CA2115142A1 - Fragmented, alpha amylase hydrolyzed amylose precipitate as fat replacer - Google Patents

Abstract

A method of preparing reduced fat foods is provided which employs a fragmented, .alpha.-amylase hydrolyzed amylose precipitate.
Amylose is precipitated and hydrolyzed with .alpha.-amylase and then fragmented to form an aqueous dispersion that is useful in replacing fat in a variety of food formulations. The amylose can be derived from a native starch which contains amylose, e.g. common corn starch and high amylose corn starch, by gelatinizing the starch followed by precipitation of the amylose.

Description

WO 93/1067:.PCr/US92/065X9 2 ~

CROSS-REFERENCE TO RELATED FAT FOODS
SThis application is a continuation-in-part ofU.S. application Serial No. 07/79&,292, filed November 26, 1991, the disclosure of which is ~; ~ incorporated herein by reference.

This invention relates to food for mulations in which at least a portion of the fat and/or oil is replaced by a ca rbohyd rate .

U . S . ~ Patent No . 4, 510,166 ~ Lench ;n et al . ) d;scloses converted- starches having a DE less than 5 and certain paste and ;gel characteristics whicl- are used as a fat~ and/or oi5~ replacement in various foods, including ice 20~ cream and ~mayonnaise. Th? ~ converted starches are descr~bed~ as~ dextrins,~ acid-converted starches (fluidity ; ;starches) ,~ enzyme-converted starches and ox;dized starches. ~lt ~is ~also disclosed that ;f the converted starches~ ~are ~not~ ~ rendered ~ cold-water soluble by the 25 conversion,-~ ~they~ are pregelatinized~ pr;or to use or cooked during use.
A ~product bullet;n ent;tled "Paselli SA2; The ~, ~Natural Alternative to Fats and Oils" (AVEBE b . a ., Foxhol, Holland,~ ~Ref. No. 05.12.31.167 EF) d;scloses the 30 use of a low-DE-hydrolysate (DE ~less than 3) made from potato starch~ as ~a replacement for fifty percent of the fat with an amount o f the low-DE-potato starch hydrolysate plus water ~starch hydrolysate at 28% dry solids) equal to t~e amount of fat replaced.

WO 93/1067~ PCI/US92/06589 _3 ~ d~ ~, U.S. Patent Nos. 3,962,465 (Richter et al. ) and 3,986,890 ~Richter et al. ) disclose the use of thermoreversible gels of a starch hydrolysate (formed by enzymatic hydrolysis) as a substitute for fat in a variety of foods, including cake creams and fillings, mayonnaise and remoulades, cream cheeses and other cheese preparations, bread~ spreads, pastes, meat and sausage products, and whipped cream.
U.S. Patent No. 4,971,723 (Chiu) discloses 10 ~ partially debranched starch prepared by enzymatic hydro!ysis of~ the ~a-1,6-D-glucosidic bonds of the starch, compris;ng amylopectin, partially debranched amylopectin and up to; 80-O ~ by ~ weight, short chain amylose and that the; partially~ debra~nched starch is useful in a variety of 5~ ways~ depending upon the degree of debranching. It ;s disclosed ~ that~ a ~ waxy maize starch ~or other waxy starch)~ can ~be~ partially debranched (~.e. to~ 25go to 70gO
short chaini ~amylose) to y;eld suff;c;ent short cha;n amyiose~to form~a thermaily~reversible gèl in an aqueous 20~ starch~ suspension.~ ~ 1t is further ~disclosed that the same degree~ of~ deb;~ranching of waxy starches is preferred for bnd~ng ~ a ~ fat-like, ~ lubricat~ng ~texture to an aqueous starch ~dispersion . ~
PGT~Pub~!ication ~ No.~ ~ WO 91/07106, published 25~ May~ 30,~: 1991, ~ d~scloses a ;~method of ~ ~preparing a food grade, ~ nsol~uble ~bulking ~agent ~ from~ ~starch that is also disclosed to~ be~us;eful as a bulking or texturizing agent in low-fat food~ formulations. The method of preparing ; the starch~ compri~ses a retrogradation process followed 30~ by enzymatic ~(e.~g.,~ a-amylasel ~or chemical (e.g., acid) hydrolysis of ~ amorphous ~ reglons in the retrograded product. ~ l~n ~this~ process, amylose is allowed to . .... . ... . .......... . ..

WO g3/1067~ PCl /US92/Q658~
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retrograde from a solution of gelatinized starch. The hydrolysis is t~en undertakèn to reduce or eliminate amorphous regions in the retrograded product.

ii ~ ::: :: : ~ -WO 93/1067~ PCI /US~2/~6589 21L~ ''i.f'.i SUMMARY OF THE INVENTION
I n one aspect, this invention relates to a food form'ulation having a reduced level of fat and/or oil comprising a mixture of a foodstuff and a particle gel as 5 a replacement for at least a substantial portion of the fat ' and/or oil of said foodstuff, said particle gel comprising a minor amount of a f ragmented, a-amylase hydrolyzed `~ ~ amylose precipitate and a major amount of an aqueous ' ~ liquid.
~; 10 In another aspect, this invention relates to a method of formulating a food containing a fat and/or oil ingredient comprising replacing at léast a substantial portion of said fat and/or oil ingredient Witt1 a particle gel as a replacement for at least a substantial portion of 15 the fat and/or oil of said foodstuff, said particle gel comprising a minor; amount of a fragmented, a-amylase hydrolyzed amyiose precipitate and a major amount of an aqueous liquid. ~
By "fragmented, a-amylase hydrolyzed amylose 20~ precipitate" Is ~ meant a starch material comprised of amylose which has~ ~been subjected to precipitation of the amylose followed ~by hydrolysis by a-amylase enzyme and then mechan;cal ~ disintegration of the hydrolyzed precipitate~ ~ into fragments.~ The ~ hydrolysis and 25 ~ disintegration will ibe sufficient to p~roduce a precipitate which will form~ an aqueous - dispersion having the characteristics of a particle gel.
I n ,another aspect, this invention r elates to a method of making` ~a composition of matter useful in 30 replacing fat and/or' oil ;n a food formulation comprising physically fragmenting; a minor~ amount of an a-amylase hydrolyzed amylose~ ~precipitate in a major amount of an aqueous liquid, the degree o~ said physically fragmenting ~::

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WO 93/1067:` PCl`~US92/06589 being sufficient to form a particle gel of said compos ition .
In another aspect, this invention relates to an aqueous dispersion useful as a replacement for fats 5 and/or oils comprising a major amount by weight of water ; ~ and a minor amount by weight of a fragmented, a-amylase hydrolyzed amylose precipitate, the degree of ~ ~ hydrolysis and fragmentation of said precipitate being ;~ ; sufficient to form a particle gel of said dispersion.
The terms ~ "foodstuff" and "food", as used herein, are intended to broadly cover nutritional and/or functional materials that are ingested by humans in the course of consuming edible fare. The term "fats and/or ~, .
oils" is intendedl ~to broadly cover edible lip;ds in 15 general, specifically the fatty acid triglycerides commonly found in foods~. The terms thus include solid fats~
plastic~ shortenings, fluid oils (and fully or partially hydrogenated ~oils)~, and the like. Common fatty acid triglycerides i nclude cottonseed oil, soybean oil, corn `20 ~ oil, peanut ~o~ canola oil, sesame oil, palm oil, palm kernel~ oil, menhaden oil, whale oil, lard, and tallow.
The~ technology ~of ~fats and/or ~o;ls ;s described generally by T ~ H~ Applewhite, "Fats and Fatty Oils", EncYclopedia of Chemical Technolo~Y, Vol. 9, pp.
25 ~ 795~ (Klrk-Othmer, eds.,~John Wiley ~ Sons, Inc., N ew York, ~ New~ York~, 3d ed.~,- 1980)~, the disclosure of which is incorporated by reference.
The use ~ of the terms "major" and "minor" in context togethe;r; ;ln this specification is meant to imply 30 that the major ~cam~ponent is p~esent in a greater amount by weight than~ the~ minor component, and no more nor less should be ~inferred therefrom unless expressly noted otherwise in; context.

WO 93/1067:~ PC~/US92/06589 211~J1f1 BRIEF DESCRIPTION OF T~IE DRAWINGS

Figure 1 shows the results of duplicate analyses of the dynamic elastic modulus (G') in kilo 5 pascals as a function of strain (m) for a particle gel of fragmented a-amylase hydrolyzed amylose precipitate at 10% precipitate solids.

