CZ262295A3 - Process of obtaining tannins and proteins from hop waste - Google Patents

Abstract

This new integrated method of producing tannins and proteins from hops used in the food industry for preparing the basic components for producing beer, soft tonizing beverages, in bakeries, producing confectioneries and also for producing prophylactic cosmetic products and in producing perfumes. The method is implemented by extracting hop waste by separating proteins and tannins on at least one filtrating stage selected from the group including filtration, ultrafiltration, nanofiltration and gel filtration. The tannins can be separated from the proteins by gel filtration. This method is environmentally friendly and all its outputs can be further used.

Description

It is an integrated process for the production of tannins and proteins from hop waste, which is used in the food industry for the preparation of essential components for brewing, soft drinks, bakery, confectionery, prophylactic cosmetics and perfume production. .

BACKGROUND OF THE INVENTION

Hops are one of the basic ingredients in brewing beer. World-wide consumption of individual hop products:

1) 20% - dried hops

2) 40% - hop granulate

3) 30% - extracts of hops

4) 10% isomerized product

Breweries (brewers) prefer extracts and granulates for easier use of alpha acids. This focus on alpha acids in all existing ways of extracting them or enriching the granulates with these acids almost completely ignored— the major essential components of the hops: proteins (12-18%) and tannins (4%). The resulting lack of proteins and tannins in the extracts and in the enriched powdered or granulated blanks is the cause of the disturbances that occur during the brewing of beer. Specifically, there are the following drawbacks in the individual hops extraction and granulation methods

- 2a their consequences:

HOP EXTRACTS:

Environmental standards are being tightened in Europe. Chemophobia was the first to touch upon agrotechnological productions. Worldwide, the following solvents were used in particular for the xCicikee hops: methyl chloride, hexane, methanol and trichlorethylene (A Porster i994, Brewing & Beverage Industry International, pp.14-18). The reasons for this were:

1) Possibility to falsify the content of valuable natural components, ie alpha acids. The extraction of hops with these carcinogenic solvents increased the extraction of hard tars (resins) and wax, which many times seemed to increase the weight of alpha acids.

2) The Act on the Protection of Beer was adopted in 1516, which states that for. Only germinated barley seeds, hops, yeast and water may be used for beer production.

3> Protein deposits and solvents were polluting waste.

4) The nutritional properties of beer deteriorated, the risk of cancer, digestive and immunological disorders increased among lovers of this excellent drink. ⁼

CO ₂ (liquid or above critical bodetri) and ethanol are among the most popular, environmentally pure extracts of hops, as solid tars, nitrates, pesticides, tannins and proteins do not enter the extract when extracted with these solvents (A Hartl 1993, Brauwelt International, Nile, 113-116, M. Biendl 1994, Brauwelt International, N4, p.

3Ϊ1-315).

HRAN GRANULATES '' '’

At temperatures below -20 ° C, the hops separate the lupulin bodies from the leaves. This phenomenon has become the basis for the preparation of enriched powders or hop granules. According to the mixes, their enrichment with lupulin bodies is distinguished:

i. Standard - Type 90, non-concentrated, simply granulated (can be modified with ascorbates, bentonite, or stabilized with magnesium salts).

2. Concentrated - type 75, type 45 and type 33, in which the lupulin bodies in the hop granules or powders are thickened 1/2, 2 or 3 times. One advantage of enriched hop granules or powders is that they contain less tannins and proteins (R.J.H. Wilson 1987, E.B.S. - Symposium on Hops, pp. 216-229).

Ignoring Protein Methods of Extraction by the Thoughts Are Not Essential (M.Moll, in and Tannins in All Old and New and Hop Granulation Was Conditional for the Success of Beer Production Procedure

Beers & Coolers 1994, p.487).

