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Fermentation production process of alkaline mycose lyase and microbe for producing the lyase

A seaweed polysaccharide and lyase technology, applied in the direction of lyase, bacteria, etc., can solve the problems of complex activity, interference, easy swelling, etc., and achieve the effect of high enzyme activity and good thermal stability

Inactive Publication Date: 2004-10-13
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The extreme enzymes of alkaliphilic microorganisms have special significance for the development of seaweed polysaccharide lyases. First, they can avoid the interference of glycosidase activity and overcome the complex defects of general seaweed polysaccharide lyase activities; It is an irreversible gel, and its molecules are easy to swell under alkaline conditions, which is also conducive to the cleavage of enzymes. Moreover, metal ion complexes are mostly formed under alkaline conditions, and the metal ions can interact with each other depending on the pH during the treatment process. Alternatives, therefore alkaline alginate lyases from alkaliphiles have special advantages

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Keep and transfer the bacterial strain N19-2 to the solid slant medium of the test tube, the agar medium consists of: (g / L) starch 5, peptone 10, yeast powder 5, K 2 HPO 4 1. MgSO 4 7H 2 O 0.15, NaCl 30, Na 2 CO 3 10. Agar powder 2%. Prepare with distilled water.

[0027] For each transfer, the agar slants were incubated at 35°C for 2 days. Then prepare the seed culture according to the following steps.

Embodiment 2

[0029] Transfer the surface growth from the agar surface to a 250ml Erlenmeyer flask containing 50ml of seed medium, culture at 35°C on a shaker at 220 rpm for 24 hours, and then transfer to four 3000mL triangle flasks containing 1000ml of seed medium In the bottle, cultivate under the above-mentioned conditions for 24 hours, and then inoculate in a 250-liter fermenter with 120 liters of fermentation medium for fermentation. The composition of the fermentation medium is (g / L) sodium alginate 12, soybean powder 4.5, monosodium glutamate 4.5, yeast powder 7, K 2 HPO 4 1.5, MgSO 4 7H 2 O 0.3, NaCl 30, Na 2 CO 3 10. 4ml soybean oil, fermentation temperature 35°C±1°C, ventilation rate 1:0.75, stirring speed 400 rpm. During the fermentation process, the bacterial cell growth delay period is about 8 hours. After entering the logarithmic phase, the bacterial cell grows rapidly, and the enzyme amount increases accordingly. The enzyme activity is the highest during the stable ph...

Embodiment 3

[0032] Strain N19-2 was inoculated from a slant into a 250ml Erlenmeyer flask containing 50ml of seed medium, and cultured on a shaker at 35°C for 24 hours as a primary seed solution. Put the primary seed solution into a 3000ml Erlenmeyer flask with 1000ml seed medium at an inoculum amount of 3%, and cultivate it on a shaker at 37° C. for 24 hours as the secondary seed solution. Put the secondary seed liquid into a 50-liter seed fermenter equipped with 25 liters of seed medium, the fermentation temperature is 35°C±1°C, the ventilation rate is 1:1.0, and the stirring speed is 450 rpm. After 24 hours of fermentation, it is Transfer to a 1 ton tank with 600 liters of fermentation medium. The composition of the fermentation medium is (g / L): sodium alginate 12, yeast powder 7, soybean powder 4.5, monosodium glutamate 4.5, K 2 HPO 4 1.5, MgSO 4 7H 2 O 0.1, NaCl 30, Na 2 CO 3 12. Soybean oil 4ml / liter, fermentation temperature 35°C, ventilation rate 1:0.75, stirring speed 24...

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Abstract

The present invention provides one kind of special basophile N19-2 and the microbe fermentation production process of alkaline mycose lyase by using sodium alginate and peptone as material in the presence of the special basophile N19-2 strain. The method is simple, the enzyme activity of the fermented liquid reaches 100 u / ml, and the post-extraction rate is over 80 %. The enzyme has optimal reaction pH value of 9.5, optimal reaction temperature of 55 deg.c and important application value in the production of oligomycose.

Description

technical field [0001] The invention relates to a novel microorganism and a method for fermenting and producing enzymes using the microorganism. Background technique [0002] Seaweed polysaccharide lyase or alginate lyase (Alginate lyase.EC4.2.2.3.) degrades seaweed polysaccharides into unsaturated uronic acid oligomers—alginic acid oilgosaccharides and Unsaturated uronic acid monomer (uronic acid), according to the action of the enzyme on the substrate, the enzyme is divided into two types, namely polyM lyase and polyG lyase. [0003] Since the 1990s, the application value of alginate lyase in the research and development of bioengineering products has attracted attention. It can not only develop new physiologically active substances, but more importantly, it has opened up a new field of industrial application. In addition to being directly used as a pharmaceutical enzyme to treat pneumonia caused by the pathogenic bacteria Pseudomonas aeruginosa, the important direction o...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/88
Inventor 马延和薛燕芬周培瑾刘颍张志钢刘洪灿
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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