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Enzymolysis preparation method of new agaro oligosaccharides

A new agar oligosaccharide and enzymatic hydrolysis technology, applied in the field of bioengineering, can solve the problems of not being absorbed and utilized by organisms, limited application of agar gel, insoluble in cold water, etc., and achieve the effect of low cost, low price and abundant sources

Inactive Publication Date: 2016-01-20
FUZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Agar has high viscosity, is insoluble in cold water, and cannot be absorbed and utilized by organisms. These shortcomings have limited the application of agar

Method used

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  • Enzymolysis preparation method of new agaro oligosaccharides
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  • Enzymolysis preparation method of new agaro oligosaccharides

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Process optimization for preparing agar by extracting asparagus by water

[0046] (1) When the ratio of liquid to material is 16:1 and the time is 60 minutes, the optimum water extraction temperature is determined to be 120°C ( figure 1 ).

[0047] (2) At a temperature of 120°C and a liquid-to-solid ratio of 16:1, determine the optimum water extraction time to be 90 minutes ( figure 2 ).

[0048] (3) At a temperature of 120°C and a time of 90 minutes, determine the optimum water-extraction liquid-material ratio of 24:1 ( image 3 ).

[0049] On the basis of single factor experiment, adopt orthogonal test to carry out the optimization of water extraction agar, according to L 9 (3 4 ) orthogonal table for temperature (100°C, 110°C, 120°C); time (60min, 90min, 120min); liquid-to-solid ratio (20:1, 24:1, 28:1) orthogonal test, the factor level table is as follows Table 1 shows.

[0050] Table 1 Factor level table

[0051]

[0052] Analysis of the re...

Embodiment 2

[0055] Enzymatic hydrolysis preparation process optimization of embodiment 2 agar oligosaccharides

[0056] (1) Under the conditions of enzymatic hydrolysis temperature of 45°C, enzymatic hydrolysis time of 2 hours, substrate concentration of 0.5%, and enzyme addition of 10U / mL, the effect of pH change on the degree of hydrolysis was studied. When the pH value is 6.5, the degree of hydrolysis is maximum ( Figure 4 ).

[0057] (2) Under the conditions of enzymatic hydrolysis time of 2 hours, substrate concentration of 0.5%, pH value of 6.5, and enzyme addition of 10U / mL, the effect of temperature change on the degree of hydrolysis was studied. When the temperature is 55°C, the degree of hydrolysis is maximum ( Figure 5).

[0058] (3) Under the conditions of enzymolysis temperature of 55℃, substrate concentration of 0.5%, pH value of 6.5, and enzyme addition of 10U / mL, the effect of enzymolysis time on the degree of hydrolysis was studied. When the enzymolysis time is 2h...

Embodiment 3

[0067] The separation and purification of embodiment 3 agar oligosaccharides

[0068] The enzymolysis product obtained by enzymolysis was separated and purified by Sephadex LH-20 gel column. Use distilled water as eluent, flow rate 0.3mL / min, collect 1.2mL in each tube, adopt phenol-sulfuric acid method to measure oligosaccharide content ( Figure 9 ).

[0069] The components were collected, concentrated and then subjected to thin-layer chromatography to determine their components and purity. The new agar-oligosaccharide mixture is well separated by the LH-20 chromatographic column, and the obtained oligosaccharides are: new agarosetrose, new agarohexose, new agaroctaose ( Figure 10 , in the figure 1 is the standard new agarose oligosaccharides, from top to bottom are new agarose, new agarotetraose, new agarosexose, new agarocatose; 2 is the new agarose in the enzymatic hydrolysis product; 3 is New agarose in the hydrolyzate; 4 is new agarose in the hydrolyzate).

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Abstract

The invention provides an enzymolysis preparation method of new agaro oligosaccharides and relates to the field of biological engineering. Asparagus serves as raw materials, agar is prepared through high-temperature and high-pressure water extraction, agarase fermentation liquid produced through spontaneous fermentation in a laboratory is used for preparing the new agaro oligosaccharides, the oligosaccharides is purified through a gel chromatography, and a set of complete new agaro oligosaccharides preparation technology is established. The agar prepared through water extraction has the advantages that the yield is high, no chemical reagents need to be consumed, and the agar is environmentally friendly and low in cost, the agar can serve as raw materials for preparing agaro oligosaccharides, and high-value utilization of asparagus is achieved; an enzymatic method is used for replacing a traditional chemical method to prepare agaro oligosaccharides, environmental pollution is reduced, and high-bioactivity agaro oligosaccharides can be obtained.

Description

technical field [0001] The invention relates to an enzymolysis preparation method of new agar oligosaccharides, which belongs to the field of bioengineering. technical background [0002] Gracilaria( Gracilarialemaneiformis ) is the genus Elephant ( GracialariaGreville ) is a seaweed plant that mainly contains polysaccharides, phycobiliproteins and other components. The planting area is wide and the annual output is high. It is mainly used as raw material for preparing agar and abalone feed. [0003] Agar (Agar) is a complex water-soluble red algae polysaccharide, which is concentrated in the medullary intercellular space of the cell wall of algal plants. It has the advantages of thermoreversibility, colorless and odorless, insensitive to pH, melting point higher than freezing point, and strong stability. Based on the many advantages of agar, it has a wide range of applications, and has applications in biology, food industry, chemical industry, medical treatment and othe...

Claims

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Application Information

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IPC IPC(8): C12P19/14
Inventor 林娟谢金盛杨捷叶秀云
Owner FUZHOU UNIV
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