U.S. patent application number 14/022316 was filed with the patent office on 2014-01-09 for preparation of soy protein isolate using calcium chloride extraction ("s703").
The applicant listed for this patent is Brandy Gosnell, Brent E. Green, Sarah Medina, Martin Schweizer, Kevin I. Segall. Invention is credited to Brandy Gosnell, Brent E. Green, Sarah Medina, Martin Schweizer, Kevin I. Segall.
Application Number | 20140010944 14/022316 |
Document ID | / |
Family ID | 43381051 |
Filed Date | 2014-01-09 |
United States Patent
Application |
20140010944 |
Kind Code |
A1 |
Segall; Kevin I. ; et
al. |
January 9, 2014 |
PREPARATION OF SOY PROTEIN ISOLATE USING CALCIUM CHLORIDE
EXTRACTION ("S703")
Abstract
A soy protein product having a protein content of at least about
60 wt % (N.times.6.25) d.b., preferably an isolate, is formed by a
procedure in which soy protein is extracted from a soy source
material using an aqueous calcium chloride solution at low pH,
generally about 1.5 to about 5, and separating the resulting
aqueous soy protein solution from residual soy protein source. The
resulting clarified aqueous soy protein solution may be diluted and
the pH adjusted within the range of 1.5-5.0. The solution may be
concentrated by ultrafiltration, diafiltered and then dried to
provide the soy protein product. The soy protein product is soluble
in acidic medium and produces transparent, heat stable solutions
and hence may be used for protein fortification of soft drinks and
sports drinks.
Inventors: |
Segall; Kevin I.; (Winnipeg,
CA) ; Schweizer; Martin; (Winnipeg, CA) ;
Green; Brent E.; (Warren, CA) ; Medina; Sarah;
(Winnipeg, CA) ; Gosnell; Brandy; (Winnipeg,
CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Segall; Kevin I.
Schweizer; Martin
Green; Brent E.
Medina; Sarah
Gosnell; Brandy |
Winnipeg
Winnipeg
Warren
Winnipeg
Winnipeg |
|
CA
CA
CA
CA
CA |
|
|
Family ID: |
43381051 |
Appl. No.: |
14/022316 |
Filed: |
September 10, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13378680 |
Feb 23, 2012 |
8557321 |
|
|
PCT/CA10/01017 |
Jan 6, 2011 |
|
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14022316 |
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61213647 |
Jun 30, 2009 |
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Current U.S.
Class: |
426/598 ;
426/431; 426/656 |
Current CPC
Class: |
A23J 1/14 20130101; A23L
2/66 20130101; A23J 3/16 20130101; A23V 2002/00 20130101; A23V
2002/00 20130101; A23V 2250/5488 20130101 |
Class at
Publication: |
426/598 ;
426/431; 426/656 |
International
Class: |
A23J 1/14 20060101
A23J001/14; A23L 2/66 20060101 A23L002/66 |
Claims
1. A method of producing a soy protein product having a soy protein
content of at least about 60 wt % (N.times.6.25) on a dry weight
basis, which comprises: (a) extracting a soy protein source with an
aqueous calcium salt solution at a pH of about 1.5 to about 5 to
cause solubilization of soy protein from the soy protein source and
to form an aqueous soy protein solution, (b) separating the aqueous
soy protein solution from residual soy protein source,
2. The process of claim 1 wherein said extraction step is effected
using an aqueous calcium chloride solution having a concentration
of less than about 1.0 M.
3. The process of claim 2 wherein said aqueous calcium chloride
solution has a concentration of about 0.10 to about 0.15 M.
4. The process of claim 1 wherein said extraction step is effected
at a temperature of about 15.degree. to about 35.degree. C.
5. The process of claim 1 wherein said aqueous soy protein solution
has a protein concentration of about 5 to about 50 g/L.
6. The process of claim 5 wherein said protein concentration is
about 10 to about 50 g/L.
7. The process of claim 1 wherein said aqueous calcium salt
solution contains an antioxidant.
8. The process of claim 1 wherein, following said separation step,
said aqueous soy protein solution is treated with an adsorbent to
remove colour and/or odour compounds from the aqueous soy protein
solution.
9. The process of claim 1 wherein following said separation step,
the pH of the aqueous protein solution, is adjusted to a different
value within the range of about 1.5 to about 5.0.
10. The process of claim 9 wherein the pH value is adjusted to
about 1.5 to about 4.4.
11. The process of claim 10 wherein the pH value is adjusted to
about 2.0 to 4.0.
12. The process of claim 1 wherein, following said separation step,
said aqueous soy protein solution is diluted to a conductivity of
less than about 80 mS.
13. The process of claim 12 wherein said aqueous soy protein
solution is diluted with about 1 to about 10 volumes of water to
provide a conductivity of said soy protein solution of about 4 to
about 29 mS.
14. The process of claim 12 wherein said water has a temperature of
about 2.degree. C. to about 70.degree. C.
15. The process of claim 14 wherein said temperature is about
10.degree. to about 50.degree. C.
16. The process of claim 15 wherein said temperature is about
20.degree. to about 30.degree. C.
17. The process of claim 12 wherein following said dilution step,
the pH of the aqueous protein solution, is adjusted to a different
value within the range of about 1.5 to about 5.0.
18. The process of claim 17 wherein the pH is adjusted to about 1.5
to about 4.4.
19. The process of claim 17 wherein the pH is adjusted to about 2.0
to about 4.0.
20. The process of claim 17 wherein said soy protein solution,
following the dilution and pH adjustment steps has a conductivity
of less than about 85 mS.
21. The process of claim 20 wherein said conductivity is about 4 to
about 34 mS.
22. The process of claim 1, wherein said aqueous protein solution
is subjected to a heat treatment step to inactivate heat-labile
anti-nutritional factors.
23. The process of claim 22 wherein the anti-nutritional factors
are heat-labile trypsin inhibitors.
24. The process of claim 22 wherein the heat treatment step also
pasteurizes the aqueous soy protein solution.
25. The process of claim 22, wherein said heat treatment is
effected at a temperature of about 70.degree. to about 160.degree.
C. for about 10 seconds to about 60 minutes.
26. The process of claim 25 wherein said heat treatment is effected
at a temperature of about 80.degree. to about 120.degree. C. for
about 10 seconds to about 5 minutes.
