Mannanase cellulase treatments

Only minor amounts of additional reducing sugars from the xylan decomposition were detected in the reaction solutions. Acid hydrolysis showed, however, that higher-molecular xylan fragments were liberated by the combined cellulase-mannanase treatment. At the end of the experi-... 

Most of the mannan in the sprucewood holocellulose was hydrolyzed by the cellulase-mannanase treatments, and only about one-third of the xylan was dissolved and partly hydrolyzed into low-molecular-weight sugars. 

Sprucewood holocellulose was treated with an endo-p-1,4-mannanase isolated from Aspergillus niger and an endo-/3-1,4-xylanase, two avicelases, and a cellobiohydrolase C isolated from Trichoderma viride. The mannanase hydrolyzed about a quarter of the mannan in 2-3 days without xylan or cellulose degradation. The xylanase hydrolyzed about half the xylan with 10% mannan solubilization. The three cellulases hydrolyzed up to 45% of the cellulose and 20% of the xylan, accompanied by 40-70% solubilization of the mannan. Combined xylanase-mannanase treatment hydrolyzed about half the xylan and mannan. Addition of mannanase to to cellulose-treated samples increased the degradation of the cellulose and mannan. Micromorphological studies of the variously treated specimens revealed a loss of substances in P/Slf T, and adjacent zones of S2 of the tracheid wall. 

The current observations confirm previous studies on beechwood and sprucewood holocellulose (7,10,19). The attack of the hemicellulose proceeds from the primary wall/Si as well as from the tertiary wall into S2 the pit chambers constitute preferred paths of enzyme diffusion into the walls. Also, substances of the middle lamella, especially in the cell corners, are removed by the xylanase and the mannanase treatments. Parallel to the removal of hemicelluloses, the fibrillar structure of the cellulose and its lamellar arrangement in transections of cell walls became obvious. In samples treated with cellulases, the cellulose fibrils were often completely hydrolyzed in the Si layer, occasionally accompanied by complete dissolution of cell-wall portions. This is also in conformity with the previous conclusion that the cellulases hydrolyze highly ordered zones of cellulose and remove hemicelluloses by hydrolysis or by detachment. 

The addition of mannanase after 48 hr of incubation with one of the three cellulases revived cellulose hydrolysis. The increase of the degradation rate was not as pronounced as in the corresponding treatments of delignified beechwood with cellulases and xylanase (JO), but the cellulose degradation reached about the same values at the end of the experiments (about 50-65% ). 

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