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Thalassospira sp. strain and method for preparing agarase

The technology of spirulina and agarase is applied in the field of preparing agarase, can solve the problems of low activity, high price, few sources of agarase, etc., and achieves simple preparation and purification process, short enzyme production time and low cost Effect

Active Publication Date: 2014-07-02
安徽桧元素生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the relatively few sources of agarase, low activity and high price, there are certain limitations in production and application
Agarase derived from microorganisms can be produced in large quantities, but generally the wild strains obtained by screening often have low enzyme production activity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Take the seawater in the coastal waters of Xiamen to take the sample liquid, absorb 1ml to dilute (10 -1 ,10 -2 ,10 -3 ,10 -4,), draw 0.5ml and spread it on the agar screening medium, culture at 25°C for 48 hours, select colonies with obvious depressions, enter the agar screening medium for streak separation and purification, and connect the pure species to the liquid medium at 25°C , 100rpm, shake the flask for 48 hours, centrifuge to take the supernatant, and use spectrophotometry to detect the agarase activity in the medium. After preliminary screening, 3 strains of bacteria capable of growing agarase were isolated and obtained, combined with the production capacity of agarase , and finally selected as the starting strain.

Embodiment 2

[0027] Through the study of morphology and physiological and biochemical characteristics, the characteristics of the agarase-producing strain are as follows:

[0028] Colony morphology: The colony is round, with smooth surface, milky white, and neat edges.

[0029] Cell morphology: This is a Gram-negative bacterium with a Vibrio-like morphology.

[0030] Physiological and biochemical characteristics: It is strictly aerobic, 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. compound as a nitrogen source.

[0031] The 16S rRNA gene of the strain was amplified and sequenced by PCR, and the length of the 16S rRNA gene fragment was found to be 1487bp. After comparison between NCBI and the ribosome database, it was identified as Helicobacter ( Thalassospira sp.), proposed to be named Hyospirum ( Thalassospira sp.) fjfst-2013007, the 16S rRNA seque...

Embodiment 3

[0033] (1) The sea rotund ( Thalassospira sp.) fjfst-2013007 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 salt 25g / L, peptone 5g / L, yeast powder 1g / L, agar 2g / L, adjusted pH to 7.4, prepared with deionized water.

[0034] (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, the fermented liquid containing agarase was obtained, and the seed culture medium was prepared by: sea salt 25g / L, peptone 5g / L, yeast powder 1g / L, agar 2g / L, pH adjusted to 7.4, and deionized water.

[0035] The activity o...

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PUM

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Abstract

The invention relates to a Thalassospira sp. fjfst-201307 strain and a method for preparing agarase by using the strain. The strain has been registered and collected at China Center for Type Culture Collection on December 25th, 2013, and the collection number is CCTCC M2013708. In a fermentation medium composed of a carbon source, a nitrogen source and inorganic salts by using the strain as the seed, the activity of the agarase is up to 11.2 U / ml under optimized culture conditions. The method for preparing the agarase comprises the following steps: salting out the fermentation liquor by ammonium sulfate, centrifugating, collecting the precipitate, dialyzing, passing the dialysate through an ion-exchange column and a gel column, and carrying out freeze-drying to obtain the agarase dry powder.

Description

technical field [0001] The invention belongs to the field of food biotechnology and relates to the screening of a strain of Helicotrium ( Thalassospira sp.) and a method for preparing agarase by culturing and fermenting the same. Background technique [0002] Agar, also known as agar, is a seaweed polysaccharide extracted from marine red algae by hot water. It is rich in resources and widely used in food, medicine, biotechnology and other fields. Agar is mainly composed of neutral Composed of agarose (agarose) and ionic sulfur agar (agar), agarose is composed of 1,3-linked β-D-galactose and 1,4-linked 3,6-inner ether-α-L - Linear polymers in which galactose residues are alternately linked repeatedly. [0003] Agarase has a wide range of sources and mainly exists in marine bacteria: Alteromonas, Cytophagium, Pseudomonas, Alteropseudomonas, Streptomyces, Microtremor, and Vibrio. It is found in the genera of animals that feed on seaweed, such as sea hares, snails, and c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/42C12N9/24C12R1/01
Inventor 张龙涛曾诚曾绍校郑宝东张怡黄旭辉欧洋
Owner 安徽桧元素生物科技有限公司
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