27. The process of claim 26 wherein said heat treatment is effected
at a temperature of about 85.degree. C. to about 95.degree. C. for
about 30 seconds to about 5 minutes.
28. The process of claim 22 wherein the heat treated soy protein
solution is cooled to a temperature of about 2.degree. to about
60.degree. C. for further processing.
29. The process of claim 28 wherein the heat treated soy protein
solution is cooled to a temperature of about 20.degree. to about
35.degree. C. for further processing.
30. The process of claim 1 wherein said soy protein solution is
dried to provide a soy protein product having a soy protein content
of at least about 60 wt % (N.times.6.25) d.b.
31. The process of claim 1 wherein said soy protein solution is
concentrated while maintaining the ionic strength thereof
substantially constant to produce a concentrated soy protein
solution having a protein concentration of about 50 to about 300
g/L and the concentrated soy protein solution is optionally
diafiltered.
32. The process of claim 31 wherein said concentrated soy protein
solution has a protein concentration of about 100 to about 200
g/L.
33. The process of claim 31 wherein said concentration step is
effected by ultrafiltration using a membrane having a molecular
weight cut-off of about 3,000 to about 1,000,000 Daltons.
34. The process of claim 33 wherein said membrane has a molecular
weight cut-off of about 5,000 to about 100,000 Daltons.
35. The process of claim 31 wherein a diafiltration step is
effected using water, dilute saline, acidified water or acidified
dilute saline on the soy protein solution before or after partial
or complete concentration thereof.
36. The process of claim 35 wherein said diafiltration is effected
using about 2 to about 40 volumes of diafiltration solution.
37. The process of claim 36 wherein said diafiltration is effected
using about 5 to about 25 volumes of diafiltration solution.
38. The process of claim 35 wherein said diafiltration is effected
until no significant further quantities of contaminants or visible
colour are present in the permeate.
39. The process of claim 35 wherein said diafiltration is effected
until the retentate has been sufficiently purified so as, when
dried, to provide a soy protein isolate with a protein content of
at least about 90 wt % (N.times.6.25) d.b.
40. The process of claim 35 wherein said diafiltration is effected
using a membrane having a molecular weight cut-off of about 3,000
to about 1,000,000 Daltons.
41. The process of claim 40 wherein said membrane has a molecular
weight cut-off of about 5,000 to about 100,000 Daltons.
42. The process of claim 35 wherein an antioxidant is present in
the diafiltration medium during at least part of the diafiltration
step.
43. The process of claim 31 wherein said concentration step and
optional diafiltration step are carried out at a temperature of
about 2.degree. to about 60.degree. C.
44. The process of claim 43 wherein said temperature is about
20.degree. to about 35.degree. C.
45. The process of claim 31 wherein the concentrated and optionally
diafiltered soy protein solution is subjected to a heat treatment
step to inactivate heat-labile anti-nutritional factors, including
heat-labile trypsin.
46. The process of claim 45 wherein said heat treatment is effected
at a temperature of about 70.degree. to about 160.degree. C. for
about 10 seconds to 60 minutes, preferably a temperature of about
80.degree. to about 120.degree. C. for about 10 seconds to about 5
minutes, more preferably about 88.degree. to about 95.degree. C.
for about 30 seconds to about 5 minutes.
47. The process of claim 46 wherein the heat treated soy protein
solution is cooled to a temperature of about 2.degree. to about
60.degree. C., preferably about 20.degree. to about 35.degree. C.,
for further processing.
48. The process of claim 1 wherein said soy protein solution is
concentrated and/or diafiltered to produce a concentrated and/or
diafiltered soy protein solution which, when dried, provides a soy
protein product having a protein concentration of at least about 60
wt % (N.times.6.25) d.b.
49. The process of claim 31 wherein said concentrated and
optionally diafiltered soy protein solution is treated with an
adsorbent to remove colour and/or odour compounds.
50. The process of claim 31 wherein said concentrated and
optionally diafiltered soy protein solution is pasteurized prior to
drying.
51. The process of claim 50 wherein said pasteurization step is
effected at a temperature of about 55.degree. to about 70.degree.
C. for about 30 seconds to about 60 minutes.
52. The process of claim 51 wherein said pasteurization step is
effected at a temperature of about 60.degree. to about 65.degree.
C. for about 10 to about 15 minutes.
53. The process of claim 52 wherein said concentrated and
optionally diafiltered soy protein solution is dried to provide a
soy protein isolate having a protein content of at least about 90
wt % (N.times.6.25) d.b.
54. The process of claim 53 wherein said soy protein isolate has a
protein content of at least about 100 wt % (N.times.6.25) d.b.
55. The process of claim 31 wherein the concentration and/or
optional diafiltration step are operated in a manner favourable to
the removal of trypsin inhibitors.
56. The process of claim 1 wherein a reducing agent is present
during the extraction step to disrupt or rearrange the disulfide
bonds of trypsin inhibitors to achieve a reduction in trypsin
inhibitor activity.
57. The process of claim 31 wherein a reducing agent is present
during the concentration and/or optional diafiltration step to
disrupt or rearrange the disulfide bonds of trypsin inhibitors to
achieve a reduction in trypsin inhibitor activity.
58. The process of claim 48 wherein a reducing agent is added to
the concentrated and optionally diafiltered soy protein solution
prior to drying and/or the dried soy protein product to disrupt or
rearrange the disulfide bonds of trypsin inhibitors to achieve a
reduction in trypsin inhibitor activity.
59. A soy protein product produced by the process of claim 1.
60. An acidic solution having dissolved therein the soy protein
product of claim 59.
61. The aqueous solution of claim 60 which is a beverage.
62. The soy protein product of claim 59 which is blended with water
soluble powdered materials for the production of aqueous solutions
of the blend.
63. The blend of claim 62 which is a powdered beverage.
64. An aqueous solution with a near neutral pH, preferably in the
pH range of about 6 to about 8, having dissolved therein the soy
product of claim 59.
65. The aqueous solution of claim 64 which is a beverage.
Description
REFERENCE TO RELATED APPLICATION
[0001] This application claims priority under 35 USC 119(e) from
U.S. Provisional Patent Application No. 61/213,647 filed Jun. 30,
2009.
FIELD OF INVENTION
[0002] The present invention is concerned with the preparation of
soy protein products.
