JP3538203B2 - Beer production method - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

TECHNICAL FIELD OF THE INVENTION   The present invention relates to a method for producing beer, in particular, passing beer through a filter medium.
And ferment so that the filter media can be reused for beer filtration.
The present invention relates to a method for washing a filter medium with element. Background of the Invention   Considering the expanded sales channels, making beer storable
To remove microorganisms (eg, bacteria) from beer
There must be. At present, microbe removal is mainly beer
It is done by pasteurization. For this, beer
Into a bottle or can, for example, and heat to a temperature of 62-69 ° C.
To kill microorganisms.   However, this pasteurization requires considerable energy consumption.
I need it. This is because the energy introduced is a chemical reaction.
These chemical reactions can trigger the product
No further disadvantage that it is difficult to control
Having. These reactions are, for example,
Adversely affect the pasteurization (“pasteurized tast
e) "), further undesired substances being formed
There is a risk. Therefore, pasteurization requires high energy consumption.
Costly and, consequently, harmful to the environment,
Relatively expensive microbial removal methods that reduce product quality
Is the law.   Another known microbial removal method is cryogenic filtration. Low
Hot filtered beer is available, for example, in the United States, Japan and Korea.
Available as so-called "draft beer". this
Beer contains technical enzymes
And is banned in Europe.   These industrial enzymes lack the inherent cryogenic-filtration methods.
Point: To prevent the filter from blocking easily,
Exist in the file. This occlusion can occur in filters, e.g. membranes
The material to be filtered from the beer, upstream of the filter
Depending on the deposits. This deposit is removed from the filter
Filtering is difficult or even impossible
The service life of the machine. This means that the membrane filter
The cost of making beer.
You.   Membrane filter to extend the service life of the filter
Manufacturer manufactures used membranes for protease, glucanase
And xylanase, eg, detergents, acids / alkali
And by treating with chemicals such as oxidants
Recommend to clean the membrane and make the used membrane reusable
I'm worried. This cleaning is, for example, in two stages, the first in which
Optionally with the enzyme described above,
This can be done by additional cleaning with chemicals.   The literature is also used for beer filtration, including various techniques.
A method for cleaning a membrane filter is disclosed. For example,
U.S. Pat.
A method for cleaning an amide microporous membrane, wherein the microporous membrane is diluted.
Discloses a method by passing a thin alkaline solution.
You. International patent application WO96 / 23579 used for beer filtration
Several different methods of membrane filters are disclosed. this
The method uses a membrane filter of β-glucanase, xylana
Treated with an enzyme-containing aqueous solution of
Washing the membrane filter with an acidic washing aqueous solution, and
Cleaning with a peroxide containing alkaline cleaning solution
It is characterized by.   For example, about 320m^TwoAssuming a filter area of
For example, the washing method is to filter 5,000 ha
After each wash, prepare an enzyme wash and filter 20,000 hectares.
Provide additional chemical cleaning after each pass. Manufacturer recommends
About 320m^TwoHas the above filter area
Filters have a typical useful life of about 100,000 hectares
This is the title.   However, the cleaning method known until now
Deposits could not be sufficiently removed, which
As the age of the filter increases, the cleaning effect
It has the drawback of reducing the rate to strength.   Yet another disadvantage is the total nitrogen content or the original wool.
Filter membrane protrusions that are not related to standards such as
Natural random blockage. Completely closed membrane filter
Can be thoroughly cleaned with state-of-the-art methods
And strongly reduce the service life of the filter. What time
The filter is too occluded to be cleaned enough
It is difficult to know if the
Is not early or timely, i.e. too early or
May be too slow to be cleaned.   In view of the above problems, an improved beer production method,
Can be cleaned especially well and can be reused
Must be able to filter beer through filter media
The necessity exists. The present invention provides such a method.
These and other advantages of the present invention, as well as other features of the present invention,
It will be apparent from the description of the invention provided herein.
Would. BRIEF SUMMARY OF THE INVENTION   The present invention is useful when the porous membrane requires cleaning.
The beer is filtered through a porous membrane with
Consisting of lulase, amylase and combinations thereof
Enzymes selected from the group, especially crystalline for 60 minutes: dissolution
Having a cellulosic cellulose activity ratio of at least about 0.1
The porous membrane is washed by contact with
Includes reuse of porous membrane and continued filtration of beer
To provide a method for producing beer. The present invention further provides a filter.
Filtration of beer through a porous membrane that gradually blocks in the middle
And porous membrane streaming or zeta potential
The char is monitored using the degree of occlusion of the porous membrane as a measure.
Judge by porous membrane runoff or zeta potential
Before the porous membrane is completely closed
Stop beer filtration, wash the porous membrane,
Including reusing the membrane and continuing filtration of the beer,
A method for producing beer is provided. BRIEF DESCRIPTION OF THE FIGURES   FIG. 1 shows an unused, ie, new, porous membrane through
Beer filtration volume (g) vs. filtration time for beer filtration
It is a graph of (second).   FIG. 2 relates to beer filtration through a closed porous membrane.
Is a graph of the amount of beer filtration (g) to be performed versus the filtration time (sec)
You.   FIG. 3 shows a previously closed and cleaned by prior art method.
Beer Filtration for Beer Filtration Through Improved Porous Membrane
5 is a graph of excess (g) versus filtration time (seconds).   FIG. 4 shows a multi-cell tube previously closed and cleaned according to the present invention.
