CN86100490A - 利用抗氨固氮菌生产富硒单细胞蛋白，维生素e和菌肥的方法 - Google Patents

利用抗氨固氮菌生产富硒单细胞蛋白，维生素e和菌肥的方法 Download PDF

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CN86100490A
CN86100490A CN198686100490A CN86100490A CN86100490A CN 86100490 A CN86100490 A CN 86100490A CN 198686100490 A CN198686100490 A CN 198686100490A CN 86100490 A CN86100490 A CN 86100490A CN 86100490 A CN86100490 A CN 86100490A
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Abstract

应用人工诱变技术培育出一组抗氨固氮菌851黄(包括01，02和03三个菌株)。该组菌具有明显抗氨固氮能力，能强力固定大气氮，在有化合态氮的培养基中，仍保持相当高的固氮酶活性，用该组菌作为生产菌株，在本发明所设计的培养基(甘露醇—酵母膏培养基，甘露醇培养基，淀粉培养基和废糖蜜培养基)中培养，可分别生产富硒、锌、钙及多种维生素的单细胞蛋白，维生素E和菌肥，该菌培养液对人类防癌延寿具有进一步的开发利用价值。

Description

本发明是关于工业发酵的一种方法，特别是关于利用一组来自维氏固氮菌（Azotobacter    vinelandii）的人工诱变菌株-抗氨固氮菌：851黄作为工业发酵的生产菌株来进行生产的方法，该抗氨固氮菌能利用大气氮生产富硒、锌的单细胞蛋白、维生素和细菌肥料的方法。
目前世界各国科学家都在研究生物固氮问题，希望由自生固氮菌来帮助人类将空气中80%的氮直接变成蛋白质和肥料，以罢脱当今世界人口和食物的矛盾。由于自然界的固氮菌不抗氨，所以不具有推广利用的价值。
本发明是为培育出一种具有抗氮固氮能力的固氮菌，要求这种菌在培养基中或周围环境里虽有化合态形式的氮存在，仍具有相当高的固氮酶活性，能强力固定大气中的氮;设计出适合于该菌生长的培养基，以达到生产出有工业价值的富硒、锌和多种维生素的单细胞蛋白，维生素E，细菌肥料及其一系列产品的方法。
本发明所涉及的抗氨固氮菌是用遗传学技术，经过人工诱变处理使维氏固氮菌（Azotobacter    vinelandii）体内的谷氨酰胺合成酶基因发生突变，而培育出一组具有明显抗氨固氮能力、能强力固定大气氮的变异菌-851黄（包括01，02及03三个菌株）。该组菌与原亲代菌株最根本的区别是在含有氮的培养基中，甚至在有硫酸铵存在的情况下仍保持相当高的固氮酶活性。该组菌不仅是生产细菌肥料的良好菌株，而且其中的01菌株能直接利用淀粉，凡是含有淀粉的物质均可以作为01菌株生长之碳源，如甘薯、马铃薯、玉米等。在本发明所设计的淀粉培养基中，用01菌株作为生产菌种，可生产出富硒、锌和多种维生素的单细胞蛋白，其培养液中全氨基酸的含量为2～20mg/ml、硒1～5ppm、锌5～20ppm，还含有其它多种维生素;851黄02菌株适合生产富含维生素E或与维生素E同样效果的营养液，它用甘露醇作为碳源，在本发明所设计的甘露醇一醇母膏培养基或甘露醇培养基中培养，其每100毫升培养液中维生素E的含量为：8.2-114毫克（试验室结果）;851黄03菌株适于利用废糖蜜作为碳源，主要用于生产富含维生素C的单细胞蛋白，其培养基中全氨基酸的含量为2～5mg/ml。
本发明所设计的培养基为：
1，淀粉培养基
干淀粉    1-2%
含有淀粉的新鲜物质（甘薯，马铃薯等）    10-20%
或
含有淀粉的干物质（玉米粉等）    1-10%
酵母膏    0.04-0.08%
磷酸氢二钾    0.05-0.1%
硫酸镁    0.02-0.04%
氯化钠    0.02-0.04%
钼酸钠    10-20ppm
硼酸    10-20ppm
亚硒酸钠    5-20ppm
硫酸锌    5-20ppm
2，甘露醇培养基
甘露醇    1%
磷酸二氢钾    0.02%
硫酸镁    0.02%
氯化钠    0.02%
硫酸钙    0.02%
碳酸钙    0.5%
3，甘露醇-酵母膏培养基
甘露醇    1%
酵母膏    0.04%
磷酸氢二钾    0.05%
硫酸镁    0.02%
氯化钠    0.02%
钼酸钠    10ppm
硼酸    10ppm
4，废糖蜜培养基
废糖蜜    2-3%
碳酸钙    0.5%
钼酸钠    10ppm
硫酸亚铁    10ppm
本发明所提供的抗氨固氮菌，克服了固氮菌在有化合态氮存在的情况下，不能固定大气氮的问题。本抗氨固氮菌不仅是生产菌肥的良好菌株，而且还可作为生产富硒、富锌、多种维生素的单细胞蛋白及维生素E的工业生产菌株，进而开发更多的产品，用于制造鳗鱼、对虾、鸡、鸭、猪各种动物饲料添加剂，它不仅介决了饲料的营养，而且还能介决饲料的粘合问题;用于食品中作为添加剂，如饼干，软糖，面包，饮料，化妆品等。
例：以抗氨固氮菌851黄01菌株作为生产菌株，培养于淀粉培养基（玉米粉），在2.5吨发酵罐中培养36-110小时（半连续发酵），通气量1∶0.6-1.2，搅拌速度100-300转/分培养温度25-30C，培养结束，每100毫升培养液中全氨基酸量为0.5-1g、硒1-5ppm、锌5-20ppm、维生素C、E及B1、B2、B12、A、K、D及烟酸等。
用此方法生产的抗氨固氮菌培养液喂养果蝇，其寿命比对照组延长一倍以上;用菌液浇作物，植株的地上部份和根部比对照组生长得旺盛，且寿命延长;用培养液喂鸡，从5日龄小鸡开始饲喂，试验组鸡健康无病，生长快。该菌液中含有抗衰老的物质，对小白鼠体内肝抗脂质过氧化作用试验结果表明，01菌株培养液与维生素E有等同效果，而02菌株培养液抗衰老作用甚至超过维生素E;用01菌株培养液对小白鼠几种组织蛋白合成试验结果表明有较好的促进作用，其作用胜过红参，其中甘薯培养液对雌性小白鼠有明显减肥效果。01菌株和02菌株对小白鼠抗化学致癌试验表明有明显的防癌效果。由此可见本发明不仅在于发明了抗氨固氮菌，介决了抗氨固氮问题，而且在于该菌培养液对人类防癌延寿具有进一步开发利用价值。
本发明不限于此实施例，其保护范围由权项要求限定。

