Perhydrolase variant providing improved specific activity
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12N-009/18
C12P-007/40
A61L-009/00
출원번호
US-0657590
(2012-10-22)
등록번호
US-8486679
(2013-07-16)
발명자
/ 주소
DiCosimo, Robert
Payne, Mark Scott
Gavagan, John Edward
출원인 / 주소
E. I. du Pont de Nemours and Company
인용정보
피인용 횟수 :
0인용 특허 :
28
초록▼
An acetyl xylan esterase variant having perhydrolytic activity is provided for producing peroxycarboxylic acids from carboxylic acid esters and a source of peroxygen. More specifically, a Thermotoga maritima acetyl xylan esterase gene was modified using error-prone PCR and site-directed mutagenesis
An acetyl xylan esterase variant having perhydrolytic activity is provided for producing peroxycarboxylic acids from carboxylic acid esters and a source of peroxygen. More specifically, a Thermotoga maritima acetyl xylan esterase gene was modified using error-prone PCR and site-directed mutagenesis to create an enzyme catalyst characterized by an increase in specific activity. The variant acetyl xylan esterase may be used to produce peroxycarboxylic acids suitable for use in a variety of applications such as cleaning, disinfecting, sanitizing, bleaching, wood pulp processing, and paper pulp processing applications.
대표청구항▼
1. An isolated polypeptide having perhydrolytic activity comprising the amino acid sequence of SEQ ID NO: 11. 2. The polypeptide of claim 1; wherein said polypeptide is characterized by a peracetic acid formation specific activity that is higher than the peracetic acid formation specific activity of
1. An isolated polypeptide having perhydrolytic activity comprising the amino acid sequence of SEQ ID NO: 11. 2. The polypeptide of claim 1; wherein said polypeptide is characterized by a peracetic acid formation specific activity that is higher than the peracetic acid formation specific activity of the Thermotoga maritima C277T acetyl xylan esterase having amino acid sequence SEQ ID NO: 5. 3. A process for producing a peroxycarboxylic acid comprising: (a) providing a set of reaction components comprising: (1) at least one substrate selected from the group consisting of: (i) one or more esters having the structure [X]mR5 whereinX=an ester group of the formula R6—C(O)O;R6 =C1 to C7 linear, branched or cyclic hydrocarbyl moiety, optionally substituted with hydroxyl groups or C1 to C4 alkoxy groups, wherein R6 optionally comprises one or more ether linkages for R6 =C2 to C7;R5=a C1 to C6 linear, branched, or cyclic hydrocarbyl moiety or a five-membered cyclic heteroaromatic moiety or six-membered cyclic aromatic or heteroaromatic moiety optionally substituted with hydroxyl groups; wherein each carbon atom in R5 individually comprises no more than one hydroxyl group or no more than one ester group or carboxylic acid group; wherein R5 optionally comprises one or more ether linkages;m is an integer ranging from 1 to the number of carbon atoms in R5; and wherein said esters have solubility in water of at least 5 ppm at 25° C.;(ii) one or more glycerides having the structure wherein R1=C1 to C21 straight chain or branched chain alkyl optionally substituted with an hydroxyl or a C1 to C4 alkoxy group and R3 and R4 are individually H or R1C(O);(iii) one or more esters of the formula: wherein R1 is a C1 to C7 straight chain or branched chain alkyl optionally substituted with an hydroxyl or a C1 to C4 alkoxy group and R2 is a C1 to C10 straight chain or branched chain alkyl, alkenyl, alkynyl, aryl, alkylaryl, alkylheteroaryl, heteroaryl, (CH2CH2O)n, or (CH2CH(CH3)—O)nH and n is 1 to 10;(iv) one or more acylated monosaccharides, acylated disaccharides, or acylated polysaccharides; and(v) any combination of (i) through (iv);(2) a source of peroxygen; and(3) an enzyme catalyst comprising the polypeptide of claim 1;(b) combining the set of reaction components under suitable reaction conditions whereby peroxycarboxylic acid is produced; and(c) optionally diluting the peroxycarboxylic acid produced in step (b). 4. The process of claim 3 further comprising the step of: d) contacting a hard surface or inanimate object with the peroxycarboxylic acid produced in step (b) or step (c); whereby said hard surface or said inanimate object is disinfected, bleached, destained or a combination thereof. 5. The process of claim 3 wherein the inanimate object is a medical instrument. 6. The process of claim 3 further comprising the step of: d) contacting an article of clothing or a textile with peroxycarboxylic acid produced in step (b) or step (c); whereby the article of clothing or textile receives a benefit. 7. The process of claim 6 wherein the benefit is selected from the group consisting of disinfecting, sanitizing, bleaching, destaining, deodorizing, and combinations thereof. 8. The process of claim 3 further comprising the step of: d) contacting wood pulp or paper pulp with peroxycarboxylic acid produced in step (b) or step (c); whereby the wood pulp or paper pulp is bleached. 