Beer filtration volume for beer filtration through porous membranes
(G) Graph of filtration time (seconds).   Figure 5 shows the measurement of the zeta potential of the filter media.
It is a schematic diagram showing a device.   Figure 6 shows the zeta potential (mV) of the filter media versus the electrolyte
Graph of solution pH, curve “a” relates to a new porous membrane
And curve "b" is partially obstructed in connection with beer filtration.
For a given porous membrane, curve "c" is related to beer filtration.
It relates to a porous membrane that is almost completely occluded.   FIG. 7 illustrates the use of the bypass system and the measuring device of FIG.
FIG. 2 is a schematic diagram showing an apparatus for filtering beer. Description of the preferred embodiment   The present invention relates to a method for producing beer, in particular low temperature filtered beer.
provide. This method involves passing through a porous membrane, a membrane filter.
Until the membrane filter needs cleaning
Filter the beer and contact the porous membrane with the enzyme
Wash the porous membrane and then reuse the porous membrane to
Continuation of the filtration.   The porous membrane is made of protease, xylanase and / or
By cellulase rather than by lucanase and / or
Better and more gentle washing with amylase
It has been surprisingly found that this can be done. Washing according to the invention
Purification depends on the useful life of the porous membrane used for beer filtration.
The use of porous membranes in beer production.
Greatly improve the commercial benefits associated with   Enzymes include cellulase, amylase and combinations thereof
Selected from the group consisting of As mentioned above, porous
Along with cellulase and / or amylase,
Use Rotase, Xylanase and / or Glucanase
It is not necessary to use it and it is preferable not to use it. Cellulase
Preferably at least about 0.1, more preferably less
Both about 0.3, preferably at least about 0.4, more preferably
Is at least about 0.5, most preferably at least about 1,
Crystalline at least about 1.2: soluble cellulose activity
Ratio (described in more detail below). Suitable cell
Lase from Aspergillus, especially Aspergillus niger
And cellulase. The preferred cellulase is Tric
hoderma, preferably Trichoderma reesei and Trichode
rma Iongibrachiatum and Thermomonospora, preferably T
Includes cellulase derived from hermomonospora fusca
I do. Other sources of cellulases are in US Patent 4,912,056
Has been described. Suitable amylases are α-amylases,
β-amylase and combinations thereof. Yo
More preferably, the method of the present invention comprises a cellulase and an amylase.
No other enzymes are used, that is, the porous membrane is treated with cellulase or
Do not come into contact with enzymes other than amylase. Most preferred
Means that the enzyme used in the method of the present invention is cellulase,
Suitably, no enzymes other than cellulase are used, i.e.
Contact the membrane with cellulase and not with other enzymes.
No.   The porous membrane can be any membrane suitable for filtering beer. Book
In connection with the invention, the porous membrane is typically a microporous membrane, i.e.
It is a porous membrane having a pore size of about 0.02 to 1 μm. porous
The membrane preferably has a pore size of about 0.1-1 μm, most preferably
It has a pore size of about 0.45 μm. Such a porous membrane
Removes bacteria and other unwanted microorganisms from beer
Preferably, beer is pasteurized
Avoid the need to germs. Porous membranes also come from beer
Used to remove yeast and other undesirable substances
be able to. Suitable porous membranes are, for example, ceramics and
An inorganic material such as a metal, preferably
, Polyether sulfone, polyolefin, polyvinyl
Manufactured from organic polymers, such as redene fluoride
Porous membranes. The porous membrane is preferably poly
Mido porous membranes, especially nylon-6,6 porous membranes.   In a preferred embodiment of the method according to the invention, the porous membrane
It is characterized by additionally contacting with an aqueous base solution.
Alternatively, in the first step, the porous membrane is brought into contact with an aqueous base solution.
In the second step, the enzyme is brought into contact with the enzyme. As an aqueous base solution
Is advantageous to use an aqueous solution of NaOH and / or KOH
It is known. The base is 0.1-1N, more preferably 0.25-1.
N, most preferably at a concentration of 0.5-1N.
New Treatment with aqueous base at a temperature of 40 to 90 ° C
Best done.   A further advantageous embodiment of the method according to the invention comprises a cell
Raise the treatment to a temperature of 40-50 ° C and a pH of 4.5-5.5.
And treatment with α-amylase at a temperature of 60 to 75 ° C.
At a pH of 4.6 to 5.8 and by β-amylase.
At a temperature of 40-60 ° C and a pH of 4.6-5.8.
It is characterized by the following.   Streaming potentia of porous membrane
l) or when the zeta potential no longer changes
It is convenient to carry out the washing until. Porous during operation
Outflow potential generated in the membrane or calculated from it
Zeta potential (see below) depends on the porous membrane
This is a good indicator that the occluded material has been removed.
You.   The present invention also ensures that the porous membrane is cleaned in a timely manner.
To extend the useful life of porous membranes
The purpose is to: Therefore, in the present invention, the filtration proceeds
Beer through a porous membrane that gradually closes as
There is provided a production of beer comprising filtering. Porous membrane
Is only partially occluded, ie completely occluded
Stop filtration at a specific point that has not yet been reached
You. The degree of occlusion is preferably determined by any suitable means,
For example, as generally described in US Patent 5,449,465
Monitor the pressure drop across the porous membrane
Can be determined by Alternatively, the present invention
Outflow potential from the filter and / or zeta
Provides confirmation of cleaning time by measuring potential.   This aspect of the invention is based on the runoff potential
Zeta potential extrapolated from recorded data
-Indicates the pH range according to the degree of blockage (beer brewing within this pH range)
Or the filtration takes place) and therefore closes
A reliable, almost quantitative indicator of occlusion status
Based on the perception of taste. Therefore, the outflow of the porous membrane
Measurement of potential and / or zeta potential
Can provide an accurate image of a particular occlusion.   It is known that porous membranes work in two ways. No.
