阅读说明：本技术 一种利用鲍鱼内脏粗酶高效制备低黏度魔芋胶的方法 (A method of low-viscosity konjac glucomannan is efficiently prepared using the thick enzyme of abalone internal organ ) 是由 孙乐常 文嘉欣 曹敏杰 张凌晶 章骞 陈玉磊 于 2019-08-22 设计创作，主要内容包括：本发明属于魔芋胶酶解技术领域,公开了一种利用鲍鱼内脏粗酶高效制备低黏度魔芋胶的方法,包括以下步骤：取鲍鱼内脏,用缓冲液抽提、离心后冷冻干燥制成粗酶,再加入魔芋胶溶液中,在一定条件下进行酶解反应,得到不同程度的低黏度魔芋胶酶解产物。本发明采用鲍鱼内脏为原料提取粗酶,并用该粗酶降解魔芋胶,以此得到低黏度目标产物,反应可控性强、条件温和、对设备无腐蚀。本发明利用低值的鲍鱼内脏,不仅减少资源浪费和环境污染,还可大幅降低生产成本。(The invention belongs to konjac glucomannan zymolysis technique fields, disclose a kind of method for efficiently preparing low-viscosity konjac glucomannan using the thick enzyme of abalone internal organ, the following steps are included: taking abalone internal organ, thick enzyme is made with freeze-drying after buffer extractions, centrifugation, it adds in konjaku sol solution, enzyme digestion reaction is carried out under certain condition, obtains different degrees of low-viscosity konjac glucomannan enzymolysis product.The present invention uses abalone internal organ to extract thick enzyme for raw material, and with the thick enzyme degraded konjak, obtains low-viscosity target product with this, and reaction controllability is strong, mild condition, corrosion-free to equipment.The present invention utilizes the abalone internal organ of low value, not only reduces the wasting of resources and environmental pollution, production cost can also be greatly reduced.)

1. a kind of method for efficiently preparing low-viscosity konjac glucomannan using the thick enzyme of abalone internal organ, which comprises the steps of:

(1) it produces the thick enzyme of abalone internal organ: abalone internal organ being cleaned with clear water, are placed on group after mixing with 20mM phosphate buffer Knit in bruisher and be homogenized, under the conditions of 4 DEG C, 10000 × g centrifugation 20min take supernatant, 3% active carbon under the conditions of 25 DEG C, 180rpm is uniformly mixed 30min removal of impurities except the freeze-drying acquisition thick enzyme of abalone internal organ after raw meat；

(2) it prepares konjaku rubber powder: the thick enzyme of abalone internal organ prepared by step (1) being even added in 2% konjaku sol solution, sufficiently Heating reaction certain time under the conditions of 50 DEG C after mixing, after completion of the reaction, enzyme deactivation 10min in boiling water bath, freeze-drying obtains enzyme Solve konjaku rubber powder；

(3) the enzymatic hydrolysis konjaku rubber powder that step (2) is prepared is dispersed in distilled water and forms enzymatic hydrolysis konjaku glue solution, Survey its viscosity.

2. a kind of method for efficiently preparing low-viscosity konjac glucomannan using the thick enzyme of abalone internal organ according to claim 1, special Sign is that the abalone internal organ are fresh abalone internal organ or the abalone internal organ frozen.

3. a kind of method for efficiently preparing low-viscosity konjac glucomannan using the thick enzyme of abalone internal organ according to claim 1, special Sign is that the abalone internal organ include the internal organ of the common abalone such as haliotis discus hannai Ino, Haliotis diversicolor, haliotis diversicolor Reeve.

4. a kind of method for efficiently preparing low-viscosity konjac glucomannan using the thick enzyme of abalone internal organ according to claim 1, special Sign is, in step (1), the pH of 20mM phosphate buffer used is 5.0-7.0, and optimal pH is 6.0.

5. a kind of method for efficiently preparing low-viscosity konjac glucomannan using the thick enzyme of abalone internal organ according to claim 1, special Sign is, in step (1), the ratio of abalone internal organ used and phosphate buffer is 1:3-1:5 (w/v), best proportion For 1:4 (w/v).

