JP2021159064A - Konjac powder multienzyme hydrolysate and preparation method thereof - Google Patents

Abstract

To provide a konjac powder multienzyme hydrolysate and a preparation method thereof.SOLUTION: Konjac powder is purified in alcohol to obtain wet konjac powder raw material; dealcoholization is carried out on the wet konjac powder to obtain purified konjac powder; water and enzyme are added to perform enzymolysis; and finally, the konjac powder multienzyme hydrolysate is obtained. Optimizing the process can provide the enzymatic hydrolysate including certain instant polysaccharose and oligosaccharide.SELECTED DRAWING: None

Description

The present invention relates to the technical field of functional food processing, and more particularly to a konjac fine powder enzymatic decomposition product by a multienzyme and a method for producing the same.

Konjac has high functionality and medical value. According to modern research, the main chemical constituents of konjac stalks are glucomannan, starch, other polysaccharides, crude proteins, essential amino acids, and the human body such as potassium, calcium, magnesium, sodium, iron, manganese, and copper. It is an essential trace element.

Konjac glucomannan is a rich renewable natural polymer resource following starch and cellulose, and has biodegradability and water solubility. Glucomannan is a natural dietary fiber with a large molecular weight and the highest viscosity, and can adsorb 200 times as much water molecules as its own volume to form a viscous solution, which can enhance the feeling of fullness. Is. Moreover, since it is not affected by digestive enzymes of the human body, it does not generate calories. That is, glucomannan is a well-known soluble and multifunctional polysaccharide, and it is also possible to produce a konjac oligosaccharide having a more enhanced function by appropriately decomposing it. However, glucomannan has a slow rate of water absorption expansion and dissolution, and usually takes 2 hours or more.

Currently, the enzymatic degradation of konjac glucomannan presents the following issues that require optimization. (1) Since the enzyme is decomposed by a single enzyme, the amount of enzyme used is large, and the enzyme decomposition requires an excessive amount of time. In addition, the utilization rate of konjac fine powder is low. (2) In some enzymatic decomposition methods, the detection of reducing sugars is the main index, and a large amount of reducing sugars does not mean a large amount of oligosaccharides. On the contrary, on the premise that the contents of soluble solids are the same, the higher the content of reducing sugars, the lower the content of oligosaccharides. Therefore, in the case of such an enzymatic decomposition technique, there is no point in producing oligosaccharides. (3) Some enzymatic decomposition methods can be applied only to laboratory research and cannot be introduced into full-scale production. (4) Since konjac fine powder has a characteristic of water absorption and expansion, a feeling of abdominal bloating is likely to occur if it is eaten without decomposition, and there is a certain safety risk. However, such a defect can be eliminated by appropriately decomposing, and it is possible to maintain the conventional functional characteristics while increasing the dissolution rate.

An object of the present invention is to provide a polyenzymatic konjac fine powder enzymatic decomposition product and a method for producing the same. According to this, the drawbacks such as long enzymatic decomposition time, incomplete enzymatic decomposition, and low utilization rate of konjac fine powder in the prior art are eliminated, and an enzyme containing a polysaccharide and an oligosaccharide having a certain rapid solubility is eliminated. Degradation products are obtained.

In order to realize the object of the above invention, the present invention provides the following technical means.

The present invention provides a method for producing a konjac fine powder enzymatic decomposition product using multiple enzymes. The production method is as follows: (1) Purifying the konjak refined raw material in alcohol to obtain konjak wet semen, and (2) Purifying the konjak moistened powder with alcohol removal treatment. (3) The step of obtaining the enzymatically decomposed konjak powder by multiple enzymes by mixing the obtained purified konjak powder with water and an enzyme and performing enzymatic decomposition is included.

Preferably, the volume concentration of alcohol in the step (1) is 55 to 75%. The solid-liquid ratio of the konjac refined powder raw material and alcohol in the step (1) is 1 g: 3 to 5 ml, and the purification time in the step (1) is 1 to 3 hours. The temperature of the alcohol removal treatment in the step (2) is 50 to 70 ° C.

Preferably, the mass-volume ratio of the purified konjac fine powder to water in the step (3) is 15 to 30 g: 100 mL. The enzyme in step (3) includes glucomannanase, cellulase and pectase. The dose of the glucomannanase was 50 to 70 u / g with respect to the purified konjac refined powder, the dose of the cellulase was 40 to 60 u / g with respect to the purified konjac refined powder, and the pectase was 0.03 to 0. It accounts for 05%. The temperature of the enzymatic decomposition is 55 to 65 ° C., and the enzymatic decomposition time is 2 to 3 hours.