First, the porous membrane acts as a sieve, in this case
Particles larger than the pores of the
You. Second, the porous membrane acts by electrostatic attraction
It has been known. Particles with a diameter much smaller than the porosity of the membrane
Is the zeta potential of the filter media and the zeta potential of the particles.
Adheres to the membrane when the polarity is opposite
(For example, Pall Filtrationstechnik GmbH, I
nformational Brochure SD 872h See G
When).   Use the zeta potential to determine the degree of blockage of the porous membrane
The fact that it can be measured
Not known.   Zeta potential of porous membrane depends on membrane chemistry
Affected by Experts say that Zeta Potentia
Filter changes at a sufficiently high rate in proportion to the degree of blockage.
There is no difficulty in selecting only Luther--the present invention
Recognize. Filter online and data acquisition
Through continuous monitoring and through the filtration process
Should be stopped in a timely manner, for example, if occlusion begins.
Can be.   Washing of filters that are not yet completely blocked is complete.
It is much easier than cleaning a completely blocked filter.
And guarantees a longer mode life. Therefore, the present invention
The preferred method is the zeta potential of the filter
Is up to 20% of the value that the filter showed in its unused state
At the time of the drop or the occlusion does not exceed 80%.
Stop the filter.   Another approach to this method is to use a porous polyamide film.
The zeta potential measured at a pH of 4.2
Filtration stops when it exceeds -5 mV.   Beer has a formal filtration, ie filtration through a porous membrane
Before, it is preferred to undergo a pre-filtration. Diatomie
Diatomite, also known as diatomite,
Used almost exclusively before prefiltration. Diatomaceous earth and thick
Combined with deep-bedfiltration
It is possible.   The invention can be used in any suitable beer production system.
Wear. Preferably, the present invention relates to U.S. Patents 5,417,101 and 5,595.
Cluster filter as described in 4,161
Used for   The invention also relates to a feeder line for beer that is subjected to filtration.
, Porous membrane, and filtered beer discharge line (run
-Off line) for filtration of beer
Also regarding units. This acts as a bypass,
Porous membrane and meter cell through which beer flows
(Meter cell) membrane filter outflow potential and
// Illustration for monitoring zeta potential
A meter cell characterized by means such as electrodes
All modules are featured.   The invention also relates to a feeder line for beer that is subjected to filtration.
, A porous membrane, and a line for unloading filtered beer.
It also has a filtration unit for filtering beer.
Treat. Unlike this paragraph, this filtration unit
Is the outflow potential when beer flows through the porous membrane
Monitor Char and / or Zeta potential
E.g. electrodes attached to a porous membrane for reading
It is characterized by the following means. In this variation,
Zeta potential is bypassed to the membrane filter
Not measured via the provided meter cell, but rather
It is measured at the membrane filter itself.   Any suitable bypass in connection with embodiments of the present invention
Forms can be used. Preferably, the invention is a US
Incorporates the apparatus and method described in Patent 5,449,465.   General condition where the zeta potential of the filter is blocked
Is discovered in beer filtration as follows:
Can be 1. The outflow potential of the porous membrane during the filtration process and / or
Or constantly observe changes in the zeta potential
Prevent unexpected or random occurrences
Therefore, the degree of membrane occlusion can be accurately identified,
Know timely measurements for filter replacement
Can be. 2. It is possible to stop filtration before the porous membrane is completely closed
it can. This facilitates easy cleaning of the filter
You. Occlusion material in a completely occluded filter should be
Removed only with extreme difficulty depending on the cleaning method
Can be removed from the filter
And shorten the useful life.   If filtration is stopped before complete occlusion, the washing process
Filters are much easier and more complete, filters are
Keeps a long life. In the case of a polyamide porous membrane,
Zeta potential is about 80% of its original value
Not lost much, ie more than 80% occluded
If the filtration is stopped when not
Successful removal of all occlusive substances can be achieved
You. 3. Success of cleaning method depends on zeta potential of cleaned membrane
Can be tested by measuring the Washing
Action reduces the zeta potential to almost its original value
To return to.   In this way, the cleaning process can be
It can be evaluated and / or optimized. 4. Track the aging of porous membranes due to repeated use
To provide a reasonable estimate of the residual service life prediction.
Can be obtained. 5. By measuring the zeta potential, the liquid
Of the system occluding material and filter material and / or filter
Assess the interaction between the shunt means and filter
Material (filter material) and shunting material (shunting ma)
terial) (for example, diatomite, bentonite,
-Light, polyvinylpyrrolidone) in beer filtration
Can be tested for compliance. 6. Until the time when occlusion starts, specific membrane load
e load) (hl / m^Two) Is recorded under it
The service life of the porous membrane
Can be estimated.   Zeta potential showing a significant change in proportion to the degree of obstruction
A porous membrane with a vial is most suitable for this method.
Those skilled in the art are aware of this. These parameters
Demonstration can be performed using the simple test method described above.
Easy enough.   The following examples further illustrate the invention, but do not
The examples are intended to limit the scope of the invention in any way
Should not be interpreted. Example 1   This example illustrates the effectiveness of the beer production method of the present invention.
