KR100699431B1 - Brewing method with maltogenic amylase - Google Patents

Abstract

The present invention relates to a liquor manufacturing method using maltogenic amylase. In particular, the present invention saccharifies starch with maltose to allow the fermentation by the yeast to proceed slowly to reduce the sugar content, as well as to form branched oligosaccharides, which can significantly increase the content of useful branched oligosaccharides in the body. It relates to a manufacturing method and to liquor produced by the above method.

Functional alcohol, low calorie alcohol, maltogenic amylase, branched oligosaccharides

Description

Brewing method using maltogenic amylase {BREWING METHOD WITH MALTOGENIC AMYLASE}

Figure 1 shows the manufacturing process (a) of the functional low calorie liquor using the maltogenic amylase of the present invention and the conventional liquor manufacturing process (b).

Figure 2 is a comparative analysis of the functional low calorie liquor prepared in the present invention and the components of commercially available alcohol using thin film chromatography.

Figures 3a to 3f is a comparative analysis of the components of the functional low calorie liquor prepared in the present invention and the components of the commercially available alcohol using high performance ion exchange chromatography.

[TECHNICAL FIELD OF THE INVENTION]

The present invention relates to a liquor manufacturing method using maltogenic amylase, and more specifically, to produce maltose and branched oligosaccharides using maltogenic amylase during casting and fermented by yeast, thereby producing a low-calorie functional liquor It is about a method.

[Private Technology]

Conventional liquor manufacturing method hydrolyzes the raw starch, such as rice, glutinous rice, barley, corn, wheat, etc. with glucoamylase of malt fungus, alpha amylase and beta amylase of malt, to produce glucose or maltose, and to use alcohol with yeast Fermentation. In the mainstream prepared in this way, several kinds of sugars, including glucose, remain, and these are mainly maltooligosaccharides in which glucose molecules are linked by alpha-1,4 bonds, and branches in which glucose molecules are linked by one or more alpha-1,6 bonds. It consists of branched oligosaccharides.

Glucose and maltooligosaccharides are digested and absorbed by the body to act as obesity factors, while branched oligosaccharides are not digested and absorbed by the body but act as active factors of the bifidus bacteria, which are intestinal useful bacteria. It is being spotlighted as a health-oriented food material that is effective in preventing and preventing tooth decay. However, liquor prepared by the conventional casting method has a low content of branched oligosaccharides and high total sugar content, including glucose, is considered to be a harmful food that not only causes obesity but also harms health when drinking excessively.

In order to solve the problems of the prior art, the present invention aims to provide a liquor production method that can lower the total sugar content in liquor and at the same time increase the branched oligosaccharide content.

It is another object of the present invention to provide a saccharifying agent that can be used to prepare a low calorie liquor having a high content of branched oligosaccharides.

In order to achieve the above object, the present invention provides a liquor manufacturing method comprising a saccharification and fermentation step, wherein the liquor manufacturing method comprises a liquor manufacturing method comprising saccharifying a liquor raw material with maltogenic amylase to produce maltose and branched oligosaccharides. to provide.

In another aspect, the present invention provides a liquor produced by the liquor manufacturing method.

In another aspect, the present invention provides a casting saccharifying agent comprising maltogenic amylase as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present inventors found that the existing liquor manufacturing method produces glucose by decomposing starch into glucoamylase, but high concentrations of glucose cause shock in yeast and are not completely utilized as a carbon source of yeast, and remain large in the final liquor. We tried to solve the problem. Thus, the inventors of the present invention, when using maltogenic amylase instead of glucoamylase, not only maltose and branched oligosaccharides are produced, but also maltose is sufficiently used by the yeast without causing an impact on the yeast, while the sugar content in the final liquor is significantly lower It was confirmed that a functional, low calorie liquor containing a large amount of branched oligosaccharides was prepared, and completed the present invention.

The main process of the liquor manufacturing method of the present invention is briefly shown in FIG. That is, maltogenic amylase according to the present invention glycosylates starch with maltose to facilitate fermentation by yeast, not only to reduce the sugar content, but also to form branched oligosaccharides, significantly increasing the content of useful branched oligosaccharides in the body.

Maltogenic amylase according to the present invention may be isolated and purified from a natural prokaryote or eukaryote, and may be prepared by artificially expressing a gene thereof by genetic recombination technology. Examples of strains producing maltogenic amylase include, but are not limited to, Thermus sp. And Bacillus stearothermophilus . Maltogenic amylase hydrolyzes starch mainly into maltose units and simultaneously transduces them to produce functional branched oligosaccharides and maltose wherein glucose molecules are linked by at least one alpha-1, 6 bond. The maltose produced is broken down by the yeast and converted to alcohol, but the branched oligosaccharides are contained in the final product without being broken down by the yeast.

