摘要
为获得高产淀粉酶菌株,丰富淀粉酶生产菌株资源,该研究以中温大曲为样品,利用淀粉水解圈法初筛、摇瓶发酵复筛得到高产淀粉酶菌株,结合菌落形态观察、革兰氏染色和16S rRNA同源序列分析对其进行菌种鉴定,通过单因素试验、Plackett-Burman试验、最陡爬坡试验和Box-Behnken试验确定其产淀粉酶的最佳培养条件。结果表明,共分离筛选得到6株产淀粉酶菌株,其中一株产淀粉酶活最高的菌株(编号为DQ47)被鉴定为枯草芽孢杆菌(Bacillus subtilis),其最佳培养条件为:可溶性淀粉60 g/L,酵母粉37 g/L,发酵温度39℃,装液量85 mL/250 mL,转速175 r/min,初始pH值6,接种量3%。在此优化条件下,淀粉酶活力为8158.23 U/mL,是优化前的14.75倍。
To obtain high amylase-producing strains and enrich the resources of amylase-producing strains,using medium-temperature Daqu as the sample,the high amylase-producing strains were screened by primary screening of starch hydrolysis circle method and re-screening of shaking flask fermentation,and then the strain was identified by colony morphology observation,Gram staining and 16S rRNA homologous sequence analysis.The optimal culture conditions were determined by single-factor tests,Plackett-Burman tests,steepest climb tests and Box-Behnken tests.The results showed that six strains of amylase-producing bacteria were isolated and screened,and one strain with the highest amylase-producing activity(No.DQ47)was identified as Bacillus subtilis.The optimal culture conditions were as follows:soluble starch 60 g/L,yeast powder 37 g/L,fermentation temperature 39℃,loading volume 85 ml/250 ml,rotation speed 175 r/min,initial pH 6,inoculum 3%.Under the optimized conditions,the amylase activity was 8158.23 U/ml,which was 14.75 times that of before optimization.
作者
胡晓龙
冯大鸿
田瑞杰
王永亮
曹满堂
魏涛
韩素娜
沈祥坤
何培新
HU Xiaolong;FENG Dahong;TIAN Ruijie;WANG Yongliang;CAO Mantang;WEI Tao;HAN Suna;SHEN Xiangkun;HE Peixin(School of Food and Bioengineering,Zhengzhou University of Light Industry,Zhengzhou 450001,China;Jiahu Distillery Co.,Ltd.,Luohe 462400,China;Postdoctoral Programme,Yangshao Distillery Co.,Ltd.,Sanmenxia 472400,China;Henan Food Industry Science Research Institute Co.,Ltd.,Zhengzhou 450053,China)
出处
《中国酿造》
CAS
北大核心
2023年第1期78-85,共8页
China Brewing
基金
国家自然科学基金项目(31801535)
中原产业创新领军人才项目(214100510009)
三门峡市引进高层次人才(团队)创新创业项目(2020111)
中原科技创新领军人才项目(224200510017)。
关键词
中温大曲
淀粉酶
枯草芽孢杆菌
培养条件优化
响应面法
medium-temperature Daqu
amylase
Bacillus subtilis
culture conditions optimization
response surface method