6. a kind of method for efficiently preparing low-viscosity konjac glucomannan using the thick enzyme of abalone internal organ according to claim 1, special Sign is, in step (1), the enzyme activity of the thick enzyme of abalone internal organ of preparation is 2000U/g or more.

7. a kind of method for efficiently preparing low-viscosity konjac glucomannan using the thick enzyme of abalone internal organ according to claim 1, special Sign is, in step (2), the amount ratio of abalone internal organ crude enzyme liquid used and konjac glucomannan is 100U:1g.

8. a kind of method for efficiently preparing low-viscosity konjac glucomannan using the thick enzyme of abalone internal organ according to claim 1, special Sign is, in step (2), the time of enzymatic hydrolysis is 30-120min.

9. a kind of method for efficiently preparing low-viscosity konjac glucomannan using the thick enzyme of abalone internal organ according to claim 1, special Sign is, in step (3), the enzymatic hydrolysis konjac glucomannan concentration of aqueous solution produced is 1%.

Technical field

It is the present invention relates to konjac glucomannan zymolysis technique field, in particular to a kind of efficiently to prepare low stick using the thick enzyme of abalone internal organ Spend the method for konjac glucomannan.

Background technique

The main active konjaku glucomannan (Konjac glucomannan, KGM) of konjac glucomannan, is a kind of water solubility Non-ionic macromolecule polysaccharide, it is to pass through β-Isosorbide-5-Nitrae and a small amount of by the molar ratio of 1:1.6 by D-Glucose and D-MANNOSE β -1,3 pyranosides bond close the linear Random copolymer RCP formed, and the acetyl group for possessing 5% to 10% replaces residue.About There is one to be attached on the position C-6 on every 19 saccharide residues with the acetyl group that ester bond combines.Konjac glucomannan cannot be by human body alimentary canal Digestion enzyme hydrolysis, be a kind of non-thermal indigestible dietary fiber.KGM has been demonstrated that enteron aisle, promotion enteron aisle can be dredged Activity, strengthen immunity, and have effects that reduce Blood Cholesterol, blood glucose level and fat content.

The highest a kind of natural polysaccharide of viscosity in the polysaccharide that konjaku glucomannan is almost studied at present, can water swelling 80 ~100 times, there are good thickening properties.Natural konjac glucomannan is not easy to store etc. since viscosity is excessive, stability is poor, degradable Disadvantage limits its application in the food industry.Currently, the main method of degradation of polysaccharide is acid degradation method and enzyme edman degradation Edman.

Summary of the invention

For above-mentioned deficiency in the prior art, low stick efficiently is prepared using the thick enzyme of abalone internal organ the present invention provides a kind of The method of konjac glucomannan is spent, it is environmentally friendly, stable and efficient.

In order to achieve the above object of the invention, the technical solution adopted by the present invention are as follows:

A method of low-viscosity konjac glucomannan efficiently being prepared using the thick enzyme of abalone internal organ, is included the following steps:

(1) it produces the thick enzyme of abalone internal organ: abalone internal organ being cleaned with clear water, are placed after being mixed with 20mM phosphate buffer Be homogenized in tissue mashing machine, under the conditions of 4 DEG C, 10000 × g centrifugation 20min take supernatant, 3% active carbon under the conditions of 25 DEG C, 180rpm is uniformly mixed 30min removal of impurities except the freeze-drying acquisition thick enzyme of abalone internal organ after raw meat；

(2) it prepares konjaku rubber powder: the thick enzyme of abalone internal organ prepared by step (1) is even added in 2% konjaku sol solution, Heating reaction certain time under the conditions of 50 DEG C after mixing well, after completion of the reaction, enzyme deactivation 10min in boiling water bath, freeze-drying obtains To enzymatic hydrolysis konjaku rubber powder；

(3) the enzymatic hydrolysis konjaku rubber powder that step (2) is prepared is dispersed in distilled water and forms enzymatic hydrolysis konjaku glue Solution surveys its viscosity.

