A self-generated biosolvent from cellulosic biorefining is used to wash/extract and optionally esterify free fatty acids from microbial oil. Free fatty acids or fatty acid esters and a hydrophobic biohydrocarbon are processed to provide a hybrid liquid fuel intermediary whereas the lipid oil is derived from cellulosic sugars and from microbial conversion of cellulose, hemicellulose, and/cellulosic sugar to lipids. In addition, higher utilization of woody biomass for the production of cellulosic sugars can be further converted to lipid oils, and the remaining hemicellulose/lignin fraction is converted to a novel hydrophobic biohydrocarbon and self-generated biosolvent in which both the processed lipid oils and biohydrocarbon can be reacted together to form a new fuel intermediary. The self-generated biosolvent comprising butanol, butanol esters, and additional biochemicals, can penetrate a microbial cell wall structure to provide higher extraction yields. The process integrates supercritical esterification of the free fatty acids using the self-generated biosolvent blend. Esterification of free fatty acids from microbial oil can be blended with a liquid biohydrocarbon to create a fuel intermediary. A phase separation process for separation of aqueous and hydrophilic materials from hydrophobic liquids is provided, wherein hydrophobic materials are the biosolvent and fatty acid esters.

The present invention relates to a method for preparing a yeast protein concentrate, said method comprising the lysis of yeast cells in a suspension that was adjusted to a particular pH prior to lysis, subsequently subjecting the soluble fraction obtained from lysis to filtration to reduce the content of molecules smaller than 30 kDa, and optionally drying the solution obtained from filtration. The present invention further relates to a yeast protein concentrate obtainable by the method of the invention. The yeast protein concentrate comprises a high amount of proteins which are still folded and are therefore capable of aggregation to form a solid protein matrix upon heating. In addition, the yeast protein concentrate of the invention will be of unobtrusive taste and is therefore particularly suited for use in the preparation of food items, such as meat substitute products.

The invention relates to a method for producing a microbial cell extract with improved functional properties. The invention further relates to a microbial cell extract obtained by or obtainable by said method. The invention further relates to the use of said microbial cell extract with improved functionality, with applications in gelation agents, thickening agents, foaming agents, emulsification agents, texturing agents and other suitable applications.

The present invention relates to a method for preparing a yeast protein concentrate, said method comprising the lysis of yeast cells in a suspension that was adjusted to a particular pH prior to lysis, subsequently subjecting the soluble fraction obtained from lysis to filtration to reduce the content of molecules smaller than 30 kDa, and optionally drying the solution obtained from filtration. The present invention further relates to a yeast protein concentrate obtainable by the method of the invention. The yeast protein concentrate comprises a high amount of proteins which are still folded and are therefore capable of aggregation to form a solid protein matrix upon heating. In addition, the yeast protein concentrate of the invention will be of unobtrusive taste and is therefore particularly suited for use in the preparation of food items, such as meat substitute products.

The invention relates to a method of cell lysis for forming at least one pore in the cell wall and/or cell membrane of a cell in an aqueous medium for release of at least one intracellular target protein and/or lipid from the cell into the aqueous medium, including subjecting the cell with the intracellular target protein and/or lipid in the aqueous medium to an electrohydraulic comminution; and separating the target protein and/or lipid from the cell. The electrohydraulic comminution is brought about by a pulse current source. The cell in the aqueous medium is subjected to electrical impulse discharges between at least two electrodes of an underwater radio path of a container at a specific operating voltage, impulse energy, and pulse frequency, and at least one pore is formed in the cell wall and/or cell membrane.

The disclosure relates to novel methods, compositions, and genetically modified microorganisms for extracting and/or isolating bioproducts from microorganisms having recalcitrant cell walls. In some aspects, the disclosure relates to solvent-free methods of extracting and/or isolating bioproducts. The disclosure further relates to bioproducts having less than 10 ppm of a solvent.

Disclosed are methods for isolating an analyte from a sample. In some aspects, the methods are for selectively isolating a microbial cell analyte, such as a nucleic acid, from a sample containing or suspected of containing mammalian cells. The selective isolation method includes selective lysis of the mammalian cells and filtration of the resulting lysate through a filter that retains intact microbial cells, followed by on-filter lysis of the retained microbial cells to release the microbial cell analyte. The released analyte is then eluted from the filter. In other aspects, the methods utilize on-filter lysis of a sample containing intact cells (e.g., microbial cells) to release the analytes, followed by elution of the released analytes from the filter by passing an immiscible liquid through the filter. The isolated analytes may be analyzed using a suitable assay depending on the type of analyte molecule. Also disclosed are fluidic systems and lysis solutions that may be used for isolating an analyte according to the disclosed methods.

Disclosed are methods for isolating an analyte from a sample. In some aspects, the methods are for selectively isolating a microbial cell analyte, such as a nucleic acid, from a sample containing or suspected of containing mammalian cells. The selective isolation method includes selective lysis of the mammalian cells and filtration of the resulting lysate through a filter that retains intact microbial cells, followed by on-filter lysis of the retained microbial cells to release the microbial cell analyte. The released analyte is then eluted from the filter. In other aspects, the methods utilize on-filter lysis of a sample containing intact cells (e.g., microbial cells) to release the analytes, followed by elution of the released analytes from the filter by passing an immiscible liquid through the filter. The isolated analytes may be analyzed using a suitable assay depending on the type of analyte molecule. Also disclosed are fluidic systems and lysis solutions that may be used for isolating an analyte according to the disclosed methods.

The present invention relates to a method for producing microbial lipids.

Disclosed are methods for isolating an analyte from a sample. In some aspects, the methods are for selectively isolating a microbial cell analyte, such as a nucleic acid, from a sample containing or suspected of containing mammalian cells. The selective isolation method includes selective lysis of the mammalian cells and filtration of the resulting lysate through a filter that retains intact microbial cells, followed by on-filter lysis of the retained microbial cells to release the microbial cell analyte. The released analyte is then eluted from the filter. In other aspects, the methods utilize on-filter lysis of a sample containing intact cells (e.g., microbial cells) to release the analytes, followed by elution of the released analytes from the filter by passing an immiscible liquid through the filter. The isolated analytes may be analyzed using a suitable assay depending on the type of analyte molecule. Also disclosed are fluidic systems and lysis solutions that may be used for isolating an analyte according to the disclosed methods.