The fragmented, a-amylase hydrolyzed amylose precipitate is made by the sequential steps of precipitation, enzymatic hydrolysis, and f ragmentation of a stàrch material containing amylose. ~tarcl1 is ~eneraliy comprised of a highly-branched glucan having a-1,4 and -1,6 linkages, denominated amylopectin, and a substantially linear glucan, having almost exclusively a-1,4 linkages, ~ denominated amylose. Methods of determining~ the amounts of each are referenced in R.L.
Whistler et al.~, Starch: Chemistry and Technology, pp.
25-35 (Academic Press, Inc., New York, New York, 1984), the disclosure of which is incorporated by reference. ~ As used herein, the term "amylose" includes native amylose ~and~, unless otherwise expressly noted in context, modified ~amylose. Examples of modified amylose include acid-modif~ied amylose,~ enzyme-modified amylose (e.g. a-amyiase, ~ amylase, isoamyiase, or pullulanase) and chemically~ substituted amylose, provided the levels of chemical substitution le. g . hydroxypropylation, ` 30 crosslinking, etc. ) are insufficient to prevent precipitation and enzymatic hydrolysis of the amylose to the desired degree. Starches having a substantial proportion (i.e. at least 15% by weight) of amylose are ' ~

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WO 93/1067~ PCr/US92/065X9 2 1 t ,~ 8-preferred and examples of these include the common non-mutant starches of cereals, tubers and legumes, e . g . corn, wheat, rice, potato, tapioca, and pea .
Preferred for use herein are starches derived from corn 5 (Zea mays) such as common corn starch - and high amylose corn starch, each of which are examples of starches containing greater than 15% amylose. Examples ; of such starches from high amylose corn include ~ Hl-SET ~) C and HYLONTM (each about 5~O amylose by ; ~ 10 weight) and HY~ONTM Vll (about 70O amylose by weight), all ava;lable from National Starch and Chemical Corporation, Bridgewater, New Jersey.
In certain embodiments, the starch is comprised of a ~ major amount of amylose. In such 15 ~ embodiments~, the~ ~starch employed is from a mutant variety of native ~starch which contains a major amount of amylose or is obtained by ~ractionation of amylose from a starch variety containlng both~ amylose and amylopectin.
Methods for the fractionation ~of amylose~ and amylopectin 20 ~ from native~ starch ~are disclosed in~, for example, U.S.
Patent No. 3,067,067 (Etheridge).
If the~ starch chosen as a starting material is not in~ pre-gelatinized ~or instant form, the starch must be gelatinized ~or~pasted prlor to~ precipitation of the 25~ ~amylose. The~gelatinization or pasting;process disrupts, at~ least in ~s~ubstantial part, the assoc~ative bonding of the starch ~molecules~ in the starch granule. This permits the amylose to associate ancl precipitate. This disruption is accomplished~ by ~heating ~a slurry~ of the starch to a 30 sufficient temperature for a~ sufficient length of .time depending upon the inherent resistance of the particular starch to gelatinization and the amount of moisture i ~ present in the~ slurry. The slurry will typically be , - :
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g comprised of a major amount of water (i.e. at least 50~
by weight) and a minor amount of the starch starting ; material (i.e. Iess than about 50~ by weight).
Preferably, the starch slurry will contain at least a~out 5 5% starch, typically between about 10% to about 25%
starch. The pH of the slurry will generally be substantially neutral, i.e. from about 3.5 to about 9 and more preferably from about 6 to 8, to minimize hydrolysis of the; starch molecules. The time, 10 temperature, slurry solids and pH should be optimized to gelatinize the starch, yet minimize hydrolysis of the starch.
The appropriate temperature, pressure and period of treatment needed to provide a starch paste is preferably obtained by processing aqueous starch slurries in equipment commonly known in the art as steam injection~ ~heaters or jet cookers . I n such equipment, ~superatmospheric steam is injected and mixed with a wateri slurry~ of starch in a throat section of a 20~ ~ jet~. ~ Upon ~ contact;~ with the ~ injected steam, the starch granules are~ ;~uniformly and thermally treated under turbu!ent condition~s whereupon ~ ~the starch ~ranules are gel~atinized and~ ` solubilized . ~Examples of steam injection heaters wherein ~thé~ temperature, pressure and feed rate 25 ~ c an ~be regulated t o provide th~e desired starch pastes are disclosed ~in~U.~S. Patent Nos. 3,197,337; 3,219,4B3;
~; ànd 3,133,B36. ~ More uniformly solubilized starch pastes are obtained by use of the steam injection heater in combination~with: a~ holding zone such as coiled tubing or 30 a pressurized~ ~ tank constructed to minimize liquid channe~ing. Other pasting equipment, e . g . heat exchangers,~ homogenizers, cookers, votators, sizeometer cookers, kettle cookers, etc., may be employed provided .

WO 93/1067~ PCI/US92/06589 ~ 1 4 ~ - l o-the pasting conditions can be adequately maintained.
The starch solution may also be treated to remove impurities therefrom. Treatment with, for example, activated carbon will remove residual proteins 5 and lipids that may contribute to off-flavors and/or colors.
The gelatinized starch is then optionally treated with a ~debranching enzyme, i . e . an enzyme capable of hydrolyz;ng the 1, 6-glucosidic bond of 10 amylopectin without~ significant capability of hydrolyzing the~ 1,4-glucosldic~ bond. Enzymes from a variety of I ~ sources are capabl'e o f debranching amylopectin. U.S.
Patent No. 3~,370,840 (Sugimoto et al.) describes sources of ~ debranching`~ enzymes, the disclosure of which is 15 incorporated ~ herein~ by reference. Examples of useful enzymes include~puliulanases derived from bacteria of the genus ~ Aerobacter~ ~ (e.g. E.C. 3.2.1.41 pullulan 6-glucanohydrolase)~ ~ ~ and ~ isoamylases derived f rom bacteria ~of~t~he~ genus Pseudo~monas (e.g. E.C. 3.2.1,68 20~ ~ glycogen~ ~ ~ 6-glucanohydrolase). Particularly useful enzymes include ~thermostable ~ enzymes, e . g . thermostable pullulanases~ às~ disclosed in PCT Publ. ~No. WO 92/026~4, published~February~20, 1992, the disclosure of which is incorporated~ by~ re~ference, and which are obtained from 25 ;~; members of the~ ~'genus Pyrococcus. The debranching enzyme may~'be~in~ solution~ during debranching or it may be immobilized~ on~ a so!id support.
The~ debranching enzyme preparation should be as specific~ as~ ~; possible for the hydrolysis of the 30 1 ,6-glucosidic ~ bond of amylopectin and amylose. Thus, the enzyme~- preparation, if it contains a mixture of enzymes, is ~ preferably essentially free of enzymes capable of ~ ~hydrolyzing a-l ,4-glucosidic bonds.

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WO 93/1067~ PCl/US92/06589 2 ~

