1) Three β-1, 4-mannanases were isolated from germinated guar bean through extraction, ammonium sulfate fractionation, column chromatography on cellulose derivatives and gel filltration on Sephadex G-100. They were designated as, β-1, 4-mannanase A.B and C, respectively, in the order of isolation.
2) These enzymes were different in several aspects such as pH optimum, effect of metal ions, adsorbability on cellulose derivatives, molecular weight, Michaelis constant toward reduced ivory nut mannan A, mode of action and extent of hydrolysis of the mannan.
3) β-1, 4-Mannanases A and C were proposed to be two different endo-enzymes of random-splitting type producing a series of oligosaccharides from β-1, 4-mannans. β-1, 4-Mannanase B was suggested to be possibly an exo-type enzyme catalyzing a stepwise splitting from the non-reducing end of β-1, 4-mannans to produce mannose.
4) Guaran was subjected to hydrolysis by the purified enzymes and the consequence was discussed in connection with structural requirements of the enzymes toward substituted β-1, 4-mannans and their role in germinating guar seeds.
|