Methods, compositions, and cells for generating acyl alcohols. Compositions comprising acyl alcohols. Methods of cleaving acyl amino acids and/or acyl alcohols to generate free fatty acids, free amino acids, and/or free alcohols.

Methods, compositions, and cells for generating acyl alcohols. Compositions comprising acyl alcohols. Methods of cleaving acyl amino acids and/or acyl alcohols to generate free fatty acids, free amino acids, and/or free alcohols.

The present invention relates to a polypeptide having lipase activity wherein the polypeptide when aligned with the polypeptide according to SEQ ID NO: 1, comprises at least an amino acid substitution L410X and optionally one or more amino acid substitutions chosen from S365Q, S365N, L413M, G414A, G414S, G414V, G414T, V534L and V534l, wherein the 10 numbering of amino acid position(s) is/are defined with reference to SEQ ID NO: 1. The invention further relates to a process for preparing a product comprising an oil or fat comprising bringing an intermediary form of the product comprising oil or fat into contact with a polypeptide as disclosed herein and the use of a polypeptide as disclosed herein to saturated fatty acids in an oil or fat.

The present invention relates to a polypeptide having lipase activity wherein the polypeptide when aligned with the polypeptide according to SEQ ID NO: 1, comprises at least an amino acid substitution L410X and optionally one or more amino acid substitutions chosen from S365Q, S365N, L413M, G414A, G414S, G414V, G414T, V534L and V534l, wherein the 10 numbering of amino acid position(s) is/are defined with reference to SEQ ID NO: 1. The invention further relates to a process for preparing a product comprising an oil or fat comprising bringing an intermediary form of the product comprising oil or fat into contact with a polypeptide as disclosed herein and the use of a polypeptide as disclosed herein to saturated fatty acids in an oil or fat.

The object of an aspect of the present invention is to provide a method for producing a long-chain fatty acid, the method making it possible to prevent a coloring component from being contained in the long-chain fatty acid and recover the long-chain fatty acid at a high yield. The above problem is solved by providing a method for producing a long-chain fatty acid, the method including the steps of: (a) with use of an enzyme, decomposing, into fatty acids, triglyceride contained in a raw vegetable oil which is derived from a vegetable-derived oil-containing waste; (b) removing, by distillation, a short-chain fatty acid contained in the raw vegetable oil which has been subjected to the step (a); and (c) recovering, by short-path distillation, a long-chain fatty acid contained in the raw vegetable oil which has been subjected to the step (b).

The present disclosure discloses a method for increasing the usage efficiency of a lipase in the enzymatic reaction, belonging to the technical field of food processing. In the present disclosure, the peroxide value of raw materials required for enzymatic synthesis of a structured lipid is reduced by a distillation method or an adsorption method, and then the raw material and a lipase undergo catalytic reaction to synthesize the structured lipid, so as to maintain the activity of the lipase and increase the reuse rate of the enzyme. The distillation method includes deodorization and molecular distillation. The present disclosure provides a method for manufacturing a structured lipid by a high efficiency and low-cost enzymatic synthesis, addressing to the issues such as large enzyme consumption, expansive cost, low enzyme utilization rate in the traditional industrial production. Therefore, the method provided by the present disclosure has certain development potential and practical significance.

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.

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.

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.