BACKGROUND TO THE INVENTION
[0003] In U.S. Provisional Patent Applications Nos. 61/107,112
(7865-373) filed Oct. 21, 2008, 61/193,457 (7865-374) filed Dec. 2,
2008, 61/202,070 (7865-376) filed Jan. 26, 2009, 61/202,553 filed
Mar. 12, 2009 (7865-383), 61/213,717 (7865-389) filed Jul. 7, 2009,
61/272,241 (7865-400) filed Sep. 3, 2009 and U.S. patent
application Ser. No. 12/603,087 (7865-415) filed Oct. 21, 2009 (US
Patent Publication No. 2010-0098818), assigned to the assignee
hereof and the disclosures of which are incorporated herein by
reference, there is described the preparation of a soy protein
product, preferably a soy protein isolate, which is completely
soluble and is capable of providing transparent and heat stable
solutions at low pH values. This soy protein product may be used
for protein fortification of, in particular, soft drinks and sports
drinks, as well as other acidic aqueous systems, without
precipitation of protein. The soy protein product is produced by
extracting a soy protein source with aqueous calcium chloride
solution at natural pH, optionally diluting the resulting aqueous
soy protein solution, adjusting the pH of the aqueous soy protein
solution to a pH of about 1.5 to about 4.4, preferably about 2.0 to
about 4.0, to produce an acidified clear soy protein solution,
which may be optionally concentrated and/or diafiltered before
drying.
SUMMARY OF THE INVENTION
[0004] It has now been surprisingly found that a soy protein
product having a protein content of at least about 60 wt %
(N.times.6.25) d.b. may be formed by a procedure involving
extraction of the soy protein source with calcium chloride at low
pH values.
[0005] In one aspect of the present invention, a soy protein source
material is extracted with aqueous calcium chloride solution at low
pH and the resulting aqueous soy protein solution is optionally
diluted, optionally adjusted in pH within the acidic range, then
subjected to ultrafiltration and optional diafiltration to provide
a concentrated and optionally diafiltered soy protein solution,
which may be dried to provide the soy protein product.
[0006] The soy protein product provided herein, having a protein
content of at least about 60 wt % (N.times.6.25) d.b., is soluble
at acid pH values to provide transparent and heat stable aqueous
solutions thereof. The soy protein product may be used for protein
fortification of, in particular, soft drinks and sports drinks, as
well as other aqueous systems without precipitation of protein. The
soy protein product is preferably an isolate having a protein
content of at least about 90 wt %, preferably at least about 100 wt
% (N.times.6.25) d.b.
[0007] In accordance with one aspect of the present invention,
there is provided a method of producing a soy protein product
having a soy protein content of at least about 60 wt %
(N.times.6.25), on a dry weight basis, which comprises: [0008] (a)
extracting a soy protein source with aqueous calcium salt solution,
generally calcium chloride solution, at low pH, generally about 1.5
to about 5.0, to cause solubilization of soy protein from the
protein source and to form an aqueous soy protein solution, [0009]
(b) separating the aqueous soy protein solution from residual soy
protein source, [0010] (c) optionally diluting the aqueous soy
protein solution [0011] (d) optionally adjusting the pH of the
aqueous protein solution to a value within the range of about 1.5
to about 5.0, preferably about 1.5 to about 4.4, more preferably
about 2.0 to about 4.0, and differing from the pH of extraction.
[0012] (e) optionally concentrating the aqueous soy protein
solution while maintaining the ionic strength substantially
constant by using a selective membrane technique, [0013] (f)
optionally diafiltering the concentrated soy protein solution, and
[0014] (g) optionally drying the concentrated and diafiltered soy
protein solution.
[0015] The soy protein product preferably is an isolate having a
protein content of at least about 90 wt %, preferably at least
about 100 wt % (N.times.6.25) d.b.
[0016] Although this specification refers mainly to the production
of a soy protein isolate, the concentration and/or diafiltration
steps described herein may be manipulated to produce a soy protein
product of lesser purity, for example, a soy protein concentrate
having a protein content of at least about 60 wt %, but which has
substantially similar properties to the isolate.
[0017] The novel soy protein product of the invention can be
blended with powdered drinks for the formation of aqueous soft
drinks or sports drinks by dissolving the same in water. Such blend
may be a powdered beverage.
[0018] The soy protein product provided herein may be provided as
an aqueous solution thereof having a high degree of clarity at acid
pH values and which is heat stable at these pH values.
[0019] In another aspect of the present invention, there is
provided an aqueous solution of the soy product provided herein
which is heat stable at low pH. The aqueous solution may be a
beverage, which may be a clear beverage in which the soy protein
product is completely soluble and transparent or an opaque beverage
in which the soy protein product does not increase the opacity. The
soy protein product also has good solubility at about pH 7. An
aqueous solution of the soy protein product, prepared at a near
neutral pH, such as a pH of about 6 to about 8, may be a
beverage.
[0020] The soy protein product produced according to the process
herein lacks the characteristic beany flavour of soy protein
isolates and are suitable, not only for protein fortification of
acidic media, but may be used in a wide variety of conventional
applications of protein isolates, including but not limited to
protein fortification of processed foods and beverages,
emulsification of oils, as a body former in baked goods and foaming
agent in products which entrap gases. In addition, the soy protein
product may be formed into protein fibers, useful in meat analogs,
and may be used as an egg white substitute or extender in food
products where egg white is used as a binder. The soy protein
product may also be used in nutritional supplements. Other uses of
the soy protein product are in pet foods, animal feed and in
industrial and cosmetic applications and in personal care
products.
GENERAL DESCRIPTION OF INVENTION
[0021] The initial step of the process of providing the soy protein
product involves solubilizing soy protein from a soy protein
source. The soy protein source may be soybeans or any soy product
or by-product derived from the processing of soybeans including but
not limited to soy meal, soy flakes, soy grits and soy flour. The
soy protein source may be used in the full fat form, partially
defatted form or fully defatted form. Where the soy protein source
contains an appreciable amount of fat, an oil-removal step
generally is required during the process. The soy protein recovered
from the soy protein source may be the protein naturally occurring
in soybean or the proteinaceous material may be a protein modified
by genetic manipulation but possessing characteristic hydrophobic
and polar properties of the natural protein.