I will tell. In particular, this example demonstrates cellulase and amylase
Porous membrane clogged in the process of beer filtration using
Enough, this porous membrane is reused for continuous beer filtration
Demonstration that it can be cleaned so that it can be
I do.   Nylon-6,6 (NB type, Pall Filtrationstechnik G
porosity manufactured from mbH, commercially available from Germany)
The membrane was used as a filter. Such a filter
Is often used in the latest technology for low-temperature filtration of beer
It is.   The so-called membrane filter test by Esser (Monatszei
tschrift fuer Brauerei (Monthly Magazine for
Breweries), 25^thYear 6, No. 145-151, 1972)
The filtration performance of the filter was evaluated. This test is filterable
Check for measures to improve performance
Reliable for you.   Measure the filtration efficiency of a new, i.e., unused, porous membrane.
Pressure filter (SM16536, 200ml capacity; Serto
commercially available from rius GmbH, Goettingen, Germany)
With a polyamide nylon having a diameter of 47 mm and a porosity of 0.2 μm.
Used for Ron-6,6 porous membrane.   Beer cooled to 0 ° C is porous under isobar conditions (1bar)
Through the membrane, the filtrate volume was weighed every 10 seconds. 200 g of filtrate
After being obtained, the test was stopped. The results are plotted in the diagram of FIG.
Shown as rough. Figure 1 shows a filter that has not been used under the above conditions.
-After about 210 seconds, 200 g of filtrate was obtained.
Show.   Under the same conditions, the closed or used porous membrane
The filtration performance was tested. The results are shown in FIG. 2, which is 720 seconds.
Only about 60 g of filtrate were obtained
And   The clogged porous membrane is washed according to prior art methods.
Was. In this method, as described below, the membrane is first
It was then chemically washed by the enzyme.   For enzyme cleaning, occluded membranes should be pH 5
Mixture of agent and acid component (P3-Ultrasil 75; from Henkel
(Commercially available) set with 0.05% aqueous solution
A mixture of β-glucanase and xylanase (P3
-1% of Ultrasil 65; commercially available from Henkel)
The solution was treated with the aqueous solution at a temperature of 50 ° C. for 1 hour. So
After this, this treatment was performed once more.   Next, the membrane was adjusted to pH 9 to 9.5 (with a surfactant and an alkali component).
(P3-Ultrasil 91; commercially available from Henkel)
Surface active with (possible) 0.15% aqueous solution)
Mixture of sexually active agent, glucanase and protease (P3
0.5% of -Ultrasil 62; commercially available from Henkel)
Treated with an aqueous solution at a temperature of 50 ° C. for 3 hours,
And rinsed with warm water (50 ° C.).   For chemical cleaning, the membrane is combined with a surfactant and an acid component.
Mixture (P3-Ultrasil 75; commercially available from Henkel
Treated with 1% aqueous solution of Noh) at 60 ° C for 30 minutes,
It was then rinsed with fresh water. Next, the membrane
Mixture of surfactant and alkali component (P3-Ultrasil 9
1; commercially available from Henkel) 1% plus surfactant
Mixture with oxygen donor (P3-Ultrasil 05; from Henkel
(Commercially available) with an aqueous solution containing 1%
Treat at a temperature of 60 ° C for 30 minutes and then with fresh water.
Rinsed. The membrane is then combined with a surfactant and an acid component.
Mixture (P3-Ultrasil 75; commercially available from Henkel
Another 30 minutes treatment with 0.5% aqueous solution of
Rinsing water with fresh water, the conductivity of fresh water
Rinsed until reaching.   Next, the filtration performance of the washed porous membrane was measured under the above conditions.
Was tested again. The results are shown in FIG. Figure 3 is about 600 seconds
After a short time, 200 g of filtrate was obtained, so that the filtration performance was slightly improved.
Indicates that it was good.   The filtration efficiency is shown in FIG.
Washed by the method of the invention. This film is C₁-And C_x−
To an aqueous solution of lulase (this solution has a pH value of 4.7)
Therefore, the treatment was performed at a temperature of 45 ° C. for 30 minutes. Next, attach the membrane
The same solution, but at a pH value of 5.0 and a temperature of 50 ° C.
And finally at a pH of 4.7 and a temperature of 60 ° C for 60 minutes.
I understood.   Subsequently, the membrane was rinsed with warm water at 50 ° C. Book
The filtration performance of the membrane washed by the method of the invention
Tested accordingly. The results are shown in FIG.   FIG. 4 shows that 200 g of filtrate was obtained after about 220 seconds.
Show. This represents a significant improvement over the prior art (Figure 3).
Represent. Therefore, the method according to the present invention is based on the prior art cleaning method.
Much better for used membranes than is possible with the method
Cleaning is possible.   According to the present invention, amylase is used instead of cellulase.
, Good results were obtained as well. like this
In addition, the cleaning method according to the present invention makes it possible to
You can extend your life. Example 2   This example is intended to assist in cleaning the porous membrane.
Outflow potential or zeta potential of porous membrane
The use of is explained. In particular, runoff potential or zeta
・ When the potential is the cleaning degree of the membrane, the membrane is most sufficient when
Demonstrated to be useful to determine if it will be washed
Have been.   Anton Paar shows zeta potential of membrane filters
  GmbH, an electrokinetic measuring system in Austria
(Electrokinetic measuring system) Measured by EKA
did. This measurement is based on the runoff potential method. Electrolytes
Flows through the filter and the counterion shears
ng-off) (potential potential)
Is detected by the electrode, and the zeta
Calculate the potential (see below).   Figure 5 shows the runoff or zeta potential
Fig. 1 schematically shows a measurement cell for which is measured. Reference number 1 is measurement
In this cell, the porous membrane 2 is made of polytetrafluur.
