It has been found that esterification of a hydroxy-fatty acid by a lipase can be coupled with oleate hydratase (OHase) generation of that hydroxy-FA from an unsaturated FA with a cis C9-C10 double bond, e.g. oleic acid, in a single aqueous buffered reaction medium at low temperature, e.g. 30° C. A simple one-pot enzymatic method to produce fatty acid estolides from one or more triglycerides, e.g. starting from a natural plant oil, is thereby enabled in which the same lipase catalyses both the initial hydrolysis of triglyceride and the final esterification step.

It has been found that esterification of a hydroxy-fatty acid by a lipase can be coupled with oleate hydratase (OHase) generation of that hydroxy-FA from an unsaturated FA with a cis C9-C10 double bond, e.g. oleic acid, in a single aqueous buffered reaction medium at low temperature, e.g. 30° C. A simple one-pot enzymatic method to produce fatty acid estolides from one or more triglycerides, e.g. starting from a natural plant oil, is thereby enabled in which the same lipase catalyses both the initial hydrolysis of triglyceride and the final esterification step.

A method for producing cis-unsaturated fatty acid includes the operations below. (i) An oil-water mixture is provided, wherein the oil-water mixture includes 1 to 10 parts by weight of oil and 1 part by weight of water. (ii) 0.002 to 0.5 parts by weight of a recombinant Candida rugosa lipase 1 (rCRL1) is added into the oil-water mixture. (iii) The oil-water mixture is emulsified. (iv) The emulsified oil-water mixture is hydrolyzed and fatty acid is generated. (v) Oil-water is separated at a temperature of 55° C. to 65° C. and an oil phase layer is extracted. (vi) The cooling and filtering step is performed to obtain cis-unsaturated fatty acid.

The purpose of the present invention is to provide an oil/lipid that contains dihomo-γ-linolenic acid at a higher purity. Provided is a microbial oil/lipid that is specified by a high content of dihomo-γ-linolenic acid contained therein and/or a reduced content of undesirable constituting fatty acid(s).

It has been found that esterification of a hydroxy-fatty acid by a lipase can be coupled with oleate hydratase (OHase) generation of that hydroxy-FA from an unsaturated FA with a cis C9-C10 double bond, e.g. oleic acid, in a single aqueous buffered reaction medium at low temperature, e.g. 30° C. A simple one-pot enzymatic method to produce fatty acid estolides from one or more triglycerides, e.g. starting from a natural plant oil, is thereby enabled in which the same lipase catalyses both the initial hydrolysis of triglyceride and the final esterification step.

It has been found that esterification of a hydroxy-fatty acid by a lipase can be coupled with oleate hydratase (OHase) generation of that hydroxy-FA from an unsaturated FA with a cis C9-C10 double bond, e.g. oleic acid, in a single aqueous buffered reaction medium at low temperature, e.g. 30° C. A simple one-pot enzymatic method to produce fatty acid estolides from one or more triglycerides, e.g. starting from a natural plant oil, is thereby enabled in which the same lipase catalyses both the initial hydrolysis of triglyceride and the final esterification step.

The disclosure relates to the field of specialty chemicals and methods for their synthesis. In embodiments, the disclosure provides viable bacterial cells which comprise heterologous dual 3-hydroxy-acyl-ACP dehydratase/isomerases, etc. The disclosure further provides monounsaturated fatty acid derivative molecules produced by the viable bacterial cells which are non-native to the bacterial cells. The disclosure further provides methods for the preparation and production of non-native monounsaturated fatty acid derivative molecules such as e.g., an ω3-monounsaturated fatty acid derivative, an ω5-monounsaturated fatty acid derivative, an ω9-monounsaturated fatty acid derivative, an ω11-monounsaturated fatty acid fatty acid derivative, etc.

Described are methods of using a lipase for hydrolysis and esterification. In a first method of producing a medium chain fatty acid by hydrolysis, the method comprises providing a polypeptide with at least 90% degree of identity to SEQ ID NO. 3, and contacting the polypeptide with a medium chain fatty acid ester and water to produce the medium chain fatty acid. In a second method of forming an ester, the method comprises providing a polypeptide with at least 90% degree of identity to SEQ ID NO. 3; and contacting the polypeptide with a long chain fatty acid, an alcohol, and water to form the ester of the long chain fatty acid and the alcohol.

The present invention relates to a lipase variant of a parent lipase, which variant has lipase activity, at least 75% but less than 100% sequence identity to SEQ ID NO: 3 and comprises a substitution at one or more positions corresponding to positions 1; 2; 3; 4; 5; 6; 7; 9; 10; 11; 12; 16; 19; 30; 31; 34; 36; 37; 39; 40; 42; 44; 51; 52; 53; 54; 56; 58; 59; 70; 71; 72; 73; 83; 84; 86; 88; 90; 92; 93; 95; 96; 100; 101; 102; 104; 106; 109; 110; 112; 117; 119; 124; 125; 127; 128; 131; 132; 133; 134; 135; 137; 158; 159; 160; 161; 162; 163; 165; 166; 167; 168; 170; 181; 182; 183; 189; 190; 192; 194; 196; 202; 210; 211; 212; 220; 225; 227; 228; 229; 230; 231; 233; 237; 238; 239; 240; 242; 246; 247; 248; 252; 259; 262; 264; 269 of SEQ ID NO: 3. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

The present disclosure provides compositions and methods for enzymatic oil degumming.