Stabilization of perhydrolases
View Patent ↗Disclosed herein are enzyme powders comprising a spray-dried formulation of at least one CE-7 esterase, at least one oligosaccharide excipient, and optionally at least one surfactant. Also disclosed herein is a process for producing peroxycarboxylic acids from carboxylic acid esters using the aforementioned enzyme powders. Further, disinfectant and laundry care formulations comprising the peracids produced by the processes described herein are provided.
1. A process to stabilize the perhydrolysis activity of an enzyme when present in a formulation comprised of said enzyme and a carboxylic acid ester, the process comprising:
(a) providing an aqueous formulation comprising:
(i) at least one enzyme structurally classified as a CE-7 enzyme and having perhydrolysis activity and a signature motif comprising:
(1) an RGQ motif at amino acid residues 118-120,
(2) a GXSQG motif at residues 179-183, and
(3) an HE motif at residues 298-299
when aligned to reference sequence SEQ ID NO:1 using CLUSTALW, said enzyme further having at least 30% amino acid identity to SEQ ID NO:1,
(ii) at least one oligosaccharide excipient, and
(iii) optionally at least one surfactant; and
(b) spray-drying the aqueous formulation of (a) to produce an enzyme powder which substantially retains the perhydrolysis activity of the at least one enzyme when present in a formulation comprised of a carboxylic acid ester and the enzyme powder.
2. The process of claim 1 , wherein the at least one oligosaccharide excipient has a number average molecular weight of at least about 1250 and a weight average molecular weight of at least about 9000.
3. The process of claim 2 , wherein the at least one oligosaccharide excipient is selected from the group consisting of maltodextrin, xylan, mannan, fucoidan, galactomannan, chitosan, raffinose, stachyose, pectin, inulin, levan, graminan, amylopectin, and mixtures thereof.
4. The process of claim 3 , wherein the at least one oligosaccharide excipient is maltodextrin.
5. The process of claim 1 , wherein the at least one oligosaccharide excipient is trehalose.
6. The process of claim 1 , wherein the carboxylic acid ester is selected from the group consisting of monoacetin, diacetin, triacetin, monopropionin, dipropionin, tripropionin, monobutyrin, dibutyrin, tributyrin, and mixtures thereof.
7. The process of claim 6 , wherein the carboxylic acid ester is triacetin.
8. The process of claim 1 , wherein the at least one surfactant is present and is polysorbate 80.
9. The process of claim 2 , wherein the at least one oligosaccharide excipient has a number average molecular weight of at least about 1700 and a weight average molecular weight of at least about 15000.
10. The process of claim 1 , wherein the at least one enzyme comprises an amino acid sequence selected from the group consisting of SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:19, and SEQ ID NO:20, wherein amino acid residue 277 of SEQ ID NO: 19 or SEQ ID NO: 20 is selected from the group consisting of alanine, valine, serine, and threonine.