[0022] Protein solubilization from the soy protein source material
is effected most conveniently using calcium chloride solution,
although solutions of other calcium salts may be used. In addition,
other alkaline earth metal compounds may be used, such as magnesium
salts. Further, extraction of the soy protein from the soy protein
source may be effected using calcium salt solution in combination
with another salt solution such as sodium chloride. Additionally,
extraction of the soy protein from the soy protein source may be
effected using water or other salt solution, such as sodium
chloride, with calcium chloride subsequently being added to the
aqueous soy protein solution produced in the extraction step.
Precipitate formed upon addition of the calcium chloride then is
removed prior to subsequent processing.
[0023] As the concentration of the calcium salt solution increases,
the degree of solubilization of protein from the soy protein source
initially increases until a maximum value is achieved. Any
subsequent increase in salt concentration does not increase the
total protein solubilized. The concentration of calcium salt
solution which causes maximum protein solubilization varies
depending on the salt concerned. It is usually preferred to utilize
a concentration value less than about 1.0 M, and, more preferably,
a value of about 0.10 M to about 0.15 M.
[0024] In a batch process, the solubilization of the protein is
effected at a temperature of from about 1.degree. C. to about
100.degree. C., preferably about 15.degree. to about 35.degree. C.,
preferably accompanied by agitation to decrease the solubilization
time, which is usually about 1 to about 60 minutes. It is preferred
to effect the solubilization to extract substantially as much
protein from the soy protein source as is practicable, so as to
provide an overall high product yield.
[0025] In a continuous process, the extraction of the soy protein
from the soy protein source is carried out in any manner consistent
with effecting a continuous extraction of soy protein from the soy
protein source. In one embodiment, the soy protein source is
continuously mixed with calcium salt solution and the mixture is
conveyed through a pipe or conduit having a length and at a flow
rate for a residence time sufficient to effect the desired
extraction in accordance with the parameters described herein. In
such a continuous procedure, the solubilization step is effected
rapidly, in a time of up to about 10 minutes, preferably to effect
solubilization to extract substantially as much protein from the
soy protein source as is practicable. The solubilization in the
continuous procedure is effected at temperatures between about
1.degree. C. and about 100.degree. C., preferably between about
15.degree. C. and about 35.degree. C.
[0026] The extraction is generally conducted at a pH of about 1.5
to about 5.0. The pH of the extraction system (soy protein source
and calcium salt solution) may be adjusted to any desired value
within the range of about 1.5 to about 5.0 for the extraction step
by the use of any convenient food grade acid, usually hydrochloric
acid or phosphoric acid.
[0027] The concentration of soy protein source in the calcium salt
solution during the solubilization step may vary widely. Typical
concentration values are about 5 to about 15% w/v.
[0028] The protein extraction step with the aqueous calcium salt
solution has the additional effect of solubilizing fats which may
be present in the soy protein source, which then results in the
fats being present in the aqueous phase.
[0029] The protein solution resulting from the extraction step
generally has a protein concentration of about 5 to about 50 g/L,
preferably about 10 to about 50 g/L.
[0030] The aqueous calcium salt solution may contain an
antioxidant. The antioxidant may be any convenient antioxidant,
such as sodium sulfite or ascorbic acid. The quantity of
antioxidant employed may vary from about 0.01 to about 1 wt % of
the solution, preferably about 0.05 wt %. The antioxidant serves to
inhibit the oxidation of any phenolics in the protein solution.
[0031] The aqueous phase resulting from the extraction step then
may be separated from the residual soy protein source, in any
convenient manner, such as by employing a decanter centrifuge,
followed by disc centrifugation and/or filtration, to remove
residual soy protein source material. The separated residual soy
protein source may be dried for disposal. Alternatively, the
separated residual soy protein source may be processed to recover
some residual protein, such as by a conventional isoelectric
precipitation procedure or any other convenient procedure to
recover such residual protein.
[0032] Where the soy protein source contains significant quantities
of fat, as described in U.S. Pat. Nos. 5,844,086 and 6,005,076,
assigned to the assignee hereof and the disclosures of which are
incorporated herein by reference, then the defatting steps
described therein may be effected on the separated aqueous protein.
Alternatively, defatting of the separated aqueous protein solution
may be achieved by any other convenient procedure.
[0033] The aqueous soy protein solution may be treated with an
adsorbent, such as powdered activated carbon or granulated
activated carbon, to remove colour and/or odour compounds. Such
adsorbent treatment may be carried out under any convenient
conditions, generally at the ambient temperature of the separated
aqueous protein solution. For powdered activated carbon, an amount
of about 0.025% to about 5% w/v, preferably about 0.05% to about 2%
w/v, is employed. The adsorbing agent may be removed from the soy
protein solution by any convenient means, such as by
filtration.
[0034] The resulting aqueous soy protein solution may be diluted
with water generally with about 0.5 to about 10 volumes, preferably
about 1 to about 2 volumes, in order to decrease the conductivity
of the aqueous soy protein solution to a value of generally below
about 90 mS, preferably about 4 to about 31 mS.
[0035] The water with which the soy protein solution is mixed may
have a temperature of about 2.degree. to about 70.degree. C.,
preferably about 10.degree. to about 50.degree. C., more preferably
about 20.degree. to about 30.degree. C.
[0036] The optionally diluted soy protein solution may be adjusted
in pH to a value different from the extraction pH but still within
the range of about 1.5 to about 5.0, preferably about 1.5 to about
4.4, more preferably about 2.0 to about 4.0, by the addition of any
suitable food grade acid, such as hydrochloric acid or phosphoric
acid, or food grade alkali, usually sodium hydroxide as
required.
[0037] The diluted and optionally pH adjusted soy protein solution
has a conductivity of generally below about 95 mS, preferably about
4 to about 36 mS.
[0038] The aqueous soy protein solution may be subjected to a heat
treatment to inactivate heat labile anti-nutritional factors, such
as trypsin inhibitors, present in such solution as a result of
extraction from the soy protein source material during the
extraction step. Such a heating step also provides the additional
benefit of reducing the microbial load. Generally, the protein
solution is heated to a temperature of about 70.degree. to about
160.degree. C., preferably about 80.degree. to about 120.degree.
C., more preferably about 85.degree. C. to about 95.degree. C. for
about 10 seconds to about 60 minutes, preferably about 30 seconds
to about 5 minutes. The heat treated acidified soy protein solution
then may be cooled for further processing as described below, to a
temperature of about 2.degree. C. to about 60.degree. C.,
preferably about 20.degree. to about 35.degree. C.
