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Alginate-lyase-generating bacilli and preparation method and application thereof

A technology for bacillus and enzyme production medium, which is applied in the field of microorganisms and can solve the problems of low fermentation efficiency, low alginate lyase enzyme activity and the like

Inactive Publication Date: 2018-07-13
YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, many of these microorganisms are pathogenic bacteria, such as marine Vibrio, which can cause sepsis in mammals, resulting in low activity of alginate lyase and low fermentation efficiency, thus limiting the application of this type of microorganisms

Method used

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  • Alginate-lyase-generating bacilli and preparation method and application thereof
  • Alginate-lyase-generating bacilli and preparation method and application thereof
  • Alginate-lyase-generating bacilli and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Screening of Bacillus (sp.W003):

[0051] (1) Enrichment culture: the kelp was rotted in water for 14 days to obtain a rotten kelp sample, and the sample was picked and placed in a fresh enrichment medium for 24 hours at 30°C.

[0052] (2) Separation and purification: the cultured bacterial liquid was diluted by 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 Spread it on the plate medium according to the coating plate method, and select the single colony that formed an obvious dissolution zone on the plate after culturing at 37°C for 48 hours, and use the conventional microbial isolation method to repeatedly isolate and purify until pure Bacillus (sp.W003) is obtained. .

Embodiment 2

[0054] Screening of Bacillus (sp.W024):

[0055] (1) Enrichment culture: the kelp was rotted in water for 14 days to obtain a sample of rotten kelp, and the sample was picked and placed in a fresh enrichment medium for 48 hours at 30°C.

[0056] (2) Separation and purification: the cultured bacterial liquid was diluted by 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 Spread on the plate medium according to the coating plate method, and select a single colony that forms an obvious dissolution zone on the plate after culturing at 37°C for 48 hours, and use conventional microbial isolation methods to repeatedly isolate and purify until pure Bacillus (sp.W024) is obtained .

[0057] Among them, the components and proportions of the culture medium used in the above-mentioned enrichment culture and separation and purification process are as follows:

[0058] Enrichment medium: peptone 5.0g / L, yeast extract powder 10.0g / L, prepared in seawater.

[0059] Plate medium: sodium alginate 10.0g...

Embodiment 3

[0061] Identification of Bacillus (sp.W003) and Bacillus (sp.W024)

[0062] Bacillus (sp.W003) and (sp.W024) obtained by screening high-yield alginate lyase, after the strains used were grown on the primary screening medium for 48 hours, a clear dissolution circle could be formed around the Bacillus (sp.W003) colony, such as figure 2 As shown, the shape of Bacillus (sp.W003) was rod-shaped, Gram staining was negative ( Figure 4 ). The colony cultured on the slope for 48 hours was light white, round slightly convex, smooth surface, moist lawn, small colony, obvious transparent circle, no flagella and spore formation. In one embodiment, the optimum temperature of Bacillus (sp.W003) is 30°C, and the optimum pH is 7.5.

[0063] One embodiment of Bacillus (sp.W024), preservation time: 2017-12-04, preservation address: China, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, preservation number: CGMCC NO.15011. In one embodiment, an obvious dis...

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Abstract

The invention provides a bacillus (sp.W003). The collection number of the bacillus is CGMCC NO.15010. The invention further provides a bacillus (sp.W024). The collection number of the bacillus (sp.W024) is CGMCC NO.15011. The invention provides a method adopting the bacilli to prepare alginate lyase. The method includes following steps: activating the bacilli, and coating activated bacteria liquidonto a slope culture medium; picking and inoculating the bacilli on the slope culture medium into a container filled with an enzyme-generating culture medium for culture, and culturing on a shaking table at temperature of 25-35 DEG C and rotating speed of 150-230r / min for 12-16h to logarithm middle and later period to obtain seed liquid; inoculating the seed liquid into a fermentation tank filledwith a fermentation enzyme-generating culture medium, and fermenting in conditions of 25-35 DEG C in temperature, 150-230r / min in rotating speed and 7-8 in PH for 2-4d to obtain alginate oligosaccharide fermention liquid. The invention further provides application of the bacilli in generating alginate lyase and fermenting a culture medium for producing alginate oligosaccharide. The alginate lyasegenerated by the bacilli is high in enzyme activity and fermentation efficiency.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a bacillus producing alginate lyase and a preparation method and application thereof. Background technique [0002] Algin oligosaccharides are low molecular weight fragments obtained by hydrolyzing algin with a small degree of polymerization and good water solubility. Because alginate oligosaccharides have various physiological activities, such as anticancer, anticoagulant, lipid-lowering, immune regulation and anti-aging effects, they can be used as health food, therapeutic drugs or fungicides, so they can be widely used in food, In the fields of medicine, agriculture and cosmetics, it has great application and development value. [0003] At present, the preparation methods of alginic oligosaccharides mainly include chemical degradation method and enzymatic hydrolysis method. The chemical degradation method has the disadvantages of product inhomogeneity, poor repeatability, and la...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/80C12P19/04C12R1/07
CPCC12N9/80C12P19/04C12N1/205C12R2001/07
Inventor 李莉莉秦松武敏刘正一焦绪栋崔玉琳
Owner YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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