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Method for producing alginate hydrolysate

A technology for producing brown algae and alginate lyase, which is applied in the field of production of alginate hydrolyzate, can solve the problems of low degradation rate, unreachable, low viscosity, etc., and achieve the effect of high hydrolysis efficiency

Inactive Publication Date: 2020-05-26
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, chemical and physical methods are often used to hydrolyze alginate to reduce the molecular weight of alginate, but there are still many defects in the existing chemical and physical methods
For example, Yang Guixia and others hydrolyzed alginate by gamma radiation to reduce its viscosity, but this method has a low degradation rate and is not suitable for industrial production (see references for details: Yang Guixia, Li Xiaoyan. Polysaccharide polymer material sodium alginate Irradiation degradation of alginate[J]. Atomic Energy Science and Technology, 2013,47(05):730-734.); Feng et al. hydrolyzed alginate by ultrasonic method to reduce its viscosity, but Feng et al. also found that Ultrasonic frequencies exceeding 28kHz will promote the covalent aggregation of the molecular weight of alginate, causing the molecular weight of alginate to increase as the frequency increases, which in turn makes the degree of hydrolysis of alginate hydrolyzate low and cannot achieve lower viscosity (see reference for details: Feng L,Cao Y,Xu D,et al.Molecular weight distribution,rheological property and structural changes of sodium alginate induced by ultrasound[J].Ultrasonics Sonochemistry,2017,34:609-615.)
[0005] The above defects greatly limit the application of alginate in many fields such as medicine, food, printing and spinning, agriculture, daily chemical industry, etc.

Method used

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Examples

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Effect test

preparation example Construction

[0057] The preparation method involved in the following examples is as follows:

[0058] The preparation method of alginate lyase is as follows:

[0059] In a sterile environment, use an inoculation loop to dip the Vibrio natriegens SK42.001 bacterial solution stored in a glycerol tube, streak it on a sodium alginate solid plate, and place it in a constant temperature incubator at 37°C for 15 hours. , to obtain a single colony; in a sterile environment, pick a single colony and inoculate it into a 50mL seed medium, and cultivate it at 28°C and 200rpm for 12h to obtain a seed liquid; ) was inoculated into 50mL fermentation medium, cultivated at 28°C and 200rpm for 24h to obtain a fermentation broth; the fermentation broth was centrifuged at 10000r / min for 5min, and the alginate-containing lyase (amino acid sequence as shown in SEQ ID NO: 1) was taken ) of the fermentation supernatant (enzyme activity 1.4U / mL).

Embodiment 1

[0060] Embodiment 1: the preparation of alginate hydrolyzate (one-step method)

[0061] Specific steps are as follows:

[0062] Firstly, the alginate lyase was mixed according to 2.5U / g 褐藻胶 , 3.75U / g 褐藻胶 , 5U / g 褐藻胶 , 7.5U / g 褐藻胶 , 10U / g 褐藻胶 The addition amount of 10g / L alginate is added to the reaction system containing 10g / L alginate (the reaction system containing 10g / L alginate is the alginate solution with a concentration of 10g / L), and the reaction is carried out at a temperature of 35°C and a pH of 7. 6h, the reaction solution 1-5 was obtained, and then the reaction solution was separated to obtain the alginate hydrolyzate 1-5; the separation was firstly adding anhydrous water and ethanol to obtain a mixed solution, and then the mixed solution was left to stand for 24 hours, then centrifuged at 8000 rpm and 25° C. for 15 minutes to obtain a precipitate to obtain alginate hydrolyzate.

[0063] The concentration of the alginate hydrolyzate in the reaction solutions 1-...

Embodiment 2

[0070] Embodiment 2: the preparation of algin hydrolyzate (batch method)

[0071] Specific steps are as follows:

[0072] Option 1: First add alginate lyase at 2.5U / g 褐藻胶 The addition amount of 10g / L alginate is added to the reaction system containing 10g / L alginate (the reaction system containing 10g / L alginate is the alginate solution with a concentration of 10g / L), and the reaction is carried out at a temperature of 35°C and a pH of 7. 6h to obtain the reaction solution 6, and then separate the reaction solution to obtain the alginate hydrolyzate 6; 反应体系 The amount of addition is to dissolve the alginate in the reaction system through a high-speed disperser, and then according to 2.5U / g 褐藻胶 Alginate lyase is added to the reaction system; the separation is to first add absolute ethanol to the reaction solution according to the volume ratio of 1:3 (reaction solution: absolute ethanol) to obtain a mixed solution, and then mix the mixed solution After standing still for 24 h...

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Abstract

The invention discloses a method for producing alginate hydrolysate, and belongs to the technical field of biology. The invention provides the method for producing low-viscosity alginate hydrolysate.The method comprises the following steps: adding alginate lyase with an amino acid sequence represented by SEQ ID NO:1 and alginate into a reaction system at one time, and reacting for 6 h to lower the viscosity of the alginate hydrolysate to 15cp. According to the method, the alginate lyase with the amino acid sequence represented by SEQ ID NO:1 and alginate are intermittently added into the reaction system, and a reaction is performed for 6 h, so that the viscosity of the alginate hydrolysate can be as low as 15cp.

Description

technical field [0001] The invention relates to a method for producing alginate hydrolyzate, which belongs to the field of biotechnology. Background technique [0002] Alginate is a polysaccharide polymer extracted from brown algae such as kelp, sargassum, and macroalgae. It is composed of β-D-1,4-mannuronic acid (Mannuronate, M) and C-5 epimer α- One or two sugar units in L-1,4-guluronate (G) are linked by β-1,4 glycosidic bonds. Due to its gel properties, alginate can be used as a stabilizer, thickener or gel agent in many fields such as medicine, food, printing and spinning, agriculture, and daily chemical industry. In addition, due to its biological activities such as anticoagulant, hypoglycemic, hypolipidemic, anti-free radical, anti-tumor, immune protection, and plant growth promotion, alginate can be widely used as an active biomacromolecule in medicine, food, printing and spinning, agriculture, etc. , daily chemicals and many other fields. [0003] Studies have sh...

Claims

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Application Information

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IPC IPC(8): C12P19/04C12N9/88C12N15/60
CPCC12N9/88C12P19/04C12Y402/02
Inventor 江波张涛陈佳薇
Owner JIANGNAN UNIV
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