9. The process of claim 3 wherein the substrate is selected from the group consisting of: monoacetin; diacetin; triacetin; monopropionin; dipropionin; tripropionin; monobutyrin; dibutyrin; tributyrin; glucose pentaacetate; β-D-galactose pentaacetate; sorbitol hexaacetate; sucrose octaacetate; xylose tetraacetate; acetylated xylan; acetylated xylan fragments; β-D-ribofuranose-1,2,3,5-tetraacetate; tri-O-acetyl-D-galactal; tri-O-acetyl-D-glucal; monoesters or diesters of 1,2-ethanediol; 1,2-propanediol; 1,3-propanediol; 1,2-butanediol; 1,3-butanediol; 2,3-butanediol; 1,4-butanediol; 1,2-pentanediol; 2,5-pentanediol; 1,6-pentanediol; 1,2-hexanediol; 2,5-hexanediol; 1,6-hexanediol; 4-acetoxybenzoic acid; and mixtures thereof. 10. The process of claim 9 wherein the substrate is triacetin. 11. The process of claim 3 wherein the peroxycarboxylic acid produced is peracetic acid, perpropionic acid, perbutyric acid, perlactic acid, perglycolic acid, permethoxyacetic acid, per-β-hydroxybutyric acid, or mixtures thereof. 12. The process of claim 3 wherein the enzyme catalyst is in the form of a microbial cell, a permeabilized microbial cell, a microbial cell extract, a partially purified enzyme, or a purified enzyme. 13. A composition comprising: (a) a set of reaction components comprising: (1) at least one substrate selected from the group consisting of: (i) one or more esters having the structure [X]mR5 whereinX=an ester group of the formula R6—C(O)O;R6=C1 to C7 linear, branched or cyclic hydrocarbyl moiety, optionally substituted with hydroxyl groups or C1 to C4 alkoxy groups, wherein R6 optionally comprises one or more ether linkages for R6=C2 to C7;R5=a C1 to C6 linear, branched, or cyclic hydrocarbyl moiety or a five-membered cyclic heteroaromatic moiety or six-membered cyclic aromatic or heteroaromatic moiety optionally substituted with hydroxyl groups; wherein each carbon atom in R5 individually comprises no more than one hydroxyl group or no more than one ester group or carboxylic acid group; wherein R5 optionally comprises one or more ether linkages;m is an integer ranging from 1 to the number of carbon atoms in R5; andwherein said esters have solubility in water of at least 5 ppm at 25° C.;(ii) one or more glycerides having the structure wherein R1=C1 to C21 straight chain or branched chain alkyl optionally substituted with an hydroxyl or a C1 to C4 alkoxy group and R3 and R4 are individually H or R1C(O);(iii) one or more esters of the formula: wherein R1 is a C1 to C7 straight chain or branched chain alkyl optionally substituted with an hydroxyl or a C1 to C4 alkoxy group and R2 is a C1 to C10 straight chain or branched chain alkyl, alkenyl, alkynyl, aryl, alkylaryl, alkylheteroaryl, heteroaryl, (CH2CH2O)n, or (CH2CH(CH3)—O)nH and n is 1 to 10;(iv) one or more acylated monosaccharides, acylated disaccharides, or acylated polysaccharides; and(v) any combination of (i) through (iv);(2) a source of peroxygen; and(3) an enzyme catalyst comprising the polypeptide of claim 6; and(b) at least one peroxycarboxylic acid formed upon combining the set of reaction components of (a). 14. A peracid generation and delivery system comprising: (a) a first compartment comprising (1) an enzyme catalyst comprising the polypeptide of claim 6;(2) at least one substrate selected from the group consisting of: (i) one or more esters having the structure [X]mR5 whereinX=an ester group of the formula R6—C(O)O;R6=C1 to C7 linear, branched or cyclic hydrocarbyl moiety, optionally substituted with hydroxyl groups or C1 to C4 alkoxy groups, wherein R6 optionally comprises one or more ether linkages for R6=C2 to C7;R5=a C1 to C6 linear, branched, or cyclic hydrocarbyl moiety or a five-membered cyclic heteroaromatic moiety or six-membered cyclic aromatic or heteroaromatic moiety optionally substituted with hydroxyl groups; wherein each carbon atom in R5 individually comprises no more than one hydroxyl group or no more than one ester group or carboxylic acid group; wherein R5 optionally comprises one or more ether linkages;m is an integer ranging from 1 to the number of carbon atoms in R5; andwherein said esters have solubility in water of at least 5 ppm at 25° C.;(ii) one or more glycerides having the structure wherein R1=C1 to C21 straight chain or branched chain alkyl optionally substituted with an hydroxyl or a C1 to C4 alkoxy group and R3 and R4 are individually H or R1C(O);(iii) one or more esters of the formula: wherein R1 is a C1 to C7 straight chain or branched chain alkyl optionally substituted with an hydroxyl or a C1 to C4 alkoxy group and R2 is a C1 to C10 straight chain or branched chain alkyl, alkenyl, alkynyl, aryl, alkylaryl, alkylheteroaryl, heteroaryl, (CH2CH2O)n, or (CH2CH(CH3)—O)nH and n is 1 to 10;(iv) one or more acylated monosaccharides, acylated disaccharides, or acylated polysaccharides; and(v) any combination of (i) through (iv); and(3) an optional buffer; and(b) a second compartment comprising (1) source of peroxygen;(2) a peroxide stabilizer; and(3) an optional buffer. 15. The peracid generation and delivery system of claim 14 wherein the substrate comprises triacetin. 16. A laundry care product comprising the polypeptide of claim 3. 17. A personal care product comprising the polypeptide of claim 3.
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