If the ' ⁴ ' * 'slope'is'this' view 'is wrong and must be corrected. The antioxidant, microbicidal, anticancerogenic physico-colloidal qualities of tannins and the nutritional and foaming-chemical-colloidal properties of hop proteins - this is just a short list of their price as a natural, enriching raw material

- beer. To confirm the exceptional importance of proteins and tannins for beer production, the following known facts are given: in beer prepared by the classic Pilsen method (100% dried hops), proteins and tannins are one of the basic components, namely 1 liter contains: 200 mg polyphenols, 40 mg f Xavonoidii , up to 55 mg of catechin, up to 24 mg of technical flax epica, 70.2 mg of proanthocyanins, up to 5 g of proteins, 1.2 g of amino acids, of which: 0.3 g essential, 0.4 ^of semiesential, 0.4 g replaceable and 70 mg of amine nitrogen (M. Mol, in Beers & Coersers 1994, p. 487; RAMussche 1994, Omnich Review Article, Staff Comm.). It should be noted that 85 wt. % of beer protein is of plant origin and the remaining 15% is from yeast (M.Moll, in: Beers &Coolers; 1994, p.487). all plant proteins of beer are highly thermostable since they pass from plant to beer after a ninety minute brewing process at 100 ° C. Considering the high protein content of hops (up to 18% by weight, 6 to 8% by weight thermostable), the question of their transition to beer is logical and confirmed - at a dose of 200 g of hops per 100 l beer finds 200 mg of hop proteins, ie 6 to 7 wt. % of the total amount of protein derived from added hops (Mol 1994). In contrast, in the beer prepared from hop extract, the content of the above-mentioned hop components is reduced to 60 to 70% by weight. % (RAMussche 1994, Omnichem Review Article. Staff Comm.). In particular, by ignoring the need for these thermostable protein substances (amino acids and peptides) and tannins to extract hops, a number of serious problems have arisen which have arisen prior to enriched hop granulate technology nitrogen content for yeast during fermentation, lack of antioxidants, stabilizing and foaming components of beer. These consequences have forced the technology (brewers) of brewing beer using extracts and powders: too low from extract, enriched granules or powder to look for substitutes for hops protein substances. The following bacterial and chemical preparations are widely used as substitutes:

1) Yestex 61 artificial formulations, Yestax ™, to improve the functional activity of yeast at the beer fermentation stage.

2) Collagen, PVPP (polyvinylpolypyrrolidone) and antioxidants & 2 ^ 2θ5 ^# CgH ₇ NaOgH ₂ O - for beer stabilization.

3) Propylene glycol ester of alginic acid (PGA) for foam formation.

PGA is insoluble at pH values above 6.7 by digestive juices and even detergents. Without commenting on this, we draw attention to the fact that human saliva has a pH higher than the mention of the critical pH (PGA) of the beer foam. as a result of the large distribution of CO ₂ extracts, less strong and weakly acidic, neutral and basic peptides and oligopeptides are found in beers in the USA compared to Europe (Yokata et al. 1993, ASBC Journal, vol. 51, N2, pp. 54-57) ). Moreover, the source of this small amount of peptides are highly thermostable bacterial preparations CONVERTASE A AG, BG, PA, GAP, MG 25, MG 30 and TSA from the following bacteria and fungi: Bacillus subtillis var.

Unfortunately, it should be noted that in the US brewing industry simultaneously with the increased use of CO 2 extraction of hops, growth in the use of bacterial-chemical additives can be observed.

According to international expert prof. A. Porstra (data from r. 1994) increases by the use of C0 ₂ extracts. In this case, the classical composition of the beer is inevitably disrupted by microbial-chemical fillers. The only way to maintain the natural plant and healthy composition of beer using hop extracts is to extract parallel alpha acids and hop-tannin hop components.

With respect to process economics and existing methods, this current problem solves the invention as described below.

SUMMARY OF THE INVENTION It is an object of the present invention to improve existing technologies to enrich the extract of hops with proteins and tannins. The waste from the production of hop granules, particularly of the type 75, 45 and 33 but also in waste from extraction of C0 ₂ with the initial contents of these substances and practically preserved intact. Therefore, the use of these substances appears to be economically and technologically advantageous.

Considering that the protein and tannins of the hops are well soluble in water, deionized water is used according to the invention for the extraction of waste.