Minimizing hydrolysis of a-1,4-glucosidic bonds will help to minimize the amounts of dextrose and soluble oligomers produced during debranching. Because these soluble saccharides are not believed to contribute to the 5 functionality of the debranched material, minimizing their production will enhance the yield of functional material.
The debranching enzyme is allowed to act upon the solubilized starch con~tain;ng amylopectin. The ` optimum concentration of enzyme and substrate in the 10 debranching medium~ will, in general, depend upon the level of activity of the enzyme which, in turn, will vary depending upon the ~ enzyme source, enzyme supplier and :~ the concentration~ of the enzyme in commercial batches.
When the isoamylase E . C . 3 . 2 .1 . 68, derived f rom 15 Pseudomonas ~ ~amyloderamosa, ~ available from Sigma 'C~hemical Co., St. :Louis, Missouri, is employed, typical condltions ~include~-the~ treatment~ of a starch solution at 5% ~to 30% ~by~ weight ;starch solids with about 50 units of enzyme,~ per ~gram ~of ~starch, for a period of about 48 20~ hours to obtai;n substantially. compiete debranching.
The ~ optimum pH and temperature of the ; debranching~ mediu'm `will also depend upon the choice of i enzyme~. The ~ debranching medium may, in~ addit;on to the water ~ used ~ to~'~ soiubilize the starch;, contain buffers 25 t o ensure thàt~the~ pH~ will be maintàined at an optimum level throughout~ the~ ~debranching. Examples of useful bufters~ incl~ude~ acetates, citrates, ~and;'the salts of other weak acids. With ~ the iisoamylase described above, the pH is preferably maintained at; about 4.C) to 5.0 and the :30~ temperature~ from: ~about 40C to about S()C. With the thermostable ~ pu~llulanase described above, the pH is preferabiy maintained ~between 5 and 7 and the optimum temperature should be ~between 85C and 115C.
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, WO g3/1067; PCl/US92~065~') 211 J~'~2 The debranching is allowed to proceed until the desired degree of debranching has been obtained.
The precise degree of debranching needed to obtain the desired particle gel of the debranched amylopectin starch 5 may vary dependlng upon the source of the starch and the precise properties desired in the resulting gel.
Preferably, the degree of debranching is sufficient to .
convert more than about 80% of the amyl~pectin in the starch to short chain amylose and, more preferably, at lO least about 90~Q of the amylopectin.
In preferred embodiments, essentially all of the amylopectin is converted to short chain amylose. The amount of short chain amylose can be measured by gel permeation ch~romatography as set forth in U. S. Patent No. 4,971,723, ~wherein short chain amylose is calculated from the relative area of the peak obtained within the molecular weight ~ ~ range of 500 to 20,000. Thus, preferably less~ ;than 20% of the amylopectin that was originally~ ~ present w;ll be pres~ent as molecular species 20~ havlng a molecular weight in excess of;20,000 gimol, and most preferably,~ essentially no amylopectin having a molecula~r we~ght~ n~ excess of ~20,000 g/mol will remain.
lt~s;hould be~;noted;~that if amylose~ Is ~present, at least a portion; thereof~may be debranched ;to produce molecules 25 ~ ~ above the 20,000 g/mol cut-off and molecules below the 20,000 g/mol ~cut-off. To~ measure how much of the material elutl~ng~ between 500 g/moi and 20,000 g/mol is debranched amylopectin and how mucll is debranched amylose, it may be necessary to fractionate the starting 30 ~ starch into ~its~amylose and amylopectin fractions and ;` then debranch and elute each f~action separately.) ~ , ::~ :

WO 93/10675 PCr/US92/065X9 2~S 5 ~ f'J

The solution of gelatinized st~rch is then allowed to form a precipitate. Generally, the solution will be cooled from the temperature at which the starch is pasted to reduce the solubility of the gelatinized 5 starch therein. The solution will typically be held at elevated temperature (e.g. 65C to 90C) until substantial equilibrium is achieved between the ~ , supernatant and the precipitate. The precipitate can be isolated from the supernatant, e. g . by centrifugation, 10 prior to fragmentation, but isolation from the supernatant is~not~necessary to form a useful product.
Heating (e~. g . to about 70C) of the particles while in contact with the aqueous medium to dissolve at least a portion of ;the mass of the particles and then 15 cooling of the suspension/solution can also be employed in forming the ~ particle gel of this invention. This heatlng to ~an elevated temperature and then reformation ;of the~ particles ~tends to make the particles resistant to melting or'dissolving when an aqueous dispersion of the 20~ partlclés is~ exposed to heat~ in processing, ë. g . in a pasteurization~ ~ ~ stép. In general, the higher the tèmperature'~to ~whlch the particles in ~the liquid medium are~ ~ heated ~ ~ta~nd~ thus the~ greater~;~ the amount of p~recipitate~ that ~ is redissolved),~ ~the higher the 25 ~ ~ temperature~ ~at which the resulting aqueous dispersion of the particles~will~be~stable. ~Repetition of the dissolving and reformation~ may improve the temperatu~re stability of the resulting a~queous dispersion.
It is~ ~also~ ~advantageous to heat the precipitate 30 to redissolve~ a su~bstantial portion of the low melting polysaccharides~ ~and then treat the heated~ suspension of precipitate ;~wit~ acid or enzyme to hydrolyze soluble ~ : ~
polysaccharides~ in~ the solution . ( It may also be : ' WO 93/1067~ 2 1 ~ ~ t ~ ~ PCr/US92/06589 advantageous to filter the slurry while hot to remove soluble polysaccharides or their hydrolysates. ) The dissolving and reprecipitation steps alone improve the stability of the aqueous dispersion by increasing the S amount of the fragmented precipitate which remains as insoluble fragments in an aqueous dispersion that is exposed to heat. Further, a slow rate of heating and/or cooling (e.g. from about 0.005C/min. to about 0~5C/min. ~for each) may be advantageous. However, 10 the remaining soluble fraction of the precipitate can associate to form relatively large particles that are present in ~the ~precipitate after fragmentation and that can contribute a~ "chalky" or "gritty" texture to the dispersion. T;reatment of the heated suspension/solution 15 of the precipitate w~th acid or enzyme to hydrolyze a substantial portion~ of ;the soluble fraction can reduce or eliminate such~ ~;large partic~les. Typical treatment conditions will invol~ve mild hydrolysis; catalyzed by acid, e.g. i n a ~ 'solutlon~ of ~ 0 1 N ~ HCI for one hour, or, 20~ preferably,~by~ enzyme, e.g. a-amylas~e~
; The~ prec~p~tated~ omylose~ s 'then treated with :an ~a-amylase~enzyme,~ ~I.e. an'~endo-enzyme capable of hydrolyzing ~the~ 4-glucosidic bond ~ of amylose and amylopectin to: yield~products having- an a configuration.
25 ~ ~ The ~enzyme~ ~ ~is~ ~allowed ~ to act~; upon the precipitated amylose and~ ~théreby hydrolyze ~ those regions in the precipitate that~ are ~susceptible ~ to hydrolysis ~ The optimum concentratlon of enzyme and substrate in the hydrolysis medium~ will, in general, depend upon the 3 0 level of acti~vity~of~the enzyme~which, in turn, will vary depending upon ~:the enzyme source, enzyme supplier and the concentration~of the~ enzyme in commercial batches.

.. , .. . ... ... ... .... . . . ... , .. , ..... .. , , ~ , W 0 93tlO67~ P ~ /US92/06589 2~! ~5 ~ `~1 2 The a-amylase can be from a variety of sources. Common sources of a-amylase are bacterial, e.g. Bacillus subtilis, or fungal, e.g. Asper~illus oryzae, or mammalian, e. 9 . human salivary, porcine 5 pancreatic, etc. The optimum pH and temperature of the hydrolysis medium will also depend upon the choice of enzyme. The hydrolysis medium may, in addition to the water used i n the hydrolysis of the starch, contain buffers to cnsure ~that the pH will be maintained at an 10 optimum level'- throughout the hydrolysis. Examples of useful buffers i nclude acetates, citrates, phosphates, and'~ the salts ~of~ other weak acids. With porcine ' pancreatic a-amylase, the pH is preferably maintained at a'bout 6.0 to 8.0~:an~d~the temperature from about 20C to 15 about 30C.~
The hyd~rolysis is allowed to proceed until the desired~ degree ~ of~-~hydroiysi 5~: has been obtained. The u ~ precise~ degree~ of~ hydrolysis ~; needed to obtain the desired; particle ~ gel o f the fragmented, a-amylase 1 20 ~ hydrolyzed amylose~ precipitate may vary depending upon the sourcei ~of the starch and the precise properties ' desired~in ~'he~resujlti~ng gel. Typically, the degree of hydroiysis~will be~sùch that fragmentation of the product will ~yield~a~ gel~ that exhibits ~a trans;tion from a region 25~ ~ of ~ ~substantially~ ~constant dynamlc elastic modulus (G') 'versus ~ shear ~strain~ to a region of decreasing G' versus shea~r strain~ said~' transit;on be;ng at a shear strain of less than about~ 5Q m;llistrain, and preferably less than about 10 mill;strain.~ The transition indicates fracture of 30 the part;cle~network~ with the~ particle gel and is typically a sharp transit;on.~ The dynamic elast~ic modulus can be measured with ~ a ~Bohlin model VOR Rheometer, from Bohlin Rheolog;, l~nc., East Brunswick~, New Jersey.