The present invention further provides a konjac fine powder enzymatic decomposition product obtained by the above-mentioned production method.

The present invention provides a konjac fine powder enzymatic decomposition product using multiple enzymes and a method for producing the same. The content of the functional component (for example, oligosaccharide) in the enzymatic decomposition product obtained by this method is 5.69 to 5.86 g with respect to 100 g of the refined powder, and 39.33 to 40.5% of the total sugar content. Occupy. In addition, it further contains a polysaccharide having a certain rapid solubility. The product has actions such as prevention of obesity, reduction of blood lipids, and prevention of hepatic fatty degeneration. In addition, it has effects such as lowering blood glucose, anti-inflammatory, and improving free radical removal in the living body, and has an improving effect on hyperglycemia in diabetic model rats.

According to the present invention, by performing enzymatic decomposition using konjac refined powder as a raw material and using a complex enzyme, the enzymatic decomposition process is accelerated, the enzymatic decomposition efficiency is improved, and the quality of the enzymatic decomposition product is improved. By optimizing the process parameters, production process parameters such as selection of enzyme to be used, enzyme dose, and enzymatic decomposition time can be rationalized, and an enzymatic decomposition product containing a polysaccharide and an oligosaccharide having a certain rapid solubility can be obtained.

The present invention provides a method for producing a konjac fine powder enzymatic decomposition product by a multienzyme, which comprises the following steps.

(1) Konjac wet fine powder is obtained by purifying the raw material of konjac fine powder in alcohol.

(2) Purified konjak fine powder is obtained by treating the wet konjak fine powder with alcohol.

(3) The obtained purified konjak refined powder is mixed with water and an enzyme and enzymatically decomposed to obtain an enzymatically decomposed konjak refined powder by multiple enzymes.

In the present invention, the konjak refined powder is preferably a commercially available high-purity konjak refined powder.

In the present invention, the volume concentration of alcohol in step (1) is preferably 55 to 75%, more preferably 60 to 70%, and even more preferably 62 to 68%.

In the present invention, the solid-liquid ratio of the konjac refined powder raw material and alcohol in the step (1) is preferably 1 g: 3 to 5 ml, and more preferably 1 g: 3.5 to 4.5 ml.

In the present invention, the purification time of the step (1) is preferably 1 to 3 hours, more preferably 1.5 to 2.5 hours.

In the present invention, in the purification, it is preferable that the raw material of konjak refined powder is stirred with alcohol to be uniformly mixed and immersed.

In the present invention, the purification is for removing _{fishy odor, pungent taste, SO 2, etc. contained in konjac fine powder.}

In the present invention, the alcohol removal treatment is preferably carried out by hot air drying as a means.

In the present invention, the temperature of the alcohol removal treatment is preferably 50 to 70 ° C, more preferably 55 to 65 ° C, and even more preferably 58 to 62 ° C.

In the present invention, the alcohol removal treatment is for removing the alcohol odor, and is based on the fact that the alcohol odor is no longer felt.

In the present invention, the mass-volume ratio of the purified konjac fine powder in step (3) to water is preferably 15 to 30 g: 100 mL, and more preferably 20 to 25 g: 100 mL.

In the present invention, the enzyme of step (3) preferably contains glucomannanase, cellulase and pectase.

In the present invention, the dose of the glucomannanase is preferably 50 to 70 u / g with respect to the purified konjac fine powder, more preferably 55 to 65 u / g with respect to the purified konjac fine powder, and further preferably the purified konjac. It is set to 60 to 62 u / g with respect to the refined powder. The dose of the cellulase is preferably 40 to 60 u / g with respect to the purified konjac powder, more preferably 45 to 55 u / g with respect to the purified konjac powder, and further preferably 47 with respect to the purified konjac powder. ~ 52u / g. The dose of the pectase is preferably 0.03 to 0.05% of the total mass of the system, and more preferably 0.035 to 0.04% of the total mass of the system.

In the present invention, the system refers to a mixture obtained by mixing purified konjac fine powder, an enzyme and water.

In the present invention, the temperature of the enzymatic decomposition is preferably 55 to 65 ° C, more preferably 58 to 60 ° C. Further, preferably, the enzymatic decomposition time is set to 2 to 3 hours.

The principle of the enzymatic decomposition in the present invention is that glucomannan is enzymatically decomposed into konjac polysaccharide and konjac oligosaccharide having a low degree of polymerization by using glucomannanase and cellulase at the same time based on the specificity of the enzyme. In addition, pectin decomposes pectin into galacturonic acid, which destroys the particle structure of konjac fine powder, thereby improving the action and effect of other enzymes.