Filter holders 3 and 4 made from polyethylene
It is clamped so as not to bend. filter
-Holders 3 and 4 have two horn pistons 5 and 6, respectively.
Of the measuring cell 1 that moves into the cylindrical portion 7 of the measuring cell 1.
Installed for.   The ends 3 and 4 of the pistons 5 and 6, respectively, are filtered
It has micropores 10 and 11 for the fluid to be
And 9 are pressed against the porous membrane 2. Electrodes 8 and 9 are pistons 5
Connect to the electrical terminals 12 and 13 extending inside the
Outflow potential generated when fluid flows through 2
Can be measured. Low polarization during passage of current
A silver electrode or a silver chloride electrode is preferred as the electrode. Fixie
6 and 7 are attached to seals 14 and 15, respectively, while
They are mobile, while on the other hand they are
Mounted so as not to allow fluid to leak.   The fluid to be filtered passes through the supply line 16 and the measuring cell
1 into the cylindrical portion 7 and through the micro holes 10 in the piston 6.
And an electric potential is generated through the electrode 8 and the porous membrane 2.
You. The filtered fluid flows through the electrode 9 and likewise
Passing through the micro holes 11 of the piston while forming the
The measurement cell exits through the discharge line 17.   Zeta potential from measured runoff potential
To calculate the supply line 16 and the discharge line 17
Measurement of differential pressure in the measuring cell during the measurement (not explained), conductivity and
And pH values are also required. The zeta potential is
It is calculated from the measured quantities as follows: Where U is the outflow potential and ΔP is the differential pressure.
, LF is the conductivity, η is the viscosity, εε₀Is invitation
The electric power.   Zeta potential of membrane filter as occlusion progresses
The change in char is shown in FIG. This figure shows the dimensions in millivolts.
Data potential is plotted on the ordinate and the zeta
The measured pH value of the potential is plotted on the abscissa
FIG. PH value of electrolyte solution (0.001N KCl aqueous solution)
Was set with 0.1N HCl or 0.1N NaOH. Specify
The resulting pressure difference was 350 mbar.   With the measuring cell described above, a new, i.e.
Polyamide porous membrane (NB type; Pall Filtrationstechni)
k GmbH, 6072 Dreieich 1, commercially available from Germany
Zeta potential at various pH values
, A diagram was obtained.   The results for the unused porous membrane are plotted as curve “a”.
To Unused porous membrane is about -18 depending on alkaline pH.
It has a zeta potential of mV and
Data potential rises to a final pH of about 3.
It is evident that the number reaches zero.   Curve “b” is plotted under the same conditions described above, except that beer
Porous after use for filtration and therefore with partial occlusion
3 shows the dependence of zeta potential on the pH value of the membrane.
As is evident, this zeta potential is partially
Somewhat increased due to mechanical occlusion, but at a pH value of about 7
It only reaches a value of about -15 mV.   Curve “c” shows the same porous membrane almost completely closed.
Is plotted with respect to. This zeta poten
The char now changes only slightly with the pH value
Not fall below about −2 mV even in the alkaline range.
It is clear that   To test the cleaning according to the invention, the membrane to be cleaned is
Cleaned membrane zeta for measuring zeta potential
・ Zeta potential of unused membrane
If the shift is as large as possible,
there were.   Its zeta potential is sufficient as a function of the degree of occlusion
The porous membrane, which varies to a large extent,
It is obvious to a person skilled in the art that they are particularly well suited for use in
Or maybe. This feature is easily tested by those skilled in the art
It can be easily determined.   The pH of the beer that is filtered by the porous polyamide membrane
The zeta potential at (about pH = 4.2) is progressive
This process is subject to severe changes due to
Particularly suitable for As can be seen from FIG.
In addition, the membrane at this particular pH at the start of the filtration is approximately -8 mV
Shows the zeta potential of A completely occluded membrane is about
It has a zeta potential of −2 mV.   FIG. 7 shows the bypass shown in FIG.
Filter comprising a filtration chamber 18 having a meter cell 22
9 shows the deformation of the excess unit. This filtration chamber 18 is
Holds candles (filter candles).   Beer to be filtered is supplied from line 20 to filtration chamber 18.
Through the filter candle (membrane filter) 19
And filtered from the filtration chamber 18 through the discharge line 21
Serve as beer.   The meter cell is shown in FIG. 7 without details. Meter cell
The actual flow through 22 is the cm of the surface of the porous membrane in filtration chamber 18.^Two
Per cm of porous membrane surface^TwoBeer per bottle
Must be controlled to the extent that is filtered.   Meter cell during filtration of filter test membrane 2 (FIG. 5)
1 Severe changes in zeta potential inside the filtration chamber
Enables evaluation of the condition of filter candles 19 in 18
I do. Example 3   This example demonstrates the use of fermentation of soluble and crystalline cellulose substrates.
Of cellulase from Aspergillus niger in elemental degradation
Explain effectiveness.   Cellulase derived from Aspergillus niger is Fluka (item
No. 22178). This enzyme has two different types
Cellulose: soluble carboxymethylcellulose (CMC,
(Available from Aldrich as item number 41927-3) and crystals
Cellulose (Avicel, item number PH-105 from FMC
(Available). The test method was (i) 18 ml CMC (1%) or Avicel (1%).