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WO 93/1067~ PCI /US92/06589 S.~

After the desired degree of hydrolysis is obtained, the a-amylase enzyme in solution is deactivated, e. g . by heating to denature the enzyme.
The hydrolysis medium may be concentrated by removal 5 of water therefrom, e.g. by evaporation, to facilitate precip;tat;on .
The isolated a-amylase hydrolyzed amylose precipitate is typically washed and then dried ~e. g . to a , ~
low moisture ~content, typically 3-12%) after isolation to 10 allow for handling~ and storage prior to further ; processing. Examples of drying techniques include spray drying, flash ~drying, tray drying, belt dry;ng, and sonic drying. The dried precipitate may be hygroscopic. ~ Thu~s, some rehydration during handling ~and storage may occur. Depending upon the precise composition ~ ~of ~ the precipitote and the conditions (including length~-of~time) of~ storage, steps to maintain the moisture at~a ~ low content may be necessary (e.g.
moi~sture barrier packaging and~or control of humidity in 20~ the storage ~environment) . ~ if the moisture content is allowed to~rise~too~ far~ ~e.g. greater than about 20%, or possibly ~greater ~ than 15%), bulk handling problems and/or~microbiological stability~problems~might arise.
The~;a-amylase ~ hydrolyzed amylose precipitate ;~ 25~ is ~subjected ~to~a~ physical fragmentation as by mechanical disintegration,~ .e~ fragmented. The degree of fragmentation ~ will be ~ sufficient to cause` the precipitate to form a ~ particle ~ gel in an aqueous medium. The mechanical; ~disintegration of the precipitate may be carried out ~ n~ ~several ways, as by subjecting it to attrition in~ a mill,~ or~ to a high speed shearing action, or to the ac~tion ~of high pressures. Disintegration is generally ca~rrled~out in~ ;the~ presence of a major amount .

WO 93/1067~i PCI/US92/065X9 2 1 1 ~ r ~

by weight of a liquid medium, preferably water.
Although tap water is the preferred liquid medium for the dispersion of fragmented starch precipitate, other liquids are suitabie provided sufficient water is present 5 to hydrate the fragmented starch precipitate and, thus, result in a dispersion having the characteristics of a particle gel. Sugar solutions, polyols, of which ~Iycerol is an examp:le, alcohols, particularly ethanol, isopropanol, and the like, are good examples of suitable 10 liquids that can be in admixture with water in the liquid medium. Typically, however, the starch precipitate will be physically fragmented in potable water.
The mechanical disintegration is preferably accomplished by subjecting an aqueous dispersion of the 15 precipitate to high shear, e.g. in a Waring blender or a homc?genizer such as that disclosed in U . S . Patent No.

4,533,254 (Cook et al. ) and commercially available as a MICROFLUIDIZERTM from Microfluidics Corporation, Newton, Massachusetts, or a homogenizer such as the 20 RANNIETM high~ pressure laboratory homogenizer, Model Mini-lab, type~ 8~.30 H, APV Rannie, Minneapolis, Minnesota. Homogenizers useful in forming suspensions or emulsions~ are~ described generally by H. Reuter, "Homogenizatlon", ~EncYclopedia of Food Science, pp.
25 ~ 374-376, (M. S. Peterson and A. H. Johnson, eds., AVI
I ~ Publ. Co., Westport, Connecticut, 1978), L. H. Rees and W. D. Pandoife, "Hcmogenizers", EncYclopedia of Food ~Enqineering, pp. 467-472 (C. W. Hall et al. eds., AVI Publ. Co., Westport, Connecticut, 1986), and W. C.
30 Griffin, "Emulsions", EncYclopedia of Chemical Technoloqy, Vol. 8, pp. 900-930 (Kirk-Othmer, eds., John Wiley ~ Sons, Inc., New York, New York, 3d ed., 1979), the disclosures of which ~re incorporated herein by reference.

WO 93/1067~ PCr/US92/065X9 2 ~ 2 The temperature of the starch precipitate during the fragmentation step should be maintained below the temperature at wh;ch a major portion of the precipitate will dissolve in the aqueous medium. Thus, it may be desirable to cool the precipitate during disintegration. Alternatively, heat produced during fragmentation may cause the precipitate to dissolve, but cooling may cause the dissolved precipitate to `reprecipitate and; form a useful product. Whatever 10 method is used, ~the ~disintegrat~on is carried out to such an extent ;that~ the ~ resulting finely-divided product is characterized by its`ability to form a particle gel in the liquid medium in which it is attrited or in which it is subsequent\y ~ dispersed .
15 ~ ; ~ The ~ ;~starch particles which make up the particle gel can~ ~be analyzed in a variety of ways.
Rheologic-l ~ measurements can ~ be made to determine the rheological~;characteristics of;~ the resulting dispersion.
Typlcaliy~,~ the~àqueo~us dis~persion of starch particles will 1 20~ ` ex~hibit a~ ~ transition in dynamic elastic modùlus ~G') versus~shear~ st~rain~at less than about 50 millistrain, and preferabiy ~ ies-s than ~ about lO milllstrain, said transition beingi~from~ a~substantiaily~ constant G' versus shear strain ~to ~a~ ~decreasing G' ~ ~versus~ shear stra;n. The 25~ transition~ indicates ~ fractu~re ~ of ~ ;the particle network withi~n the~ particle~ gel and ~is typically a sharp transition . ~
Ana!ysis of the starch particles formed after dissolution~ shows~ that the starch has a measurable 3 0 crystall jnity. ~ The ~ crystalline ~ regions of particles derived from fully debranched waxy ;maize starch (essentially no ~amylose compone~nt) exhibit a diffraction pattern characteristic of a starch material consisting ,~

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W O 93/1067~ PC~r/US92/065X~
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_19_ essentially of A-type starch crystals. The crystalline regions of particles derived from substantially fully debranched common corn starch (about 28% amylose) exhibit a diffraction pattern characteristic of a starch 5 material consisting essentially of B-type starch crystals.
It should also be noted that mechanical disintegration may be sufficient to produce an aqueous dispersion having the desired particle gel characteristics, but still leave a sufficient number of 10 particles of suffîcient size to exhibit a "particulate" or "chalky" mouthfeel when ingested. Such chalkiness can be reduced by the mild hydrolysis discussed above or by reducing the particle size of the starch precipitate before, during or after mechanical disintegration so that 15 substantially all (typically at least about 95~, preferably at least 99%) of the precipitate will pass a U . S . #325 mesh sieve (i.~e. ;~substantially all particles are less than 45 microns). An example of a milling device suitable for such size ~reduction is a TROSTTM Air Impact Mill from 20 Garlock, Inc., Newton, Pennsylvania.
The~ use of the fragmented, a-amylase hydrolyzed amylose precipitate allows for the replacement of a substantial portion (e.g. from 10% to 100~ by weight) of t~he~ fat and/or oil in a food formulation. The 25 precise level ~ of replacement that is possible without significantly ~ decreasing the organoleptic quality of the food will generally vary with the type of food. For ~' example, in a French-style salad dressing, it is generally possible to completely replace the oil component 30 that is normally ~ present. I n other types of foods, e . g .
frostings, icings, cream fillings, ice cream, margarine, etc., a maJor amount of the fat and/or oil (e. 9. about 50% to about 80%) can be replaced with little effect on WO 93/1067;~ PCI /US92/0658~

2 1. ~ 'J5 ~

the organoleptic desirability of the food. Examples of typical foods in which fat and/or oii can be replaced include frostings (e.g. icings, glazes, etc. ), creme fillings, frozen desserts (e.g. ice milk, sherbets, etc. ), 5 dressings (e.g. pourable or spoonable salad and/or sandwich dressings), meat products (e. g. sausages, processed meats, etc. ), cheese products (e.g. cheese sp reads, p roces sed cheese foods ), ma rga ri ne, f ru it butters, other imitation dairy products, puddings (e.g.
10 mousse desserts), candy (e . 9 . chocolates, nougats, etc. ), and sauces, toppings, syrups and so on.
Generally, it will be desirable to remove sufficient fat from a gi~en food formulation to achieve a reduction in calories of at least one-third per customary 15 serving or make a label claim of "cholesterol-free". (In this regard, see, for example, the list of standard serv;ng sizes for various foods published in Food Labelling; Serving Sizes, 55 Fed. Reg. 29517 ~1990) (to be codified at ~1 C. F. R. 101 .12), the disclosure of 20 which is incorporated herein by reference, and the restrictions on labelling "cholesteroi-f ree" at Food Labelling; Definitions of the Terms Cholesterol Free, Low Cholesterol and~ Reduced Cholesterol, 55 Fed. Reg. 29456 (1990)). It shs:~uld also be ~noted that the fat removed 2 5 from a particular~ formulation may be replaced with an equal amoun t by weight o~ an aqueous dispersion of fragrnented starch precipitate, but that such equality ~, may not be necessary or desirable in all instances.
Further, it may~ be desirable to remove fat and add 30 another ingredient (e. g . a gum, polydextrose, a protein, etc. ) along with the aqueous dispersion of starch precipitate.