[0039] The resulting aqueous soy protein solution may be directly
dried to produce a soy protein product. In order to provide a soy
protein isolate having a decreased impurities content and a reduced
salt content, the aqueous soy protein solution may be processed
prior to drying.
[0040] The aqueous soy protein solution may be concentrated to
increase the protein concentration thereof while maintaining the
ionic strength thereof substantially constant. Such concentration
generally is effected to provide a concentrated soy protein
solution having a protein concentration of about 50 to about 300
g/L, preferably about 100 to about 200 g/L.
[0041] Prior to the concentration step, the aqueous soy protein
solution may be subjected to a polishing operation to remove any
residual soy source material fines not removed in the separation
step discussed above. Such polishing step may be effected in any
convenient manner, such as by filtration.
[0042] The concentration step may be effected in any convenient
manner consistent with batch or continuous operation, such as by
employing any convenient selective membrane technique, such as
ultrafiltration or diafiltration, using membranes, such as
hollow-fibre membranes or spiral-wound membranes, with a suitable
molecular weight cut-off, such as about 3,000 to about 1,000,000
Daltons, preferably about 5,000 to about 100,000 Daltons, having
regard to differing membrane materials and configurations, and, for
continuous operation, dimensioned to permit the desired degree of
concentration as the aqueous protein solution passes through the
membranes.
[0043] As is well known, ultrafiltration and similar selective
membrane techniques permit low molecular weight species to pass
therethrough while preventing higher molecular weight species from
so doing. The low molecular weight species include not only the
ionic species of the food grade salt but also low molecular weight
materials extracted from the source material, such as
carbohydrates, pigments, low molecular weight proteins and
anti-nutritional factors, such as trypsin inhibitor, which
themselves are low molecular weight proteins. The molecular weight
cut-off of the membrane is usually chosen to ensure retention of a
significant proportion of the protein in the solution, while
permitting contaminants to pass through having regard to the
different membrane materials and configurations.
[0044] The concentrated soy protein solution then may be subjected
to a diafiltration step using water or a dilute saline solution.
The diafiltration solution may be at its natural pH or at a pH
equal to that of the protein solution being diafiltered or at any
pH value in between. Such diafiltration may be effected using from
about 2 to about 40 volumes of diafiltration solution, preferably
about 5 to about 25 volumes of diafiltration solution. In the
diafiltration operation, further quantities of contaminants are
removed from the aqueous soy protein solution by passage through
the membrane with the permeate. This purifies the aqueous protein
solution and may also reduce its viscosity. The diafiltration
operation may be effected until no significant further quantities
of contaminants or visible colour are present in the permeate or
until the retentate has been sufficiently purified so as, when
dried, to provide a soy protein isolate with a protein content of
at least about 90 wt % (N.times.6.25) d.b. Such diafiltration may
be effected using the same membrane as for the concentration step.
However, if desired, the diafiltration step may be effected using a
separate membrane with a different molecular weight cut-off, such
as a membrane having a molecular weight cut-off in the range of
about 3,000 to about 1,000,000 Daltons, preferably about 5,000 to
about 100,000 Daltons, having regard to different membrane
materials and configuration.
[0045] Alternatively, the diafiltration step may be applied to the
aqueous protein solution prior to concentration or to the partially
concentrated aqueous protein solution. Diafiltration may also be
applied at multiple points during the concentration process. When
diafiltration is applied prior to concentration or to the partially
concentrated solution, the resulting diafiltered solution may then
be additionally concentrated. The viscosity reduction achieved by
diafiltering multiple times as the protein solution is concentrated
may allow a higher final, fully concentrated protein concentration
to be achieved. This reduces the volume of material to be
dried.
[0046] The concentration step and the diafiltration step may be
effected herein in such a mariner that the soy protein product
subsequently recovered contains less than about 90 wt % protein
(N.times.6.25) d.b., such as at least about 60 wt % protein
(N.times.6.25) d.b. By partially concentrating and/or partially
diafiltering the aqueous soy protein solution, it is possible to
only partially remove contaminants. This protein solution may then
be dried to provide a soy protein product with lower levels of
purity. The soy protein product is still able to produce clear
protein solutions under acidic conditions.
[0047] An antioxidant may be present in the diafiltration medium
during at least part of the diafiltration step. The antioxidant may
be any convenient antioxidant, such as sodium sulfite or ascorbic
acid. The quantity of antioxidant employed in the diafiltration
medium depends on the materials employed and may vary from about
0.01 to about 1 wt %, preferably about 0.05 wt %. The antioxidant
serves to inhibit the oxidation of any phenolics present in the
concentrated soy protein solution.
[0048] The concentration step and the optional diafiltration step
may be effected at any convenient temperature, generally about
2.degree. C. to about 60.degree. C., preferably about 20.degree. C.
to about 35.degree. C., and for the period of time to effect the
desired degree of concentration and diafiltration. The temperature
and other conditions used to some degree depend upon the membrane
equipment used to effect the membrane processing, the desired
protein concentration of the solution and the efficiency of the
removal of contaminants to the permeate.
[0049] There are two main trypsin inhibitors in soy, namely the
Kunitz inhibitor, which is a heat-labile molecule with a molecular
weight of approximately 21,000 Daltons, and the Bowman-Birk
inhibitor, a more heat-stable molecule with a molecular weight of
about 8,000 Daltons. The level of trypsin inhibitor activity in the
final soy protein product can be controlled by manipulation of
various process variables.
[0050] As noted above, heat treatment of the aqueous soy protein
solution may be used to inactivate heat-labile trypsin inhibitors.
The partially concentrated or fully concentrated soy protein
solution may also be heat treated to inactivate heat labile trypsin
inhibitors.
[0051] In addition, the concentration and/or diafiltration steps
may be operated in a manner favorable for removal of trypsin
inhibitors in the permeate along with the other contaminants.
Removal of the trypsin inhibitors is promoted by using a membrane
of larger pore size (such as about 30,000 to about 1,000,000 Da),
operating the membrane at elevated temperatures (such as about
30.degree. C. to about 60.degree. C.) and employing greater volumes
of diafiltration medium (such as about 20 to about 40 volumes).
