CAJ | 학술논문

从上部低次烟叶中筛选出耐高温淀粉酶产生菌GW 2和耐高温蛋白酶产生菌GW 3，通过对菌落的形态观察、16S rRNA的鉴定和同源性分析，初步确定GW 2为解淀粉芽胞杆菌，GW 3为枯草芽胞杆菌。对上部低次烟叶进行双菌种固态发酵，并优化发酵条件，最佳发酵条件：GW 2菌悬液（ OD600＝1）接种量为7?5 mL／g，GW 3菌悬液（ OD600＝1）接种量为10 mL／g，物料填充度为500 mL三角瓶中装50 g烟叶干基，当烟叶发酵水分质量分数为45％时，48℃发酵5 d。在该发酵条件下，上部低次烟叶的淀粉降解率为22?33％，蛋白质降解率为16?74％。烟叶淀粉和蛋白质的质量分数较大幅度下降，其含量指标接近优等烟叶的含量。该发酵方式比传统的贮存式烟叶发酵大大缩短了发酵时间。
종상부저차연협중사선출내고온정분매산생균GW 2화내고온단백매산생균GW 3，통과대균락적형태관찰、16S rRNA적감정화동원성분석，초보학정GW 2위해정분아포간균，GW 3위고초아포간균。대상부저차연협진행쌍균충고태발효，병우화발효조건，최가발효조건：GW 2균현액（ OD600＝1）접충량위7?5 mL／g，GW 3균현액（ OD600＝1）접충량위10 mL／g，물료전충도위500 mL삼각병중장50 g연협간기，당연협발효수분질량분수위45％시，48℃발효5 d。재해발효조건하，상부저차연협적정분강해솔위22?33％，단백질강해솔위16?74％。연협정분화단백질적질량분수교대폭도하강，기함량지표접근우등연협적함량。해발효방식비전통적저존식연협발효대대축단료발효시간。
In this study,two thermotolerant strains were isolated from tobacoo leaves?One is an amylase producing bacterial strain named GW?2 and another is a protease producing bacterial strain named GW?3?By morphological observation of colonies and 16S rRNA sequence homology analysis, they were identified as Bacillus subtilis and Bacillus amyloliquefaciens? Then the two strains were used for tobacco solid?state fermentation,and the condition was optimized by monofactorial experiments?The experimental results show that the best conditions for solid?state fermentation were that:inoculum of bacterial suspension( OD600=1) of GW?2 and GW?3 were 7?5 mL/g and 10 mL/g,respectively,charge amount of tobacoo was 50 g ( 500 mL flask ) , fermentation temperature was 48 ℃, moisture content of the tobacco was 45% and fermentation time was 5 d?Under these conditions of solid?state fermentation,the starch and protein decomposition rates in the upper leaves of flue?cured tobacco were 22?33% and 16?74%, respectively?The content of starch and protein in the upper leaves of flue?cured tobacco was close to the standard of the top quality tobacco?In this way, the fermentation time was greatly shortened than traditional storage fermentation.