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WO 93/1067~ PCI/US92/06589 2:1 1 5 1 ,1 ;

While this invention is generally directed to the replacement of fat and/or oil in a food formulation, it is of course within the contemplation of this invention that a fragmented, a-amylase hydrolyzed amylose ; 5 precipitate wiil be used in an entirely new formulation to which it contributes fat-like organoleptic qualities but is ~; ~ not, in the~ strictest sense, replacing a pre-existing fat or oil ingredient. Moreover, it is contemplated that the ` fragmented, a-amylase hydrolyzed amylose precipitate will 10 ~ have utility as a~ thickener, bodying agent, or the like in foods that normally~ do not have a significant fat or oil component. ~ ~
In general, the fragmented, a-amylase hydrolyzed amylose~ precipitate is incorporated into the 15 ~ food ~as an aqueous~dispersion, typically comprised of a major ~amount ~(i.e~ ~greater than 50~ by weight) of water o r other liquid ~medium and ;~a; minor amount (i.e. Iess than 50% by weight, typically 1Q% to 40%) of starch precipitate ~solids. ~ Alternatively, the isolated precipitate 20 ~ ~can be~ ~mlxed~ ~with the ~ood along ~with water and then - subjected~ to~ ;disintegration in those instances when the other ~ogredlents~of~the~food~ are capable of withstanding the condition of disintegratlon, e~.;g. a salad dressing or u i mitation sour cream. ~ ~ ~
; 25; ~ ; It ~is:: contempl~ated that commercial production and ~ ~ ~;use ~ may ~ involve hydrolysis, mechanical disintegration, ~ and ~ dryin g (e.g. spray drying) of the :: : :
, ~ ~ fragmented starch precipitate to produce an item of commerce. ~ This item of commerce; will then be purchased 30 ~ ~by a~; food~ processor for use ~as an ingredient. To incorporate the~ dried, fragmented, a-amylase hydrolyzed amylose precipitate~ into a food p~roduct, it may be useful and/or necessary to further mechanically disintegrate the :

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WO 93/1067~ PCr/US92/065X~
X ~ i 2 starch precipitate while dispersing it into the foodstuff in which it will be employed. However, the techniques employed for such mechanical disintegration should not need to be nearly as vigorous as the ori`ginal mechanical disintegration prior to drying.
As noted above, the terms "food" and foodstuffs" are intended broadly, as relating to both nutritional and/or functional food ingredients. It is contemplated that one or more food ingredients may be mixed with the~ ~ aqueous dispersion of fragmented, a-amylase hydrolyzed amylose precipitate, or even dry mixed with the~ a-amylase hydroiyzed amylose precipitate prior to mechanical dis~ntegration.
Among ~ the food ingredients which may be included in ~ the food formulations of this invention are flavors, thickeners~ (e.g. starches ~ and hydrophilic colloids) ,; nutrierrts ~ (e. g. carbohydrates, proteins, I ipids,~ etc.~ antioxldants, antimicroblal ~agents, non-fat milk so!ids,~egg~:solids,~acidulants, and~so~on.
20~ Hydrophllic ~ colloids can `include natural gum material such~ ~as ~xanthan ~gum, gum tragacanth, locust bean gum,~ ;guar~ ;gum,~ algin, alginates, gelatin, !rish moss,~ pectin,~gum~àrabic, ~gum ghatti, gum karaya and plant~ hemicelluloses, e.g. c orn ~ hull gum. ~ Synthetic 25~ ~ gums~ such ~ ~as ~ ~water-soluble salts of carboxymethyl celluiose~ càn~; also~be used~. ;IStalches can al~o ~ be added to the food.- ~;Examples of suitable~ starches include corn, waxy maize, ~vheat, rice, potato, and tapioca starches.
Non-fat milk solids which can be used in the 30 compositlons of th~ls invention are ~the solids of skim milk and include proteins, mineral matter and milk sugar.
Other proteins such ~as casei~n, sodlurn caseinate, c:alcium ~: : : : : :
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WO 93/tO675 PCr/US92/065X9 " .

~ ~J~

caseinate, modified casein, sweet dairy whey, modified whey, and whey protein concentrate can also be used herein .
For many foods, it is accepted practice for the user to add the required amount of eggs in the course of preparation ~ and this practice may ~e followed just as well herein. If desired, however, the inclusion of egg solids, in particularj ~egg albumen~ and dried yolk, in the food are allowable ~alternatives. Soy isolates may also be used herein in place;of the egg albumen.
Dry; or liquid flavoring agents may be added to the formulation~ These include cocoa, vanilla, chocolate, ~ , ~
coconut, peppermint, pineapple, cherry, nuts, spices, salts, flavor;~enh~ancers, among others.
lS ~ Aci~ulants~ commonly added to foods include lactic acid, citric acid, tartaric acid, malic acid, acetic acid,~ phosphoric~acid, and hydrochloric~acid.
Generally, the other components of the various types of food ~formu~lations wi~il be conventional, although ~precise amou`nts~ of individual components and the presence of ~ some ~ of the conventional components may m ~ well be unconventional in a given formulation. For examp!e, the~ conventional~ other components for foods such as~ frozen~desserts~ and ~dressings, are described in 25~ European Pàtent~ Publication ~ No. 0 340 035, published November 2, ~ 1989~ (the ~ pe~t~nent disclosure of which is incorporated herein b:y reference), and the components and processing~ of table spreads is disclosed in U . S .
Patent No. 4~,869,919 (Lowery), the disclosure of which i5: incorporated:~by~reference.
- ~ ~ A ~ particularly advantageous use of the , ~ ~
fragmented starch ~precipitates described herein may be the use thereof ~ to replace~ a portion of the shortening , . . .

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WO 93/1067~ PCI/US92/06589 2 1 1 ~ ~ 4 ~ -24-used in a layered pastry article . I n layered pastry articles (Danish, croissants, etc. ), layers of a bread dough are assembled with a "roll-in" placed between the layers. The roll-in commonly contains a "shortening"
S (i.e. a fat and/or oil component) from an animal (e.g.
butter~ or vegetable (e. g . partially hydrogenated soybean oil) source. The assembled article, optionally containing a filling ~or topping, is then baked to form a finished pastry. At least a portion of the shortening of 10 an otherwise conventional roll-in can be replaced with an aqueous dispersion of fragmented, a-amylase hydrolyzed amylose precipitate, ~ preferably in admixture with an emulsifier (e.g.~ mono- and/or d;-glycerides), and used t~o make a layered pastry.
5 ~ The following examples will illustrate the invention and variations thereof within the scope and spirit of the invention will be~ apparent therefrom. All parts, percentages,~ ratios and the like are by weight throughout this~ specification and the appended claims, 20 ~ unless ~otherwise noted in context.