[0052] Extracting and/or membrane processing the protein solution
at a lower pH (1.5-3.0) may reduce the trypsin inhibitor activity
relative to processing the solution at higher pH (3.0-5.0). When
the protein solution is concentrated and diafiltered at the low end
of the pH range, it may be desired to raise the pH of the retentate
prior to drying. The pH of the concentrated and diafiltered protein
solution may be raised to the desired value, for example pH 3, by
the addition of any convenient food grade alkali such as sodium
hydroxide. If it is desired to lower the pH of the retentate prior
to drying, this may be done so by the addition of any convenient
food grade acid such as hydrochloric acid or phosphoric acid.
[0053] Further, a reduction in trypsin inhibitor activity may be
achieved by exposing soy materials to reducing agents that disrupt
or rearrange the disulfide bonds of the inhibitors. Suitable
reducing agents include sodium sulfite, cysteine and
N-acetylcysteine.
[0054] The addition of such reducing agents may be effected at
various stages of the overall process. The reducing agent may be
added with the soy protein source material in the extraction step,
may be added to the clarified aqueous soy protein solution
following removal of residual soy protein source material, may be
added to the concentrated protein solution before or after
diafiltration or may be dry blended with the dried soy protein
product. The addition of the reducing agent may be combined with a
heat treatment step and the membrane processing steps, as described
above.
[0055] If it is desired to retain active trypsin inhibitors in the
concentrated protein solution, this can be achieved by eliminating
or reducing the intensity of the heat treatment step, not utilizing
reducing agents, operating the concentration and diafiltration
steps at the higher end of the pH range (3.0.to 5.0), utilizing a
concentration and diafiltration membrane with a smaller pore size,
operating the membrane at lower temperatures and employing fewer
volumes of diafiltration medium.
[0056] The concentrated and optionally diafiltered protein solution
may be subject to a further defatting operation, if required, as
described in U.S. Pat. Nos. 5,844,086 and 6,005,076. Alternatively,
defatting of the concentrated and optionally diafiltered protein
solution may be achieved by any other convenient procedure.
[0057] The concentrated and diafiltered aqueous protein solution
may be treated with an adsorbent, such as powdered activated carbon
or granulated activated carbon, to remove colour and/or odour
compounds. Such adsorbent treatment may be carried out under any
convenient conditions, generally at the ambient temperature of the
concentrated protein solution. For powdered activated carbon, an
amount of about 0.025% to about 5% w/v, preferably about 0.05% to
about 2% w/v, is employed. The adsorbent may be removed from the
soy protein solution by any convenient means, such as by
filtration.
[0058] The concentrated and diafiltered aqueous soy protein
solution may be dried by any convenient technique, such as spray
drying or freeze drying. A pasteurization step may be effected on
the soy protein solution prior to drying to reduce the microbial
load. Such pasteurization step may be effected under any desired
pasteurization conditions. Generally, the concentrated and
optionally diafiltered soy protein solution is heated to a
temperature of about 55.degree. to about 70.degree. C., preferably
about 60.degree. to about 65.degree. C., for about 30 seconds to
about 60 minutes, preferably about 10 minutes to about 15 minutes.
The pasteurized concentrated and diafiltered soy protein solution
then may be cooled for drying, preferably to a temperature of about
25.degree. to about 40.degree. C.
[0059] The dry soy protein product has a protein content in excess
of about 60 wt % N.times.6.25) d.b. Preferably, the dry soy protein
product is an isolate with a high protein content, in excess of
about 90 wt % protein, preferably at least about 100 wt %
(N.times.6.25) d.b.
[0060] The soy protein products produced herein are soluble in an
acidic aqueous environment, making the product ideal for
incorporation into beverages, both carbonated and uncarbonated, to
provide protein fortification thereto. Such beverages have a wide
range of acidic pH values, ranging from about 2.5 to about 5. The
soy protein products provided herein may be added to such beverages
in any convenient quantity to provide protein fortification to such
beverages, for example, at least about 5 g of the soy protein per
serving. The added soy protein product dissolves in the beverage
and does not impair the clarity of the beverage, even after thermal
processing. The soy protein product may be blended with dried
beverage prior to reconstitution of the beverage by dissolution in
water. In some cases, modification of the normal formulation of the
beverages to tolerate the composition of the invention may be
necessary where components present in the beverage may adversely
affect the ability of the composition to remain dissolved in the
beverage.
EXAMPLES
Example 1
[0061] This Example illustrates the preparation of transparent,
heat stable protein solutions utilizing extraction with calcium
chloride solution at low pH.
[0062] Soy white flakes (10 g) were combined with 0.15M calcium
chloride solution (100 ml) and the pH of the samples adjusted
immediately to 4.8 and 1.5 with HCl. The samples were extracted at
room temperature for 30 minutes using a magnetic stirrer. The pH of
the samples was monitored and adjusted two times during the 30
minute extraction. The extract was separated from the spent meal by
centrifugation at 10,200 g for 10 minutes and the centrates further
clarified by filtration using 25 .mu.m pore size filter paper. The
clarity of the filtrates was measured using a HunterLab ColorQuest
XE operated in transmission mode to supply a percentage haze
reading. The samples were then diluted with one volume of reverse
osmosis purified water and the haze level measured again. The pH of
the diluted samples was then adjusted to 3 using either HCl or NaOH
as necessary. The haze level of the pH adjusted samples was then
analyzed. The samples were then heat treated to 95.degree. C. for
30 seconds, immediately cooled to room temperature in ice water and
the haze level re-assessed.
[0063] The haze values determined for the various samples are shown
in Tables 1 and 2.
TABLE-US-00001 TABLE 1 Haze values for the treatment of samples
from extraction with calcium chloride solution at pH 1.5 sample
haze (%) filtrate 27.8 diluted filtrate 17.1 diluted filtrate at pH
3 16.8 diluted filtrate at pH 3 after heat treatment 10.4
TABLE-US-00002 TABLE 2 Haze values for the treatment of samples
from extraction with calcium chloride solution at pH 4.8 sample
haze (%) filtrate 36.2 diluted filtrate 99.1 diluted filtrate at pH
3 8.4 diluted filtrate at pH 3 after heat treatment 6.0
[0064] As may be seen from the results presented in Tables 1 and 2,
the initial filtrates were somewhat hazy, however improved clarity
may have been obtained by utilizing a finer filter. Dilution with
one volume of water improved the clarity of the pH 1.5 sample but
introduced precipitation in the pH 4.8 sample. Adjusting the pH of
the diluted samples to 3 gave good clarity to the sample that was
originally at pH 4.8, while the sample that was originally at pH
1.5 had perhaps a slight haze. After heat treatment both samples
were considered clear.
