WO2012051745A1 - 一种蛋白核小球藻破壁提取生物活性物质的方法 - Google Patents

一种蛋白核小球藻破壁提取生物活性物质的方法 Download PDF

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Publication number
WO2012051745A1
WO2012051745A1 PCT/CN2010/002172 CN2010002172W WO2012051745A1 WO 2012051745 A1 WO2012051745 A1 WO 2012051745A1 CN 2010002172 W CN2010002172 W CN 2010002172W WO 2012051745 A1 WO2012051745 A1 WO 2012051745A1
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nucleus
frozen block
chlorella pyrenoidosa
bioactive substances
chlorella
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PCT/CN2010/002172
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English (en)
French (fr)
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汤永强
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Tang Yongqiang
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Publication of WO2012051745A1 publication Critical patent/WO2012051745A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/02—Algae
    • A61K36/05—Chlorophycota or chlorophyta (green algae), e.g. Chlorella
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/10—Preparation or pretreatment of starting material
    • A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding

Definitions

  • the invention relates to the field of biotechnology, in particular to a method for extracting biologically active substances by breaking the wall of the nucleus. ⁇ Background technique ⁇
  • Chlorella is a high-energy plant that is rich in nutrients such as amino acids, proteins, and vitamins. It contains a special bioactive growth factor C.GF (Chlorella Growth Facter), which has a high utilization value.
  • C.GF Chlorella Growth Facter
  • the present invention is directed to the above-mentioned drawbacks of the prior art, and provides a method for extracting a biologically active substance by nucleus nucleus to extract a biologically active substance in chlorella.
  • a method for extracting a biologically active substance by a nucleus nucleus comprising the following steps: A. placing a bacterium of the nucleus nucleus Freezing at a temperature of minus 20 to 40 ° C, then placing the frozen frozen block into a container, and rapidly extruding the frozen block by physical extrusion to instantly break and dissolve the frozen block; B. The broken wall liquid is placed in a stirrer to stir; (When the temperature is 0 ⁇ 5 ° C, the membrane breaking technology is used to enrich the broken wall liquid to obtain a biological active substance.
  • the frozen block is extruded using the gravity of 100 to 150 kilograms of the object.
  • the present invention freezes the living body of Chlorella pyrenoidosa, so that the cytosol in the chlorella chlorella is frozen and expanded to form a squeeze on the cell wall, and then the physical block is used to make the frozen block. Rapid rupture, the combination of these two means can fully rupture the cell wall of chlorella, allowing cytosol to flow out, thereby effectively extracting biologically active substances in the cytosol of Chlorella pyrenoidosa.
  • the whole extraction process of the present invention is carried out by physical methods at a low temperature, and does not cause any chemical changes and influences on the biologically active substance, and fully ensures the activity of the active substance.
  • the method for extracting a biologically active substance from the nucleus of the nucleus of the present invention comprises the following steps:
  • Example 1 First, the living nucleus of Chlorella chlorella is frozen at minus 3 (TC temperature to obtain a frozen block, and then the frozen block is placed in a container, and the frozen block is quickly pressed by physical pressure to instantly break the frozen block. Because the frozen block is subjected to rapid impact, the frozen block is dissolved by heat to obtain a broken wall liquid; the obtained broken wall liquid is placed in a high-speed stirrer to fully overflow the cytosol in the cell wall of Chlorella pyrenoid; then the temperature is 2 ° C In the environment, the membrane-separating technique is used to enrich the agitated wall-breaking liquid to obtain a biologically active substance. The membrane separation technique is a conventional prior art and will not be described in detail herein.
  • Example 2 First, the living nucleus of the chlorella nucleus is frozen at a temperature of minus 20 ° C to obtain a frozen block, and then the frozen block is placed in a container, and the frozen block is quickly squeezed by the gravity of 150 kg of the object. Instantly break the frozen block, because the frozen block is fast Impact, the frozen block is dissolved by heat to obtain a broken wall liquid; the obtained broken wall liquid is placed in a high-speed stirrer to fully overflow the cytosol in the cell wall of Chlorella pyrenoid; then, in the environment of temperature °C, The membrane separation technique enriches the agitated liquid after the agitation to obtain a biologically active substance.
  • Example 3 First, the living bacterium of Chlorella chlorella is frozen at a temperature of minus 40 ° C to obtain a frozen block, and then the frozen block is placed in a container, and the frozen block is quickly squeezed by gravity of 100 kg of the object to make water The frozen block is instantly broken and dissolved to obtain a broken wall liquid; the broken liquid liquid is used to fully overflow the cytosol in the cell wall of Chlorella chlorella, and then the membrane is separated by a membrane separation technique at a temperature of 5 ° C. Afterwards, the broken wall liquid is enriched to obtain a biologically active substance.
  • the bioactive substance obtained above can be added as an additive to a drug, or can be used as a drug for epidermal growth, and can also be used as a neuromodulator and an immune-regulating nutrient for health care, and has a very high application value.
  • the above-mentioned living bacterium of Chlorella pyrenoidosa refers to a slurry obtained by separating the living body of the nucleus nucleus and the culture solution. Since the chlorella leaves the culture and cannot survive, it is necessary to retain a certain amount of the culture solution, so that the mixture in which the chlorella is separated from the culture solution is a slurry.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biotechnology (AREA)
  • Immunology (AREA)
  • Dermatology (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Botany (AREA)
  • Medical Informatics (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Biomedical Technology (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Description

