Method for synthesizing acarbose through microbial fermentation

A technology for microbial fermentation and acarbose, applied in the direction of microorganism-based methods, microorganisms, biochemical equipment and methods, etc., can solve the problems of low acarbose production, achieve the promotion of biomass formation, increase production, The effect of promoting the consumption of carbohydrate substrates

Active Publication Date: 2012-04-04
ZHEJIANG UNIV OF TECH +2
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0014] Aiming at the problem of low yield of acarbose synthesized by microbial fermentation, the present invention provides an improved method for fermenting and synthesizing acarbose, that is, adding SAM in a timely and appropriate amount to the fermentation medium to increase the yield of acarbose

Method used

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  • Method for synthesizing acarbose through microbial fermentation
  • Method for synthesizing acarbose through microbial fermentation
  • Method for synthesizing acarbose through microbial fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] (1) Preparation of medium:

[0041] Prepare solid plate culture medium, its medium composition is as follows: sucrose 25g / L, peptone 2g / L, tyrosine 1g / L, K 2 HPO 4 0.1g / L, KCl 0.5g / L, MgSO 4 ·7H 2 O 0.5g / L, FeSO 4 ·7H 2 O 0.1g / L, agar 20g / L, prepared with tap water, initial pH 7.0. Sterilize at 121°C for 30 minutes.

[0042] Prepare seed culture medium, its culture medium composition is as follows: cornstarch 15g / L, soybean cake powder 40g / L, glycerol 20g / L, K 2 HPO 4 0.1g / L, CaCO 3 2g / L, prepared with tap water, initial pH 7.0. Sterilize at 121°C for 30 minutes.

[0043] Prepare the fermentation medium, the composition of which is as follows: maltose 80g / L, glucose 20g / L, soybean cake powder 10g / L, corn steep liquor 5g / L, CaCO 3 6g / L, FeCl 3 0.1g / L, CaCl 2 2g / L, prepared with tap water, pH 7.0. Sterilize at 121°C for 30min.

[0044] (2) Preparation of SAM aqueous solution:

[0045] SAM solutions with SAM concentrations of 0.25mM, 0.5mM, 1mM, 2mM, 4mM, 7mM ...

Embodiment 2

[0053] According to the results of Example 1, the concentration of SAM in the fermented liquid was selected to be 20 μM and 100 μM respectively, and according to the culture method of Example 1, the impact of adding SAM on the acarbose output in the fermented liquid was investigated at different times when the fermentation was carried out, At the same time, the shake flask fermentation without adding SAM solution but adding an equal volume of sterile water was used as a control, and the concentration of acarbose in the fermentation broth was detected by HPLC, and the results are shown in Table 3.

[0054] Table 3: Effect of SAM addition time on acarbose yield

[0055]

[0056] It can be seen from the above table that adding an appropriate amount of SAM in the early stage of fermentation, that is, from 0h to 60h, can significantly increase the fermentation yield of acarbose, especially adding an appropriate amount of SAM between 0h and 36h of fermentation can significantly in...

Embodiment 3

[0058] According to the results of Examples 1 and 2, SAM was added to the fermentation broth at the 12th hour of fermentation, and the SAM concentration in the fermentation broth was selected to be 10 μM and 20 μM respectively. According to the cultivation method of Example 1, the effect of adding SAM on Aka The influence of wave sugar fermentation process and yield, fermentation carries out 168 hours altogether, meanwhile, with the shaking flask fermentation that does not add SAM solution, but adds equal volume sterile water as contrast, HPLC detects the concentration of acarbose in the fermented liquid, the result As shown in Table 4.

[0059] Table 4: Effects of adding / not adding SAM on the process and yield of acarbose formed by fermentation

[0060]

[0061] It can be seen from the above table that adding an appropriate amount of SAM to the culture medium at the 12th hour of fermentation can promote the fermentation yield of acarbose in the whole fermentation process. ...

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Abstract

The invention discloses a method for synthesizing acarbose through microbial fermentation, which comprises the following steps that acarbose producing strains CCTCC NO: M 209022 are inoculated to a fermentation culture medium containing carbon sources, nitrogen sources and inorganic salt applicable to all strains, the fermentation culture is carried out for 96 to 192hours at 20 to 32 DEG C, afterthe fermentation is completed, the obtained fermentation liquid is extracted and separated, and the acarbose is obtained. The method is characterized in that the fermentation culture is carried out for 0 to 60h, and water solution of ademetionine is added, so the ademetionine concentration in the fermentation culture medium is 1 to 300 mu mol / L. Through the supplementary addition of the ademetionine in the acarbose fermentation process, the fermentation level of the acarbose is improved.

Description

(1) Technical field [0001] The invention belongs to the field of biochemical industry, and relates to a method for microbial fermentation and synthesis of acarbose, in particular to a method for increasing the synthetic yield of acarbose by adding adenosylmethionine during the fermentation process. (2) Background technology [0002] Acarbose (Acarbose) is an effective α-glucosidase inhibitor, which delays the breakdown and digestion of carbohydrates by competitively inhibiting the activity of α-glucosidase involved in carbohydrate degradation on the small intestine wall, delaying and It is an oral drug clinically used to treat type II diabetes by reducing the absorption of glucose so as to control the blood sugar level after meals in the human body. [0003] The molecular formula of acarbose is C 25 h 43 NO 18 , molecular weight 645.6, pKa1 value 5.1. In the early 1970s, Frommer et al. first isolated acarbose from the culture solution of actinomycete Actinoplanes utahens...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/00C12R1/045
Inventor 郑裕国孙丽慧李明刚王远山沈寅初
Owner ZHEJIANG UNIV OF TECH
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