title: Cloning, purification, and characterization of galactomannan-degrading enzymes from Myceliophthora thermophila.
abstract: Genes of β-mannosidase 97 kDa, GH family 2 (bMann9), β-mannanase 48 kDa, GH family 5 (bMan2), and α-galactosidase 60 kDa, GH family 27 (aGal1) encoding galactomannan-degrading glycoside hydrolases of Myceliophthora thermophila C1 were successfully cloned, and the recombinant enzymes were purified to homogeneity and characterized. bMann9 displays only exo-mannosidase activity, the K(m) and k(cat) values are 0.4 mM and 15 sec(-1) for p-nitrophenyl-β-D-mannopyranoside, and the optimal pH and temperature are 5.3 and 40°C, respectively. bMann2 is active towards galactomannans (GM) of various structures. The K(m) and k(cat) values are 1.3 mg/ml and 67 sec(-1) for GM carob, and the optimal pH and temperature are 5.2 and 69°C, respectively. aGal1 is active towards p-nitrophenyl-α-D-galactopyranoside (PNPG) as well as GM of various structures. The K(m) and k(cat) values are 0.08 mM and 35 sec(-1) for PNPG, and the optimal pH and temperature are 5.0 and 60°C, respectively.
authors.affiliation: Chemical Faculty, Lomonosov Moscow State University, Moscow, 119991, Russia. info@rector.msu.ru
authors.forename: G S
authors.initials: GS
authors.lastname: Dotsenko
authors.forename: M V
authors.initials: MV
authors.lastname: Semenova
authors.forename: O A
authors.initials: OA
authors.lastname: Sinitsyna
authors.forename: S W A
authors.initials: SW
authors.lastname: Hinz
authors.forename: J
authors.initials: J
authors.lastname: Wery
authors.forename: I N
authors.initials: IN
authors.lastname: Zorov
authors.forename: E G
authors.initials: EG
authors.lastname: Kondratieva
authors.forename: A P
authors.initials: AP
authors.lastname: Sinitsyn
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D020037:Sordariales / Q000201:enzymology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D005656:Fungal Proteins; D011994:Recombinant Proteins; D006026:Glycoside Hydrolases