CN1047695A - 一种营养液的生产方法 - Google Patents

一种营养液的生产方法 Download PDF

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CN1047695A
CN1047695A CN89104728A CN89104728A CN1047695A CN 1047695 A CN1047695 A CN 1047695A CN 89104728 A CN89104728 A CN 89104728A CN 89104728 A CN89104728 A CN 89104728A CN 1047695 A CN1047695 A CN 1047695A
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Abstract

应用一株命名为YZH菌的菌株，用该株菌作为生产菌株，在本发明所设计和使用的培养基中培养，可分别生产出富含多糖、多肽、双糖、低聚糖、Y因子、多种微量元素及多种维生素的单细胞蛋白的营养液，该营养液对人类不仅具有健全人体免疫、增强代谢的功能，而且对癌细胞有明显的抑制与杀伤作用，这对于防治人类癌症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，特别是关于利用假单孢菌属的一种命名为YZH菌的菌株作为生产菌株进行工业发酵来生产单细胞蛋白营养液的方法。
在当今世界，人们通过各种尝试采用生物发酵方法生产单细胞蛋白取得成功。杨振华利用具有特殊功能的851R菌及设计出适合于该菌生长的以大豆为主要原料的培养基，运用常规的工业发酵的生产方法生产出851R型超级营养液。大豆是蛋白质丰富的食物，但由于大豆中含有蛋白酶抑制剂，所以大豆蛋白很难被人体全部吸收，然而，大豆蛋白经过851R菌利用后，转化为单细胞蛋白营养液，不但成为易被人体吸收的蛋白源，而且该营养液具有健全中枢神经系统、提高机体免疫功能、调节内分泌的功效，该营养液对癌细胞的生长有某些抑制作用。（中国发明专利申请号        88107615.5，1988年10月31日）。
本发明的目的是通过具有特殊功能的YZH菌使用适合于该菌生长的含氮源培养基，以达到用YZH菌作为生产菌株，运用常规的工业发酵的生产方法生产出Y型营养液，以及运用多种离子交换树脂、有机溶剂提取浓缩发酵液中有效营养成分。用YZH菌生产出的Y型营养液不仅具有健全中枢神经系统、提高机体免疫功能、调节内分泌的功效，而且对癌细胞有明显的抑制与杀伤作用，甚至还能使癌细胞逆转为正常细胞。
本发明所涉及的微生物是假单孢菌属变异菌YZH菌，其主要特征为：菌体极端鞭毛，鞭毛数1～2根，该菌体内含有YZH片段，可用已克隆的YZH片段运用DNA分子杂交技术可检测出该片段的存在（该菌种已送交武汉中国典型培养物保藏中心保藏，保藏号为CCTCC        No.M89049）。
在本发明所设计或使用的含氮源培养基中，用YZH菌作为生产菌种，可生产出富含多糖、多肽、双糖、低聚糖、Y因子、多种微量元素及维生素的单细胞蛋白营养液。该营养液中含有二十余种氨基酸。每100毫升这种营养液中含有：氨基酸350～800毫克，维生素E0.2～0.5mg，维生素B20.05～0.1mg，维生素PP0.7～1.0mg，硒0.04～0.08mg，锌0.2～0.5mg，钼0.08～0.24mg，钴0.08～0.24mg，锰0.05～0.06，铜0.07～0.2mg，铁0.1～0.7mg等。干物质1.21～1.38g，蛋白质1.21～2.2g，脂肪0.11～0.13g，脂肪酸中肉豆蔻7.1～8.3%，棕榈14.3～15.8%，硬脂74.4～75.5%，油酸0.1～0.8%。
本发明所设计及使用的培养基为：
1.含氮源培养基
大豆粉        5～10%
或豆浆（以大豆量计）        5～15%
或牛肉汤        5～15%
酵母膏        0.02～0.15%
或酵母粉        0.02～0.3%
或蛋白胨、牛肉膏        0.02～0.4%
碳酸钙        0.05～0.25%
磷酸氢二钾        0.02～0.1%