The present invention further provides a konjac fine powder enzymatic decomposition product obtained by the above-mentioned production method.

Hereinafter, the technical means provided by the present invention will be described in detail in combination with examples, but it should not be understood as limiting the scope of protection of the present invention.

Step 1: Purification: The konjac fine powder raw material was purified in alcohol having a volume concentration of 55%. The solid-liquid ratio of the konjak refined powder raw material to alcohol was 1 g: 3 ml, and the konjak wet fine powder was obtained by purifying for 1 hour.

Step 2: Alcohol removal: Konjac wet powder was treated with alcohol removal. The alcohol removal temperature was set to 50 ° C., and the purified konjac fine powder was obtained by removing the alcohol.

Step 3: Enzymatic decomposition: 15 g of purified konjac fine powder was taken, 100 mL of water was added, and glucomannanase, cellulase and pectase were further added for enzymatic decomposition. The dose of glucomannanase was 50 u / g with respect to purified konjac fine powder, and the dose of cellulase was 40 u / g with respect to purified konjac fine powder. The dose of pectase was 0.03% of the total mass of the system. By enzymatically decomposing for 2 hours under the condition of 55 ° C., a konjac fine powder enzymatically decomposed product by multiple enzymes was obtained.

The content of the functional component (oligosaccharide) in the obtained enzymatic decomposition product was 5.86 g with respect to 100 g of the refined powder, accounting for 40.5% of the total sugar content.

Step 1: Purification: The konjac fine powder raw material was purified in alcohol having a volume concentration of 60%. The solid-liquid ratio of the konjac fine powder raw material to alcohol was 1 g: 4 ml, and the konjak wet fine powder was obtained by purifying for 1.5 hours.

Step 2: Alcohol removal: Konjac wet powder was treated with alcohol removal. The alcohol removal temperature was 65 ° C., and the purified konjac fine powder was obtained by removing the alcohol.

Step 3: Enzymatic decomposition: 25 g of purified konjac fine powder was taken, 100 mL of water was added, and glucomannanase, cellulase and pectase were further added for enzymatic decomposition. The dose of glucomannanase was 60 u / g with respect to purified konjac fine powder, and the dose of cellulase was 50 u / g with respect to purified konjac fine powder. The dose of pectase was 0.04% of the total mass of the system. By enzymatically decomposing for 2.5 hours under the condition of 60 ° C., a konjac fine powder enzymatic decomposition product by multiple enzymes was obtained.

The content of the functional component (oligosaccharide) in the obtained enzymatic decomposition product was 5.78 g with respect to 100 g of the refined powder, accounting for 39.95% of the total sugar content.

Step 1: Purification: The konjac fine powder raw material was purified in alcohol having a volume concentration of 75%. The solid-liquid ratio of the konjak refined powder raw material to alcohol was 1 g: 5 ml, and the konjak wet fine powder was obtained by purifying for 3 hours.

Step 2: Alcohol removal: Konjac wet powder was treated with alcohol removal. The alcohol removal temperature was set to 70 ° C., and the purified konjac fine powder was obtained by removing the alcohol.

Step 3: Enzymatic decomposition: 30 g of purified konjac fine powder was taken, 100 mL of water was added, and glucomannanase, cellulase and pectase were further added for enzymatic decomposition. The dose of glucomannanase was 70 u / g with respect to the purified konjac powder, and the dose of cellulase was 60 u / g with respect to the purified konjac powder. The dose of pectase was 0.05% of the total mass of the system. By enzymatically decomposing for 3 hours under the condition of 65 ° C., a konjac fine powder enzymatically decomposed product by multiple enzymes was obtained.

The content of the functional component (oligosaccharide) in the obtained enzymatic decomposition product was 5.69 g with respect to 100 g of the refined powder, accounting for 39.33% of the total sugar content.

Animal experiments were carried out on the enzymatic decomposition products of konjac fine powder produced by the polyenzymes produced in Examples 1 to 3.