%), (Ii) 5 ml sodium acetate buffer (50 mM, pH 4.
8) and (iii) sodium acetate buffer (50
5 ml of enzyme solution in mM, pH 4.8)
Preparation of the solution was required. Next, incubate 1.4ml
0.1 ml glucose solution (0.15%) and 1.5 ml
3,5-dinitrosalicylic acid (DNS) reagent from Sigma
(Available as eye number D-0550).
Thus, a test solution was prepared. Boil the test solution for 15 minutes
Was. Total μmol glucose equivalents as a function of time (minutes) /
mg enzyme using two parallel samples, Miller, Anal.
According to the method described in Chem. 31, 426-28 (1959),
Using linear calibration with glucose standard
And measured by spectrophotometry (575 nm). Protein
The mass is described in Bradford, Anal. Biochem. 72, 248-64 (1976).
Bovine serum albumin (BSA) standard
It was measured using the standard.   Enzymatic degradation of cellulose leads to glucose production
The total μmol glucose equivalent / mg enzyme is measured for a specific species.
A class of celluloses such as soluble (CMC) or crystalline (Avi
cel) is a measure of enzyme activity for cellulose.   From Aspergillus niger
Table 1 shows the results of this evaluation of the induced cellulase.
You. 0.8mg test solution of soluble (CMC) cellulose substrate
Contains enzyme / 28 ml culture (approximately 17.6 μg protein).
0.35 mg test solution of crystalline (Avicel) cellulose substrate
Contains enzyme / 28 ml culture (approximately 7.7 μg protein).   Crystalline cellulose groups compared to soluble cellulose substrates
These enzymes, which have relatively strong activity on quality,
Especially effective for cleaning porous membranes used in filtration of steel
It was found that Generated on crystalline cellulose substrates
Glucose equivalents and soluble cellulose substrates
The ratio of glucose equivalents formed depends on the presence of the enzyme in the present invention.
It is an index of efficacy, crystalline: soluble cellulose activity ratio and
To be described. The crystalline: soluble cellulose activity ratio is
Time range of the test protocol described in the examples of
30 minutes, 60 minutes and / or 90 minutes, especially above 60 minutes
It is desirable to have the following values.   As is clear from the data shown in Table 1,
Cellulase from Gils niger crystallizes in 60 minutes
Quality: soluble cellulose having an activity ratio of 0.11, such a cellular
Regarding the purpose of cleaning porous membranes used in beer filtration
It is a moderately effective enzyme. Example 4   This example demonstrates the use of an enzymatically degradable soluble cellulose substrate.
Trichoderma reesei on crystalline cellulose substrates
Of cellulase derived from Trichoderma reesei
Shows sex.   Cellulase derived from Trichoderma reesei
Obtained from Fluka (item no. 22173). Perform this enzyme
Evaluation was performed in the same manner as in Example 3.   Cellulase derived from Trichoderma reesei
Table 2 shows the results of the evaluation. Soluble (CMC) cellulo
The test solution of the substrate is 0.37mg enzyme / 28ml culture solution (about 128μ
g protein). Crystalline (Avicel) cellulose
The test solution for the substrate was 0.08 mg enzyme / 28 ml culture solution (about 25.6 μl
g protein).   As is evident from the data in Table 2,
Crystalline Cellulase Derived from Sauce at 60 Minutes:
The soluble cellulose activity ratio is 0.54, such a cellular
For the purpose of cleaning porous membranes used by Beer for filtration of beer
It is an excellent enzyme. Example 5   This example demonstrates the use of a soluble cellulose substrate that is
Subtilis on crystalline and crystalline cellulose substrates
Of β-cellulase derived from Bacillus subtilis
Shows effectiveness.   Cellulase derived from Bacillus subtilis
Obtained from a (item number 49106). This enzyme was used in Example 3
Was evaluated in the same manner as described above.   Β-cellulase derived from Bacillus subtilis
Table 3 shows the results of the evaluation. Soluble (CMC) cellulo
The test solution of the substrate is 14.4mg enzyme / 28ml culture solution (about 8.3μ
g protein). Crystalline (Avicel) cellulose
The substrate test solution was 16.6 mg enzyme / 28 ml culture solution (about 8.8 μg
Protein).   As is evident from the data shown in Table 3, Bacillus
  Β-cellulase from subtilis crystallizes in 60 minutes
Quality: soluble cellulose having an activity ratio of 0.10, such a cellular
Regarding the purpose of cleaning porous membranes used in beer filtration
It is a moderately effective enzyme. Example 6   This example demonstrates the use of an enzymatically degradable soluble cellulose substrate.
Thermomonospora fuss on crystalline cellulose substrates
Exocell derived from mosquito (Thermomonospora fusca)
Shows the efficacy of exocellulase.   Exocellular derived from Thermomonospora fusca
Rose E3 was obtained from Cornell University. At 50 ° C
Shake (i) 18 ml CMC (1%) or Avicel (1
%), (Ii) 9 ml sodium acetate buffer (50 mM, pH 5.
6), and (iii) sodium acetate buffer (50 mM, pH 5.6)
In addition to containing 1ml of medium enzyme solution (about 960μm protein)
Evaluated this enzyme in the same manner as in Example 3. test
The solution should be stained rather than the DNS test described in Example 3.