According to the invention, the waste is extracted with a polar solvent, and the extract thus obtained is separated in at least one filtration step. · - '. ', - · - = ~

Preferably, the polar solvent is water containing ethanol at a concentration of 0.1 to 2.0% by weight, preferably at a concentration of 1 to 1.5% by weight, and the extraction is preferably carried out at ambient temperature, which is preferably between 10 and 30 ° C.

The separation of proteins and tannins is preferably carried out on at least one filtration stage selected from the group consisting of filtration, ultrafiltration, filtration and gel filtration.

preferably, ultrafiltration and nanofiltration are used, s = is used in ultrafiltration of a membrane which does not permeate substances of molecular weight above a value selected from 5 to 100 kDa, preferably 20 to 30 kDa, even more preferably 8 to 12 kDa, and then the filtrate obtained is subjected to a nanofiltration in which the proteins together with the tannins are separated on a membrane which is impermeable to substances having a molecular weight above the value chosen in the range of 0.1 to 10 kDa, preferably 0.3 to 0.7 kDa.

In a particularly preferred embodiment of the present invention, ultrafiltration and nanofiltration are utilized, employing ultrafiltration of a membrane which is impermeable to substances of molecular weight above 10 kDa, and then subjecting the filtrate to nanofiltration in which the proteins together with tannins are separated on a membrane which does not permeate substances with molecular weight above 0.5 kDa.

The mixture of proteins and peptides retained in the nanofiltration step of the last stage is optionally further treated according to the invention by gel filtration using deionized, degassed water as eluent. In this case, the elution area is separated in the range. 150 to 2000 Pa, which is a solution of tannins, whereupon preferably another fraction, which is a solution of peptides, is collected.

Preferably, the at least one filtration stage, in particular ultrafiltration, is tangential to the concentration of crumbs and tannins after gel filtration.

In particular, according to the invention, for example, the powder or pulverized hop waste material from the above-mentioned products is placed in an extractor and washed out of the waste by proteins and tannins. this is preferably carried out for 2 hours. After separating the liquid from the cuttings, the remaining cuttings can still be used for feed preparation. The separation can be effected preferably by centrifugation or mechanically. Proteins and tannins are separated from the extract obtained in the stages shown in the diagram in Fig. 1.

In the first stage, high molecular weight proteins are separated from the liquid mass by tangential ultrafiltration.

In the second stage, the hop extract remaining after ultrafiltration is divided into two fractions by tangential nanofiltration:

(a) fractions of tannins and peptides with an average molecular weight (0,5 to 10 kDa); and

(b) truncating the ecumenic fraction containing trace elements, vitamins and amino acids.

After concentration by tangential nanofiltration on an MGA-100 membrane, the low molecular weight fraction can be used in yeast fermentation in beer production.

In the third stage, the concentrated tanino-peptide fraction is separated by gel filtration into tannins and peptides.

After freeze-drying or vacuum dehydration, the obtained proteins, tannins and peptides can be stored in the form of hygroscopic powders under anaerobic conditions in the dark, preferably in a high molecular weight molecular fraction, preserved by the addition of ethanol (up to 20%).

One of the main advantages of the process according to the invention is the possibility of its use in all existing hops extraction or granulation processes. With its introduction, production costs increase by no more than 0.01%, while its profitability is increased significantly (by 35 to 40%) due to the acquisition of highly valuable organic products. At the same time, a number of tasks in the field of brewing and agrobiotechnology, namely:

1) The plant raw material of hops is used considerably more rationally and becomes, together with alpha acids, a source of highly valuable proteins, tannins, vitamins and trace elements.

2) New hops organic products can also be used outside the brewing industry in toning and prophylactic drinks, in the manufacture of perfume and cosmetics, in the manufacture of confectionery, in bakery, etc.

3) Without prejudice to the laws of protection of beer purity, the products obtained according to the invention can be clarified and the organoleptic properties and structure can be stabilized, increased in value, influenced by the appearance of consumers, etc. as desired.

