Production of peracids using an enzyme having perhydrolysis activity
View Patent ↗A process is provided for producing peroxycarboxylic acids from carboxylic acid esters. More specifically, carboxylic acid esters are reacted with an inorganic peroxide, such as hydrogen peroxide, in the presence of an enzyme catalyst having perhydrolysis activity. The present perhydrolase catalysts are classified as members of the carbohydrate esterase family 7 (CE-7) based on the conserved structural features. Further, disinfectant formulations comprising the peracids produced by the processes described herein are provided.
1. A process to disinfect a hard surface or inanimate object using an enzymatically-produced peroxycarboxylic acid composition, said process comprising:
a) providing a set of reaction components, said components comprising:
1) a substrate selected from the group consisting of:
i) esters having the structure
wherein R 1 ═C1 to C7 straight chain or branched chain alkyl optionally substituted with an hydroxyl or a C1 to C4 alkoxy group and R 2 ═C1 to C10 straight chain or branched chain alkyl, alkenyl, alkynyl, aryl, alkylaryl, alkylheteroaryl, heteroaryl, (CH 2 CH 2 —O) n H or (CH 2 CH(CH 3 )—O) n H and n=1 to 10;
ii) glycerides having the structure
wherein R 1 ═C1 to C7 straight chain or branched chain alkyl optionally substituted with an hydroxyl or a C1 to C4 alkoxy group and R 3 and R 4 are individually H or R 1 C(O);
iii) acetylated saccharides selected from the group consisting of acetylated monosaccharides, acetylated disaccharides, and acetylated polysaccharides;
2) a source of peroxygen; and
3) an enzyme catalyst having perhydrolysis activity, wherein said enzyme catalyst comprises an enzyme comprising a signature motif that aligns with a reference sequence SEQ ID NO:2 using CLUSTALW, said signature motif comprising:
i) an RGQ motif at amino acid positions 118-120 of SEQ ID NO:2;
ii) a GXSQG motif at amino acid positions 179-183 of SEQ ID NO:2; and
iii) an HE motif at amino acid positions 298-299 of SEQ ID NO:2;
b) combining said reaction components under suitable aqueous reaction conditions whereby a peroxycarboxylic acid product is formed;
c) optionally diluting said peroxycarboxylic acid product; and
d) contacting said hard surface with the peroxycarboxylic acid produced in step b) or step c) whereby said hard surface or said inanimate object is disinfected.
2. The process of claim 1 wherein the signature motif of the enzyme further comprises an LXD motif that aligns with amino acid positions 267-269 of SEQ ID NO:2 using CLUSTALW.
3. The process of claim 2 wherein the enzyme catalyst having perhydrolysis activity comprises an enzyme having a contiguous signature motif with an amino acid sequence selected from the group consisting of SEQ ID NO:61 and an amino acid sequence at least 50% identical to SEQ ID NO:61.
4. The process of claim 1 wherein the enzyme comprises an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14 1 SEQ ID NO:16, and SEQ ID NO:32 or a substantially similar enzyme having perhydrolase activity derived by substituting, deleting or adding one or more amino acids to said amino acid sequence.
5. The process of claim 4 wherein the substantially similar enzyme having perhydrolase activity is at least 95% identical to one or more amino acid sequences selected from the group consisting of SEQ ID NO:2, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, and SEQ ID NO:32.
6. The process of claim 1 wherein the substrate is selected from the group consisting of methyl lactate, ethyl lactate, methyl glycolate, ethyl glycolate, methyl methoxyacetate, ethyl methoxyacetate, methyl 3-hydroxybutyrate, ethyl 3-hydroxybutyrate, monoacetin, diacetin, triacetin, monopropionin, dipropionin, tripropionin, monobutyrin, dibutyrin, tributyrin, glucose pentaacetate, xylose tetraacetate, acetylated xylan, β-D-ribofuranose-1,2,3,5-tetraacetate, tri-O-acetyl-D-galactal, and tri-O-acetyl-glucal, and mixtures thereof.
7. The process of claim 1 wherein the hard surface or the inanimate object is selected from the group consisting of tanks, conveyors, floors, drains, pipes, coolers, freezers, equipment surfaces, walls, valves, belts, joints, windows, water treatment facilities, pools, spas, fermentation tanks, pharmaceutical or biopharmaceutical manufacturing equipment, pharmaceutical or biopharmaceutical ingredients, pharmaceutical or biopharmaceutical excipients, clothing, medical equipment, dental equipment, surgical equipment, endoscopes, laparoscopes, scrubs, shoes, cooking surfaces, food preparation surfaces, bathroom surfaces, toilets, livestock, cattle, dairy cows, goats, horses, pigs, beef, poultry, pork, vegetables, fruits, seafood, and combinations thereof.
8. The process of claim 1 wherein the enzyme catalyst is a recombinant E. coli strain lacking catalase activity.