol is covalently joined to LONG-CHAIN ACYL COA to form triglyceride. It was formerly categorized as EC 2.3.1.124. Acyl-Coenzyme A Diacyl-glycerol Acyltransferase,DAGAT Enzyme,Diacylglycerol Acyltransferase,Diacylglycerol O-Acyltransferase 1,Diglyceride Acyltransferase,Diglyceride Acyltransferase 1,Palmitoyl-CoA-sn-1,2-Diacylglycerol Acyltransferase,Palmitoyl-Coenzyme A-sn-1,2-Diacylglycerol Acyltransferase,A-sn-1,2-Diacylglycerol Acyltransferase, Palmitoyl-Coenzyme,Acyl Coenzyme A Diacyl glycerol Acyltransferase,Acyltransferase 1, Diglyceride,Acyltransferase, Diacylglycerol,Acyltransferase, Diglyceride,Acyltransferase, Palmitoyl-CoA-sn-1,2-Diacylglycerol,Acyltransferase, Palmitoyl-Coenzyme A-sn-1,2-Diacylglycerol,Diacylglycerol O Acyltransferase,Diacylglycerol O Acyltransferase 1,O-Acyltransferase 1, Diacylglycerol,O-Acyltransferase, Diacylglycerol,Palmitoyl CoA sn 1,2 Diacylglycerol Acyltransferase,Palmitoyl Coenzyme A sn 1,2 Diacylglycerol Acyltransferase
D060847 Metabolic Engineering Methods and techniques used to genetically modify cells' biosynthetic product output and develop conditions for growing the cells as BIOREACTORS. Engineering, Metabolic
D025062 Yarrowia A genus of ascomycetous yeast in the family Dipodascaceae, order SACCHAROMYCETALES. Yarrowia lipolytica,Yarrowias

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