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Method for producing catalpol by utilizing suspension-cultured rehmannia glutinosa cambium stem cells

A technology of suspension culture and cambium, applied in the field of medical bioengineering, can solve the problems of inability to stabilize active secondary metabolites for a long time and low content of catalpol in dedifferentiated cells of Rehmannia glutinosa, to overcome changes in physiological and biochemical characteristics, strong shear resistance Effects of shearing force and genetic information stabilization

Active Publication Date: 2018-05-15
XINXIANG MEDICAL UNIV
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

The catalpol content of rehmannia dedifferentiated cells obtained by existing plant cell culture technology is low, and cannot be used for long-term stable industrial production of active secondary metabolites

Method used

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  • Method for producing catalpol by utilizing suspension-cultured rehmannia glutinosa cambium stem cells
  • Method for producing catalpol by utilizing suspension-cultured rehmannia glutinosa cambium stem cells

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preparation example Construction

[0043] The preparation process of the MS liquid medium and the MS solid medium is the same as the preparation process of the above-mentioned WPM liquid medium and WPM solid medium.

[0044] The formula of table 3 anti-browning preservation solution

[0045]

[0046]

[0047] Below we will describe and illustrate the technical solutions of the present invention in detail in the form of specific examples.

Embodiment 1

[0049] In this embodiment, a method for producing catalpol using suspension-cultured rhizoma rhizome cambium stem cells specifically comprises the following steps:

[0050] S1: Obtaining the culture system of rehmannia cambium stem cells

[0051] In the spring morning, collect the newly grown young stems of the three-year-old wild Rehmannia genus Rehmannia piasezkii Maxim, cut them into 3-5 cm sections, rinse them with flowing sterile distilled water for 6 hours, dry the surface moisture, cut into 1 cm of xylem, phloem, and cambium containing vascular bundles 2 Small pieces of as explants;

[0052] Under sterile conditions, rinse the explants with flowing sterile distilled water continuously for 2 hours, then soak them in 70% ethanol solution for 2.0 minutes, transfer them to 0.2% sodium hypochlorite solution for 5 minutes, and then sterilize them with 0.01% mercuric chloride solution. Bacteria treatment for 2 minutes, and finally rinsed with flowing sterilized distilled wat...

Embodiment 2

[0064] In this embodiment, a method for producing catalpol using the stem cells of the rhizome of Rehmannia glutinosa stem cells cultured in suspension, the acquisition of the stem cell culture system of the rhizoma rehmanniae root is the same as in Example 1, the only difference being:

[0065] At the 8th day of culture, 120 mmol / L methyl-β-cyclodextrin and 120 μmol / L methyl jasmonate were added to the bioreactor to stimulate the accumulation of catalpol, and the co-culture time was 16 days. The final obtained catalpol pure product is 16.18g.

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Abstract

The invention discloses a method for producing catalpol by utilizing suspension-cultured rehmannia glutinosa cambium stem cells. The method specifically comprises the following steps: taking rehmanniaglutinosa plant stems as explants and establishing a rehmannia glutinosa cambium stem cell culture system capable of stable growth through induced culture, isolation culture and subculture; then adding a combined inductor beta-CD and methyl jasmonate in a bioreactor for suspension culture and refining through ultrasound extraction, macro-porous resin absorption and a high-performance liquid-phasepreparation column to obtain the high-purity hinokitiol. The method disclosed by the invention has the benefits that the utilized plant stem cell culture system has the excellent characteristics of synchronous growth, stable genetic character, high growth speed, stable catalpol synthesis and the like and provides a novel method for catalpol preparation as a raw material for catalpol preparation;in addition, compared with a chemical synthesis method, the method for producing the catalpol by utilizing a bioreactor culture technology combined with a plant stem cell culture technology has the advantages of being short in production period, high in safety, low in cost, high in yield, low in pollution and the like.

Description

technical field [0001] The invention belongs to the technical field of medical bioengineering, and in particular relates to a method for producing catalpol by using suspension cultured rehmannia plant stem cambium stem cells. Background technique [0002] Rehmannia glutinosa is one of the most famous Chinese herbal medicines in my country. According to statistics, there are more than 80 kinds of Chinese patent medicines that use raw rehmannia glutinosa as the main raw material, such as Liuwei Dihuang Wan, Zhibai Dihuang Wan, Wuji Baifeng Wan, Xiaotang Wan, etc. Catalpol is one of the main active ingredients in Rehmannia glutinosa, which has a wide range of pharmacological activities, such as hypoglycemic, antitumor, anti-inflammatory, antiviral, neuroprotective, etc. On January 25, 2017, "Calbinol Tablets" independently developed by Qinghai Yangzong Pharmaceutical Co., Ltd. obtained the approval for clinical trials of Class I traditional Chinese medicines from the State Food...

Claims

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Application Information

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IPC IPC(8): C12P19/60C12N5/04
CPCC12N5/04C12P19/60
Inventor 周鹏飞刘友才张伟李海华闫思涵谢永生张毅敏史世强
Owner XINXIANG MEDICAL UNIV
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