Starting from price data (Wilson 1987), alpha acids increase their own costs in the production of SS ^ Hnar granules by 23% and in organic solvent extraction to 24 * (even more in CO ₂ extraction), while in the integrated waste-free process according to of the invention can be increased with any method, the balance gain can be increased 2 to 2.5 times due to the high market prices of new organic products (100 tW ts SC <A.AA ^ O, w.

1994, and the thermostable, very valuable hops proteins / peptides useful for foaming are, according to experts, even 3 to 5 times higher than tannins).

obtained that by

Taking into account the long tradition of using hops for beer production and the harmlessness of the products obtainable according to the invention for consumers, there is no doubt about their competitiveness in comparison with bacterial-chemical and other grate products, in particular halotanin. The possibility of marketing new products by the method according to the invention can be stated by an independent expert K.E.N. E. Tovni (Ministry of Economy of Georgia) is practically unlimited in both the internal and world markets. Moreover, in his opinion, meeting the needs of the Czech brewing market with new bio-products of hops will ensure a considerable profit of foreign currencies with regard to the growth of popularity and competitiveness of products of Czech breweries. This creates foreign exchange reserves for the purchase of all antioxidants, stabilizers, foaming agents, etc.

DETAILED DESCRIPTION OF THE INVENTION

Example 1 kg of waste from the production of granulated hops of type 45 is placed in an extractor and a 1: 5 solution of 1% ethanol (diluted with deionized water) is added thereto. After two hours of intensive extraction at 25 ° C, the extract was separated from the residue by centrifugation and the separated pellet was washed twice with 5% methanol. The washed pellet, consisting predominantly of cellulose and membrane proteins, is granulated or preserved by

Caλ -X Ul (Laccisil from Medipharm, Czech Republic, at a rate of 1 g per 100 kg of cuttings) and used as feed or as a component in compound feed. All extractions were combined and divided into three basic fractions:

1) Protein fraction - up to 10 kDa.

2) Tanino-peptide fraction - from 10 to 0.5 kDa.

3) Fractions of vitamins and trace elements - below 0.5 kDa

The protein fraction is separated from the extracts on a membrane tangential ultrafiltration device utilizing polymeric ultrafiltration membranes of the type UPM-10, (or GR8lpp) produced on the basis of the polysulfonamide Sulfon 4T on the warp of nonwoven Lavsan and polypropylene. Proteins do not pass through this membrane and are concentrated separately in the reservoir. The protein fraction is lyophilized and the thus obtained white hygroscopic powder is hermetically packaged and stored in a desiccator. protein yield (generally N x 6,2S) zl kg waste from the production of · hop type 45 granulates ranges from 10 to 150 g, depending on the quality of the starting hop raw material. of which is 25 to 30% protein thermostable at 45 ° C, 3 to 3.5% Protein highly thermostable ^ ^ when "teplbtě ^ 75 ^ ⁰ No" ^ a "" 2 "to ^_ 2T? % of super-thermostable proteins that can withstand 95-100 ° C for 10 minutes.

The tannin-peptide fraction along with the smaller particles lightly

12 passes through the above-mentioned membrane type and then separates from the vitamin and trace element fraction on a second nanofiltration apparatus using nanofiltration membranes of the MGA-70 (or GA865pp) type, which are polymeric films based on modified cellulose acetate on non-woven Lavsan and polypropylene matrices. A fraction of vitamins and trace elements easily pass through a nanofiltration membrane of this type, but the tanino-peptide fraction does not progress and is concentrated more than 30 times.

To separate and purify tannins and peptides, concentrated extract is applied to a gel filtration column packed with Fractogel TSK HW 40 (F). The separation is effected by elution with deionized and degassed water (conductivity 0.5 to microsiemens) at 50 ° C.

Hop tannins are eluted from the gel filtration unit in peaks ranging from 1500 to 2000 Da. The tan-brown fractions of the tannins are collected and dehydrated by lyophilization or vacuum drying, and can then be stored in the dark. From 1 kg of waste from the production of hop granulates of type 45, 15-25 g of a high molecular weight tannin preparation having a purity of 85-90% are obtained.