Was evaluated using the test.   Cellulase derived from Thermomonospora fusca
Table 4 shows the results of the evaluation.   As is clear from the data shown in Table 4,
Exocellulase from Nospora fusca in 60 minutes
Crystalline: soluble cellulose having an active ratio of 1.33,
Purification purpose of porous membrane used by cellulase in beer filtration
Is an excellent enzyme with respect to Example 7   This example demonstrates the use of an enzymatically degradable soluble cellulose substrate.
Bacillus subtilis (Ba
of α-amylase derived from Cillus subtilis)
Shows sex.   Α-amylase derived from Bacillus subtilis
Obtained from Fluka (item number 10069). At 30 ° C
Shake (i) 18 ml CMC (1%) or Avicel (1
%), (Ii) 5 ml sodium acetate buffer (50 mM, pH 6.
9), and (iii) sodium acetate buffer (50 mM, pH6.9)
In addition to containing 5ml of medium enzyme solution (about 8.5μm protein)
Evaluated this enzyme in the same manner as in Example 3. test
The solution should be stained rather than the DNS test described in Example 3.
Was evaluated using the test.   Α-amylase derived from Bacillus subtilis
Table 5 shows the results of the evaluation.   As is evident from the data shown in Table 5, Bacillus
  Α-amylase from subtilis is crystallized at 60 minutes
Quality: Soluble cellulose activity ratio about 0 (<0.1 μmol detection limit
), Wherein the amylase is of the cellular type described above.
About the purpose of cleaning porous membranes used in beer filtration
Is not an effective enzyme. Example 8   This example demonstrates the use of an enzymatically degradable soluble cellulose substrate.
Effectiveness of various cellulases on crystalline cellulose substrates
Shows sex.   Cellulase preparation: (a) C_x-Cellulase (powder, goods
Eye number VP0945 / 2), (b) from Trichoderma reesei
C₁Cellulase (powder, item number VP0965 / 2), (c)
C₁Cellulase (liquid, item number Cleanzym SB1),
(D) C₁Cellulase (liquid, item number VP0976 / 4),
(E) cellulase (liquid, item number VP0971 / 1), and
(F) Cellase (liquid, item number VP0971 / 4) is Erbslo
h Obtained from Company. The culture solution is (i) sodium acetate
23 ml CMC (1%) or A in buffer buffer (50 mM, pH 4.8)
vicel (1%) and (ii) sodium acetate buffer (50m
M, pH 4.8)
The enzyme was evaluated in the same manner as in Example 3. Powdered enzyme tone
0.5 mg / ml (5 mg / ml) and liquid enzyme preparation (5 μl / ml)
% Stock solutions were prepared. The solution was shaken at 30 ° C. Trial
The test solution was diluted 1: 5 before the DNS test described in Example 3.
Was evaluated using a color test rather than a test.   Table 6 shows the evaluation results for various cellulases. G
Lucose equivalent data is average μmol glucose equivalent / min
(All time intervals) and (the data conditions described in Tables 1 to 5).
Not standardized per mg enzyme). Of course,
Crystalline: soluble as long as the units are allocated when calculating the ratio
Activity ratio varies with units of glucose equivalent
No (ie, the ratio has no units).   As is apparent from the data shown in Table 6, various cells
Lase is crystalline in the range of 0.4-1.0 in 60 minutes: soluble cellulo
Have the same activity ratio as the porosity used for beer filtration.
A superior enzyme for the purpose of cleaning membranes
ing. Example 9   This example demonstrates the effectiveness of the method of the present invention for making beer.
This is to explain the nature in detail. In particular, this embodiment
Uses only cellulase (ie uses other enzymes)
No) and a porous membrane for use in continuous beer filtration
Clogged while filtering the beer for the purpose of returning
It shows that the porous membrane is very excellent in cleaning.   Beer with various characteristics is equipped with a cluster filter
Nylon with 0.45 μm pore rating
−6,6 porous membrane (about 300m^Two) (PALL-CFS, Pall Filtrati
onstechnik GmbH, Germany). Specific bee
At the filtration interval, the porous membrane is subjected to the cleaning step of the present invention.
Was.   The cleaning process consists of a circulation of 0.5% NaOH solution for 15 minutes, followed by 6 minutes.
Included 0 minutes soaking. Then the porous membrane is washed with water
Backflushed. Cluster fill with 38 ° C water
An internal loop was installed through the heater device. Add lactic acid to water
To adjust the pH to 4.2 ± 0.3, then the Erbsloh Company
Trichoderma Longibrachiatum (Tric
hoderma longibrachiatum)
6 l of enzyme preparation containing (item number VP0945 / 1)
Added to water via pump. Enzyme preparation in water (about 20-40
g enzyme / 100kg filter housing fluid volume concentration)
Circulate for about 15 minutes, then soak for 30 minutes, then another 15 minutes
And soaked for a final 6 hours. Then porous
The membrane was backflushed with water.   After filtering about 90,000hl of all beer with a porous membrane,
The porous membrane was cleaned and then the porous membrane was returned to the facility. That is,
Beer filtration was continued. About 100,000hl, about 140,000hl and
After filtering about 160,000hl of whole beer through porous membrane
At the same time, the porous membrane was similarly cleaned and returned to the facility. About 190,00
0hl total beer filtered through porous membrane
Later, the porous membrane was broken mechanically.   The above data shows that the beer
Can be produced. In particular, the porous material according to the invention
This practice of effectively cleaning the membrane and returning it to the facility
The results of the example show that the beer production process
Indicates that the working time can be extended.   Cited herein, including patents, patent applications and publications
All references herein are incorporated by reference in their entirety.
