Purification and characterization of a low molecular weight of beta-mannanase from Penicillium occitanis Pol6.
The highest beta-mannanase activity was produced by Penicillium occitanis Pol6 on flour of carob seed, whereas starch-containing medium gave lower enzymes titles. The low molecular weight enzyme was purified to homogeneity by ammonium sulfate precipitation, gel filtration, and ion-exchange chromatography procedures. The purified beta-mannanase (ManIII) has been identified as a glycoprotein (carbohydrate content 5%) with an apparent molecular mass of 18 kDa. It was active at 40 degrees C and pH 4.0. It was stable for 30 min at 70 degrees C and has a broad pH stability (2.0-12.0). ManIII showed K (m), V (max), and K (cat) values of 17.94 mg/ml, 93.52 U/mg, and 28.13 s(-1) with locust bean gum as substrate, respectively. It was inhibited by mannose with a K (I) of 0.610(-3) mg/ml. ManIII was activated by CuSO4 and CaCl2 (2.5 mM). However, in presence of 2.5 mM Co2+, its activity dropped to 60% of the initial activity. Both N-terminal and internal amino acid sequences of ManIII presented no homology with mannanases of glycosides hydrolases. During incubation with locust bean gum and Ivory nut mannan, the enzyme released mainly mannotetraose, mannotriose, and mannobiose.
Unite Enzyme and Bioconversion, Ecole Nationale d'Ingénieurs de Sfax, Route Soukra km 3.5 B.P.W, 3038 Sfax, Tunisia.
Monia
M
Blibech
Raoudha Ellouz
RE
Ghorbel
Ines
I
Fakhfakh
Patricia
P
Ntarima
Katheleen
K
Piens
Abir Ben
AB
Bacha
Semia Ellouz
SE
Chaabouni
D000595:Amino Acid Sequence; D002413:Cations, Divalent / Q000494:pharmacology; D005685:Galactans / Q000378:metabolism; D007700:Kinetics; D008351:Mannans / Q000378:metabolism; D008361:Mannosidases / Q000302:isolation & purification* / Q000378:metabolism*; D008970:Molecular Weight; D010407:Penicillium / Q000201:enzymology*; D053149:Plant Gums / Q000378:metabolism
D002413:Cations, Divalent; D005685:Galactans; D008351:Mannans; D053149:Plant Gums; D008361:Mannosidases; C041867:endo-1,4-beta-D-mannanase; C017471:locust bean gum