Solazyme, Inc.

Phycoil Biotechnology International. Inc.

Corbion Biotech, Inc.

Terravia Holdings, Inc.

Chung-Soon Im

Anna Coragliotti

Scott Franklin

Anthony G Day

Donald Eugene Trimbur

Harrison F Dillon

Rika Regentin

Jane Kim

Diana Vincent

Bioreactors and methods for cultivating microalgae under heterotrophic growth conditions using fruit wastewater as a carbon source are provided herein. The heterotrophic growth conditions include low irradiance of light sufficient to initiate light-activated metabolism, but not photosynthesis. The methods provide a cost effective and environmentally friendly solution for converting wastewater from dried fruit and wine industries into products like biofuels.

Utilization of wastewater for microalgal cultivation

The invention provides methods of manufacturing alkanes from triglyceride oils produced through fermentation of oil-bearing microbes. The processes provided herein can utilize a variety of carbohydrate feedstocks including cane bagasse, sugar beet pulp, corn stover, glycerol, corn starch, sorghum, molasses, waste glycerol, and other renewable materials. These processes further comprise hydrotreating, hydrocracking, isomerization, distillation, and other petrochemical processes for use with oil-bearing microbes and products derived therefrom to manufacture fuels. Particular embodiments include the manufacture of ASTM D975 and ASTM D1655 compliant fuels. Genetically engineered microbes provided herein can be used in the manufacture of renewable diesel and renewable jet fuel.

Renewable diesel and jet fuel from microbial sources

Bioreactors and methods for cultivating microalgae are provided herein. The bioreactor and methods include features and modifications to improve heterotrophic growth efficiency by providing a light signal.

Microalgae fermentation using controlled Illumination

The invention discloses novel methods of producing hydrocarbons through heterotrophic cultivation of. Also provided are novel hydrocarbon compositions. A preferred species for engineering is the microalgae species. Additional methods of cultivation include providing certain nutrient sources.

Heterotrophic cultivation of hydrocarbon-producing microalgae

The invention provides methods of cultivating oil-bearing microbes using xylose alone or in combination with other depolymerized cellulosic material. Also provided are microorganisms comprising an exogenous gene encoding a polysaccharide degrading enzyme, such as a cellulase, a hemicellulase, a pectinase, or a driselase. Some methods of microbial fermentation are provided that comprise the use of xylose and depolymerized cellulosic materials for the production of oil-bearing microorgansims.

Chlorella species containing exogenous genes

The invention provides methods of cultivating oil-bearing microbes using cellulosic material. Also provided are microorganisms containing one or more exogenous genes that facilitate the use of cellulosic materials as a feedstock. Also provided are microorganisms and methods for manufacturing non-alcohol-based fuels and fuel feedstocks through a process of converting cellulosic materials into oils.

Production of oil in microorganisms

The invention provides methods of cultivating oil-bearing microbes using cellulosic material. Also provided are microorganisms that manufacture non-alcohol-based fuels and fuel feedstocks through a process of converting cellulosic materials into oils. Also provided are compositions comprising depolymerized cellulosic materials and oil-bearing microbes. Some methods of microbial fermentation are provided that comprise combining depolymerized cellulosic materials with other non-cellulosic feedstocks to enhance the economics of renewable fuel manufacturing. Particular advantages of the processes provided herein include production of oils rather than alcohols through cellulosic processes. Additional advantages include methods for manufacturing high nutrition oils from non-edible feedstocks such as wood chips, switchgrass, and bagasse.

Use of cellulosic materials for cultivation of microorganisms

The invention provides methods of manufacturing biodiesel and other oil-based compounds using glycerol and combinations of glycerol and other feedstocks as an energy source in fermentation of oil-bearing microorganisms. Methods disclosed herein include processes for manufacturing high nutrition edible oils from non-food feedstock materials such as waste products from industrial waste transesterification processes. Also included are methods of increasing oil yields by temporally separating glycerol and other feedstocks during cultivation processes. Also provided herein are oil-bearing microbes containing exogenous oil production genes and methods of cultivating such microbes on glycerol and other feedstocks.

Glycerol feedstock utilization for oil-based fuel manufacturing

The invention provides methods of modifying the lipids produced by microbial organisms through genetic engineering. The invention also provides genetically engineered microbes and methods of fermenting microbes for oil production. Also provided are oils, fuels, oleochemicals, chemical precursors, and other compounds manufactured by such modified microorganisms. Exemplary oil-bearing organisms include organisms containing one or more exogenous genes encoding a fatty acyl-ACP thioesterase, fatty acyl-CoA/aldehyde reductase, fatty acyl-CoA reductase, fatty aldehyde reductase, fatty aldehyde decarbonylase, and/or an acyl carrier protein.

Lipid pathway modification in oil-bearing microorganisms

The invention provides methods of manufacturing oils and oil-based products such as transportation fuels, industrial chemicals, edible oils, lubricants and plastics using sucrose feedstocks from sugar cane, sugar beets, and molasses for bioproduction processes. The disclosed processes utilize oleaginous microbes as a conversion technology to convert chemical energy produced by sugar cane, molasses and sugar beets into energy-containing oils and oil derivatives. Also provided herein are oleaginous microbes containing one or more exogenous sucrose utilization genes.

The invention provides methods of manufacturing oils and oil-based products such as transportation fuels, industrial chemicals, lubricants and plastics using sugar cane, sugar cane juice, corn steep liquor, corn starch or depolymerized cellulosic material as a feedstock for bioproduction processes. Oils are manufactured using microalgae expressing sucrose invertase, which allows the microalgae to produce oil using sucrose-based feedstocks, along or in combination with other feedstocks.

Heterotrophic microalgae expressing invertase

The invention provides methods of manufacturing oils and oil-based products such as transportation fuels, industrial chemicals, edible oils, lubricants and plastics using sugar cane, sugar beets, and cane/beet agricultural processing byproducts as a feedstock for bioproduction processes. The disclosed processes utilize oil-bearing microbes as a conversion technology to convert chemical energy produced by sugar cane and sugar beets into energy-containing oils and oil derivatives. Also provided herein are oil-bearing microbes containing one or more exogenous sucrose utilization genes.

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Palo Alto, CA, United States of America

Chung-Soon Im