The liquor manufacturing method of the present invention, in the liquor manufacturing method comprising the step of saccharification and fermentation, comprises liquifying the liquor raw material with maltogenic amylase to produce maltose and branched oligosaccharides. The liquor manufacturing method may be a conventional liquor method, and further includes a liquefaction step using alpha amylase and a saccharification step using maltogenic amylase, in particular, prior to the fermentation step of the liquor raw material, and not performing a saccharification step using existing glucoamylase. It is characterized by not. The saccharification step using the existing glucoamylase is carried out using Baekkukyun, Nuruk, coenzyme, purified enzyme, Hwangkuk, Black Bacillus or malt. That is, the liquor manufacturing method of the present invention is (a) liquefaction step of liquefaction by processing the alpha amylase to the liquor raw material, (b) saccharification step of producing maltose and branched oligosaccharides by treating maltogenic amylase to the liquefied liquor And (c) a fermentation step of fermenting the maltose using yeast.

The raw material of the liquor may be all kinds of foods used in conventional casting, for example, may be a starch raw material. Starch ingredients include white rice, glutinous rice, brown rice, barley (covered barley, rice barley, spring barley), sweet potato, corn, potato, soybean, sorghum, millet, crude, wheat flour, sugarcane, juniper fruit, herbal medicine, herbal medicine, rye and agave (Blue mezcal), but is not limited thereto, and includes all kinds of raw materials that can be easily implemented by those skilled in the art. The choice of starch raw material is preferably appropriately selected according to the liquor to be prepared, such as beer, whiskey, rum, gin, tequila, schnapps, vodka, takju, medicinal sake, sake, and soju.

In the step (a), alpha amylase commercially available in the liquor raw material may be used, and the alpha amylase treatment method and the amount of usage may employ a conventional method. For example, alpha amylase can be liquefied by treating 10 to 5000 U per gram of liquor raw material, and the liquefaction temperature (20 to 100 ° C.) and time are in accordance with conventional methods. In the liquefaction step, pullulanase may be additionally treated during alpha amylase treatment, and the amount of pullanase may be 1 to 1000 U per gram of liquor raw material. In addition, the step (a) can be omitted by purchasing and using a raw material liquefied using a commercially available alpha amylase.

Step (b) is a step of producing maltose and branched oligosaccharides by treating maltogenic amylase to the liquefied raw material, for example, 1 to 5000 U maltogenic amylase can be added per gram of liquefied raw material, but preferably Preference is given to conventional maltogenic amylase treatment levels. The saccharification step may be performed at 10 to 80 ° C. for at least 1 day, preferably at 15 to 60 ° C., but more preferably according to conventional conditions.

Step (c) is a step of inoculating fermented yeast, such as Saccharomyces cerevisiae in the saccharified raw material, for example, inoculated with yeast bacteria of 0.0001 to 10 g (dry weight) per 1 g of liquefied raw material 20 It may be fermented for 1 day or more at 35 ℃.

In the steps (b) and (c), maltogenic amylase and yeast may be simultaneously added to the liquefied raw material to simultaneously perform the saccharification and fermentation steps.

The liquor manufacturing method of the present invention can be applied by adopting a wide variety of well-known methods according to the liquor, it can be easily implemented by anyone skilled in the art to which the present invention belongs.

The present invention also provides a casting saccharifier comprising maltogenic amylase as an active ingredient. The saccharifying agent is used in the saccharification step during casting, the amount may be 1 to 5000 U per g of liquor raw materials, but is not limited thereto. Since the saccharifying agent has excellent activity efficiency at 15 to 60 ° C., it is preferable to maintain the temperature at the time of saccharification, and the reaction time may be 1 day or more.

The saccharifying agent according to the present invention may further include food additives other than maltogenic amylase. The food additive may be any substance added to the usual liquor.

Hereinafter, examples of the present invention will be described. The following examples are only for illustrating the present invention and the present invention is not limited to the following examples.

Example 1

Starch was used liquefied corn starch (manufactured by Samyang Genex), and maltogenic amylase transformed the maltogenic amylase gene cloned from Bacillus stearothermophilus into Escherichia coli MC1061 to express maltogenic amylase therefrom. And purified by column chromatography (Cha et al ., 1998, Eur. J. Biochem. 253, 251-262).

To 180 ml of liquefied corn starch, 170 U / mg of maltogenic amylase and 500 mg of yeast ( Saccharomyces serevisiae ) were added and left at 27 ° C. for 16 days.

Example 2

Starch was used as rice starch (Rice starch S-7260), and maltogenic amylase was transformed from the maltogenic amylase gene cloned from Bacillus stearothermophilus into Escherichia coli MC1061, thereby producing maltogenic Amylase was expressed and then purified by column chromatography (Cha et al., 1998, Eur. J. Biochem. 253, 251-262).

To 15 g (30% by weight) of rice starch, alpha amylase 23600 U (5451 U / mg) and flulanase 2271 U (24 U / mg) were added and liquefied at 90 ° C, followed by maltogenic amylase 11066 U (170 U / mg) ( Saccharomyces serevisiae ) 150 mg was added and left at 27 ° C. for 16 days to liquefy rice starch and alcohol fermentation due to yeast.

