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Artificial bear bile powder and quality evaluation method thereof

A kind of bear bile powder, artificial technology, applied in the field of biology

Inactive Publication Date: 2018-11-16
SHANGHAI UNIV OF T C M
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved in the present invention lies in a kind of artificial bear bile powder and its quality evaluation method

Method used

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  • Artificial bear bile powder and quality evaluation method thereof
  • Artificial bear bile powder and quality evaluation method thereof
  • Artificial bear bile powder and quality evaluation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] 1 The effect of the secondary seed medium on the growth curve

[0097] 1) Apply the engineering bacteria stored in the -80°C refrigerator to the LB solid plate medium containing 100 mg / L ampicillin by streaking method, and activate and culture at 37°C for 12 hours;

[0098] The composition of the LB solid medium is as follows: tryptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L, agarose 15.0g / L.

[0099] 2) Pick a single colony from the LB solid plate medium, inoculate it in the LB liquid medium supplemented with 100mg / L ampicillin, and culture it with shaking at 225rpm and 37°C for 9 hours to obtain the first-grade seeds;

[0100] The composition of described LB liquid medium is as follows: tryptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L;

[0101] 3) Inoculate the above culture into 2-YT liquid medium and the same LB liquid medium as described in 2) at a ratio of 1:20, culture with shaking at 225rpm / min and 37°C, and measure both Growth curve;

[010...

Embodiment 2

[0107] Optimization experiment of M9-GY optimized medium

[0108] 1) Apply the design method of Box-Behnken Design in the Design-Expert.V8.0.6.1 software, based on the composition of the M9 basic medium, design a test with glucose, glycerol, yeast extract and tryptone as four factors, Glucose, yeast extract, and tryptone all choose the three levels of 0, 10, and 20g / L, and glycerol chooses the three levels of 0, 15, and 30ml / L. The OD600 and conversion rate are used as the investigation indicators at the same time. The 29 groups of programs designed by the software and The results are as follows:

[0109]

[0110] 2) The specific fermentation process is as follows: Obtain first-grade seeds according to the operation in Example 1, inoculate the first-grade seeds into liquid 2-YT medium at a ratio of 1:20, and shake and culture at 225rpm and 37°C for 4 hours to obtain 2-YT. Grade seeds; the second grade seeds are inoculated into the above-mentioned various liquid mediums in ...

Embodiment 3

[0114] Optimization of induction culture phase

[0115] 1 Effect of IPTG concentration on conversion rate

[0116] 1) According to the operation in Example 1, the first-grade seeds were obtained, and the first-grade seeds were inoculated into the liquid 2-YT medium at a ratio of 1:20, and cultured with shaking at 225 rpm and 37° C. for 4 hours to obtain the second-grade seeds;

[0117] 2) Inoculate the secondary seeds into the M9-GY optimized medium liquid medium at a ratio of 1:20, in triplicate, and shake and culture at 225rpm and 37°C for 5 hours;

[0118] 3) Add IPTG to the final concentration of 0.1, 0.3, 0.6, 0.8, 1.0mM respectively, shake and culture at 225rpm, 30°C for 3 hours; Shake culture at 25°C for 6 hours for fermentation; the results are as follows figure 1 as shown in a:

[0119] 2. Combination optimization of induction conditions

[0120]Select the pre-log (4 hours), mid-log (5.5 hours), and late log (7.5 hours) of the growth curve of the recombinant engi...

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Abstract

The invention provides artificial bear bile powder and a quality evaluation method thereof and belongs to the field of biotechnology. The artificial bear bile powder is prepared through simple processes, is qualified, utilizes cheap and easily available raw materials and is a breakthrough in the preparation of artificial bear bile powder by biotechnology means.

Description

technical field [0001] The invention relates to an artificial bear bile powder and a quality evaluation method thereof. Specifically, it is a process of carrying out large-scale fermentation and cultivation in a fermentation tank with recombinant engineering bacteria, and purifying and preparing artificial bear bile powder, and evaluating the quality of the artificial bear bile powder produced by the process, which belongs to the field of biotechnology. Background technique [0002] Bear bile powder is the powder product of black bear or brown bear gall bladder. Clinically, it is often used for heat convulsions, epilepsy, convulsions, and jaundice in children; external use is used to treat carbuncles, hemorrhoids, red eyes, and cloudy eyes. Bear bile powder mainly contains bile acid, and its main active ingredient is tauroursodeoxycholic acid (TUDCA). The Chinese Pharmacopoeia stipulates that the content of TUDCA should not be less than 23%. [0003] At present, bear bile ...

Claims

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Application Information

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IPC IPC(8): G01N30/02A61K35/57
CPCA61K35/57G01N30/02
Inventor 赵淑娟王峥涛许盈芃杨莉周吉燕胡之璧张金家
Owner SHANGHAI UNIV OF T C M
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