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Process for preparing ophiopogonamidase by fermentation

A technology of asparaginase and fermentation tank, which is applied in the direction of hydrolytic enzymes and bacteria, which can solve the problems of high production cost, high fermentation titer, separation and purification technology that cannot meet the requirements, and achieve the effect of reducing the cost of raw materials

Inactive Publication Date: 2005-02-16
广州高新广微生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are many deficiencies in the domestic existing production of asparaginase technology, mainly 1. the fermentation titer is low (10 units / ml), the production cost is too high and not suitable for the requirements of the market, 2. the high fermentation titer is only limited to genetic engineering Bacteria, and the separation and purification technology does not meet the requirements

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] (1) plate culture

[0016] Prepare 10 plates, wrap them in kraft paper, and sterilize them for later use. According to the formula of beef extract 3.5%, NaCl 1%, agar 1.2%, pH 6.8, mix 300 milliliters of culture medium, be packed in 500 milliliters of triangular flasks, sterilize, cool to 70 ℃, pour petri dish (30 milliliters / dish), Solidified, ready to use. Dissolve the lyophilized Escherichia coli wild strain with 200 microliters of sterile water, take 100 microliters to spread on a plate, and incubate at 39° C. for 14 hours.

[0017] (2) Seed tank cultivation

[0018] Fermentation culture formula: beef extract 3.5%, corn steep liquor 2%, yeast powder 1%, monosodium glutamate 1.5%, pH 6.8.

[0019] A 100-liter tank is used for the seed tank, and the filling capacity is 60 liters, so 60 liters of culture medium should be prepared according to the above formula (actually only add water to 55 liters, and the steam condensed water after sterilization is estimated to be...

Embodiment 2

[0023] (1) plate culture

[0024] Prepare 10 plates, wrap them in kraft paper, and sterilize them for later use. According to the formula of beef extract 4%, NaCl 1%, agar 1.5%, pH 7.0, mix 300 milliliters of culture medium, be packed in 500 milliliters Erlenmeyer flask, sterilize, cool to 70 ℃, pour petri dish (30 milliliters / dish), Solidified, ready to use. The lyophilized Escherichia coli strains mutated by ultraviolet (UV) were dissolved in 200 microliters of sterile water, 100 microliters were taken to spread on a plate, and cultured at 39° C. for 14 hours.

[0025] (2) Seed tank cultivation

[0026] Fermentation culture formula: beef extract 4%, corn steep liquor 2%, yeast powder 1.5%, monosodium glutamate 1.8%, pH 7.0.

[0027] A 100-liter tank is used for the seed tank, and the filling capacity is 60 liters, so 60 liters of culture medium should be prepared according to the above formula (actually only add water to 55 liters, and the steam condensed water after ster...

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PUM

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Abstract

A process for preparing ophiopogonamidase features the colibacillus as initial one is fermented in proper culture medium and in proper condition to obtain efficient expression of ophiopogonamidase (50-80 unit / ml). It can be used to prepare ophiopogonamidase,

Description

technical field [0001] The invention belongs to the field of biological fermentation, and more specifically relates to a method for fermenting and producing asparaginase. Background technique [0002] Asparaginase is an important drug for the treatment of acute lymphoblastic leukemia and plays an irreplaceable role in the treatment of children with acute lymphoblastic leukemia. There are many deficiencies in the domestic existing production of asparaginase technology, mainly 1. the fermentation titer is low (10 units / ml), the production cost is too high and not suitable for the requirements of the market, 2. the high fermentation titer is only limited to genetic engineering bacteria, and the separation and purification technology did not meet the requirements. Contents of the invention [0003] The invention overcomes the shortcomings of the prior art and provides a method for high-efficiency expression and production of asparaginase. [0004] The method of the present i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/82
Inventor 梁淑娃彭中健杨冠东杜少平
Owner 广州高新广微生物技术有限公司