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摘 要：以宋河中高温大曲为试材共分离出3株芽孢菌编号分别为Sp1、Spm1、Spm2，采用形态学和生理生化特性及16sRNA经鉴手段，并采用透明圈法、固态培养和液态发酵法研究优势芽孢菌Sp1的生长与产酶、产酸、产香功能特性。结果表明：Sp1、Spm1、Spm2均鉴定为地衣芽孢杆菌。菌株Sp1最适生长温度为40℃、pH为5.5-6.5，能利用5%-6%的葡萄糖生长，能耐受6%-8%的乙醇，在该条件下产香效果良好，产酸较稳定。30℃、pH 5.5、糖度6%时，产淀粉酶活力达到最高57.35 U/mL，同时产纤文素酶活力达到最高21.84 U/mL。30℃、pH 4.5、糖度6%时，产果胶酶活力达到最高49.23 U/mL。30℃、pH 6.5、糖度3%时，产蛋白酶活力达到最高98.62 U/mL。35℃、pH 4.5、糖度6%时，产酯化酶活力达到最高26.67 U/mL。36947
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毕业论文关键词：宋河大曲；芽孢菌；分离鉴定；优势菌；功能特性
Study on Isolation and Functional Properties of Bacillus in Daqu
Abstract: Three spore strains numbered Sp1, Spm1, Spm2. were isolated from Songhe high temperature and identified by morphology observing and physiological and biochemical properties test and 16s RNA, the growth, acid- producing, enzyme-producing, aroma-producing characteristics of preponderant strain Sp1 in Daqu were studied by observing transparent circle, solid-state culturing and liquid-state fermentation. The results showed that: strain Sp1, Spm1, Spm2 were all identified as bacillus licheniformis. The optimum conditions of Strains Sp1 to grow，produce aromatic matter and acid were 40℃，pH 5.5 - 6.5, 5%-6% glucose，6%-8% ethanol. The activity of amylase reached the highest 57.35 U/mL under the conditions of 30, pH 5.5 and 6% sugar, while the activity of cellulase reached the highest 21.84 U/mL. The activity of pectinase reached a maximum of 49.23 U/mL under the conditions of 30, pH 4.5 and 6% sugar. The activity of protease reached the highest 98.62 U/mL under the conditions of 30, pH 6.5 and 3% sugar. The activity of the enzyme activity reached the highest 26.67 U/mL under the conditions of 35, pH 4.5 and 6% sugar.
Key words: Songhe Daqu; Bacillus; Isolation and identification; Preponderant strain; Functional properties
目 录
摘要..1
引言..2
1 材料与方法 2
1.1 材料 2
1.1.1 试材 2
1.1.2 主要培养基 2
1.2 主要仪器与设备 3
1.3 主要方法 4
1.3.1 大曲芽孢菌的富集培养与分离 4
1.3.2 大曲芽孢菌的鉴定 4
1.3.2.4 种属鉴定 6
1.3.3 功能特性研究 6
1.3.4 数量优势芽孢菌的生长特性研究 7
1.3.5 数量优势芽孢菌功能特性的研究 7
1.3.6 产酶活力测定 8
2 结果与分析 8
2.1 芽孢菌的分离与鉴定 8
2.1.1 形态学特征 8
2.2 生理生化鉴定 10
2.3 分子生物学鉴定 10
2.3 功能特性 11
2.3.1 产淀粉酶功能 11
2.3.2 产纤文素酶功能 11
2.3.3 产蛋白酶功能 12
2.3.4 产果胶酶功能更 12
2.3.5 产酯化酶鉴定 12
2.4 优势菌株的功能特性研究 13
参考上述实验结果，最终选定数量优势菌株Sp1进行功能特性研究。 13
2.4.1 标准曲线 13
2.4.2 温度对Sp1菌株生长状况及产酶能力的影响 14
2.4.3 pH对Sp1菌株生长状况及功能特性的影响 17
2.4.4 糖度对Sp1菌株生长状况及功能特性的影响 21