US2015141680A1PendingUtilityA1

Enzymatic removal of steryl glycosides

38
Individually held — no corporate assignee on recordPriority: Mar 16, 2012Filed: Mar 14, 2013Published: May 21, 2015
Est. expiryMar 16, 2032(~5.7 yrs left)· nominal 20-yr term from priority
Inventors:Hugo MenzellaSalvador PeiruLeandro Vetcher
A23D 9/04C12N 9/20C12N 9/2428C12N 9/2408C12Y 302/01021C10L 2200/0476C11B 3/003C10L 2270/026C12N 9/2402C12P 7/64C12P 19/02C12Y 302/01104C11C 3/003C12P 19/14C10L 1/026C12N 9/2445Y02E50/10C12P 7/649C12P 7/6458
38
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Claims

Abstract

The present invention provides compositions and methods related to the production and use of enzymes suitable for reducing the amount of steryl glycosides or saturated monoacyl glycerols in a lipid mixture.

Claims

exact text as granted — not AI-modified
1 . A method for reducing steryl glycoside in a sample, comprising:
 In a reaction mixture, mixing a thermostable enzyme with a sample comprising steryl glycoside under a condition suitable for said thermostable enzyme for a suitable period of time to degrade said steryl glycoside, thereby reducing steryl glycoside in said sample to obtain a processed sample.   
     
     
         2 . The method of  claim 1 , wherein said sample comprises oil, fat, or biofuel. 
     
     
         3 . The method of  claim 2 , wherein said biofuel comprises biodiesel. 
     
     
         4 . The method of  claim 1 , wherein said steryl glycoside comprises steryl glucoside. 
     
     
         5 . The method of  claim 1 , wherein said steryl glycoside has a solubility that is more than 50 ppm. 
     
     
         6 . The method of  claim 1 , wherein said steryl glycoside has a solubility that is more than 80 ppm. 
     
     
         7 . The method of  claim 1 , wherein said steryl glycoside has a solubility that is more than 100 ppm. 
     
     
         8 . The method of  claim 1 , wherein said thermostable enzyme is capable of hydrolyzing the glycosidic bond of a steryl glucoside or acylated steryl glucoside. 
     
     
         9 . The method of  claim 1 , wherein said thermostable enzyme comprises a glycosidase enzyme. 
     
     
         10 . The method of  claim 1 , wherein said thermostable enzyme comprises a glucosidase enzyme. 
     
     
         11 . The method of  claim 1 , wherein said thermostable enzyme comprises a variant of an enzyme selected from Table 1. 
     
     
         12 . The method of  claim 1 , wherein said thermostable enzyme comprises a variant having an amino acid sequence that has at least 95% identity to a sequence selected from the sequences of Table 1. 
     
     
         13 . The method of  claim 1 , wherein said thermostable enzyme comprises an amino acid sequence selected from the sequences of Table 1. 
     
     
         14 . The method of  claim 1 , wherein said sample comprises 0.1% to 30% of water. 
     
     
         15 . The method of  claim 1 , wherein said mixing is carried out at a temperature that is between about 50° C. and about 110° C. 
     
     
         16 . The method of  claim 15 , wherein said mixing is carried out at a temperature that is above about 65° C. 
     
     
         17 . The method of  claim 15 , wherein said mixing is carried out at a temperature that is above about 70° C. 
     
     
         18 . The method of  claim 15 , wherein said mixing is carried out at a temperature that is above about 85° C. 
     
     
         19 . The method of  claim 1 , wherein said mixing is carried for about 30 minutes to 24 hours. 
     
     
         20 . The method of  claim 1 , wherein the amount of said steryl glycoside is reduced by at least 20%. 
     
     
         21 . The method of  claim 1 , wherein the amount of said steryl glycoside is reduced by at least 80%. 
     
     
         22 . The method of  claim 1 , wherein said processed sample comprises less than about 20 ppm of said steryl glycoside. 
     
     
         23 . The method of  claim 1 , wherein said reaction mixture comprises an enzyme selected from the group consisting of beta-glucosidases, sterol-esterases, amyloglucosidases, and pectinases. 
     
     
         24 . The method of  claim 1 , further comprising collecting said processed sample. 
     
     
         25 . An oil produced by the method of  claim 1 . 
     
     
         26 . An isolated thermostable enzyme that is capable of hydrolyzing the glycosidic bond of steryl glycosides or acylated steryl glycosides. 
     
     
         27 . The thermostable enzyme of  claim 26 , wherein said thermostable enzyme has a hydrolytic activity of at least 5 g of steryl glycoside per gram of enzyme per hour at a temperature that is between about 50° C. and about 99° C. 
     
     
         28 . The thermostable enzyme of  claim 26 , wherein said thermostable enzyme has an activity of at least 5 g of steryl glycoside per gram of enzyme per hour at a temperature that is above about 65° C. 
     
     
         29 . The thermostable enzyme of  claim 26 , wherein said thermostable enzyme has an activity of at least 5 g of steryl glycoside per gram of enzyme per hour at a temperature that is above about 70° C. 
     
     
         30 . The thermostable enzyme of  claim 26 , wherein said thermostable enzyme has an activity of at least 5 g of steryl glycoside per gram of enzyme per hour at a temperature that is above about 75° C. 
     
     
         31 . The thermostable enzyme of  claim 26 , wherein said thermostable enzyme comprises a variant of one of the enzymes listed in Table 1. 
     
     
         32 . The thermostable enzyme of  claim 26 , wherein said thermostable enzyme comprises a variant having an amino acid sequence that is at least 95% identical to the sequence of one of the enzymes listed in Table 1. 
     
     
         33 . The thermostable enzyme of  claim 26 , wherein said thermostable enzyme comprises an amino acid sequence selected from the sequences of Table 1. 
     
     
         34 - 46 . (canceled)

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