WO2019110818A1

WO2019110818A1 - Method for manufacturing a fermented liquor with low purine content

Info

Publication number: WO2019110818A1
Application number: PCT/EP2018/084026
Authority: WO
Inventors: Jongho Woo; Jumi SEO
Original assignee: Anheuser-Busch Inbev S.A.
Priority date: 2017-12-08
Filing date: 2018-12-07
Publication date: 2019-06-13
Also published as: KR20200113189A; CN111448303A; BE1025860A1; BE1025860B1

Abstract

A method of producing fermented liquor; whereby the alcohol fermentation broth is treated with a multistage filtration process including a reverse osmosis filtration whereby the purine in the fermentation broth is reduced to a concentration less than 1.0 ppm.

Description

Method for manufacturing a fermented liouor with low purine content

Technical part

This invention is directed to a method for manufacturing a fermented liquor with low purine content in the fermentation broth by reducing the content of purine contained in fermented broth.

Technology as the Background of the Invention

Generally, the fermented alcoholic beverage is produced through the process of generating alcohol fermentation by action of the yeast on the saccharified liquid obtained through the saccharification process after saccharification using an active enzyme such as malt; or raw materials including malt and starch such as wheat, rice, barley, corn, sorghum, potato, starch sugar. This resulting fermented liquor contains a large amount of purine component.

Purine is alkaline compound with a purine ring (purine nucleus), as an organic compound composed of carbon atoms and nitrogen atoms. This purine ring has a form in which a five-membered ring of imidazole is bonded to a six- membered ring of pyrimidine. In nature, that exists in such as adenine, guanine, hypoxanthin, and xanthine as a constituent of nucleic acid (DNA, RNA) and coenzyme (NAD, FAD). When this purine is ingested into the body through food, It is converted to uric acid form via xanthine.

When the content of uric acid is excessively increased in the blood, it becomes hyperosmolemia, and when these uric acid is crystallized and accumulate in the joints, gout occurs.

In general, gout is metabolic disease that causes various symptoms by being deposited In the cartilage of the joints of the body and surrounding tissues of tendons caused by increased concentration of uric acid in the blood and the crystals of uric acid can not be discharged to the outside of the body. The deposition of urate crystals induces joint inflammation leading to recurrent seizures with severe pain and deposition of gout nodule (tophi), occurs deformation and disability of the joint. In addition to joint abnormalities, it causes various kidney diseases and the uric acid may cause kidney stones (nephrolithiasis, nephrolithiasis) in the kidneys.

As mentioned above, there is a large amount of purine in the fermented liquor. Especially, it is known that beer, which is a representative of effervescent fermented liquor, contains more purine than wine or makgeolli which is another fermented wine. This is because not only purine is produced from yeast during fermentation but also purine exists in malt, which is the main ingredient of beer.

Therefore, In line with the present invention, it is therefore an objective to produce a fermented liquor or a beverage having reduced purine content.

Various methods have been described in the prior art . For example, methods to i ) to reduce the amount of malt used in the material or replace the nitrogen source required for fermentation with a other proteins besides malt, ii ) methods to digest purine bases of the purine nucleotides generated in the wort using enzymes and reduce purine by degrading purine base magnetized in yeast during subsequent fermentation .Also, methods iii) whereby purine contained in the wort or the fermentation broth is adsorbed and removed by filtering the fermentation liquid obtained through the wort or fermentation process.

However, removalof the purine using the above-mentioned method, is not efficient to remove sufficient purine, especially towards low levels.

The present invention proposes a process for producing fermented liquor having reduced purine content through removing purine effectively from the fermentation broth in an efficient way and towards concentration levels even below 1 ppm .

The process steps of the present invention for achieving the above-mentioned objective comprise i); a mixing step which mixes a raw material and water to produce a mash; ii) a saccharification step which saccharifies the mash to produce malt wort; iii) A fermentation step which adds yeast to the malt wort and subsequently fermenting to produce a fermentation broth; and iv) filtering step which produces the stock solution of the fermentation broth in which the solid component and the purine are reduced and/or removed by multi-stage filtration of the fermentation broth. The above process steps are such that it is possible to provide a fermented alcoholic beverage in which the purine content is reduced more effectively than the conventional technique.

Preferably after the filtration step, a blending step may be added which homogeneously mixes Into the fermentation broth a further component at least one selected from the group consisting of fragrance, sugar, sweetener, acidulant and coloring matter.

