Studies On The Use Of Biotechnology For Flue-Crued Tobacco Leaves During Fermentation

To explore the feasibility of adding enzyme to hydrolyze starch and protein of flue-cured tobacco leaves during fermentation, this paper had a lot of orthogonal matrix experiments and used the effectual means of biotechnology screened a bacterial strain to ferment tobacco leaves (cut tobacco).There were conclusion as below:1 In order to hydrolyze the starch of tobacco leaves, this paper selected a-amylase and glucose-amylase, the result showed that adding two kinds of amylase respectively also could decrease the starch content of B2F leaves by 21.88% and 24.17% on average; adding a-amylase and glucose-amylase respectively could decrease the starch content of C3F leaves by 33.23% and 34.68% on average, but the main influential factors were not identical; adding glucose-amylase could effectively hydrolyze the starch of X2L leaves, the average decreasing rate was 33.83%. Mixed two kinds of amylase, added it to reduce the starch content of tobacco leaves, B2F, C3F, X2L, and analyzed the results by means of orthogonal matrix. The preferable treatment conditions were as follows: For B2F ,4x of mixture of a-amylase and glucose-amylase (1:1,w/w),temperature 45℃ and relative humidity 70% for 30×; For C3F ,8× of mixture of a-amylase and glucose-amylase (1:5,w/w),temperature 50℃ and relative humidity 60% for 30×; For X2L ,8× of mixture of a-amylase and glucose-amylase (1:5,w/w),temperature 35℃ and relative humidity 60% for 30×.2 This experiment selected neutral protease to hydrolyze the protein of tobacco leaves ,and it could decrease the protein content of B2F, C3F and X2L by 5.20% ,4.71% and 9.28% respectively. But we added the mixture of a-amylase and glucose-amylase at that preferable treatment conditions first , and then added the neutral protease . The starch content of B2F, C3F and X2L were decreased respectively by 26.77%, 29.54%, 35.00%; The protein content were decreased respectively by 13.36%, 3.10%, and 11.79%; The amino acid content of three grade were increased remarkablely , the rates of increase were 7.11%,11.45%,9.52%; The changes of RS, TN and nicotine were not obvious.3 This experiment screened a new strain named G₁ , it producted a-amylase which could hydrolyze the starch of tobacco leaves . The optimal medium consisted the contents of tobacco leaves and other material such as cornmeal, the initial pH of the...