Preferably, the abalone internal organ are fresh abalone internal organ or the abalone internal organ frozen.

Preferably, the abalone internal organ include the internal organ of the common abalone such as haliotis discus hannai Ino, Haliotis diversicolor, haliotis diversicolor Reeve.

Preferably, the pH of 20mM phosphate buffer used is 5.0-7.0, and optimal pH is in step (1) 6.0。

Preferably, the ratio of abalone internal organ used and phosphate buffer is 1:3-1:5 (w/ in step (1) V), best proportion is 1:4 (w/v).

Preferably, the enzyme activity of the thick enzyme of abalone internal organ of preparation is 2000U/g or more in step (1).

Preferably, the amount ratio of abalone internal organ crude enzyme liquid used and konjac glucomannan is 100U:1g in step (2).

Preferably, the time of enzymatic hydrolysis is 30-120min in step (2).

Preferably, the enzymatic hydrolysis konjac glucomannan concentration of aqueous solution produced is 1% in step (3).

The invention has the benefit that abalone internal organ is used to extract thick enzyme for raw material, and with the thick enzyme degraded konjak, with This obtains low-viscosity target product, and reaction controllability is strong, mild condition, corrosion-free to equipment, has not only widened traditional konjac glucomannan Application range, and abalone internal organ can be efficiently used, reduce the wasting of resources and environmental pollution, there is significant environmental protection effect.

Detailed description of the invention

Fig. 1 is the result figure that the efficiency of konjac glucomannan is decomposed using different enzymes.

Specific embodiment

In order that the present invention can be more clearly and readily understood, below by technical solution in the embodiment of the present invention It is clearly and completely described.

A method of low-viscosity konjac glucomannan efficiently being prepared using the thick enzyme of abalone internal organ, is included the following steps:

(1) it produces the thick enzyme of abalone internal organ: abalone internal organ being cleaned with clear water, are placed after being mixed with 20mM phosphate buffer It is homogenized in tissue mashing machine, the pH of phosphate buffer used is 5.0-7.0, and optimal pH is 6.0；

The ratio of abalone internal organ and phosphate buffer is 1:3-1:5 (w/v), and best proportion is 1:4 (w/v)；

Then under the conditions of 4 DEG C, 10000 × g centrifugation 20min takes supernatant, and 3% active carbon is under the conditions of 25 DEG C, 180rpm It is uniformly mixed freeze-drying after 30min removal of impurities removes raw meat and obtains the thick enzyme of abalone internal organ, the enzyme activity of the thick enzyme of abalone internal organ of preparation is 2000U/g or more；

(2) it prepares konjaku rubber powder: the thick enzyme of abalone internal organ prepared by step (1) is even added in 2% konjaku sol solution, The amount ratio of abalone internal organ crude enzyme liquid and konjac glucomannan is 100U:1g, heats reaction after mixing well under the conditions of 50 DEG C, digests 30- 120min, after completion of the reaction, enzyme deactivation 10min in boiling water bath, freeze-drying obtain enzymatic hydrolysis konjaku rubber powder；

(3) the enzymatic hydrolysis konjaku rubber powder that step (2) is prepared is dispersed in distilled water and forms enzymatic hydrolysis konjaku glue Solution, the enzymatic hydrolysis konjac glucomannan concentration of aqueous solution produced are 1%, finally survey its viscosity.

Preferably, abalone internal organ are fresh abalone internal organ or the abalone internal organ that freeze, including it is haliotis discus hannai Ino, variegated The internal organ of the common abalone in the In Fujian Provinces such as Bao, haliotis diversicolor Reeve；Abalone abalone internal organ are the leftover bits and pieces of abalone processing, are usually directly lost Abandon or used as animal feed, but nutritional ingredient rich in is natural polymorphic type glycosidase library, with it is at low cost, The high feature of enzymolysis efficiency.

Enzyme activity is defined as: under conditions of 37 DEG C, every min decomposes 1% sodium carboxymethylcellulose and generates 1mmol/L reproducibility The required enzyme amount of sugar is defined as 1 enzyme activity unit.