Example 2
[0065] This Example illustrates the preparation of a soy protein
isolate in accordance with one embodiment of the invention.
[0066] 20 kg of defatted, minimally heat treated soy flour was
added to 200 L of 0.15M calcium chloride solution at ambient
temperature and agitated for 30 minutes to provide an aqueous
protein solution. Immediately after the flour was dispersed in the
calcium chloride solution, the pH of the system was adjusted to 3
by the addition of dilute HCl. The pH was monitored and corrected
to 3 periodically over the course of the 30 minute extraction. The
residual soy flour was removed by centrifugation to yield 174 L of
protein solution having a protein content of 3.37% by weight. The
protein solution was then combined with 174 L of reverse osmosis
purified water and the pH corrected to 3. This solution was then
polished by filtration to yield 385 L of filtered protein solution
having a protein content of 1.21% by weight.
[0067] The filtered protein solution was reduced in volume to 25 L
by concentration on a PVDF membrane having a molecular weight
cutoff of 5,000 daltons. The concentrated protein solution was then
diafiltered with 125 L of reverse osmosis purified water. The
resulting diafiltered, concentrated protein solution had a protein
content of 14.51% by weight and represented a yield of 81.3 wt % of
the filtered protein solution. The diafiltered, concentrated
protein solution was then dried to yield a product found to have a
protein content of 99.18% (N.times.6.25) d.b. The product was
termed S005-A13-09A S703.
[0068] A 3.2 wt % protein solution of S005-A13-09A S703 was
prepared in water and the colour and clarity assessed using a
HunterLab Color Quest XE instrument operated in transmission mode.
The pH of the solution was measured with a pH meter.
[0069] The pH, colour and clarity values are set forth in the
following Table 3:
TABLE-US-00003 TABLE 3 pH and HunterLab scores for 3.2% protein
solution of S005-A13-09A S703 sample pH L* a* b* haze (%) S703 3.12
87.31 0.67 18.99 43.9
[0070] As may be seen from Table 3, the solution of 5703 in water
was semi-transparent, not transparent. The relatively high level of
haze in this sample resulted in the L* value being somewhat lower
than expected.
[0071] The colour of the dry powder was also assessed with the
HunterLab Color Quest XE instrument in reflectance mode. The colour
values are set forth in the following Table 4:
TABLE-US-00004 TABLE 4 HunterLab scores for S005-A13-09A S703 dry
powder sample L* a* b* S703 85.67 0.05 10.57
[0072] As may be seen from Table 4, the dry product was very light
in colour.
Example 3
[0073] This Example contains an evaluation of the heat stability in
water of the soy protein isolate produced by the method of Example
2 (S703).
[0074] A 2% w/v protein solution of S005-A13-09A 5703 in water was
produced and the pH adjusted to 3. The clarity of this solution was
assessed by haze measurement with the HunterLab Color Quest XE
instrument. The solution was then heated to 95.degree. C., held at
this temperature for 30 seconds and then immediately cooled to room
temperature in an ice bath. The clarity of the heat treated
solution was then measured again.
[0075] The clarity of the protein solution before and after heating
is set forth in the following Table 5:
TABLE-US-00005 TABLE 5 Effect of heat treatment on clarity of
S005-A13-09A S703 solution sample haze (%) before heating 43.6
after heating 30.7
[0076] As can be seen from the results in Table 5, it was found
that the initial solution of 5005-A13-09A 5703 was quite hazy.
However, the solution was heat stable, with the haze level actually
reduced somewhat by the heat treatment.
Example 4
[0077] This Example contains an evaluation of the solubility in
water of the soy protein isolate produced by the method of Example
2 (S703). Solubility was tested based on protein solubility (termed
protein method, a modified version of the procedure of Morr et al.,
J. Food Sci. 50:1715-1718) and total product solubility (termed
pellet method).
[0078] Sufficient protein powder to supply 0.5 g of protein was
weighed into a beaker and then a small amount of reverse osmosis
(RO) purified water was added and the mixture stirred until a
smooth paste formed. Additional water was then added to bring the
volume to approximately 45 ml. The contents of the beaker were then
slowly stirred for 60 minutes using a magnetic stirrer. The pH was
determined immediately after dispersing the protein and was
adjusted to the appropriate level (2, 3, 4, 5, 6 or 7) with diluted
NaOH or HCl. A sample was also prepared at natural pH. For the pH
adjusted samples, the pH was measured and corrected two times
during the 60 minutes stirring. After the 60 minutes of stirring,
the samples were made up to 50 ml total volume with RO water,
yielding a 1% w/v protein dispersion. The protein content of the
dispersions was measured using a Leco FP528 Nitrogen Determinator.
Aliquots (20 ml) of the dispersions were then transferred to
pre-weighed centrifuge tubes that had been dried overnight in a
100.degree. C. oven then cooled in a desiccator and the tubes
capped. The samples were centrifuged at 7800 g for 10 minutes,
which sedimented insoluble material and yielded a clear
supernatant. The protein content of the supernatant was measured by
Leco analysis and then the supernatant and the tube lids were
discarded and the pellet material dried overnight in an oven set at
100.degree. C. The next morning the tubes were transferred to a
desiccator and allowed to cool. The weight of dry pellet material
was recorded. The dry weight of the initial protein powder was
calculated by multiplying the weight of powder used by a factor of
((100-moisture content of the powder (%))/100). Solubility of the
product was then calculated two different ways:
[0079] 1) Solubility (protein method) (%)=(% protein in
supernatant/% protein in initial dispersion).times.100
[0080] 2) Solubility (pellet method) (%)=(1-(weight dry insoluble
pellet material/((weight of 20 ml of dispersion/weight of 50 ml of
dispersion) x initial weight dry protein powder))).times.100
[0081] The natural pH value of the protein isolate produced in
Example 1 in water (1% protein) is shown in Table 6:
TABLE-US-00006 TABLE 6 Natural pH of S703 solution prepared in
water at 1% protein Batch Product Natural pH S005-A13-09A S703
3.36
[0082] The solubility results obtained are set forth in the
following Tables 7 and 8:
TABLE-US-00007 TABLE 7 Solubility of S703 at different pH values
based on protein method Solubility (protein method) (%) Batch
Product pH 2 pH 3 pH 4 pH 5 pH 6 pH 7 Nat. pH S005- S703 95.8 100
81.7 0.0 71.7 100 100 A13-09A
TABLE-US-00008 TABLE 8 Solubility of S703 at different pH values
based on pellet method Solubility (pellet method) (%) Batch Product
pH 2 pH 3 pH 4 pH 5 pH 6 pH 7 Nat. pH S005- S703 95.9 95.9 83.8
11.9 69.2 96.0 95.2 A13-09A
[0083] As can be seen from the results of Tables 7 and 8, the 5703
product was highly soluble at pH values 2, 3 and 7 as well as at
the natural pH. The solubility was slightly lower at pH 4.