说 明 书 一种蛋白核小球藻破壁提取生物活性物质的方法
【技术领域】
本发明涉及生物技术领域，尤其涉及一种蛋白核小球藻破壁提取 生物活性物质的方法。 【背景技术】
小球藻是一种高能量植物，其含有丰富的氨基酸、蛋白质及维生 素等营养物质， 尤其含有特有生物活性生长因子 C.GF ( Chlorella Growth Facter ), 具有艮高的利用价值。
但是， 目前还没有能够有效地提取小球藻中生物活性物质的方 法。
【发明内容】
本发明针对现有技术的上述缺陷，提供一种蛋白核小球藻破壁提 取生物活性物质的方法， 以提取小球藻内生物活性物质。
为了解决上述技术问题， 本发明是通i以下技术方案实现的： 一种蛋白核小球藻破壁提取生物活性物质的方法，其包括以下步 骤： A、将蛋白核小球藻活体浆液置入零下 20〜40°C温度冷冻， 然后 将冷冻得到的冰冻块放入容器内，釆用物理挤压方法快速对冰冻块进 行挤压， 使冰冻块瞬间破碎和溶解； B、 将挤压得到的破壁液放入 搅拌器中搅拌； （、 在温度为 0~5°C的环境下， 采用膜分离技术将破 壁液富集得到生物活性物质。 优选地， 步骤 A中采用 100〜150千克物体的重力对冰冻块进行 挤压。
从以上技术方案可以看出， 本发明对蛋白核小球藻活体进行水 冻，使蛋白核小球藻内的胞液冷冻膨胀对胞壁形成挤压，再釆用物理 挤压方法使冰冻块快速破裂，这两种手段结合后能够充分使小球藻的 胞壁破裂，使胞液流出， 从而有效提取蛋白核小球藻胞液内的生物活 性物质。 本发明整个提取过程均在低温下采用物理方法进行， 不会对 生物活性物质造成任何化学变化和影响， 充分保证活性物质的活性。
【具体实施方式】
下面结合具体实施方式对本发明作进一步详细的说明。
本发明蛋白核小球藻破壁提取生物活性物质的方法包括以下步 骤：
实施例一： 首先将蛋白核小球藻活体浆液在零下 3(TC温度下进 行冷冻得到冰冻块，然后将冰冻块放入容器中采用物理压力快速对水 冻块挤压， 使冰冻块瞬间破碎， 由于冰冻块受到快速沖击， 冰冻块受 热溶解得到破壁液；将得到的破壁液放入高速搅拌器使蛋白核小球藻 细胞壁内的胞液充分溢出； 然后在温度为 2°C的环境下， 采用膜分离 技术将搅拌过后的破壁液富集得到生物活性物质。其中的膜分离技术 为常规的现有技术， 这里不作详细介绍。
实施例二： 首先将蛋白核小球藻活体浆液在零下 20°C温度下进 行冷冻得到冰冻块，然后将冰冻块放入容器中釆用 150千克物体的重 力快速对水冻块进行挤压，使冰冻块瞬间破碎， 由于水冻块受到快速 沖击，冰冻块受热溶解得到破壁液； 将得到的破壁液放入高速搅拌器 使蛋白核小球藻细胞壁内的胞液充分溢出； 然后在温度为 o°c的环境 下， 釆用膜分离技术将搅拌过后的破壁液富集得到生物活性物质。
实施例三： 首先将蛋白核小球藻活体浆液在零下 40°C温度下进 行冷冻得到冰冻块，然后将冰冻块放入容器中采用 100千克物体的重 力快速对冰冻块进行挤压，使水冻块瞬间破碎和溶解得到破壁液； 将 破壁液^高速搅拌器使蛋白核小球藻细胞壁内的胞液充分溢出；然 后在温度为 5°C的环境下， 采用膜分离技术将搅拌过后的破壁液富集 得到生物活性物质。
上述得到的生物活性物质可作为添加剂添加到药物当中，也可作 为表皮速生长的药物，还可作为神经调节剂和保健用的免疫力调节营 养液， 具有非常高的应用价值。
上述蛋白核小球藻活体浆液是指蛋白核小球藻活体与培养液分 离后得到的浆液。 因为小球藻离开培养^不能存活， 所以必须存留 一定量的培养液， 因此称小球藻活体与培养液分离后的混合物为浆 液。
以上所述实施例仅表达了本发明的几种实施方式，其描述较为具 体和详细，但并不能因此而理解为对本发明专利范围的限制。应当指 出的是，对于本领域的普通技术人员来说， 在不脱离本发明构思的前 提下， 还可以做出若干变形和改进， 这些都属于本发明的保护范围。 因此， 本发明专利的保护范围应以所附权利要求为准。