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WO 93/10~7~ PCr/VS9~/065X9 EXAMP LES

E~CAMPLE 1 Into a 4-liter beaker was placed 118 grams of potato amylose (A0512 Sigma Chemical Co., St. Louis, Missouri) and 2242 grams deionized water to give a 5%
slurry. The slurry was heated to 98C in ~ water bath to solubilize the amylose and this solution was placed in ; ~ ~ 10 a refrigerator overnight to promote precipitation/
crystaliization . ~ ~ The~ resulting slurry was centrifuged at about 6000 x g using an IEC model 1~-22 centrifu~e and the supernatant was discarded. The wet sediment was , resuspended in~ deionized water to about 2000 ml volume 15 and heated to 98C in a water bath to solubilize that port~on of amylose susceptible to solubilization at that temperature ~ then~ the mlxture was placed in a refrigerator o vernight to promote precipitation/
crystallization. ~ The resulting slurry was again 20~ centrifuged~ as~ ~before ~ and the sup rnatant discarded.
The~ sediment was~resuspended in water, heated to 98C
in; ~a water~ ~bath ~ to solubilize some amylose, cooled to promote precipitatlo`n/crysta~llizatlon and centrifuged one n ~ last time. The~ resulting centrifuged we~ sediment 25 containing 8~.1%~dry substance was placed in a sample jar : and stored in~a refrigerator.~ ~ ~
Int o a;~ 5-liter 3-neck~ round bottom flask equipped witlh a stirrer and temperature controlled water bath was placed 7~2 grams ~ of the wet freshly 30 ~ precipitated potato~ amylose above, ~ 2356 grams deionized water and 64; grams of molar phosphase buffer at pH

6.9. The suspension was stirred and the pH maintained ; ~ at 6.9. To thissuspension was added 320 units ::: : ~ : : :

: :
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7~ PCr/US~2/065X9 2 ~

(5 units/gram amylose) of porcine pancreatic ~-amylase enzyme (A6255, Sigma Chemical Co., St. Louis, Missouri). The mixture was allowed to react with stirring at 25C for 24 hours then one half of the 5 mixture was added to 8: volumes of ethanol (formula 3A) with stirring in a 4-liter beaker. The resulting aqueous alcoholic mixture was centrifuged as above at about 6000 x g and the sedîment dried in a vacuum oven overnight at 50C.
:: 10A 10~6 dry: solids slurry of the dried product above was sheared ~ using a small Waring blender at controlled cond~tions ~(120 volts, 60C, 8 1/2 minutes) and left to stand 3 hours before the sample was sent for analysis for yleld~ stress, 170 NMR wate~r immobilization, 15: molecular weight by: GPC~ and ,old-water solubles. The : :results of these a~nalyses of the resulting creme are as follows. An anal~ysls of ~ the molecular weight (by gel permeation chromatography) showed ~ a weight average (Mw): of ~SS,900, a :number average (Mn): of: 9,100 and a 20 ~ péak molecula r ~:~weight of 3 2,000. Thè water immobîlizatîon ~by~ ~7O: NMR) ~exhibited by the creme was ., ; ~ . .
177~ sec ' and~the~yleld: stress was 531 pascals. The : cold-water solubles: of: the~ powder~ were: 13% by weight.
It~ is~ specu~1ated: that~ further enzyme hydrolysis 25: :to:: give:~ a level-off DP~ of about 65 will result in even greater o17 NMR ~water immobillzation values.

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":' WO 93/1067~ PCI/US92/06~8g 2 ~15 l /æ ~
-27- -:

SPOONABLE SALAD DRESSING :

5 A spoonable dressing can be prepared from the creme of Example 1 as follows.

I n~redients %, wt .

Part A
:
Water ~ 22 . 00 ISOSWEET (~) 100 high fructose corn syrup (Staley) 17.0n Vinegar, white, 100 grain 10.00 SWEETOSE (~) 4300~ corn syrup (Staley) 5.00 $TAR-DRI ~ 35R~ corn syrup solids (Staley) 3.20 DELTA 7393 starch ~Staley) 2.85 ~:
Salt ~: 1.80 Carboxymethyl cèllulose 7MF (Aqualon) ().20 : Mustard ~ powder (McCormick) 0.10 Titanium~ dioxide~ ~(Warner-Jenkinson) 0.0~
Garlic: powder: : - ~ : 0.05 Onion powder ~ 0. 05 Paprika ~ 0.0125 ~ Calclum disodium: E:DTA 0.0075 : ~ : Pa rt B ~
Creme of Example 1 (10% dry solids~ 27.40 Soybean oil 8.00 Egg yolk, fresh: ~ 2.00 : : : Lemon juicei sin~le strength 0.25 Tota l100 . 00 . . .

WO 93/1067:. PCI /US92/065~9 ~ } 1 P roced u re ~..
1. Place water, vinegar, ISOSWEET, and SWEETOSE
into a steam jacketed, swept surface cooker.
5 2. Thoroughly blend the dry ingredients of Part A, then disperse them into the water/vinegar/syrup mixture with agitation. Continue mixing while heating with steam to 190F. Maintain this temperature for 5 minutes (with mixing); then immediately cool to 90F.
3. Transfer Part A to a Hobart bowl; add egg yolk, creme, and lemon juice. Mix at low speed for 5 minutes with paddle.
4. Slowly add soybean oil and mix for another 5 minutes.
5. Process through a colloid mill at a 0.026" setting.
Pack off finished product.

20;

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., WO 93/1 067:. PCI /USg2/06s89 -29- 2 1 ~

BUTTERMILK DRESSING

- 5 A buttermilk dressing can be prepared from the creme of Example 1 as follows.

In~redients ~, wt.

Buttermilk, liquid, 1% fat 30.00 : Water 23 . 94 Creme of Example 1 23.50 STAR-DRI (É~) 10 maltodextrin ~Staley) 8.00 Vinegar, white, 100~ grain 5.40 : S~asoning mix #962-2489 (Griffith Labs) 5.00 Buttermilk solids, Beatreme #983 (Beatrice) 1.00 Instant TENDER-JEL-(~) C starch (Staley) 1.00 Salt 0 75 Sugar ~ ~ 0.65 ~; Carboxymethyl ceilulose, 7 MF (Aqualon) 0.40 Titanium dioxide (Wàrner-Jenkinson) 0.10 Potassium ~ sorbate ~ 0 . 08 ;Sodium benzoate i 0.0725 Garlic powder 0.05 ~Onion powder 0 05 Calcium disodium E:DTA 0.0075 Total 100.00 ' :

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WO 93tl067:. PCI /US92/0658 211S ~ 30-P roced u re 1. Blend all dry ingredients together.
2. Place water and buttermilk into a Hobart mixing bowl. Disperse the dry ingredient blend int~ this water/buttermilk mixture; mix with a Hobart paddle for 10 minutes at low speed.
3. Add the creme and mix for 10 minutes at medium ` ~ speed .
10 4. Add vinegar and mix for 1 minute.
5. Process through a col!oîd mill at a 0.02" setting.
Pack off finished product.
.

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WO g3/10675 ' PCI /US92/06589 , EXAMPLE 4 ~ t~

FRENCH DRESSING

5 A French dressing can be prepared from the creme of Example 1 as follow,s.

In-qredlents ~ %, wt.

~Creme of Example 1 35.00 I:SOSWEET (g) ~I~OO high fructose ~
corn syrup (Staley) ~ 25 . 00 ' ~ Water 23.98 Vine~ar, white~ IOO~:~graln 10.00 ,` I5~ Tomat~ paste, 24-26% solids ~ 3.50 Sa~t; ~ 1 . 50 Seasoning mix~ #912-0135 (Griffith Labsl 0.30 MIRA-THIK~ :468~ starch (Staley) 0.30 : Seasoning mix #F34037 (McCormick) 0.10 :;20~ ~Xanthan~:gum,~Kelt~rol~: TF (Kelco): 0.10 G:u:ar gum #8/22~ TIC ~Gums) ~ 0.05 E~ Mustard powder: :~ ~ : 0.05 ' "' ~
Potass i um~ sorbah~ 0 . 05 ' ~ ' ::Titanium~ dioxide (~Wa~rner-Jenkinson) 0.04 :"
25~ `Paprika ~ 0.0225 Calcium disodium :EDTA : : 0.0075 Color, yellow ~FD~C #5/#6 ~ to suit Total 100.00 ,",:

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, - .:
:: : ',-WO 93/1067~ P~/US92/(~6589 2 ~ 1 Ll 2 P rocedu re 1. Place water and iSOSWEET into a Hobart bowl.
Blend all dry ingredients together and disperse into the water and ISOSWEET mixture. Mix at low speed and allow to hydrate for 5 minutes.
2. Add tomato paste, creme and vinegar. Mix for 5 mi n utes .
3. Process through a colloid mill at a û.02" setting.
Pack off finished product.

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WO 93/1067: PCl`/US92/06589 2 1 ~ c `~

DIJON DRESS I NG

5 A Dijon vinaigrette dressing can be prepared from the creme of Example 1 as fo!lows.

I n~redients ~, wt .