Claims (9)

1、一种利用固氮菌作为生产菌株进行工业发酵生产的新方法，其特征是该生产菌株是采用遗传学技术，人工诱变处理维氏固氮菌(Azotobacter  vine-landii)而得到的一组抗氨固氮菌，命名为851黄(包括01、02及03三个菌株)，该组菌具有明显的抗氨固氮能力，能强力固定大气氮；在含有化合态氮的培养基中(甚至在含有硫酸铵的培养基中)仍保持固氮酶活性，利用该组菌作为生产菌株在本发明所设计的不同培养基中培养，可由培养液中分别获得富硒、锌及多种维生素的单细胞蛋白、维生素E或与维生素E同样效果的营养液和菌肥，其培养液中还含有维生素C、B1、B2、B12、A、K、D及烟酸等，该培养液可供药品、食品、饲料、化妆品、粘合剂等二次开发利用。
2、如权利要求1所说的生产方法，其特征是采用人工诱变处理得到的能直接利用淀粉作为碳源的人工诱变处理得到的抗氨固氮菌851黄中的01菌株作为生产菌株，在本发明所设计的淀粉培养基中培养，其培养液含氨基酸、硒、锌，还含有其它多种维生素，主要用于生产单细胞蛋白和菌肥。
3、如权利要求1所说的生产方法，其特征是采用人工诱变处理得到的抗氨固氮菌851黄02菌株作为生产菌株，在本发明所设计的甘露醇-酵母膏培养基（或甘露醇培养基）中培养，主要用于富含维生素E或与维生素E同样效果的营养液的生产。
4、如权利要求1所说的生产方法，其特征是采用人工诱变处理得到的抗氨固氮菌851黄中的03菌株作为生产菌株，在本发明所设计的废糖蜜培养基中培养，其培养液主要用于生产富含维生素C的单细胞蛋白。
5、如权利要求1所说的生产方法，其特征是采用人工诱变处理得到的抗氨固氮菌851黄（包含01、02、03三个菌株）作为生产菌株生产的菌液在药品、食品、饲料、化妆品、粘合剂等方面的二次开发利用。
6、如权利要求2所说的淀粉培养基，其特征在于含有干淀粉1-2%，含有淀粉的新鲜物质（甘薯、马铃薯等）10-20%，或含淀粉的干物质1-10%，酵母膏0.04-0.08%，及无机盐（磷酸氢二钾、硫酸镁、氯化钠、钼酸钠、硼酸及亚硒酸钠、硫酸锌）。
7、如权利要求3所说的甘露醇酵母膏培养基，其特征在于含有甘露醇1%，酵母膏0.04%及无机盐（磷酸二氢钾、硫酸镁、氯化钠、钼酸钠及硼酸）。
8、如权利要求3所说的甘露醇培养基，其特征在于含有甘露醇1%及无机盐（磷酸二氢钾、硫酸镁、氯化钠、硫酸钙、碳酸钙）。
9、如权利要求4所说的废糖蜜培养基，其特征在于含有废糖蜜2-3%及无机盐（碳酸钙、钼酸钠及硫酸亚铁）。
CN86100490A 1986-04-08 1986-04-08 利用抗氨固氮菌生产富硒单细胞蛋白, 维生素e和菌肥的方法 Expired CN86100490B (zh)

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CN86100490A CN86100490B (zh) 1986-04-08 1986-04-08 利用抗氨固氮菌生产富硒单细胞蛋白, 维生素e和菌肥的方法
EP86114048A EP0241583A3 (en) 1986-04-08 1986-10-10 Antiammonia azotobacter and the use thereof
JP61247258A JPS62239987A (ja) 1986-04-08 1986-10-17 アンチアンモニア　アゾトバクター
US07/252,578 US4877739A (en) 1986-04-08 1988-09-30 Antiammonia azotobacter and the use thereof

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CN103342593A (zh) * 2013-07-05 2013-10-09 长沙学院 一种有机硒作物营养剂及其制备方法

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CN1023899C (zh) * 1989-07-17 1994-03-02 杨振华 一种营养液的生产方法
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CN103342593A (zh) * 2013-07-05 2013-10-09 长沙学院 一种有机硒作物营养剂及其制备方法
CN103342593B (zh) * 2013-07-05 2015-01-14 长沙学院 一种有机硒作物营养剂及其制备方法

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EP0241583A2 (en) 1987-10-21
JPS62239987A (ja) 1987-10-20

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