Novel vibrionaceae vibrio bacterial strain and application thereof

A technology of Vibrio and Vibrio, applied in the field of microorganisms, can solve the problems of limited application of alginate oligosaccharides, poor substrate specificity, low enzyme production and the like

Inactive Publication Date: 2007-02-28
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Dozens of alginate lyases have been isolated from bacteria and invertebrates, but the disadvantages of low enzyme production and poor substrate specificity generally limit the application of alginate oligosaccharides

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Example 1 Vibrio CCTCC NO: Isolation of M 205030

[0012] Take 1 g of the eroded part of kelp, soak it in 100 ml of sterilized seawater, stir for 1 hour, take 1 ml of the suspension and put it into 100 ml of liquid medium, and culture it statically at 25°C for one week. The culture solution was streaked on a solid plate to isolate single clones, and the single clones were purified and cultivated for 5 consecutive generations. Pick a single clone and inoculate it into a test tube containing 3 ml of liquid medium, and culture at 25°C for 12 hours at 200 r / min. The culture solution was transferred into a 250 ml Erlenmeyer flask containing 50 ml of liquid medium at a ratio of 1%, and cultured at 25° C. at 200 r / min for 3 days. The culture solution was centrifuged at 12,000 r / min at 4°C for 10 minutes to remove bacteria, the enzyme activity of the supernatant was measured, and the strain with the highest enzyme activity was selected. One of the bacterial strains produced t...

Embodiment 2

[0013] Example 2 Identification of Vibrio CCTCC NO: M 205030

[0014] 1. Morphological characteristics

[0015] The colonies of the obtained strains cultured for 24 hours on the conventionally used Zobell 2216E plate were round, translucent, smooth, with neat edges and a diameter of 2.5 mm; on the conventionally used TCBS plate, the colonies were yellow and 3.2 mm in diameter. This strain is Gram-negative, rod-shaped, and motile.

[0016] 2. Growth conditions

[0017] Strain CCTCC NO: M 205030 is a facultative anaerobic bacterium. CCTCC NO: M 205030 grows well in the range of 1% to 10% NaCl concentration, the optimum NaCl concentration is 4%, the optimum growth temperature is 25°C, and the optimum growth pH is 7.0.

[0018] 3. Physiological and biochemical characteristics

[0019] Routine biochemical experiments showed that the strain CCTCC NO: M 205030 had a positive oxidase reaction, fermented glucose to produce acid and did not produce gas, and was resistant to Vibrio i...

Embodiment 3

[0025] The preparation of embodiment 3 alginate lyase

[0026] (1) Cell culture

[0027] A single clone of Vibrio CCTCC NO: M 205030 was picked and inoculated into a test tube containing 3 ml of liquid medium, and cultured at 25° C. at 200 r / min for 12 hours. The culture solution was transferred into a 250 ml Erlenmeyer flask containing 50 ml of liquid medium at a ratio of 1%, cultured at 25° C. and 200 r / min for 3 days, and the culture solution was collected. The liquid medium formula used is (g / L): sodium alginate 3.0g, KH 2 PO 4 3.0g, K 2 HPO 4 7.0g, (NH 4 ) 2 SO 4 2.0g, MgSO 4 0.1g, FeSO 4 0.1g, NaCl 30.0g.

[0028] (2) Separation and purification of enzymes

[0029] The culture solution was centrifuged at 12,000 r / min at 4°C for 10 minutes to remove bacteria, and the supernatant was collected. Slowly add ammonium sulfate to the supernatant to 80% saturation, let stand at 4°C for 3h, centrifuge at 13000g at 4°C for 30min, discard the supernatant, dissolve ...

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Abstract

The invention relates the vibrionaceae vibrio strain and its application. The invention relates the vibrionaceae vibrio strain from sea-tangle, and the output of algin lyase made by the vibrionaceae vibrio strain is 2-5 times than that of algin lyase made with now technology. The algin lyase can be used to make algin oligosaccharide.

Description

technical field [0001] The present invention relates to the field of microorganisms. Specifically, the present invention relates to a new Vibrio sp.QY1 strain of the genus Vibrio, the preservation number of which is CCTCC NO: M 205030, and the application of the strain to prepare alginate lyase and alginate oligosaccharides. Background technique [0002] Alginate is an acidic marine polysaccharide with rich sources and wide uses, which is derived from brown algae and certain bacteria (including Pseudomonas, nitrogen-fixing bacteria, etc.), composed of β-D-mannuronic acid and α-L-guro Uronic acid is formed by β-1,4 glycosidic bonds, and can be divided into polymannuronic acid, polyguluronic acid and alternating blocks of mannuronic acid and guluronic acid. Recently, the biological activities of alginate oligosaccharides have been continuously discovered, such as anti-tumor, anti-virus, anti-bacterial, anti-cardiovascular disease, etc., and its broad application prospects hav...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/42C12P19/00C12R1/63
Inventor 于文功韩峰
Owner OCEAN UNIV OF CHINA
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