Shall be.   The invention has been described with emphasis on the preferred embodiments.
However, various modifications of the preferred embodiment are possible for those skilled in the art.
The present invention may be practiced other than as specifically described in the present specification.
It will be clear. Accordingly, the present invention provides the following
Within the spirit and scope of the present invention as defined by
All modifications shall be included.

   ────────────────────────────────────────────────── ─── Continuation of front page    (72) Inventor Tsunker, Geralt               Republic of Austria-8075 Hull               To Bei Graz, Schurgasse 1 (72) Inventor Zello, Walter               Republic of Austria Ar-2440 Reise               Nberg, Upras 5 (72) Inventor Ribish, Volker               Republic of Austria-8010 Gra               Tu, Heinrichstrasse 28 (72) Inventor Landhahn, Horst               Federal Republic of Germany Day 64297 Darm               Stadt, Frankensteiner               Strase 58 (72) Inventors Degen, Peter Jay               United States of America New York 11748,               Huntington, Grays Way 24                (56) References JP-A-1-228509 (JP, A)                 JP-A-4-267933 (JP, A)

Claims (28)

(57) [Claims] 1. The method of claim 1, wherein the beer is filtered through the porous membrane until the time when the porous membrane has to be cleaned, and comprises cellulase, amylase and a combination thereof in the absence of a protease or glucanase to clean the porous membrane. Contacting an enzyme selected from the group with the porous membrane,
A method of making beer, comprising then reusing the porous membrane to continue filtering beer. 2. The method according to claim 1, wherein the porous membrane is not contacted with an enzyme other than the cellulase or the amylase. 3. The method according to claim 1, wherein the porous membrane is contacted with the cellulase. 4. Filtering the beer through the porous membrane until the time the porous membrane must be cleaned and having a crystalline: soluble cellulose activity ratio of at least about 0.1 in 60 minutes to clean said porous membrane. A process for producing beer comprising contacting cellulase with the porous membrane and then reusing the porous membrane to continue filtration of the beer. 5. The method according to claim 3, wherein the porous membrane is brought into contact with the cellulase but not with other enzymes. 6. The cellulase having a crystalline: soluble cellulose activity ratio of at least 0.1 in 60 minutes.
6. The method according to any one of 3 or 5. 7. The method of claim 6, wherein said cellulase has a crystalline: soluble cellulose activity ratio of at least 0.3 in 60 minutes. 8. The method of claim 7, wherein said cellulase has a crystalline: soluble cellulose activity ratio of at least 0.4 in 60 minutes. 9. The method of claim 8, wherein said cellulase has a crystalline: soluble cellulose activity ratio of at least 0.5 in 60 minutes. 10. The method of claim 9 wherein said cellulase has a crystalline: soluble cellulose activity ratio of at least 60 minutes. 11. The method according to claim 11, wherein said cellulase is at least 1.2 minutes in 60 minutes.
11. A crystalline: soluble cellulose activity ratio of
The method described in. 12. The method according to claim 12, wherein said cellulase is Trichoderma (Trichoderma).
12. The method according to any one of claims 1 to 11, wherein the method is derived from derma). 13. The method of claim 12, wherein said Trichoderma is Trichoderma reesei or Trichoderma longibrachiatum. 14. The method according to claim 14, wherein the cellulase is thermomonospora (Th
emomonospora). 15. The thermomonospora fusca according to claim 1, wherein said thermomonospora is Thermomonospora fusca.
The method described in 4. 16. The method according to claim 1, wherein said porous membrane is contacted with said amylase. 17. The method according to claim 17, wherein the amylase is α-amylase, β-amylase.
17. The method according to claim 16, wherein the method is selected from the group consisting of amylase and combinations thereof. 18. The method according to claim 1, wherein the porous membrane is further contacted with a basic aqueous solution before reuse.
The method described in the section. 19. The method according to claim 18, wherein the porous membrane is contacted with the basic aqueous solution before contacting with the enzyme. 20. The method according to claim 19, wherein the basic aqueous solution is NaOH and / or KO.
20. The method according to claim 18 or 19, wherein the method is an aqueous solution of H. 21. The method according to claim 21, wherein the base is 0.1 to 1 in the basic aqueous solution.
21. The method according to any one of claims 18 to 20, wherein the method is present at a concentration of N. 22. The method according to claim 18, wherein the porous membrane is contacted with the basic aqueous solution at a temperature of 40 to 90 ° C. 23. The method according to claim 19, wherein the porous membrane has a temperature of 40 to 50 ° C. and a pH of 4.5 to 4.5.
23. The method of any one of claims 1-22, wherein the method is contacted with the cellulase at 5.5. 24. The method according to claim 19, wherein the porous membrane has a temperature of 60 to 75 ° C. and a pH of 4.6 to 65.
24. The method of any one of claims 1-3 and 6-23, wherein the method is contacted with an α-amylase at 5.8. 25. The method according to claim 25, wherein the porous membrane has a temperature of 40 to 60 ° C. and a pH of 4.6 to 40.
24. The method of any one of claims 1-3 and 6-23, wherein the method is contacted with a β-amylase at 5.8. 26. The method according to claim 1, wherein the porous membrane is cleaned until the zeta potential of the porous membrane has been changed. 27. The method according to claim 1, wherein the time during which the porous membrane has to be cleaned is determined by the pressure drop across the porous membrane. 28. The method according to claim 1, wherein the time during which the porous membrane has to be cleaned is determined by the outflow or zeta potential of the porous membrane.