Production of Fatty Acid Alkyl Esters by Use of Two Lipolytic Enzymes

Abstract

A method for producing fatty acid alkyl esters, wherein a solution comprising triglyceride and alcohol is contacted with a first lipolytic enzyme having a relatively higher activity on free fatty acids than on triglyceride and a second lipolytic enzyme having a relatively higher activity on triglyceride than on free fatty acids.

1 - 15 . (canceled) 
     
     
         16 . A method for producing fatty acid alkyl esters, comprising making a reaction mixture by contacting a solution comprising a substrate and an alcohol, which substrate comprises triglyceride, with a first lipolytic enzyme having a ratio of activity on triglyceride to activity on FFA below 0.2 and a second lipolytic enzyme having a ratio of activity on triglyceride to activity on FFA above 0.5. 
     
     
         17 . The method of  claim 16 , wherein the substrate further comprises free fatty acids in the range of 0.01-95% per weight free fatty acids. 
     
     
         18 . The method of  claim 16 , wherein the triglyceride is derived from one or more of vegetable oil feedstock, rapeseed oil, soybean oil, mustard oil, sunflower oil, canola oil, coconut oil, hemp oil, palm oil, tall oil, animal fats including tallow, lard, poultry and fish oil. 
     
     
         19 . The method of  claim 16 , wherein the molar ratio between alcohol and fatty acid residues is least 0.1 and maximum 10. 
     
     
         20 . The method of  claim 16 , wherein the molar ratio between alcohol and fatty acid residues is in the range 0.3-5. 
     
     
         21 . The method of  claim 16 , wherein the molar ratio between alcohol and fatty acid residues is in the range 0.4-2. 
     
     
         22 . The method of  claim 16 , wherein the alcohol is methanol or ethanol. 
     
     
         23 . The method of  claim 16 , wherein the reaction mixture further comprises water. 
     
     
         24 . The method of  claim 16 , wherein the first lipolytic enzyme has a ratio of activity on triglyceride to activity on FFA in the range of 0.01-0.2 and the second lipolytic enzyme has a ratio of activity on triglyceride to activity on FFA in the range of 0.5-20. 
     
     
         25 . The method of  claim 16 , wherein the first lipolytic enzyme is 60% identical with a lipolytic enzyme selected from the group consisting of Lipase B from  Candida antarctica, Hyphozyma  sp. lipase and  Candida parapsilosis  lipase. 
     
     
         26 . The method of claim  235  wherein the first lipolytic enzyme is Lipase B from  Candida antarctica, Hyphozyma  sp. lipase or  Candida parapsilosis  lipase. 
     
     
         27 . The method of  claim 16 , wherein the second lipolytic enzyme is 60% identical with a lipolytic enzyme selected from the group consisting of  T. lanuginosus  lipase,  H. insolens  cutinase,  C. antarctica  lipase A, lipases from  Candida rugosa, Pseudomonas cepacia, Geotricum candidum, Rhizomucor miehei, Crytococcus  spp. S-2,  Candida parapsilosis  and  Humicola lanuginosus.    
     
     
         28 . The method of  claim 27 , wherein the second lipolytic enzyme is one of  T. lanuginosus  lipase,  H. insolens  cutinase,  C. antarctica  lipase A, lipases from  Candida rugosa, Pseudomonas cepacia, Geotricum candidum, Rhizomucor miehei, Cytococcus  spp. S-2,  Candida parapsilosis  and  Humicola lanuginosus.    
     
     
         29 . The method of  claim 16 , wherein the process is proceeding in a batch mode. 
     
     
         30 . The method of  claim 16 , wherein the process is proceeding in a continuous mode. 
     
     
         31 . The method of  claim 16 , further comprising mixing solution phases in the reaction mixture using a high shear mixer. 
     
     
         32 . The method of  claim 16 , wherein the process is conducted in a counter-current mode.