Step 1: Preparation and grouping of animal models 50 healthy female mice were selected and fed with a general diet for 2 weeks, followed by 10 blank control groups (fed with a general diet) and 40 model control groups (high fat). The feed was randomly divided into two groups (feeding). Next, a mouse obesity model was prepared by feeding unlimited feeds in both groups and breeding them for 4 weeks. Then, the model control group was divided into a high-lipid model group (fed with a high-fat feed) and a low-dose group of enzyme-degraded konyaku fine powder (fed with a high-fat feed, and the enzymatic decomposition product was 15 mg / 100 g ^-1 (fed). BW) · d ^-1 was administered intragastrically), and the dose group in the enzyme degradation product of Konnaku semen (fed a high-lipid feed, and the enzymatic degradation product 30 mg · 100 g ^-1 (BW) · d ^-1 was intragastrically administered. ), And the enzyme-degraded product high-dose group (feeding a high-lipid feed and intragastrically administering the ^{enzyme-degraded product 60 mg, 100 g -1} (BW), d ^-1). Subsequently, the mice in the blank control group were continuously fed with a general diet, while the mice in the high-fat model group and the low-, medium-, and high-dose groups of konjac fine powder enzyme decomposition products were continuously fed with a high-fat diet. In addition, the enzymatic decomposition product of konjac fine powder was intragastrically administered for 1 consecutive month. The breeding temperature was 24 to 27 ° C., the relative humidity was 45 to 50%, and intermittent lighting with a day / night ratio of 11h: 13h was used.

Step 2: Sample collection 10 weeks after feeding, mice in each group were fasted for 12 hours and weighed before slaughter. Then, after anesthesia, the eyeball was quickly removed, blood was collected, and the blood was killed by dislocation of the cervical spine. Next, the liver, heart, spleen, kidneys, lungs, perineal fat and subcutaneous fat in the inguinal region were quickly separated, the organs were washed with salt water, the water was removed with filter paper, and the weights of the organs and tissues were weighed. ..

Step 3: Measurement of indicators During the experimental period, mouse growth status (mouse coat color, feeding, defecation and activity) was recorded daily. In addition, the daily food intake and water intake of the mice were measured on Wednesday and Thursday, respectively. That is, the total amount of feed and the total amount of water supplied were measured on Wednesday and Thursday, and the remaining amount of feed and the remaining amount of water were measured at the same time on Thursday and Friday (that is, the amount of food intake = the total amount of feed-the remaining amount of feed). Drinking amount = total amount of water-remaining amount of water). In addition, the body weight was measured once every Saturday.

The abdomen was incised, all the adipose tissue in the mouse body (including the adipose tissue around the kidney and the subcutaneous adipose tissue in the inguinal region) was removed, the fat was measured, and the fat index was calculated based on the formula (1).

Fat index /% = m ₁ / m x 100 (1)

In the formula, m is the body weight / g of the mouse before the killing treatment, and m ₁ is the mass / g of each adipose tissue in the mouse body.

Step 4: Statistical analysis was performed with the data processing software SPSS20 and expressed as mean ± standard deviation. One-way ANOVA was used for group-to-group comparison. P <0.05 indicates that there is a significant difference between the two groups.

As is clear from Table 1, there was a significant difference between the high-lipid model group and the low-, medium-, and high-dose groups of konjac enzymatic degradation products (P <0.05). This means that konjac enzymatic degradation products can reduce the amount of water consumed by mice to some extent. The effect of the high dose group of konjac enzymatic decomposition products was the most remarkable.

Animal experiments were carried out on the enzymatic decomposition products of konjac fine powder produced by the polyenzymes produced in Examples 1 to 3.

Step 1: Creating a diabetic rat model 60 healthy female rats were successfully bred for 1 week. In addition, during the period, the amount of water consumed and the amount of food consumed by the experimental animals were measured regularly. One week later, the body weight and fasting blood glucose level of the experimental animals were measured and recorded, and 10 animals were randomly grouped (normal control group and 5 model groups). Rats in the model group were intraperitoneally injected with STZ-containing citrate-sodium citrate buffer to induce diabetes (pre-cooled citrate buffer pH = 4.5 was used and a concentration of 1). After dissolving STZ in%, it was sterilized by filtering with a microfiltration membrane of 0.22 mol / L. Since STZ tends to lose its activity, it was quickly weighed and then wrapped in tin foil to shield it from light. Was used on the spot and was always kept waiting in the ice bath). The concentration of STZ to be injected was 45 mg / kg, and the injection amount was 2%. On the other hand, rats in the normal group were intraperitoneally injected with an equal volume of citrate-sodium citrate buffer. In addition, during the STZ injection process, it was necessary to disinfect the skin of the abdominal cavity of the rat to prevent infection. After the injection was completed, each group continued to maintain their previous eating and drinking. Then, 24 hours later, blood was collected from the tail of the rat, and the fasting blood glucose level was measured. When the fasting blood glucose level of the rats was measured using a glucose meter and a blood glucose test paper kit, the fasting blood glucose level of the rats in the model group was> 16.7 mmol / L (it was possible to measure repeatedly 2-3 times). However, the blood sugar level was stable). Moreover, since polydipsia and polyuria and weight loss occurred as symptoms, it was considered that the model was successfully created.