硫酸镁        0.01～0.05%
氯化钠        0.01～0.04%
钼酸钠        5.0～30ppm
亚硒酸钠        2.5～15ppm
硫酸锌        2.5～40ppm
氯化钴        5～20ppm
还可以增加其他人体必需的微量元素，其限度为1.5～100ppm，也可增加苯甲酸或苯甲酸钠。
2.1640培养基
培养细胞的1640培养基也适用于YZH菌的生长。
该菌种可以用一般发酵工业中的发酵罐进行通气培养，这种罐它带有搅拌和通气装置，可以依需要对所培养的培养物进行搅拌，使其所处的条件一致，也可以按需要通入经过滤的无菌空气，以满足细菌等微生物在发酵过程中对氧气的需要。将YZH菌作为生产菌株，经过在种子培养基（本发明所设计和或使用的培养基）中繁殖。即由小种、中种到大种的繁殖培养，再接种到装有预先经过消毒灭菌的生产用培养基（本发明所设计和/或使用的培养基）中培养24～72小时，在培养过程中通入过滤的无菌空气，通气量为1∶0.6～1.2，搅拌速度180～260转/分，培养温度28～38℃，培养结束后经高温灭菌收获培养液。培养液可直接分装而成产品，也可采用乙醇、醋酸乙酯、丙酮等有机溶剂及大孔树脂柱、717阴离子交换柱等树脂柱对发酵的培养液进行提取、萃取及吸附有效营养成分的方法制成Y型营养液的精萃。
本发明不限于以下实施例，其保护范围由权项要求限定。
实施例1
在5吨发酵罐中培养，种子罐为0.5吨，投料量为40%。采用10%大豆培养基，其中大豆含量为200kg（大豆先经过挑选，洗净，磨成浆，去渣），酵母膏800g，磷酸氢二钾1000g，硫酸镁400g，碳酸钙5kg，氯化钠400g，钼酸钠20g，亚硒酸钠5g，硫酸锌20g，氯化钴20g，及苯甲酸1kg。接菌量按1%，种子罐为0.5吨，投料为200kg，培养基如上所述量缩小10倍。接菌量1%，（小种→中种→大种，培养24小时作为种子培养物）培养温度30℃，搅拌速度220转/分，通气量1∶0.8。种子在种子罐中培养结束后接入5吨大罐，培养温度30℃，搅拌速度220转/分，通气量1∶0.8，培养48小时结束培养。终止培养后采用高温灭菌，培养液经流水线装瓶，再经高温高压灭菌即成为产品。每100毫升培养液中，干物质含量为2.2%蛋白质含量为1.38%，脂肪含量0.13%，碳水化合物为0.09%及维生素、无机盐等详见下表：
实施例二
用实施例一生产所得的培养液，经过离心去渣后再装瓶而成产品。
实施例三
用实施例二生产所得离心去渣后的培养液，通过薄膜过滤浓缩后再装瓶成产品。
实施例四
用实施例三生产所得的浓缩培养液，用95%乙醇提取、浓缩成精萃。
实施例五
用实施例一生产所得的培养液，用大孔树脂D4006吸附后用水洗脱，再经中性氧气铝柱浓缩成产品。
实施例六
Y型营养液的上清液对各种离体癌细胞均有较明显的抑制生长与杀伤作用，甚至对癌细胞有逆转作用，促使癌细胞变成正常细胞。例含20%的Y型营养液的上清液对人胃癌腺细胞MGC803培养12～24小时内出现抑制与杀伤效果，没有被杀伤的癌细胞继续培养7～9天内逐渐逆转为与正常细胞相似，见以下图片的说明。
图1-2相差显微镜活细胞观察，图1示MGC80-3细胞，图2为经15%"851"处理的MGC80-3细胞形态。
图3-6扫描电镜观察，示MGC80-3细胞边缘丝状伪足增多，表面具有丰富的微绒毛。
图7-10扫描电镜观察，示经20%"851"处理的MGC80-3细胞表面微绒毛消失，细胞边缘有片状伪足，并出现长胞质突起。一些细胞表面有皱痕状结构及少量小泡状突起。
图11-14透射电镜观察，示经20%"851"处理的MGC80-3细胞中可出现细胞核形规则（N），核仁（Nu）缩小，异染色质减少，常染色质增多，细胞质内高尔基体（G），线粒体（M）发达，粗糙型内质网（REG）增多等与正常细胞相似的显著变化。
图15-18透射电镜观察，示经20%"851"处理的MGC80-3细胞中可出现细胞核（N）内常染色质丰富（图15），细胞质内可见环形辫状体（图16）和正常分泌颗粒（图17SG），以及发达的粗糙型内质网（图18，RER）等与正常细胞相似的超显微结构特征。
Y型培养液（上清）检测结果报告表

