Hydrolases, nucleic acids encoding them and methods for...

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 15/55 (2006.01) A01H 5/00 (2006.01) A01H 5/10 (2006.01) A01K 67/027 (2006.01) A23C 9/12 (2006.01) A23D 7/00 (2006.01) A23D 9/00 (2006.01) A23K 1/165 (2006.01) A23L 1/305 (2006.01) A61K 38/46 (2006.01) A61K 48/00 (2006.01) C07H 21/00 (2006.01) C07K 16/40 (2006.01) C07K 19/00 (2006.01) C08C 19/36 (2006.01) C10M 177/00 (2006.01) C11D 3/386 (2006.01) C12N 5/10 (2006.01) C12N 5/16 (2006.01) C12N 9/14 (2006.01) C12N 9/16 (2006.01) C12N 9/48 (2006.01) C12N 9/99 (2006.01) C12N 11/00 (2006.01) C12N 15/63 (2006.

Patent

CA 2601472

The invention provides hydrolases, polynucleotides encoding them, and methods of making and using these polynucleotides and polypeptides. In one aspect, the invention is directed to polypeptides, e.g., enzymes, having a hydrolase activity, e.g., an esterase, acylase, lipase, phospholipase (e.g., phospholipase A, B, C and D activity, patatin activity, lipid acyl hydrolase (LAH) activity) or protease activity, including thermostable and thermotolerant hydrolase activity, and polynucleotides encoding these enzymes, and making and using these polynucleotides and polypeptides. The hydrolase activities of the polypeptides and peptides of the invention include esterase activity, lipase activity (hydrolysis of lipids), acidolysis reactions (to replace an esterified fatty acid with a free fatty acid), transesterification reactions (exchange of fatty acids between triglycerides), ester synthesis, ester interchange reactions, phospholipase activity and protease activity (hydrolysis of peptide bonds). In another aspect, the invention provides methods for hydrolyzing steryl esters and triglycerides (e.g., in a paper pulp), into sterols, glycerol and free fatty acids, using enzyme(s) of the invention. The invention provides enzymes and methods for decreasing the amount of lipophilic extracts ("pitch") in a pulp- comprising composition. The polypeptides of the invention can be used in a variety of pharmaceutical, agricultural and industrial contexts, including the manufacture of cosmetics and nutraceuticals.

L'invention concerne des hydrolases, des polynucléotides codant pour ces hydrolases ainsi que des méthodes de fabrication et d'utilisation de ces polynucléotides et polypeptides. Selon un aspect, l'invention concerne des polypeptides, notamment des enzymes, à activité hydrolase telles qu'estérase, acylase, lipase, phospholipase (par exemple phospholipase à activité A, B, C et D, activité patatine, activité lipide acyle hydrolase) ou à activité protéase, dont une activité hydrolase thermostable et thermotolérante, des polynucléotides codant pour ces enzymes, ainsi que la fabrication et l'utilisation de ces polynucléotides et polypeptides. Les activités hydrolase des polypetides et peptides de l'invention englobent une activité estérase, une activité lipase (hydrolyse des lipides), des réactions d'acidolyse (visant à remplacer un acide gras estérifié par un acide gras libre), des réactions de transestérification (échange d'acides gras entre des triglycérides), synthèse d'ester, réactions de transestérification, activité phospholipase et activité protéase (hydrolyse de liaison peptidiques). Selon un autre aspect, l'invention concerne des méthodes permettant d'hydrolyser des stéryl esters et des triglycérides (par exemple dans une pâte à papier) en stérols, glycérol et acides gras libres au moyen du ou des enzymes de l'invention. Sont décrites des enzyme set des méthodes permettant de réduire la quantité d'extraits lipophiles ("poix") dans une composition renfermant de la pâte à papier. Les polypeptides de l'invention peuvent s'utiliser dans divers contextes pharmaceutiques, agricoles et industriels, notamment pour la fabrication de cosmétiques et de nutraceutiques.

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