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Clitopilus prunulus preserving method

A technology of oblique mushroom and sodium alginate, which is applied in the field of bioengineering, can solve the problems of liquid paraffin being toxic to bacterial cells, bacterial species death, etc., and achieve the effects of promoting production and research and prolonging preservation time.

Active Publication Date: 2016-11-09
宁夏泰瑞制药股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] 1. Liquid paraffin has a toxic effect on the bacteria, and it is easy to cause the death of the bacteria

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] The method of embedding and preserving the purified water of Clitopilus prunulus is as follows:

[0018] 1) Collect mycelium:

[0019] Inoculate the mushroom Clitopilus prunulus, batch number TMI14-469 (the strain is provided by Ningxia Tairui Pharmaceutical Co., Ltd., seed room), in the liquid PDA medium, 24~26℃, 220~240r / min, shaking culture for 4d (At this time, Clitopilus prunulus is at the end of logarithmic growth phase). Take 30mL of the above seed solution in a 50mL sterile centrifuge tube, centrifuge at 3500r / min for 10min. The supernatant was discarded, and the precipitate was mycelia with a mass of 8-10 g.

[0020] 2) Preparation of embedding reagents:

[0021] Sodium alginate solution: 13.69% (w / v) sucrose + 18.4% (w / v) glycerin + 9% (w / v) sodium alginate, prepared in 50 mL.

[0022] Calcium chloride solution: sucrose 13.69% (w / v) + glycerol 18.4% (w / v) + calcium chloride 1.11% (w / v), prepare 1 bottle, 400mL.

[0023] The above two solutions were adjust...

Embodiment 2

[0033] The method of embedding and preserving the purified water of Clitopilus prunulus is as follows:

[0034] 1) Collect mycelium:

[0035] Inoculate the mushroom Clitopilus prunulus, batch number TMI14-469 (the strain is provided by Ningxia Tairui Pharmaceutical Co., Ltd., seed room), in the liquid PDA medium, 24~26℃, 220~240r / min, shaking culture for 4d (At this time, Clitopilus prunulus is at the end of logarithmic growth phase). Take 28mL of the above seed solution in a 50mL sterile centrifuge tube, centrifuge at 3000r / min for 10min. The supernatant was discarded, and the precipitate was mycelia with a mass of 8-10 g.

[0036] 2) Preparation of embedding reagents:

[0037] Sodium alginate solution: 13.69% (w / v) sucrose + 18.4% (w / v) glycerin + 11% (w / v) sodium alginate, prepared in 50 mL.

[0038] Calcium chloride solution: sucrose 13.69% (w / v) + glycerin 18.4% (w / v) + calcium chloride 1.5% (w / v), prepare 2 bottles, each bottle 200mL.

[0039] The above two solution...

Embodiment 3

[0049] The method of embedding and preserving the purified water of Clitopilus prunulus is as follows:

[0050] 1) Collect mycelium:

[0051] Inoculate the mushroom Clitopilus prunulus, batch number TMI12-1096 (the strain is provided by Ningxia Tairui Pharmaceutical Co., Ltd., seed room), in the liquid PDA medium, 24-26°C, 220-240r / min, shaking culture for 3d (At this time, Clitopilus prunulus is at the end of logarithmic growth phase). Take 30mL of the above seed solution in a 50mL sterile centrifuge tube, centrifuge at 3000r / min for 15min. The supernatant was discarded, and the precipitate was mycelium with a mass of 9-10 g.

[0052] 2) Preparation of embedding reagents:

[0053] Sodium alginate solution: 13.69% (w / v) sucrose + 18.4% (w / v) glycerol + 12% (w / v) sodium alginate, prepared in 50 mL.

[0054] Calcium chloride solution: sucrose 13.69% (w / v) + glycerol 18.4% (w / v) + calcium chloride 3.0% (w / v), prepare 2 bottles, each bottle 250mL.

[0055] The above two solut...

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PUM

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Abstract

The invention relates to a clitopilus prunulus preserving method. The process includes the steps that clitopilus prunulus mycelia and a sodium alginate solution are mixed, the mixed solution is dripped into a calcium chloride solution to prepare small balls, and finally the small balls are put into pure water and preserved at the temperature of 0-4 DEG C. The clitopilus prunulus preserving method can greatly prolong the clitopilus prunulus preserving time while ensuring strain survival and has a positive acceleration effect on pleuromutilin and derivatives thereof.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, in particular to a preservation method of Clitopilus prunulus. Background technique [0002] Clitopilus prunulus belongs to Basidiomycetes, Agaricaceae, Agaricaceae, and Stropharia. The pleuromutilin produced by this fungus can be used to synthesize tiamulin. Tiamulin is a new type of antibiotic, which has a significant inhibitory effect on Gram-positive bacteria and mycoplasma. It has strong antibacterial ability, wide antibacterial spectrum, low toxicity and side effects, and is not easy to produce drug resistance. [0003] At present, liquid paraffin preservation method and low-temperature slope preservation method are often used for domestic preservation of Clitopilus prunulus. The shortcomings of these two methods mainly include the following points: [0004] 1. Liquid paraffin has a toxic effect on the bacteria, and it is easy to cause the death of the bacteria. [0005] 2. Liqui...

Claims

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Application Information

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IPC IPC(8): C12N1/04C12R1/645
CPCC12N1/04
Inventor 杨智孙金刚张萍
Owner 宁夏泰瑞制药股份有限公司
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