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Vampirovibrio sp. and method for preparing agarase

A bat vibrio, agarase technology, applied in the field of food biology, can solve the problem that agarase has not yet been adapted to large-scale industrial production and the like

Active Publication Date: 2014-06-18
SHANDONG XINGQIANG CHEM IND TECH RES INST CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the agarase prepared by it has not been adapted to large-scale industrial production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Take fresh abalone sold in Fuzhou, immediately immerse it in 70% alcohol for 1 minute to disinfect, then wash it with sterile water several times, use a sterile scalpel to separate it from the shell, then wash it with sterile water for several times, use tweezers and a scalpel Remove the intestinal tract on the shell, place it in a pre-sterilized mortar, add sterilized 1 ml of seawater to grind for 10 minutes, and then add 9 ml of sterilized seawater to mix well. Aspirate 1ml for dilution (10 -1 ,10 -2 ,10 -3 ,10 -4 ,), draw 0.5ml and spread it on the agar screening medium, cultivate for 48 hours at 25°C, select the colonies with obvious depressions, and put them into the agar screening medium for streak separation and purification. , 100rpm, shake flask for 48h, centrifuge to take the supernatant, and detect the agarase activity in the medium by spectrophotometry. , and finally selected B-13 as the starting strain.

Embodiment 2

[0026] After morphological and physiological and biochemical characteristics, the characteristics of B-13 strain are as follows:

[0027] Colony Morphology: The colonies are round, smooth, milky white and neatly edged.

[0028] Cell shape: This is a gram-negative bacteria with a rod-shaped shape.

[0029] Physiological and biochemical characteristics: It is a facultative anaerobic bacteria, which can use starch, glucose, sucrose, sodium acetate, D-galactose and other compounds as carbon sources, and can also use peptone, yeast extract, ammonium nitrate, ammonium sulfate, etc. compounds as nitrogen sources.

[0030] The 16S rRNA gene of the strain was amplified by PCR and sequenced, and it was found that the length of the 16S rRNA gene fragment was 1484 bp. After the comparison between NCBI and the ribosome database, it was identified as bat Vibrio ( Vampirovibrio sp.), to be named bat Vibrio ( Vampirovibrio sp.) fjfst-2013001, the 16S rRNA sequence of the strain can be foun...

Embodiment 3

[0032] (1) Vibrio batatum ( Vampirovibrio sp.) fjfst-2013001 in the seed medium, use 250mL Erlenmeyer flask to fill 30mL seed medium, sterilize according to the conventional method, cool down, and inoculate the colony, after inoculation, culture at 25°C, 140rpm shaker for 24h to obtain the seed fermentation liquid , the seed medium is: sea crystal 25g / L, peptone 5g / L, yeast powder 1g / L, agar 2g / L, adjusted pH to 7.4, prepared with deionized water.

[0033] (2) Fill 30mL fermentation medium in a 250mL Erlenmeyer flask, sterilize according to conventional methods, cool down, and insert the seed fermentation liquid into the fermentation medium according to the inoculum size of 4%, and cultivate it on a shaker at 25°C and 140rpm after inoculation After 48 hours, agarase-containing fermentation broth was obtained. The fermentation medium was prepared by: sea crystal 25g / L, peptone 5g / L, yeast powder 1g / L, agar 2g / L, adjusted pH to 7.4, and deionized water.

[0034] The activity of...

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PUM

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Abstract

The invention relates to a vampirovibrio sp. fifst-2013001 which has been registered and collected in the China Center for Type Culture Collection on December 8, 2013, with a collection number of CCTCC M 2013638, and a method for preparing an agarase from the vampirovibrio sp. fifst-2013001. The vampirovibrio sp. fifst-2013001 is taken as a strain and a fermentation culture medium composed of a carbon source, a nitrogen source and an inorganic salt is adopted, and under optimized culture conditions, the activity of the agarase reaches up to 4.17U / ml. A fermentation liquor is salted out by virtue of ammonium sulfate, a precipitate is collected after centrifuging, next, dialysis is carried out, a dialysate is put through an ion exchange column and a gel column, and then freeze-drying is carried out to obtain the dry powder of the agarase.

Description

technical field [0001] The invention belongs to the field of food biotechnology, and relates to screening a strain of Vibrio bat ( Vampirovibrio sp.) and a method for preparing agarase by culture and fermentation. Background technique [0002] Agar, also known as agar, is a kind of seaweed polysaccharide obtained by hot water extraction from marine red algae. It is rich in resources and is widely used in food, medicine, biotechnology and other fields. Agar is mainly composed of neutral polysaccharides. Agarose and ionic sulfur agarose are composed of 1,3-linked β-D-galactose and 1,4-linked 3,6-endether-α-L-galactose residues repeatedly Alternately linked linear polymers. [0003] Agarases come from a wide range of sources and are mainly found in marine bacteria: Alteromonas, Cytophaga, Pseudomonas, Alterpseudomonas, Streptomyces, Vibrio, and Vibrio. It is found in the genera of animals that feed on seaweed, such as Aplysia, Shore snail, Crown sea Zhan and so on. [0...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/24C12R1/01
Inventor 张龙涛欧洋曾绍校郑宝东张怡黄旭辉曾诚
Owner SHANDONG XINGQIANG CHEM IND TECH RES INST CO LTD
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