Example 5
[0084] This Example contains an evaluation of the clarity in water
of the soy protein isolate produced by the method of Example 2
(S703).
[0085] The clarity of the 1% w/v protein solution prepared as
described in Example 3 was assessed by measuring the absorbance at
600 nm, with a lower absorbance score indicating greater clarity.
Analysis of the samples on a HunterLab ColorQuest XE instrument in
transmission mode also provided a percentage haze reading, another
measure of clarity.
[0086] The clarity results are set forth in the following Tables 9
and 10:
TABLE-US-00009 TABLE 9 Clarity of S703 solution at different pH
values as assessed by A600 A600 Batch Product pH 2 pH 3 pH 4 pH 5
pH 6 pH 7 Nat. pH S005- S703 0.098 0.152 1.381 >3.0 1.876 0.155
0.192 A13-09A
TABLE-US-00010 TABLE 10 Clarity of S703 solution at different pH
values as assessed by HunterLab analysis HunterLab haze reading (%)
Batch Product pH 2 pH 3 pH 4 pH 5 pH 6 pH 7 Nat. pH S005- S703 12.0
20.8 86.3 91.6 90.0 19.7 29.8 A13-09A
[0087] As can be seen from the results of Tables 9 and 10, the
solutions of 5703 were clear to slightly hazy at pH 2-3. A slightly
hazy solution was also obtained at pH 7.
Example 6
[0088] This Example contains an evaluation of the solubility in a
soft drink (Sprite) and sports drink (Orange Gatorade) of the soy
protein isolate produced by the method of Example 2 (S703). The
solubility was determined with the protein added to the beverages
with no pH correction and again with the pH of the protein
fortified beverages adjusted to the level of the original
beverages.
[0089] When the solubility was assessed with no pH correction, a
sufficient amount of protein powder to supply 1 g of protein was
weighed into a beaker and a small amount of beverage was added and
stirred until a smooth paste formed. Additional beverage was added
to bring the volume to 50 ml, and then the solutions were stirred
slowly on a magnetic stirrer for 60 minutes to yield a 2% protein
w/v dispersion. The protein content of the samples was analyzed
using a LECO FP528 Nitrogen Determinator then an aliquot of the
protein containing beverages was centrifuged at 7800 g for 10
minutes and the protein content of the supernatant measured.
[0090] Solubility (%)=(% protein in supernatant/% protein in
initial dispersion).times.100
[0091] When the solubility was assessed with pH correction, the pH
of the soft drink (Sprite) (3.39) and sports drink (Orange
Gatorade) (3.19) without protein was measured. A sufficient amount
of protein powder to supply 1 g of protein was weighed into a
beaker and a small amount of beverage was added and stirred until a
smooth paste formed. Additional beverage was added to bring the
volume to approximately 45 ml, and then the solutions were stirred
slowly on a magnetic stirrer for 60 minutes. The pH of the protein
containing beverages was measured and then adjusted to the original
no-protein pH with HCl or NaOH as necessary. The total volume of
each solution was then brought to 50 ml with additional beverage,
yielding a 2% protein w/v dispersion. The protein content of the
samples was analyzed using a LECO FP528 Nitrogen Determinator then
an aliquot of the protein containing beverages was centrifuged at
7800 g for 10 minutes and the protein content of the supernatant
measured.
[0092] Solubility (%)=(% protein in supernatant/% protein in
initial dispersion).times.100
[0093] The results obtained are set forth in the following Table
11:
TABLE-US-00011 TABLE 11 Solubility of S703 in Sprite and Orange
Gatorade no pH correction pH correction Solubility Solubility
Solubility (%) in Solubility (%) in (%) in Orange (%) Orange Batch
Product Sprite Gatorade in Sprite Gatorade S005-A13-09A S703 94.8
100 99.0 93.6
[0094] As can be seen from the results of Table 11, the 5703 was
highly soluble in the Sprite and the Orange Gatorade. As 5703 is an
acidified product, protein addition had little effect on beverage
pH.
Example 7
[0095] This Example contains an evaluation of the clarity in a soft
drink and sports drink of the soy protein isolate produced by the
method of Example 2 (S703).
[0096] The clarity of the 2% w/v protein dispersions prepared in
soft drink (Sprite) and sports drink (Orange Gatorade) in Example 6
were assessed using the methods described in Example 5. For the
absorbance measurements at 600 nm, the spectrophotometer was
blanked with the appropriate beverage before the measurement was
performed.
[0097] The results obtained are set forth in the following Tables
12 and 13:
TABLE-US-00012 TABLE 12 Clarity (A600) of S703 in Sprite and Orange
Gatorade no pH correction pH correction A600 in A600 in A600 Orange
A600 Orange Batch Product in Sprite Gatorade in Sprite Gatorade
S005-A13-09A S703 0.460 0.404 0.471 0.539
TABLE-US-00013 TABLE 13 HunterLab haze readings for S703 in Sprite
and Orange Gatorade no pH correction pH correction haze (%) haze
(%) haze (%) in Orange haze (%) in Orange Batch Product in Sprite
Gatorade in Sprite Gatorade no protein 0.0 44.0 0.0 44.0
S005-A13-09A S703 58.5 74.3 55.6 79.5
[0098] As can be seen from the results of Tables 12 and 13, the
good solubility results obtained for the 5703 in the Sprite and the
Orange Gatorade did not translate to clarity in these beverages. In
fact, the resulting solutions were quite hazy.
SUMMARY OF THE DISCLOSURE
[0099] In summary of this disclosure, the present invention
provides a method of producing a soy protein isolate which is
soluble in acid media, based on extraction of a soy protein source
material using aqueous calcium chloride solution at low pH.
Modifications are possible within the scope of this invention.
* * * * *