The peptide fraction was also collected and lyophilized. It should be pointed out that the peptide fraction has shown high thermostability and some of them have rapperone properties.

Examples 2-7 '''l kg of waste from the hops by extraction of C0 ₂ is placed in the extractor and proceeding as in Example 1 except that using the extraction conditions ⁴ and filtration, listed in Table I in which the abbreviations used have the following meanings:

t. e. is the extraction temperature of the hop waste kVe;

k.p ..... is the concentration of ethanol in the wash p, p., ... is the number of washes

u. f ..... is the amount of protein in the ultrafiltrate (mg / kg of hops) n.f. ... is the amount of protein in the nanofiltrate (mg / kg of hops)

Table i

Example t.e. k.e. k.p. p.p. u.f. n.f. C. [° C] hm. % hm. % mg / kg mg / kg 2 20 May 0.5 5 2 60 tracks 3 20 May 1.5 5 2 62 tracks 4 10 0.5 5 2 52 tracks , 5 30 1.5 5 2 58 tracks 6 30 0.5 5 2 62 tracks 7 25 1.5 5 2 62 tracks Protein yield substances (generally N x 6.25) from 1 kg of waste from production hops extraction CO ₂ se move e from 120 to 140

g / kg, depending on the quality of the starting hop raw material, from 1 kg of hop production waste by CO ₂ extraction, 12 to 18 _____________, g of tannin preparation _____ s, mol. masses. , 2000 Da about purity

90%.

The examples are illustrative only and are not intended to limit the scope of the invention which is defined

Other variations and variations are apparent to those skilled ^{in the} art ^{from the} claims given to those skilled ^{in the} art on the basis of the above data, which do not depart from the scope of the invention or its protection.

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Claims (9)

A method for recovering tannins and proteins from hop waste, characterized in that the hop waste is extracted with a polar solvent, but with 4.0X1. and at least one filtration step. Method according to claim 1, characterized in that the polar solvent is water containing from 0.1 to 2.0% by weight of ethanol, preferably from 1 to 1.5% by weight, of ethanol and the extraction is carried out at ambient temperature, preferably at a temperature of 10 to 30 ° C. The method of claim 1, wherein separating the at least one filtration step, including filtration, ultrafiltration, filtration. characterized by protein and tannins performed on selected from the group of nanofiltration and gel according to any one of the claims and nanofiltration, i to 3, is utilized in the claims by taking a non-permeable membrane of a molecular value selected from a range of 5 to 100 kDa 4. A method of ultrafiltration by ultrafiltration of the mass above and then the resulting filtrate is subjected to nanofiltration, in which the proteins together with tannins are separated on a membrane which is impermeable to substances having a molecular weight above the value of &quot; selected &quot; OTi ^ áž ^ ío kĎaT “” ^ ““ ““ “' Method according to claim 4, characterized in that ultrafiltration and nanofiltration are used, in which the impermeable membrane is used in ultrafiltration, The molecular weight is selected from the range of 8 to 12 kDa, and then the resulting filtrate is subjected to nanofiltration, whereby the proteins together with tannins are separated on a membrane which is impermeable to the molecular weight above the value selected in range 0.3 to 0.7 kDa. Method according to claim 5, characterized in that it is used in the ultrafiltration of a membrane which is impermeable to substances of molecular weight above 10 kDa, and then the resulting filtrate is subjected to nanofiltration in which the proteins together with tannins are separated on the membrane that does not permeate substances with a molecular weight above 0.5 kDa. 7. The method of claim 9, further comprising treating the protein mixture in the nanofiltration step of the last gel filtration step, preferably at an elevated temperature in the range of 45-50 [deg.] C., using as the eluent of deionized, degassed water. A method according to claim 7, characterized in that the elution region in the range of 1500 to 2000 is separated Da, which is a solution of tannins, whereupon preferably another fraction, which is a solution of proteins, is collected. Method according to at least one of Claims 1 to 8, characterized in that the at least one filtration stage, preferably ultrafiltration, is tangential.