Water 38.90 : Creme of Example 1 25.00 Vinegar, white, 100 grain 14.00 Sugar 6. 50 :~ Dijon mustard (McCormick) 6.00 15: Lemc>n juice, single strength 6.00 S~lt : 2 . 35 Spice blænd #F3037~:~(McCormick) 1.00 Xanthan gum, Keltrol TF ( Kelco) 0.14 R~d be!l pepper, :dried (McCormick) 0.05 :
Potassium~ sorbate 0.04 Annatto extract~ (Warner-Jenkinson) 0.0125 ~: ~ : ; Caicium disodium EDTA 0.0075 ~
Total100.0Q :.

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~' WO 93/1067:- PCI/US92/065X9 21 ~t h~ 2 .

Procedu re 1. Place water in Hobart bowl.
2. Blend together all dry ingredients except the spice blend and red pepper. Disperse the dry ingredient blend into the water and~ mix with a paddle at low speed for S minutes.
. ~ :
3. Add the creme, mustard, vinegar and lemon juice.
Mix for 5 minutes at low speed.
10 4. Process through:~a~colloid mill at a 0;.02" setting.
5.~ Blend in spice blend~ and red pepper. Then pack off product. ~
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WO 93/10675 PCl /US92/06589 2~ i S.l ~ .

SOVR CREAM

5 A sour cream can be prepared from the creme of Example as fol lows .
.
In~redients %, wt.
.
Creme of Example 1 39.79 ~: ~ Sour cream, 18-20% fat 29.83 Water 23 . 41 Non-fat dry milk, low heat (Land O'Lakes)5.97 Lactic acid, 8830 0 - 40 15~ :Xanthan~gum, Rhodigel (Rhone-Poulenc) 0.20 Salt : : 0 . 20 Sodium citrate, hydrous, ~:
fine granular (Pfizer) 0.20 Total 100.00 ~ : ~
P ro~ed ~l re 1:.: Add lactic acid~:to wa:ter, thoroughly mix at Speed 1 25~ with; a Hobart ~ mixer equipped with ~ a ~ wire whip .
2:. Mix in sour cream~ and then all dry ingrediénts; mix until the mi~xture is uniform .
: 3. Blend in the creme to form a semi-homogeneous :
mixtu re .
30 ~ 4. Homogenize for smoothness.
5. Pack off and refrigerate until ready to use.

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WO 93/1067:~ PCl/US92/06~X9 TABLE SPREAD

5 A table spread can be prepared from the dry precipitate of Example 1 as follows.

I n~redients %, wt .

::. 10 Part A
` Water 25.00 :
Xanthan gum, Rhodigel (Rhone-Poulenc) o. 1n :
Pa rt B
Water 32 . 7306 Dry precipitate of;:Example 1 12.90 STAR-DRI (~) ~ 15. maltodexrin (Staley) 7.00 Whey powder sweet, #~7231 :(Land O'Lakes)1.00 Salt :; ~ 0 90 : 20 Potassium sorbate 0.10 ::
Calcium~ disodium:~ EDTA 0.0075 Artiflcial color, egg shade #08038 0.0()04 -~
(Warner-Jenkinson) Pa rt C ~ ; ~
STALE`~ 400-0300 partially hydrogenated ~:
corn oii (Staley) 19.83 Monoglycerides, :Dimodan LSK (Grinsted) 0.24 Leci.hin, M-C-Thin AF1/SB (Lucas Meyer) 0.15 ~:: 30 Flavor #57.752iA (Firmenich) 0.03 -: Antioxidant, Tenox 2 (Eastman) 0.01 B-Carotene, 30% in vegetable oil (Hoffmann-LaRoche) 0.0015 Total 100.00 ; ~
.

WO g3/1067~ P~/l 'S92/06SX9 37 2 ~ 3 ,~

Mixin~ Procedure , .
1. Vse a high speed mixer to mix Part A ingredients until xanthan gum is dissolved - 5 2. Comb;ne Par t A and ~ Part B ingredients and mix at slow speed until uniform. :-.
3. While mixing, heat Part; A and Part B mixture to 120-t30F.
4. Mix Part C i ngredients together and heat to ~ 120-130F.
5. Pour aqueous; pha~s~e (Parts A and B) into oil phase (Part C) while~ stirring vigorously .
6. Homogenize the entire mixture at a pressure of :

8,000~ to 15,000;~ psi~ and an~ output temperature of 140-150F.`
7. ~ ~ Immediately cool to~ 50F while stirring.
8.:~ : ; Pack ~off and~refrigerate.;

20 `:

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WO 93/1067~ PCI/US92/06589 2 ~ 4 i~.-ALPHA-AMYLASE HYDROLYSIS OF HEAT TREATED, DEBRANCHED 55% Ah~YLOSE CORN STARCH
A 2% solids slurry of 559~ high amylose corn starch ~HI-SET C) was prepared by mixing 452.3 grams (400 grams dry basis) of Hl-SET C corn starch with deionized water to give a total volume of 20 liters. The 10 suspension was heated in 2-liter batches up to 160C in a pressure reactor then cooled to about 30C to 50C by passing the hot solution through a cooled heat exchanger tube. The pH of the solution was adjusted to approximately ~ 4.5 'and the solution was placed in two 5 12-liter round bottom flasks equipped with agitation, condensers, and ~heat controlled water baths. The temperature was adjusted to 45C and 400 units per gram dry basis starch of isoamylase enzyme (f rom Hayashibara Co~. and~ containing~ 865,000 units/gram) was added to 2 0~ ~ each~ solution.~ The~ solutions were allowed to react 20 hours ~ then~ the~ Z%~ solids solutions/dispersions (the debranched starch~ ~tends to precipitate with time) were heated~ to 160C in~the pressure reactor as before to :: ` :: :
~ ~ ,comptétely dissolve t he precipitated starch and make it 25 ~ more readily ava~ilable for isoamylase enzyme attack. The solutions were cooled, the pH again checked and found to be~ approximate,ly 4 . 5, then 400 units per gram dry basis starch of isoamylase enzyme was again added and ; ~ the reaction was~allowed to proceed 18 hours at 4SC for 3() a total reaction time of 38 hours.

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WO 93/1067~; PCr/US92/06589 .

39 2 :~ 1 J 1 ~ L~

The 2~ solution/suspension after 38 hours of isoamylase digestion was heated to approximately 95C to inactivate enzyme then concentrated by rotary evaporation over a 2-day period (stored in a refrigerator 5 overnight) to approxlmately 15~ solids. This slurry was dried on stainless steel trays at 60C in a forced air oven overnight and the dried material ground to pass through a US #60 mesh sieve.
To 380 grams (350 grams dry basis) of the 10 above dried, screened material was added 1,720 grams of deionized water to give ~a 20~ solids slurry. The slurry was heat treated byi ;controlled heating in a tèmperature controlled water bath from 50C up to 100C at the rate of 0.05C per m;nute followed by controlled cooling from 100C down to 50C ~at the rate of ().OSC per minute.
The heat treated slurry ;~was poured onto a stainiess steel tray~ and dried in ~a~forced air oven~ at 50C then ground ~ . .
to pass through a~ US. #6~ mesh sieve. This sample served as ~a substrate~for a-amylase hydrolysis treatment 20~ to~improve the ease of ~creme formation on sheari`ng.
The above ~ starch substrate was enzyme hydroiyzed at 20%~ solids ~(375~ grams total ~slurry wt. ) at ; 25C with 15 units/gram starch of porcine pancreatic a-amylase (Sigma~ Chemical ~ ~ Company) . Sampîes of 25 ~ ~ reaction slurry were~withdrawn (125 grams each) after 8 hours~, 21 hours~ and 48 hours~ of hydrolysis and the pH
adjusted to 3.5 to Ina~ctivate ~enzyme. ~ The 8 hour and 21 hour samples were ;filtered on a Buchner funnel followed by wash;ng with about 25U ml each with deionized water.
30 The sample hydrolyzed 48 hours would not filter (very, very slow) and ~was centrifuged (7000 x g), the supernatant discarded, then deionized ~.~ater added back :
: ' ~ ~ . ; , ' ' WO 93/1067~ P~/US92/06~8~

21~ ~; 1 L~ ~ -40- ~

to the original sample weight. After stirring to give a homogenous mixture, this slurry was centrifuged as above and the supernatant discarded. This procedure was repeated a final time. All wet cakes or sediment S (from filtration or from~centrifugation) were mixed with 8 volumes of ethanol (formula 3A) to denature any remaining enzyme. They were then either filtered or centrifuged one last time. The wet cakes and sediment were dried in a 50C forced air oven and ground to pass 0 through a US #60 mesh sieve. The products were welghed ~ and ~yields ~ were calculated for each . Yield stress ~ values were~ o btained on 20~ solids cremes . .
prepared by shearing at 120 volts, 60C for 8 1/2 minutes ~with a Waring~ blender~ equipped with a small 5~ jacketed j ar. ~ The~ ~ yield stress values were measured :`
using~ a ~ B~rookfield~ viscometer after the ~ cremes stood at least; 3 hQurs at~ room temperature. The analytical ; r esults are reported~ ~below .

