摘要:乙醛脱氢酶(Aldehyde Dehydrogenase,ALDH)是催化乙醛氧化为乙酸的反应的酶。是人体内代谢的关键酶之一。肝中的乙醇脱氢酶负责将乙醇氧化为乙醛,生成的乙醛在乙醛脱氢酶催化下转变为乙酸,最终分解为二氧化碳和水。已知人类的乙醛脱氢酶由三个基因所编码:ALDH1A1、ALDH2及最近发现的ALDH1B1(亦称ALDH5)。从动物、植物体内能够分离得到乙醛脱氢酶,但其过程复杂成本高,并且提取量少。微生物有培养周期短、易于培养、繁殖快等优点,所以从微生物中提乙醛脱氢酶有较大的优势。62132
本实验通过研究了酿酒酵母发酵法产乙醛脱氢酶。利用单因素法和正交发对发酵条件进行优化,实验结果显示:得出最优培养基为:10g/L葡萄糖,10g/L酵母浸膏,MgSO4 3g/L,温度30℃、Ph6.0、接种量8%。
毕业论文关键词:乙醛脱氢酶;发酵条件;工艺优化
Aldehyde dehydrogenase fermentation process optimization Summary:Aldehyde dehydrogenase (Aldehyde Dehydrogenase, ALDH) is a key enzyme in the human body metabolism. Responsible for catalyzing the oxidation of acetaldehyde to acetic acid reaction, liver alcohol dehydrogenase is responsible for ethanol (alcohol component) is oxidized to acetaldehyde, the resulting acetaldehyde as a substrate in the further converted into harmless aldehyde dehydrogenase catalyzed acetic acid (vinegar that is a component). CH3CHO + NAD + CoA → acetyl-CoA + NADH + H known human aldehyde dehydrogenase encoded by the three genes: ALDH1A1, ALDH2 and ALDH1B1 recently discovered (also known ALDH5).In this study, single-factor method and steamed dumplings made experiments,Vinegar aceti yield aldehyde dehydrogenase fermentation conditions were optimized obtain the optimal culture conditions are as follows: 10g / L glucose, 10g / L yeast extract, temperature 30 ℃, pH5.0 , inoculum 5%, liquid volume 100 mL.
Keywords: aldehyde dehydrogenase; fermentation conditions; process optimization
1绪论..1
1.1 乙醛脱氢酶的概述..1
1.1.1 乙醛脱氢酶的分类1
1.1.2 乙醛脱氢酶的结构与性质1
1.2乙醛脱氢酶与疾病的防治...2
1.2.1 乙醛脱氢酶与肿瘤... 2
1.2.2 乙醛脱氢酶与心血管疾病2
1.2.3 乙醛脱4氢酶与神经系统3
1.2.4 乙醛脱氢酶与酒精中毒3
1.2.5 乙醛脱氢酶与白血病3
1.3 乙醛脱氢酶的研究进展.. 3
1.4 乙醛脱氢酶分离现状...4
1.4.1 盐析沉淀法.4
1.4.2 凝胶层析法.4
1.4.3 亲和层析法.5
1.4.4 有机溶剂沉淀法.5
1.4.5 双水相萃取法.5
1.5 本课题的研究目的、意义和内容6
2材料与方法....7
2.1 试剂与材料.7
2.1.1 试剂.....7
2.1.2 设备仪器.....8
2.2 实验方法....8
2.2.1 培养基的配置.8
2.2.2 种子液培养.....8
2.2.3 菌体量的测定方法..8
2.2.4 生长曲线的测定..9
2.2.5 酵母菌菌体培养及保存.....9
2.2.6 菌浓度的测定..9
2.2.7 NADH标准曲线绘制..9
2.2.8 乙醛脱氢酶酶活测定.10
2.3 实验步骤10
2.3.1 单因素法优化菌种发酵培养基.11
2.3.2 单因素法优化菌种发酵条件.12