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Iota-carrageenan degrading bacterium as well as fermentation method and application thereof

A technology for carrageenan and degrading bacteria, which is applied in the field of bacteria belonging to the genus Cellulophile, can solve the problems of excessive molecular weight of carrageenan, influence on solubility and absorption, and limited application, and achieves high commercial application value and production capacity. improved effect

Active Publication Date: 2014-05-14
THIRD INST OF OCEANOGRAPHY STATE OCEANIC ADMINISTATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Existing research results have shown that carrageenan has a variety of biological activities, such as anticoagulant, anti-virus, immune regulation, anti-oxidation, etc. its application in the field of medicine

Method used

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  • Iota-carrageenan degrading bacterium as well as fermentation method and application thereof
  • Iota-carrageenan degrading bacterium as well as fermentation method and application thereof
  • Iota-carrageenan degrading bacterium as well as fermentation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Isolation and purification of ι-carrageenan-degrading bacteria (Cellulophaga sp.) KL-A.

[0033] After collecting fresh live seaweed leaves from the sea area near Dongshan Island, Fujian Province, they were washed with sterile seawater, put into the primary screening medium, and cultured in shake flasks for 2 to 3 days. The culture solution was streaked on the agar plate containing the primary screening medium, and cultivated for 3 days. Transfer the colony that formed a transparent circle on the agar plate to a new agar plate for culture, and repeat streaking until a pure culture is obtained. Primary screening medium composition (mass volume ratio): iota-carrageenan 0.2%, NaCl 1.5%, NaNO 3 0.2%, MgSO 4 ·7H 2 O0.05%, CaCl20.01%, iron phosphate 0.005%, pH value 7.5.

[0034] Inoculate the pure culture isolated from the primary screening into the secondary screening medium and culture on a shaking table. Then the culture solution was streaked on the re-screened solid...

Embodiment 2

[0037] Single factor screening of carbon and nitrogen sources

[0038] 1. Select several different carbon sources respectively, with a concentration of 10g / L, use 5g / L peptone as a fixed nitrogen source, add 2g / L ι-carrageenan and 15g / L NaCl, prepare a culture medium, and inoculate The amount is 3%, the culture temperature is 30°C, the shaker speed is 200rpm, cultivated for 48h, the enzyme activity is measured by DNS method, and the influence of carbon source on the fermentation of ι-carrageenan hydrolase is shown in Table 1, and glucose is the best carbon source .

[0039] Table 1

[0040] carbon source

glucose

Glycerin

Galactose

sucrose

sodium acetate

starch

Enzyme activity (U / mg)

287.6

201.3

234.7

168.2

171.1

168.2

[0041]2. Take 15g / L glucose as the fixed carbon source, select several different nitrogen sources respectively, the concentration is 5g / L, add ι-carrageenan of 2g / L and NaCl of 15g / L in a...

Embodiment 3

[0045] Preparation of iota-carrageenan oligosaccharides by using ι-carrageenan degrading bacteria

[0046] 1) Inoculate the strain on iota-carrageenan solid medium and culture at 30°C for 3 days;

[0047] 2) Inoculate a single colony on the solid medium into 250mL shake flask medium, culture at 30°C, 200rpm for 3 days.

[0048] 3) Put the seed medium into 1L of fermentation medium at a volume ratio of 3%, and culture at 30°C and 200rpm for 3 days to obtain the fermented product.

[0049] 4) Centrifuge the fermentation broth at 12000g for 15min at 4°C to remove bacteria, and keep the supernatant for later use.

[0050] 5) Gradually add ammonium sulfate to the supernatant to 40% saturation (add solid ammonium sulfate according to the solubility of ammonium sulfate at 4°C), stir slowly to avoid the generation of air bubbles and cause enzyme inactivation, after standing at 4°C for overnight precipitation, refrigerate and centrifuge at 10,000rpm Remove impurities. Then gradually...

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Abstract

The invention provides an iota-carrageenan degrading bacterium as well as a fermentation method and an application thereof, and relates to cytophaga bacteria. The collection number of the iota-carrageenan degrading bacterium (Cellulophaga sp.) KL-A is CGMCC No. 8694. A production method of iota-carrageenan hydrolytic enzyme comprises the following steps: coating an iota-carrageenan solid medium with the activated iota-carrageenan degrading bacterium (Cellulophaga sp.) KL-A, picking and inoculating a single colony into a seed medium and culturing so as to obtain a seed solution; and inoculating the seed solution into fermentation liquid according to 1%-5% of inoculation quantity, and carrying out shake cultivation so as to obtain fermented supernatant; and carrying out salt fractionation by using ammonium sulfate and DEAE (diethyl-aminoethanol)-cellulose chromatography on the obtained fermented supernatant at the temperature of 4 DEG C, dialyzing so as to remove salt, and carrying out vacuum freeze drying so as to obtain the iota-carrageenan hydrolytic enzyme. Iota-carrageenan oligosaccharides can be applied in the preparation of antibacterial agents, antiviral drugs, immunomodulators, antioxidants and the like.

Description

technical field [0001] The invention relates to a bacterium of the genus Cellulophilus, in particular to a strain of iota-carrageenan degrading bacteria and its fermentation method and application. Background technique [0002] Carrageenan (Carrageenan), also known as staghorn gum, carrageenan, etc., is a high-molecular hydrophilic polysaccharide extracted from red algae, with repeated α-(1→4)-D-galactopyranose -β-(1→3)-D-galactopyranose (or 3,6 inner ether-D-galactopyranose) disaccharide units are connected as the basic skeleton. According to whether it contains 3,6 inner ether-D-galactopyranose, the content of sulfate group and the position of sulfate group in the molecule, carrageenan is mainly divided into κ-, λ-, and ι-3 groups. [0003] Carrageenan is widely used in the food industry because of its excellent thermoreversible gelation, anti-protein gel, hydrophilic and non-toxic properties. At present, carrageenan has been widely used in jelly, soft candy, ice cream, ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/24C12P19/14A61K31/715A61K31/7016A61K31/702A61P31/04A61P31/12A61P37/02A61P39/06C12R1/01
Inventor 邵宗泽单大鹏李旭古铮应见喜
Owner THIRD INST OF OCEANOGRAPHY STATE OCEANIC ADMINISTATION
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