20~ a-Amylase Hydrolysis ~ Prodùct ~Yield Stress, Time,~ hr. ~ Yield, 96 db ~ Pascals 8 ~ 87 . 0 ~ 288 25~ 21 ~ 84.2 ~ 348 -~
48 ~ 80. 9 365 It was noted that the texture of the 8 hour creme was the most gritty~ while the 48 hour sample was 30~ ~more~ creamy and~ ~;less~ gritty. ~ It ~ ~s~ speculated that continued enzyme~ ~hydrolysis~ would continue to improve texture (less ;gritty~) ~and Increase yie1d ~stress values.

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WO 93/1067:` PCI/US92/065X9 ~113~2 It was noted that the pH of the 48 hour hydrolyzed sample declined to 5.7 ~from 6.9). It is likely that this was caused by undesirable fermentation to form organic acids. Such a large decline in pH likely 5 also causecl reduced enzyme hydrolysis activity compared to what may have occurred if the pH had remained at 6 . 9 .
It is concluded that the enzyme hydrolysis gave considerable improvement in the shearability of the heat treated,~ debranched high amylose starch. In :
addition, it ~also ~resulted in improved smoothness of textu re .

` ~ EXAMPLE 9 ~ ' DEBRANCHED;~'HIGH; AMYLOSE CORN STARCH

A 55% ~amylose corn starch (HI-SET C) is made 2~ up ~to 25%~ sol~ds~'~then~ jet ~cooked~ at ;160C wlth a retention time o f n~10 minutes at 160C then cooled to 100C. ~The~: pH :~of~ the~ solution if ~ adjusted: to pH 6Ø
- Novo~ thermostablel'~ ~pullulanase enzyme as described in WQ~ 92/02614 ~s~ added~ at 50; units per gram of starch and 25~ ~ the~:reaction ls ;allowed to proceed at 100C for 24 hours at which~ time ~GPC~ana~iysis ~;will show that less than 10%
'~ ~ of ~ the remaining''amylose or amylopectin molecules are above about 100,(100 molecular weight.
The~ ~debranched ~solution is treated with 3%
30 w/w (weight; ~by~ ~weight ~ basis)~ of decolorizing carbon (based on starch~dry ~substance weight) at 90C. The coiorless carbon~ treated ~solution is cooled to 5C for 16 :
::

:: :
.
, WO 93/1067~ PCl /US92/065X') 2 1 1 j 1 '~ 2 -42-hours to bring about crystallization. The crystallized mass is dried in a spray drier at 15% solids after dilution with water.
The spray dried material is made up to 20~
S solids and heated from 50C to 100C at 0.û5C/minute then cooled to 100C at 0.05C/minute. The heat treated material is adjusted to pH 6.9 and treated at 20% solids at 25C for 24 hours using approximately 50 units per gram dry starch of porcine pancreatic -amylase enzyme.
The resulting slurry is adjusted to pH 3.5, with 10% HCI
heated to 60C and held at that pH and temperature about 1 hours~ to inactivate enzyme then microfiltered at 60C to reduce the soluble saccharide content to less tha~n 10% (measured at room temperature). The retentate 1S slurry is spray dried at about 15% solids to give a heat stable, shearable starch based fat replacer having a yield stress at 20~ solids greater than 400 pascals.

: ~ :

: : ;
~ .

Claims (17)

What is claimed is:

1. A foodstuff having a reduced level of fat and/or oil comprising a mixture of a foodstuff and a particle gel as a replacement for at least a substantial portion of the fat and/or oil of said foodstuff, said particle gel comprising a minor amount of a fragmented, .alpha.-amylase hydrolyzed amylose precipitate and a major amount of an aqueous liquid.

2. A foodstuff of Claim 1 wherein said particle gel exhibits a transition in dynamic elastic modulus versus shear strain from substantially constant dynamic elastic modulus to decreasing dynamic elastic modulus, said transition being exhibited at a shear strain of less than about 50 millistrain.

3. A foodstuff of Claim 1 wherein said fragmented, .alpha.-amylase hydrolyzed amylose precipitate is derived from starch from a variety of Zea mays.

4. A foodstuff of Claim 1 wherein said fragmented, .alpha.-amylase hydrolyzed amylose precipitate is derived from a starch having at least about 15% by weight amylose.

5. A method of formulating a foodstuff containing a fat and/or oil ingredient comprising replacing at least a substantial portion of said fat and/or oil ingredient with a particle gel comprising a minor amount of a fragmented, .alpha.-amylase hydrolyzed amylose precipitate and a major amount of an aqueous liquid.

6. A method of Claim 5 wherein said particle gel exhibits a transition in dynamic elastic modulus versus shear strain from substantially constant dynamic elastic modulus to decreasing dynamic elastic modulus, said transition being exhibited at a shear strain of less than about 50 millistrain.

7. A method of Claim 5 wherein said fragmented, .alpha.-amylase hydrolyzed amylose precipitate is derived from starch from a variety of Zea mays.

8. A method of Claim 5 wherein said fragmented, .alpha.-amylase hydrolyzed amylose precipitate is derived from a starch having at least about 15% by weight amylose.

9. A method of making a composition of matter useful in replacing fat and/oil in a food formulation comprising physically fragmenting a minor amount of an .alpha.-amylase hydrolyzed amylose precipitate in a major amount of an aqueous liquid, the degree of said physically fragmenting and the degree of hydrolysis being sufficient to form a particle gel of said composition.

10. A method of Claim 9 wherein said particle gel exhibits a transition in dynamic elastic modulus versus shear strain from substantially constant dynamic elastic modulus to decreasing dynamic elastic modulus, said transition being exhibited at a shear strain of less than about 50 millistrain.

11. A method of Claim 9 wherein said .alpha.-amylase hydrolyzed amylose precipitate is derived from starch from a variety of Zea mays.

12. A method of Claim 9 wherein said .alpha.-amylase hydrolyzed amylose precipitate is derived from a starch having at least about 15% by weight amylose.

13. An aqueous dispersion useful as a replacement for fats and/or oils comprising a major amount by weight of water and a minor amount by weight of a fragmented, .alpha.-amylase hydrolyzed amylose precipitate, the degree of hydrolysis and fragmentation of said precipitate being sufficient to form a particle gel of said dispersion.

14. An aqueous dispersion of Claim 13 wherein said particle; gel exhibits a transition in dynamic elastic modulus versus shear strain from substantially constant dynamic elastic modulus to decreasing dynamic elastic modulus, said transition being exhibited at a shear strain of less than about 50 millistrain.

15. An aqueous dispersion of Claim 13 wherein said fragmented, .alpha.-amylase hydrolyzed amylose precipitate is derived from starch from a variety of Zea mays.

16. An aqueous dispersion of Claim 13 wherein said fragmented, .alpha.-amylase hydrolyzed amylose precipitate is derived from a starch having at least about 15% by weight amylose.

17. A method of making a composition of matter useful in replacing fat and/or oil in a food formulation comprising:
(a) gelatinizing a starch having an amylose content of at least about 15% by weight in an aqueous medium;
(b) precipitating amylose from an aqueous medium to form an amylose precipitate from said aqueous medium;
(c) treating said precipitated amylose with an .alpha.-amylase enzyme to hydrolyze a portion of the 1,4-glucosidic bonds in said precipitated amylose to form an .alpha.-amylase hydrolyzed amylose precipitate;
(d) fragmenting said .alpha.-amylase hydrolyzed amylose precipitate in an aqueous medium to form a particle gel having a minor amount of fragmented, .alpha.-amylase hydrolyzed amylose precipitate dispersed in a major amount of an aqueous medium, said particle gel exhibiting a transition in dynamic elastic modulus versus shear strain from substantially constant dynamic elastic modulus to decreasing dynamic elastic modulus, said transition being exhibited at a shear strain of less than about 50 millistrain.