Strain for producing alginate lyase and use thereof

A technology of alginate lysing enzyme and bacterial strain, which is applied in the field of marine biology, can solve the problems of poor hydrolysis effect of alginate, achieve remarkable fermentation effect, reduce production cost, and simple ingredients

Active Publication Date: 2014-07-09
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The second object of the present invention is to provide the application of a strain producing alginate lyase, the Halomonas elongata SH-19 strain that will be...

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  • Strain for producing alginate lyase and use thereof
  • Strain for producing alginate lyase and use thereof
  • Strain for producing alginate lyase and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Screening and Identification of Example 1 Bacterial Strains

[0035] 1. Screening of enzyme-producing strains

[0036] The present invention uses the ground sea urchin intestinal tract to screen the production strain of alginate lyase, dilutes the ground sample of sea urchin intestinal tract, and then cultures it in the screening medium with sodium alginate as the only carbon source. The strains surviving in the base were used for the fermentative production of alginate lyase, and the strains were identified and stored frozen in glycerol tubes. Specific steps are as follows:

[0037] Dilute the ground sea urchin intestine samples in multiple ratios, spread them on the selection medium with sodium alginate as the only carbon source, and culture them at 25°C to 30°C for 1 to 7 days. Select the ones that can grow under these conditions Colonies, the obtained fermentation strains were selected and the one with the highest enzyme activity was identified as Halomonas sp. by...

Embodiment 2

[0058] Example 2 Experimental determination of the optimum living conditions for fermenting alginate lyase on the bacterial strain SH-19 obtained by screening:

[0059] 1) The selection and determination experiment of the carbon source in the fermentation medium:

[0060] First, five experimental groups were set up, in which, except for the carbon source, the fermentation medium was fermented under the conditions that the nitrogen source was ammonium sulfate, the pH value was 6.5, the mass percent concentration of NaCl was 3%, and the temperature was 30°C. The carbon source is respectively glucose, sucrose, starch, sodium alginate or xylose with a concentration of 0.5% by mass.

[0061] After the above-mentioned five experimental groups were fermented and cultured, the enzyme activity of the fermentation liquid of each experimental group was measured respectively, and the influence of different carbon sources on the enzyme production was tested. Such as figure 2 As shown, t...

Embodiment 3

[0070] The method for producing alginate lyase by the bacterial strain producing alginate lyase described in embodiment 3 comprises the following steps:

[0071] 1) Preparation of seed liquid: Inoculate the screened strain SH-19 on the seed medium, ferment at 26-30°C for 24-48 hours to reach the logarithmic growth phase, stop the fermentation, and obtain the seed liquid;

[0072] 2) Large-scale propagation of bacterial strains: after the seed liquid is inoculated in a primary fermentation medium, the primary fermentation medium is continuously fermented for 36 to 60 hours at a pH of 4.5 to 9.5 and a temperature of 26 to 30°C, Thereby, described bacterial strain carries out mass propagation, provides condition for later stage alginate lyase;

[0073] 3) Inducing a large amount of alginate lyase to synthesize: after the large-scale propagation of the bacterial strain SH-19, add sodium alginate and 1-3% sodium alginate and 1-3% chloride to the primary fermentation medium Sodium ...

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Abstract

The invention discloses a strain for producing alginate lyase and a use thereof. The strain is preserved in the China general microbiological culture collection center on October 9, 2013, has an accession number of CGMCC No.8312 and is named as Halomonas elongata SH-19. A 16S rDNA polynucleotide sequence of the strain is shown in the formula of SEQ ID No.1. The strain SH-19 can efficiently express alginate lyase and can be used for high-efficiency production of alginate lyase. A strain SH-19 fermentation medium comprises simple ingredients commonly used in a laboratory, has substantial fermentation effects, greatly reduces an alginate lyase production cost and can be used for large-scale production.

Description

technical field [0001] The invention belongs to the technical field of marine organisms, and in particular relates to a bacterial strain for producing alginate lyase and an application thereof. Background technique [0002] Algae are known as "energy rookies". Compared with other terrestrial plants, algae can obtain higher biomass per unit of cultivation area, and are easier to cultivate in large areas and mechanized harvesting. Therefore, algae has gradually become a modern bioenergy material, which has significant advantages in yield, production capacity and total cost. [0003] Brown algae contain a large amount of algin. The structure of brown algae itself is complex, and its cell wall composition, alginate, is difficult to degrade. Various polysaccharides are tightly combined into an organic whole. The complex structure makes it difficult to use algae. This virtually increases the cost of using algae bioenergy production. In addition, oligosaccharides of different len...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/88C12R1/01
Inventor 李德茂陈树林张藩高峰
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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