Claims (2)

1、一种利用细菌生产单细胞蛋白营养液的工业生产方法，其特征是采用假单胞菌属(pseudomonas)的一株命名为YZH的菌株(CCTCCNO：89049)作为生产菌株，在本发明所设计的以大豆为主的含氮源培养基或本发明所使用的培养基中发酵培养生产培养液，还包括用多种浓缩法精制培养液的方法。
2、如权利要求1所说的以大豆为主得含氮源培养基，其特征在于含有大豆粉5～10%，或豆浆（以大豆量计）5～15%，或牛肉汤5～15%，酵母膏（或酵母粉、蛋白胨、牛肉膏）0.02～0.4%，碳酸钙0.05～0.25%，磷酸氢二钾0.02～0.1%，硫酸镁0.01～0.05%，氯化钠0.01～0.04%，钼酸钠5～30ppm，亚硒酸钠1.5～15PPM，硫酸锌2.5～40PPM，氯化钴5～20PPM，还可以加入适量人体所需的其他微量元素，其限度为1.5～100PPM及苯甲酸或苯甲酸钠。
CN89104728A 1989-07-17 1989-07-17 一种营养液的生产方法 Expired - Fee Related CN1023899C (zh)

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CN89104728A CN1023899C (zh) 1989-07-17 1989-07-17 一种营养液的生产方法
DE69013425T DE69013425T2 (de) 1989-07-17 1990-06-25 Pseudomonasmutant, yzh-Stamm und Verfahren zur Herstellung der 851yzh-Nährlösung mittels dieses Stammes.
EP90112043A EP0408933B1 (en) 1989-07-17 1990-06-25 A mutant of pseudomonas, a strain yzh, and a process for producing 851yzh nutrient solution by application of the strain
CA002020633A CA2020633C (en) 1989-07-17 1990-07-06 Mutant of pseudomonas, a strain yzh, and a process for producing 851 yzh nutrient solution by application of the strain
AU58929/90A AU631663B2 (en) 1989-07-17 1990-07-12 A mutant of pseudomonas, a strain yzh, and a process for producing 851 yzh nutrient solution by application of the strain
KR1019900010736A KR0151858B1 (ko) 1989-07-17 1990-07-16 슈도모나스 변이주 yzh 주를 이용하여 851 yzh 영양액을 제조하는 방법
JP2189252A JP2621100B2 (ja) 1989-07-17 1990-07-17 プソイドモナスの変異株ｙｚｈ株とこの株の応用により８５１ｙｚｈ栄養液の製造方法

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JP2600115B2 (ja) * 1994-11-28 1997-04-16 工業技術院長 脂肪族ポリカーボネート系樹脂の酵素的分解方法
US7070965B1 (en) 1998-04-14 2006-07-04 Zhenhua Yang Small molecule anticancer compounds and related production process
US6214875B1 (en) 1998-04-14 2001-04-10 Zhenhua Yang Anticancer effects of specific branched-chain fatty acids and related production process
CN112753513B (zh) * 2021-02-05 2022-12-23 湖北恩施中国南方马铃薯研究中心 一种富硒叶菜型甘薯的水培快繁方法
CN113615827A (zh) * 2021-07-08 2021-11-09 武汉万德瑞生物技术股份有限公司 一种具有抗癌功能的色氨酸缺乏组合食品及其制备方法

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CN86100490B (zh) * 1986-04-08 1987-09-30 相振华 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

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CA2020633C (en) 1997-08-19
DE69013425D1 (de) 1994-11-24
DE69013425T2 (de) 1995-12-14
EP0408933B1 (en) 1994-10-19
KR910003089A (ko) 1991-02-26
AU631663B2 (en) 1992-12-03
JPH04144678A (ja) 1992-05-19
JP2621100B2 (ja) 1997-06-18
EP0408933A3 (en) 1991-09-04
CA2020633A1 (en) 1991-01-18
KR0151858B1 (ko) 1998-10-01
AU5892990A (en) 1991-01-17
CN1023899C (zh) 1994-03-02
EP0408933A2 (en) 1991-01-23

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