Isoptericola halotolerans capable of highly producing alginate lyase and application method for isoptericola halotolerans

A technology of alginate lyase and termite fungus, which is applied in the biological field to achieve the effects of high substrate conversion rate, high enzyme activity, and efficient degradation

Active Publication Date: 2012-05-02
JIANGNAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Isoptericola halotolerans capable of highly producing alginate lyase and application method for isoptericola halotolerans
  • Isoptericola halotolerans capable of highly producing alginate lyase and application method for isoptericola halotolerans

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Embodiment 1

[0018] Example 1 Isolation and identification of halophilic termite fungus (Isoptericola halotolerans) WX and preservation of bacterial strains

[0019] (1) Isolation and screening of Isoptericola halotolerans WX

[0020] The present invention obtains soil samples and waste samples from domestic chemical plants producing sodium alginate, weighs 1.0-2.0 g of soil samples in a 250 mL triangular flask containing 0.5% (w / v) sodium alginate, and ferments at 30° C. and 200 rpm After 48 hours, transfer 10mL of the culture solution to another 250mL Erlenmeyer flask filled with fresh medium for further cultivation, and perform gradient dilution after continuous acclimatization for three times. Take 0.2mL of the diluted solution and spread it on a screening solid plate, at 25-30°C After culturing for 20-24 hours, pick a single colony that can grow on a sodium alginate plate and inoculate it into a fresh homemade liquid medium. Put 30mL of fermentation medium in a 250mL Erlenmeyer flask...

Embodiment 2

[0024] Example 2 Production of alginic acid lyase by fermentation of halophilic termite fungus WX

[0025] (1) Preparation of cell liquid culture of halophilic termite (Isoptericola halotolerans) WX strain

[0026] Pick a ring of halophilic termite (Isoptericola halotolerans) WX strain on the solid self-made medium, inoculate it in a 250mL Erlenmeyer flask containing 50mL self-made medium, and place it on a shaker under the condition of 25°C-30°C. Cultivate at a rotational speed of 200r / min for 20-24h to the mid-logarithmic growth phase, and then obtain a cell liquid culture of the halophilic termite strain WX.

[0027] (2) The composition and ratio of the fermentation medium are: sodium alginate 6g / L, peptone 5-20g / L, yeast powder 4-10g / L, NaCl 20-30g / L, MgSO 4 0.2mM, Cacl 2 0.05mM, KH 2 PO 40.1mM, FeSO4 0.2mM, MnSO4 0.3mM, agar powder 10-20g / L; add distilled water to 1L, adjust the pH of the medium to 7.5-8.0 before sterilization, and sterilize under high-pressure stea...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to isoptericola halotolerans WX capable of highly producing alginate lyase and an application method for the isoptericola halotolerans WX. The isoptericola halotolerans WX capable of highly producing the alginate lyase is obtained through screening for the first time; and the strain WX is identified as the isoptericola halotolerans according to morphological characteristics and physiological-biochemical characteristics. The strain was collected in China General Microbiological Culture Collection Center on October 12th, 2011, and the collection number of the strain is CGMCC No.5336. The strain can produce the alginate lyase to degrade sodium alginate so as to produce alginate oligosaccharide, and the enzymatic activity can reach 432U/ml. The alginate lyase produced by the strain can efficiently degrade algin so as to produce the alginate oligosaccharide which has multiple bioactivities.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a halophilic termite fungus (Isoptericola halotolerans) with high yield of alginic acid lyase and an application method thereof. Background technique [0002] Fucoidan oligosaccharide has a variety of biological activities, and its medicinal value has become a new hot spot. It has good antitumor activity and anticoagulant activity, has few side effects, can lower blood sugar and blood lipids, and has obvious effects on human hereditary hyperglycemia. It can also prevent the formation of hypercholesterolemia, reduce fat and maintain normal cardiovascular function. At present, the methods for producing alginate oligosaccharides can be classified into: chemical degradation method, physical degradation method and enzymatic method. However, the degradation speed of the first two is slow, which will destroy the unique structure of polysaccharides, and requires high temperature...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/88C12R1/01
Inventor 史劲松许正宏魏丹张晓梅许鸿瑜窦文芳陆震明钱建瑛张旦旦李恒李会耿燕
Owner JIANGNAN UNIV
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