Recombinant alpha-L-rhamnosidase from Aspergillus terreus in selective trimming of rutin

0382285 - MBÚ 2013 RIV GB eng J - Článek v odborném periodiku
Gerstorferová, Daniela - Fliedrová, Barbora - Halada, Petr - Marhol, Petr - Křen, Vladimír - Weignerová, Lenka
Recombinant alpha-L-rhamnosidase from Aspergillus terreus in selective trimming of rutin.
Process Biochemistry. Roč. 47, č. 5 (2012), s. 828-835. ISSN 1359-5113. E-ISSN 1873-3298
Grant CEP: GA ČR(CZ) GAP301/11/0662; GA ČR GD305/09/H008; GA ČR(CZ) GAP301/11/0767; GA MŠMT(CZ) 7E11010
Klíčová slova: alpha-L-Rhamnosidase * Aspergillus terreus * Isoquercitrin
Kód oboru RIV: CE - Biochemie
Impakt faktor: 2.414, rok: 2012

Alpha-L-Rhamnosidase (EC 3.2.1.40) is a biotechnologically important enzyme used for derhamnosylation of many natural compounds. The extracellular alpha-L-rhamnosidase was purified from the culture of Aspergillus terreus grown on L-rhamnose-rich medium. This enzyme was found to be thermo- and alkali-tolerant, able to operate at 70 degrees C and pH 8.0. The alpha-L-rhamnosidase cDNA was cloned from A. terreus, sequenced, and expressed in the yeast Pichia pastoris as a fully functional protein. The recombinant protein was purified to apparent homogeneity and biochemically characterized. Both the native and the recombinant alpha-L-rhamnosidases catalyzed the conversion of rutin into quercetin-3-glucopyranoside (isoquercitrin), a pharmacologically significant flavonoid usable in nutraceutics. This procedure has high volumetric productivity (up to 300 g/L) and yields the product void of unwanted quercetin. The significant advantage of our expression system consists in shorter production times, up to fourfold increase in enzyme yields and the absence of unwanted beta-D-glucosidase as compared to the native production system. Thanks to its unique properties, this enzyme is applicable in a selective synthesis/hydrolysis of various rhamnose containing structures. (C) 2012 Elsevier Ltd. All rights reserved
Trvalý link: http://hdl.handle.net/11104/0212548