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摘 要：以实验室保藏的从宋河酒曲中分离出的产酯酵母、产酯霉菌、产酯芽孢菌、乳酸菌为复配菌种，探索酿造酒酯化酶复合菌剂的制备条件。采用透明圈比值法筛选产酯化酶酵母菌、霉菌以及芽孢菌。以酯化酶活性为参考指标，选择混菌比例、接种量、初始pH值以及生长温度为影响因素，通过产酯化酶多元菌株混合发酵产酯酶试验和正交试验确定最优混合固态产酯酶发酵条件。结果表明：单菌、二元菌、三元菌发酵时酯化酶活力最高分别为16.72 U/mL、18.97 U/mL、13.28 U/mL。选取二元菌作为混合产酯发酵菌种，优化发酵条件为芽孢菌和酵母菌混菌比例为4:1、混合菌种接种量为4%、发酵温度为26℃、初始pH值为4.0，酶活力达到最高为22.38 U/mL。7537
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关键词：酯化酶；复合菌剂；培养条件
The Exploration of Productive High-yielding Ester Enzyme
by Compound Microbial Agent
Abstract: Taking the ester-producing yeast, mold, bacillus and lactic acid bacteria isolated from Songhe Daqu as trail materials, the producing esterified enzyme ability was studied by multivariate mixed strains during fermenting. The yeast, mold and bacillus were screened by the enzyme activity with the method of transparent circle ratio. The esterifying enzyme activity as a reference index, select the proportions of fungus, inoculation quantity, initial pH value and growth temperature as the influence factors, the optimal fermentation conditions of mixed solid producing esterase was determined by producing esterifying enzyme multi strain mixed fermentation esterase test and orthogonal test. The results showed that the highest fermentation esterifying enzyme activity of single bacteria, binary, ternary bacteria was 16.72 U/mL, 16.72 U/mL, 13.28 U/mL respectively. Select binary bacterium, the optimal fermentation conditions for spore bacteria and yeast mixed bacteria ratio was 4:1, inoculation amount was 4%, the growth temperature was 26 ℃ and initial pH value was 4.0, the highest enzyme activity reached 22.38 U/mL.
Key Words: Esterifying enzyme；Composite agents；Culture conditions目 录
摘 要 1
Abstract 1
引言 2
1．材料与方法 2
1.1 材料 2
1.1.1 菌株来源 2
1.1.2 主要培养基 2
1.2 主要仪器与设备 3
1.3 主要实验方法 3
1.3.1 菌株的活化 3
1.3.1.1 芽孢菌的活化 3
1.3.1.2 酵母菌的活化 4
1.3.1.3 霉菌的活化 4
1.3.1.4 乳酸菌的活化 4
1.3.2 高产酯化酶菌种的筛选 4
1.3.2.1 产酯芽孢菌的筛选 4
1.3.2.2 酵母菌的筛选 4
1.3.2.3 霉菌的筛选 4
1.3.3 种子菌悬液的制备 5
1.3.4 产酯化酶多元菌株混合发酵产酯酶试验 5
1.3.4.1 最佳二元菌混菌比例的确定 5
1.3.4.2 最佳三元菌混合比例的确定 5
1.3.4.3 最佳混菌发酵接种量的确定 5
1.3.4.4 最佳混菌发酵初始pH值的确定 5
1.3.4.5 最佳混菌发酵生长温度的确定 5
1.4 多元菌混合发酵条件的优化 6
1.5 酯化酶活力的测定 6
1.5.1 粗酶液的制备 6
1.5.2 酯化酶活力测定 6
2．结果与分析 6
2.1 酯化酶的分离与筛选 6
2.1.1 芽孢菌酯化酶的分离与筛选 6
2.1.2 酵母菌酯化酶的分离与筛选 7