Development Of Celluloses And Amylases Using To Process Tobacco Leaves At Yuxi

Appropriate degradation of starch and cellulose in flue-cured tobacco leaves is the key to improve the quality of tobacco leaves. Using the enzymes which are produced by microorganism can degrade cellulose and starch in flue-cured tobacco leaves.Therefore, under the artificial control, the treatment of tobacco leaves with enzymes has become one of the key processes to improve the quality of tobacco.Bacterial strains Bacillus amyloliquefaciens Al and B. Pumilus Cl isolated from the surface of the tobacco leaves can effectively produce amylase and cellulase respectively. Two stains were cultivated and the enzymes were isolated and concentrated from the fermentation metabolites. The two enzymes are secreted as extracellular enzymes. Research of enzymatic properties showed that the strain A1has the best catalytic activity for soluble starch, followed by soluble starch,and the weakest is amylopectin, for which the activity was7×105U/mL,, with an optimum activity of55℃and pH7.0; while cellulase which was produced by strain C1has the best catalytic action on microcrystalline cellulose.followed by carboxymethyl cellulose sodium salt.It has the weakest catalysis for salicin, for which the activity was6×103U/mL, with an optimum activity of50℃and pH5.0.Study of two kinds of enzyme preparations on tobacco conditions, the cost of enzyme, enzyme reaction time, humidity on improving tobacco total sugar and reducing sugar percent content have a significant influence, and the influence of temperature is not significant. The optimum conditions for enzyme are:temperature40℃, humidity70%,96h, the total sugar and reducing sugar of tobacco leaves treated by4×10'U/kg amylase from strain A1increased12.78%and12.03%, respectively;as well as treated by4×105U/kg cellulase from strain C1increased13.87%and18.07%, respectively. Processed tobacco by A1amylase and C1cellulase, total sugar, reducing sugar content were increased obviously, in which fructose increased rates were62.50%and95.45%, respectively; glucose increased rates were78.23%and92.97%, respectively; however, treatment of tobacco leaves with commercial a-amylase and cellulase, total sugar and reducing sugar content has improved, but the glucose increased the rate of only21.90%and32.60%. respectively, and fructose content did not change basically.In terms of scale enlarging experiment, the treatment of tobacco leaves by compound A1amylase and C1cellulase, the results showed that treatment of tobacco total sugar and reducing sugar contents of both improved, plus enzyme group sugar alkali and sugar to nitrogen ratio higher than that in the control group, smoking results show that the sensory quality of the sample after the enzyme treatment improved significantly.Therefore, from the use of tobacco leaf surface screening with indigenous microbial strains preparation of enzyme, not only has the excellent safety, at the same time, it has strong pertinence to degrade the starch, cellulose and other substances in tobacco leaves, and when compared with the existing commercial enzyme treatment effect is good and can generate more and improve the quality of tobacco leaves material.