The raw materials for producing the fermented alcoholic beverage by the reduced purine content of this invention may contain, malt; at least one nitrogen source selected from the group consisting of soybean protein, wheat protein, barley, corn embryo, yeast extract, amino acid and inorganic salts; and at least one starch raw material selected from the group consisting of wheat, rice, barley, corn, sorghum, potato, starch and sugar.

Preferably, the saccharifying phase should comprise of the following phases: A liquefaction phase by introducing the enzyme; a saccharification phase liquefying the mash; a preparation of malt juice by heating following the filtration of the saccharified mash.

In the fermentation step, the fermentation broth can be prepared by adding oxygen and the upper surface yeast or lower surface yeast to the malt juice and then fermenting at 10 to 20 ' C for 1 5 to 20 days. The filtering step comprises: a first filtration step where the fermentation broth is filtered using a microfilter to remove solid matter or foreign matter in the fermentation broth; and a second filtration step through microfiltration to remove purine in the fermentation broth that has passed through the first filtration step.

More preferably, in the first filtration step, the fermentation broth can be centrifuged to take a supernatant and then filtered by using a diatomaceous earth filter. In the second filtration step, the fermentation broth having passed through the first filtration step is subjected to at least one filtration process selected from reverse osmosis filtration, an ultrafiltration and a nanofiltration to remove the purine by passing through multiple stages.

In the second filtration step, the fermentation liquid being passed through the first filtration step can be passed through the nano filtration process and the reverse osmosis filtration process sequentially in order to remove the purine.

On the other hand, another embodiment of the present invention includes a low purine fermented alcoholic beverage with a purine concentration of less than 0.5 ppm produced by the above-mentioned production method.

According to the present invention, by efficiently removing purine contained in the fermented alcoholic beverage via a multistage filtration in the manufacturing process, a low purine fermented alcoholic beverage can be produced.

In addition, through a multi-filtering process, various types of RTD (Ready To Drink) can be produced by mixing various types of additives while using a low-purine liquor with clear and non-potent flavor as a cocktail base to satisfy varying preferences of consumers.

Brief description of the drawings

Figure 1 shows the physical properties and performance of a filtration process which may be used in the second filtration step of the present invention.

Fig. 2 shows the schematic filtration process of the nanofiltration process and the reverse osmosis filtration process.

Detailed Description

Prior to describing the examples of the implementation of the present invention as follows, we would like to first clearly state that the terms and words used in the specification and the range of this claim should not be construed as limited to ordinary meanings or standard dictionary terms and, rather, they should be interpreted as meanings and concepts consistent with the technical ideas incorporated within the present invention.

As used herein and unless otherwise noted, the term "%" used herein to denote the concentration of a particular substance are as follows: For solid/solid, % is expressed in weight/weight; for solid/liquid, % is expressed in weight/volume; for liquid/liquid, % is expressed in volume/volume units.

Hereinafter, we will describe in detail the production method of a fermented alcoholic beverage with reduced purine content and the low-purine fermented product produced using the method.

First, the implementation of the present invention is a method for producing a fermented alcoholic beverage with reduced purine content by effectively removing purine contained in the fermented alcoholic beverage and ideally comprises different process steps or f the following phases: A mixing phase where a mash is produced by mixing the raw materials and water; a saccharification phase in which the mash is saccharified to produce malt juice; a fermentation phase in which yeast is added to the malt juice and then fermented to produce the fermentation broth; and a filtration phase where such malt juice is processed through a multi-filtration process to produce a stock solution from which the solid content and the purine are removed.

In the mixing phase, raw materials and water may be mixed to produce a mash for preparing the fermented alcoholic beverage. Preferred mixing weight ratio in the range of 1 : 3 to 1 :6 may be used, preferably 1 :5.

Generally, in order to produce a mash, the malt contained In the raw materials will be more than 50 wt. % and the purine content contained in a typical beer (a fermented alcoholic beverage) produced using such malt would range anywhere between 4-12 mg/100 ml.

However, in the present invention, In order to reduce the amount of purine produced in the fermentation broth, the content of malt containing purine in the raw material may be reduced to less than 25 wt. % of the total weight of the raw materials. Preferably, the malt may be used at levels below 10 wt. % of the total weight.

