枯草芽孢杆菌培养基的优化研究

毕业论文

课 题 名 称 枯草芽孢杆菌培养基的优化研究

2014年5月18日

邵阳学院2014届毕业设计（论文）

目 录

摘要 ..................................................................................................................... Ⅰ Abstract ············································································································ Ⅱ 1 前 言 ············································································································· 1 2 材料与方法 ···································································································· 3 2.1 材料以及试剂 ················································································································ 3 2.2 实验主要仪器设备 ······································································································· 4 2.3 试验方法 ·························································································································· 4 3 结果与分析 ···································································································· 6 3.1 单因素试验结果 ··········································································································· 7 3.2 正交试验结果 ··························································································· 11 4 结论及讨论 ·································································································· 13 参考文献 ········································································································· 15 致谢 ················································································································· 16

邵阳学院2014届毕业设计（论文）

枯草芽孢杆菌培养基的优化研究

摘要

本文选取BPG液体培养基研究该培养基配方对枯草芽孢杆菌生长的影响，通过相关资料的查询，选取葡萄糖、蛋白胨、牛肉膏、磷酸二氢钾这四个因素变量。先通过单因素实验确定这四个因素的最适含量，再进行L9(3)正交试验确定BPG液体培养最适培养基组合。最终得到最佳培养基配方含量是牛肉膏4.0g，蛋白胨3.8g，NaCl

4

3.3g，葡萄糖4.0g，淀粉3.3g，KH2PO4 3.4g，MnSO4·7H2O 0.2g，蒸馏水

1000mL，pH7.0，其枯草芽孢杆菌生长量为3.631×107 CFU/mL。 关键词：枯草芽孢杆菌；葡萄糖；蛋白胨；牛肉膏；磷酸二氢钾；正交实验

I

邵阳学院2014届毕业设计（论文）

Bacillus subtilis culture medium optimization research

Abstract

This article selected BPG liquid medium to study the culture medium formula of the effect on bacillus subtilis growth, through the relevant information query, glucose, peptone and beef extract, potassium dihydrogen phosphate this four factors variables. First determined by single factor experiment, the optimum content of these four factors, then L9 (34) orthogonal test to determine the BPG liquid culture combination optimal medium. Finally it is to know the best culture medium formula content is 0.40 g of glucose, peptone 0.38 g, 0.36 g beef paste, potassium dihydrogen phosphate 0.34 g.

Keywords: Glucose, peptone and beef extract, potassium dihydrogen

phosphate, single factor experiment, orthogonal experiment

II

邵阳学院2014届毕业设计（论文）

1前言

芽孢杆菌是人类最早发现的细菌之一[1]。枯草芽孢杆菌（Bacillus subtilis），为芽孢杆菌属的革兰氏阳性菌，芽孢大小为0.6～0.9×1.0～1.5 μm，外形为椭圆或柱状，一般处在菌体中央部位，形成芽孢后细菌体不会膨胀。菌落外观粗劣并且不通透，颜色为白色和微黄色，在液体培养基中生长时，会形成皱褶，生长需要氧气。

枯草芽孢杆菌是一种重要的α-淀粉酶的生产细菌，它能合成α-淀粉酶及多种酶类，与消化道和动物体内的消化酶类共同发挥作用。它在成长的过程中产生的枯草菌素、多粘菌素等活性物质，对致病菌和内源性感染的致病菌有着非常显著的抑制作用，而且作为好氧菌，可以迅速消耗肠道环境中的氧气，引起肠道缺氧，促进细菌的生长，并产生乳酸和其他有机酸，降低肠道pH值，对其他病原菌的生长起到不可忽视的作用[2]。同时枯草芽孢杆菌是一种多功能、高效、安全和极具开发潜力的微生物菌种，已经广泛应用于各类科研领域，跟随着科研水平的提高和经济的发展，枯草芽孢杆菌与人们的日常生活越来越密切，随之而来的就是对枯草芽孢杆菌越来越多的培养研究，为了在工业生产中提高枯草芽孢杆菌的产量和降低生产成本，因此我们需要研究出更好的枯草芽孢杆菌培养基[3]。

到目前为止，对枯草芽孢杆菌的国内外研究不在少数。1945年约翰逊报告了枯

草芽孢杆菌防治植物病害的控制作用。此后，枯草芽孢杆菌制备的生防制剂用来防治植物病害的研究成为了国内外研究的热门话题。1980年Papavizas G C报导，枯草芽孢杆菌可以用来防治水稻等作物受多种真菌的病害[4]。1992年Hwang等报导，枯草芽孢杆菌可以防治豌豆的根腐病[5]。20世纪90年代后，国外已有了多种枯草芽孢杆菌制剂投入市场。美国Agraquest公司用枯草芽孢杆菌QST 713菌株和QST

2808菌株分别研制出活菌杀菌剂SerenadeTM和Souata AS，已在美国登记使用，

叶面喷施能防止蔬菜，樱桃，葡萄，葫芦和核桃，霜霉病，白粉病，灰霉病，细菌和真菌疾病。GBO3和MBI 600分别由古斯塔夫森和微生物公司美国发展公司，根施或种子处理可以防止霉菌，镰刀菌，链格孢和丝核菌引起的大豆，小麦，棉花和花生根腐病。2001古斯塔夫森将淀粉芽孢杆菌和枯草芽孢杆菌的生物防治剂混合在一起，称为BioYield，解淀粉芽孢杆菌变种植物生长促进剂taensa商品生产，商品

taegrotm，用于灌木、树苗和装饰植物根部的室内培养，用来防治根腐病和枯萎病[6]。

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