Comparative example

The same procedure as in Example 1, except that glucoamylase was used instead of maltogenic amylase.

Experimental Example

The alcohol content, the total acid content, the reducing sugar content, the pH and the free amino acid content of the products prepared in Examples 1 and Comparative Examples were measured, respectively, and the results are shown in Table 1 below.

In Table 1, in Example 1 using maltogenic amylase it was confirmed that the alcohol, total acid and reducing sugar content is lower than the comparative example using glucoamylase, the pH and the content of free amino acid is somewhat higher.

Alcohol(%) Total% Reducing Sugar (%) pH Free amino acid (mg%) Comparative example 14 0.395 8.77 4.18 6.03 Example 1 12.4 0.342 5.05 4.41 6.73

In addition, the sugar content of the products prepared in Example 1 and Comparative Examples was analyzed. As a result, as shown in Table 2, it was found that Example 1 contains more functional branched oligosaccharides than the comparative example.

Party content(%) Example 1 Comparative example Glucose 0.13 8.26 maltose 2.67 0.14 Isomaltose 0.39 0.41 Maltotrios 0.22 - Pannos 2.38 - Branched tetraos 0.46 - Per total 6.25 8.81 Total Branch Oligosaccharides 3.23 0.41

In addition, the product of Example 1 and the components of commercially available alcohol were compared and analyzed. The liquor used was Hansan Sogokju (Yakju, hereinafter referred to as "A"), Gyeongju Beopju Gallery (Yakju, hereinafter referred to as "B"), Baekseju of Kuksun Party (Yakju, hereinafter referred to as "C"), and OB Beer OB beer (beer, hereinafter referred to as "D").

2 is a result of analyzing the components of Example 1 and commercially available alcohol (A to D) by thin layer chromatography, and FIGS. 3A to 3F are examples of Comparative Example 1 and commercially available alcohol (A to D). The result is analyzed by high performance ion exchange chromatography. The results are shown numerically in Table 3 below.

Party alcohol Example 1 A company Company B Company C Company D Glucose 0.13 9.39 6.20 3.74 - maltose 2.67 0.05 1.43 0.03 - Isomaltose * 0.39 0.44 0.13 0.02 0.04 Maltotrios 0.2 - 0.08 - - Isopanos * - - 0.17 - 0.01 Pannos * 2.38 - 0.39 - 0.02 Maltotetraos - - - - 0.22 Branched Tetraose * 0.46 - 0.63 - 0.14 Maltopentaos - - - - 0.05 Basin Pentaos * - - 0.25 - 0.06 Maltohexaos - - - - - Branch Hexaose * - - - - 0.03 Total Branch Oligosaccharides 3.23 0.44 1.57 0.02 0.3 Per total 6.25 9.85 9.28 3.79 0.57 Total sugar-branched oligosaccharide 3.02 9.41 7.71 3.77 0.27 *: Branched oligosaccharide

In Table 3, the content of the branched oligosaccharide was the highest in Example 1 at 3.23% by weight, and the content of sugars available in the body except the content of the branched oligosaccharide from the total sugar content was 3.02% by weight in Example 1 and 9.41% in A. %, B was 7.71 wt%, C was 3.77 wt%, and D was found to be 0.27 wt%.

As described above, the liquor manufacturing method of the present invention by using maltogenic amylase, it is possible to produce a liquor having a low sugar content and a high branched oligosaccharide content at the same time. Liquor prepared by the liquor manufacturing method of the present invention is softer than maltose content compared to glucose, and the branched oligosaccharides contained in the proliferation of bifidobacteria, which are useful intestinal bacteria, and rarely occurs tooth decay to alleviate the health side effects of drinking.

Claims (7)

Liquefaction step of liquefying the liquor raw material alpha-amylase; And Adding maltogenic amylase and yeast, which degrades starch to produce maltose and branched oligosaccharides, into the liquefaction to simultaneously perform saccharification and fermentation. A method of producing a low calorie liquor comprising. The method of claim 1, Performing the saccharification and fermentation at the same time is a low-calorie liquor production method characterized in that carried out at 27 ℃ 16 days. The method of claim 1, Performing the saccharification and fermentation at the same time is a method for producing low-calorie liquor characterized in that the saccharification is glycosylated using Bacillus stearothermophilus derived maltogenic amylase. According to claim 1, wherein the liquor raw materials are white rice, glutinous rice, brown rice, soybean, sweet potato, corn, potato, soybean, sorghum, millet, crude, wheat flour, sugar cane, juniper fruit, herbal medicine, herbal medicine, rye, agave ( Blue mezcal) and mixtures thereof. The method of claim 1, wherein the liquor is selected from the group consisting of beer, whiskey, rum, gin, tequila, schnapps, vodka, takju, yakju, sake, and soju. delete delete

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