Meanwhile, in order to facilitate yeast fermentation in subsequent processes, at least one or more of the following can be used as a substitute(s) as the nitrogen source replacing the reduced malt: soybean protein, wheat protein, barley, com germ, yeast extract, amino acid and inorganic salt.

In addition, in order to produce alcohol during the fermentation process, one or more of the following materials can be used: Wheat, rice, barley, corn, sorghum, potato, starch and sugar.

In the present invention, malt contained in the raw materials was less than 25% and soy protein, yeast extract, and etc. used in the raw materials amounted to than IX (preferably 0.1 to 1%) and the remainder contained starch raw materials.

The prepared mash may be converted into malt juice in the saccharification phase. Preferably, the saccharifying phase may comprise of the following phases: A liquefaction phase by introducing the enzyme; a saccharification phase liquefying the mash; a preparation of malt juice by heating following the filtration of the saccharified mash.

To further explain in detail, the prepared mash contains nitrogen source and/or starch raw materials. Therefore, enzyme may be added to hydrolyze the fore-mentioned nitrogen source and the starch raw materials. At this time, it is preferred that the following enzymes are used: Amylase, an enzyme that hydrolyzes polysaccharides; Protease, an enzyme that hydrotyzes protein and peptide substrate; Cellulase, an enzyme that hydrolyzes the cellulose component; or a combination of these enzymes. When using such a combination, the recommended range of usage of a-amylase, β-glucanase, cellulase, protease and glucoamylase is preferably at a ratio of 300 to 1 ,000 ppm each based on the total weight of the raw materials.

The hydrolyzing phase above may include hydrolyzing the protein contained in the mash and leaving it at a temperature of 45 to 52^* C for 30 to 120 minutes (ideally 50-70 minutes) after the enzyme is added.

For saccharfication, it is preferable that the protein is sufficiently hydrolyzed and allowed to stand at 62 to 68* C for 80-120 minutes or for 20-40 minutes at 70 to 76 * C for 20 to 40 minutes. At this time, amylase and glucoamylase act to saccharify the mash with hydrolyzed starch.

The saccharified mash may be separated into spent grain and malt juice using a filtration system and then can be converted into malt juice by heating the fore-mentioned malt juice for 30 to 90 minutes. Proteins in the fore-mentioned malt juice coagulates and form a precipitate and the precipitate may be removed by filtration. Then, the malt juice may be cooled to a temperature of 10-20^* C using a cooler. At this time, cooling the malt juice to the temperature of 10 to 20 ^* C is to smoothly ferment the malt juice by inoculating the yeast in the subsequent process. If the malt juice temperature is out of the ideal range, the activity of the yeast is slowed or destroyed, making normal fermentation impossible.

The cooled malt juice may then be fermented in the fermentation phase by adding yeast to it and leaving it stored for a predetermined time to produce a fermentation broth. Ideally, the fermentation phase may include adding oxygen and Saccharomyces cerevisiae and/or Saccharomyces pastorianus to the malt juice, and then fermenting the fermentation broth at a temperature of 10 to 20 ^* C. More preferably, in the fermentation phase, the malt juice may be fermented using Saccharomyces pastorianus yeast capable of achieving a true fermentation rate of 9096 or more.

At this time, the oxygen injected into the malt juice is preferably injected in the form of sterilized air so that the level of dissolved oxygen contained in the malt juice is 8-12 ppm.

For example, the cooled malt juice may be fermented by storing the yeast at 10 to 20 ^* C after simultaneously adding Saccharomyces pastorianus capable of true fermentation rate of 90% or more and enough oxygen to activate the yeast. At this time, the malt juice may be fermented for 10 to 1 5 days so that the alcohol is generated by the activity of the yeast while reducing the odor of fermentation.

Preferably, the fermentation broth produced through the fermentation step should have an alcohol concentration of 10 to 15 v / vX. However, the concentration of the alcohol produced in the fermentation broth is not limited to this, and can be appropriately adjusted to the desired concentration depending on the intended usage of the prepared fermented broth.

The fermentation broth produced through the fermentation step can pass through the filtration step in multiple stages to remove the remaining yeasts, yeast metabolites and solids in the malt juice, and purine components in the fermentation broth can also be efficiently adsorbed and removed. A fermented liquor with reduced content can be produced.

