摘要
为获得产淀粉酶系菌株,该研究利用碘染法初筛、酶活测定复筛,从酱香型大曲中筛选产淀粉酶菌株,通过形态学观察及16S r DNA基因序列分析进行菌种鉴定,并对其全基因组进行测序。以淀粉酶活力为评价指标,利用单因素试验和正交试验优化菌株的产淀粉酶培养基组分及发酵条件。结果表明,从酱香型大曲中筛选到9株产淀粉酶菌,其中菌株L9的淀粉酶活力最高,为403.25 U/m L。经鉴定,该菌株为地衣芽孢杆菌(Bacillus licheniformis),其基因组中具有α-淀粉酶基因与麦芽糖淀粉酶基因。地衣芽孢杆菌L9产淀粉酶的最佳发酵培养基组分为:麸皮10.0 g/L、胰蛋白胨2.0 g/L、氯化钠0.6 g/L,最佳发酵条件为:发酵温度35℃、种龄8 h、接种量3%、装液量20%、转速180 r/min,发酵时间33 h。在此优化条件下,地衣芽孢杆菌L9的淀粉酶活力为2 300.24 U/m L,为优化前的5.7倍。地衣芽孢杆菌L9具有良好的分解可溶性淀粉的能力,具备潜在的开发价值和应用前景。
In order to obtain the amylase-producing strain,the strains of amylase production from sauce-flavor(Jiangxiangxing)Daqu was preliminarily screened by iodine staining method and rescreened by enzyme activity determination.The strains were identified by morphological observation and 16S rDNA gene sequence analysis,and the whole genome of the strain was sequenced.Using amylase activity as the evaluation index,the medium components and fermentation conditions of the amylase-producing strain were optimized by single factor and orthogonal tests.The results showed that 9 amylase-producing strains were screened from sauce-flavor Daqu,among which strain L9 had the highest amylase activity(403.25 U/ml).The strain L9 was identified as Bacillus licheniformis,withα-amylase gene and maltose amylase gene in genome.The optimal medium components were as follows:bran 10.0 g/L,tryptone 2.0 g/L,NaCl 0.6 g/L.The optimal fermentation conditions were as follows:fermentation temperature 35℃,seed age 8 h,inoculum 3%,liquid loading volume 20%,rotation speed 180 r/min,and fermentation time 33 h.Under those optimized conditions,the amylase activity of B.licheniformis L9 was 2300.24 U/ml,which was 5.7 times that of before optimization.B.licheniformis L9 had a good ability to decompose soluble starch,which had potential development value and application prospect.
作者
李红霞
王雄
王新叶
张璋
唐佳代
吴远琴
刘永梅
LI Hongxia;WANG Xiong;WANG Xinye;ZHANG Zhang;TANG Jiadai;WU Yuanqin;LIU Yongmei(Department of Brewing Engineering,Moutai Institute,Zunyi 564507,China)
出处
《中国酿造》
CAS
北大核心
2024年第12期124-130,共7页
China Brewing
基金
遵义市科技计划项目(遵市科合HZ字[2021]312号)
茅台学院高层次人才科研启动经费项目(mygccrc[2022]024)
茅台学院高层次人才科研启动经费项目(mygccrc[2022]023)
茅台学院高层次人才科研启动经费项目(mygccrc[2022]003)
国家自然科学基金地区科学基金项目(32260229)。
关键词
酱香型大曲
淀粉酶系
筛选
鉴定
产酶条件优化
sauce-flavor Daqu
amylases
screening
identification
enzyme-producing conditions optimization