Claims

权利要求书
1、 一种蛋白核小球藻破壁提取生物活性物质的方法， 其特征在 于， 包括以下步骤：
A、 将蛋白核小球藻活体浆液置入零下 20~40°C温度冷冻， 然后 将冷冻得到的水冻块放入容器内，采用物理挤压方法快速对冰冻块进 行挤压， 使冰冻块瞬间块破碎和溶解得到破壁液；
B、 将挤压得到的破壁^ ^搅拌器中搅拌；
C、 在温度为 0~5°C的环境下， 采用膜分离技术将破壁液富集得 到生物活性物质。
2、 根据权利要求 1所述的蛋白核小球藻破壁提取生物活性物质 的方法， 其特征在于， 步骤 A中釆用 100〜150千克物体的重力对冰 冻块进行挤压。
PCT/CN2010/002172 2010-10-20 2010-12-27 一种蛋白核小球藻破壁提取生物活性物质的方法 WO2012051745A1 (zh)

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CN102304168A (zh) * 2011-08-17 2012-01-04 华南理工大学 小球藻蛋白质的低温超高压制备方法
JPWO2013105617A1 (ja) * 2012-01-11 2015-05-11 株式会社サン・クロレラ 血清レジスチン濃度低下のためのクロレラ細胞壁破砕物の使用
CN106821851B (zh) * 2017-02-20 2019-08-02 西安科艺诗生物技术有限公司 一种白梅花的鲜加工方法
CN107007646A (zh) * 2017-03-14 2017-08-04 郑笃达 一种辅助防治癌症的营养复合物
CN109480147A (zh) * 2018-11-27 2019-03-19 北海生巴达生物科技有限公司 利用浓缩及澄清技术制备饮料或螺旋藻生长因子粉的工艺
CN112110996A (zh) * 2020-10-29 2020-12-22 青岛科海生物有限公司 一种蛋白核小球藻中提取小球藻生长因子的方法

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JP2009057397A (ja) * 2007-08-29 2009-03-19 Sanyo Chem Ind Ltd 難接着基材用ホットメルト接着剤

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