At this time, it is preferred that the fermentation solution is in a liquid state at 0^*C or lower. This is to prevent the fermentation solution that has undergone the fermentation step from being affected by the yeast any longer, and to accelerate the coagulation and precipitation of substances causing turbidity of the fermentation solution will be.

If the temperature of the fermentation broth exceeds 0 ^* C, the substances to be filtered in the filtration step are not removed and are likely to be transferred to a later stage.

A primary filtration step of filtering the fermentation solution using a micro filter to remove yeast, solid matter or foreign matter in a fermentation solution; and a secondary filtration step of performing micro filtration to remove purine in a fermentation solution that has passed through the primary filtration step.

The primary filtration step may be performed using a micro filter to remove yeast, solid matter, or foreign matter remaining in the fermentation solution. Preferably, the fermentation solution is centrifuged to take the filtrate, and then the filtrate can be filtered using a diatomite filtration, which is a micro filter having a pore size of about 0.5 urn or less.

Meanwhile, in the fermentation solution which has passed through the primary filtration step, yeast, solid matter or foreign substances are removed, but the purine produced in fermentation still remains.

Thus, this invention, it is preferable to perform the secondary filtration step in order to effectively remove the purine causing the ventilation as described above in the fermentation solution.

In the second filtration step, the fermentation liquid that has passed through the first filtration step passes through at least one filtration step in multiple stages including the reverse osmosis filtration step, the ultrafiltration step and the reverse osmosis filtration step of the nanofiltration step, the purines can be removed efficiently and effectively.

The physical properties and performance of the filtration process that can be used in the second filtration step are shown in FIG. 1 , but the recovery rate is significantly lower than that of the microfiltration (MF), so that it can not be used in the conventional fermentation process.

FIG. 2 shows the schematic filtration process of nano-filtration process and reverse osmosis filtration process, in nano-filtration process (NF) the penetration level of respective water and alcohol flow is almost 100%, 99% each but for organics dozens of %, in the reverse osmosis filtration process (RO) water and alcohol flow is approximately 100%, 70% each but for organics some %.

The permeation fluxes of NF and RO used in this invention were maintained at 5% and 20%, respectively. When the fermentation broth was supplied at 100 L/hr, the filtrate was operated at 95 L/hr and 80 L/hr respectively.

More specifically, for example, in the second filtration step, the fermentation broth which has passed through the first filtration step is first subjected to nano-filtration using a nanofilter (nanofiltration membrane, NF membrane) (RO membrane) can be used to perform secondary reverse osmosis filtration.

At this time, the nanofilter has micro-pores having a size of several nanometers, so that monovalent ions are permeated and bivalent ions are blocked, and the purine component in the fermentation broth can be removed. Also, the osmotic membrane having ultra-fine pores in a size of about several nanometers is smaller than the nanofilter so that the salt including substances with sucrose (10 A) or ions is filtered to manufacture undiluted solution of fermented alcohol with reduced the purine efficiently.

Thus, the stock solution of the fermentation stock prepared through the second filtration step is maintained at a concentration of 10 to 1 5 v / v% as compared with the fermentation broth which has passed through the first filtration step. Whereas the purine component is efficiently and effectively removed, a characteristic of this process is a potential loss of flavour. Therefore, fermentation stock solution is preferably used as the base of the mixture. The raw material of the fermentation broth prepared through the filtration step may be prepared with a ready-to-drink (RTD) through a blending step where at least one of the following are homogeneously mixed: fragrance, sugar, sweetener, acidulant and pigment.

Hereinafter, an embodiment of the present invention will be described. However, the scope of the present invention is not limited to the following preferred embodiments, and various modifications of the present invention can be made by those skilled in the art within the scope of the present invention.

Example

Measurement of degree of fermentation bv material

Malt juice was prepared from 500 L sized brewing equipments with different contents of malt content (50%, 24%, 936), starch raw materials, yeast types and nitrogen source.

For the glucose source, 69 Brix glucose solution was used and sucrose was used as 65 Brix. For the nitrogen source, soybean protein isolate, yeast extract and inorganic salts (ammonium phosphonate) were used. Water and 1 kg of brewer 's yeast was fermented to produce malt juice.

Table 1.

The results of Table 1 indicate that the fermentation of malt juice 'Β' which has a malt content of less than 25% in the whole material was completed normal without delayed fermentation or abnormal fermentation.