Step 2: The diabetic rats that succeeded in grouping the animals and creating a dosing model were grouped and adjusted into a konjac fine powder enzyme decomposition product high / medium / low dose group, a positive administration group, and a diabetes model group, respectively. In addition, rats with the same blood glucose level as before model preparation were defined as the normal group. Next, intragastric administration was performed for 6 consecutive weeks. During the experiment, rats in each group were allowed to drink and feed freely as usual. The time of intragastric administration was 4 pm daily. Regarding the gastric dose to the rats in each group, 2.500 g / (kg · d), 1.250 g / (kg · d), and 0. It was 625 g / (kg · d) and 0.140 g / (kg · d) in the positive administration group. Moreover, an equal amount of physiological saline was administered to the normal group and the hyperglycemia model group.

Step 3: Measurement of indicators During the period of intragastric administration to rats, the weight of rats in each group was measured and recorded weekly. The results are shown in g, and the difference in body weight of rats in each group was analyzed and compared. The fasting blood glucose level of the experimental rats in each group was measured, the results were shown in mmol / L, and the changes in the blood glucose level of the rats in each group were analyzed and compared. In addition, the amount of water and food intake of the experimental animals was measured and recorded, and changes in the amount of water and food intake of rats in each group during the gastric administration period were analyzed.

Step 4: Statistical method Statistical analysis was performed with the software SPSS20 and expressed as mean ± standard deviation. One-way ANOVA was used for group-to-group comparison. P <0.05 indicates that there is a significant difference between the two groups.

As is clear from Table 2, on day 0 of administration, the amount of water consumed in the model group was significantly higher than that in the normal group (p <0.01). In addition, 21 days after the administration, the amount of drinking water in the positive administration group and each konjac enzyme decomposition product dose group decreased to a different degree. Then, on the 42nd day of administration, the amount of water consumed by the model group showed a very significant difference from that of the normal group (p <0.01). In addition, the amount of water consumed by the positive administration group and each konjac enzymatic decomposition product dose group showed a very significant difference from the model group (p <0.01). Moreover, the amount of drinking water in the high / medium dose group of konjac enzymatic degradation product was not significantly different from that in the positive dose group. As is clear from the above, the konjac enzymatic degradation product was able to alleviate the polydipsia symptom of diabetic rats. In addition, a fixed dose of konjac enzymatic degradation product had an effect similar to that of metformin in alleviating polydipsia symptoms in diabetic patients.

As is clear from the above examples, the present invention provides a method for producing a konjac fine powder enzymatic decomposition product using multiple enzymes. From the results of animal (white rat) experiments, it was found that the product has actions such as prevention of obesity, reduction of blood lipids, and prevention of hepatic fatty degeneration. It was also found that it has effects such as lowering blood glucose, lowering lipid peroxidation, anti-inflammatory, and improving free radical removal in the living body, and has an improving effect on hyperglycemia in diabetic model rats.

The above is only a preferable embodiment of the present invention. Those skilled in the art can make slight improvements and supplements on the premise that they do not deviate from the principles of the present invention, and these improvements and supplements should also be regarded as the scope of protection of the present invention.

Claims (4)

It is a method for producing konjac fine powder enzymatic decomposition products by multiple enzymes.
(1) The step of obtaining konjac wet powder by purifying the raw material of konjac powder in alcohol, and
(2) A step of obtaining purified konjac fine powder by alcohol-removing the konjak wet fine powder, and
(3) A method comprising a step of obtaining the enzymatic decomposition product of the konjac fine powder by a multienzyme by mixing the obtained purified konjac fine powder with water and an enzyme and performing enzymatic decomposition. The volume concentration of the alcohol in the step (1) was 55 to 75%.
The solid-liquid ratio of the konjak refined powder raw material and the alcohol in the step (1) was 1 g: 3 to 5 ml.
The purification time in the step (1) was set to 1 to 3 hours.
The production method according to claim 1, wherein the temperature of the alcohol removal treatment in the step (2) is 50 to 70 ° C. The mass-volume ratio of the purified konjac fine powder to the water in the step (3) was 15 to 30 g: 100 mL.
The enzyme in step (3) comprises glucomannanase, cellulase and pectase.
The dose of the glucomannanase is 50 to 70 u / g with respect to the purified konjac refined powder, the dose of the cellulase is 40 to 60 u / g with respect to the purified konjac refined powder, and the pectase is 0.03 to 0.03 to the total mass of the system. Occupies 0.05%
The production method according to claim 1, wherein the enzymatic decomposition temperature is 55 to 65 ° C., and the enzymatic decomposition time is 2 to 3 hours. A konjac fine powder enzymatic decomposition product obtained by the production method according to any one of claims 1 to 3.