Degree of reduction of purine content bv filtration method

The amount of malt juice 'A' and malt juice 'Β' with malt content of less than 25% in total material was 50% in the whole material, and the filtration method, the degree of reduction of the fructus content in malt juice according to the number of filtration were measured.

The purine content of malt juice was analyzed by the japan Brewing

Association using the method of "quantitative analysis of total purine content using

HPLC-UV method in beer, low malt beer, and third-category beer".

The purine content in malt juice was expressed as mg / 100 ml or ppm In total of four bases: adenine, guanine, hypoxantine and xantine.

First, 'A' malt juice and 'B' malt juice were each centrifuged, and the supernatant was taken and did the primary filtration using a diatomaceous filter having micro-sized pores.

The 'B' malt juice, which had been primary filtered, was passed through a nano filtration membrane (HYPERSHELL membrane, DOW) and a reverse osmotic filtration membrane (DAIRY AF membrane, CE) sequentially which is the secondary filtration. That created the fermentation liquor stock solution 'B' and RTD

(Ready To Drink) was created by using the fermentation liquor stock solution 'Β' as a cocktail base.

Table 2

As shown in Table 2, when the diatomite filtration was only carried out, the amount of purine was measured as 7.6 mg/ 100ml in the case of 'Α' malt juice, while the amount of purine was measured as 2.21 mg/ 100ml in case of 'Β' malt juice which is reduced by 70% or more of purine compared to the case of Ά' malt juice.

In case of Ά' malt juice, malt juice contains 50% of malt content, so that the amount of purine from raw malt was higher than that of 'Β' malt juice when it comes to manufacturing malt juices.

On the other hand, in case of 'Β' malt juice, the purine content in the malt juice was 2.21 mg / 100 ml - 0,06 mg / 100 ml-less than l ppm in fermentation broth of alcohol which is significantly reduced by performing a diatomite filtration and then the nano/reverse osmosis filtration sequentially as a secondary filtration. And the RTD with 8.2% alcohol content made by blending of fermented liquor was confirmed to have the purine content of less than 0.5 ppm.

Thus from the above it is made clear that the present invention minimizes the amount of producing purine by reducing malt content that can create purine during the primary fermentation to make a fermented liquor which contains a less purine content. Also, the present Invention provides a fermented liquor that contains reduced amount of purine content passing through a diatomite filtration and then after a nano / reverse osmosis in order to remove purine effectively and efficiently.

In addition, Making RTD (Ready To Drink) through blending of various additives (fragrance, sugar, sweetener, acidulant or coloring matter) with a fermented liquors stock solution having the reduced purine content through the filtration process as a cocktail base allows fermented liquor product with purine content less than 1 ppm to be provided.

Claims

Claim 1 A method of producing fermented liquor; whereby the alcohol fermentation broth is treated with a multistage filtration process including a reverse osmosis filtration whereby the purine in the fermentation broth is reduced to a concentration less than 1 .0 ppm.

Claim 2

A method according to claim 1 whereby the method comprises different process steps including a mixing phase where a mash is produced by mixing the raw materials and water; a saccharification phase in which the mash is saccharified to produce malt juice; a fermentation phase in which yeast is added to the malt juice and subsequently fermented to produce the fermentation broth.

Claim 3

A method according to claim 2 whereby in the mixing phase, raw materials and water are mixed to produce a mash for preparing the fermented alcoholic beverage in weight ratio in the range of 1 : 3 to 1 :6 , preferably 1 :5.

Claim 4

A method according to claim 2-3 whereby the saccharifying phase comprise a liquefaction phase by introducing the enzyme; a saccharification phase liquefying the mash; a preparation of malt juice by heating following the filtration of the saccharified mash.

Claim 5

A method according to claims 2-4 whereby the fermentation phase includes adding oxygen and Saccharomyces cerevisiae and/or Saccharomyces pastorianus to the malt juice, and then fermenting the fermentation broth at a preferred temperature of 10 to 20 ^* C

Claim 6

A method according to claims 1 -5 further comprising a primary filtration step of filtering the fermentation solution using a micro filter to remove yeast, solid matter or foreign matter in a fermentation solution; and a secondary filtration step of performing micro filtration to remove purine in a fermentation solution which has passed through the primary filtration step

Claim 7

A method according to claim 6 whereby in the second filtration step, the fermentation broth which has passed through the first filtration step is first subjected to nano-filtration prior to the reverse osmosis filtration.