JP2540204B2 - Chemically stable starch emulsifier with storage stability - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to modified starch. The starch is useful in shelf-stable emulsions, especially in systems that require stable oil-in-water emulsions, especially gum arabic in beverages flavored with flavor oils such as citrus oil (citrus oils). Useful as an alternative. Further, the present invention relates to a method for producing a modified starch and a chemical composition, a food and a beverage containing the modified starch.

Various chemical compositions are used in the food, cosmetics, paint, pharmaceutical and polymer industries as well as textile and leather processing, flotation,
Used as an emulsifier in oil drilling and agricultural spraying operations. In many of its uses, the emulsifier also acts as a stabilizer of continuous phase viscosity or fluidity. Often this use requires that the emulsion be shelf stable for long periods of time.

Typical compositions which act as aqueous emulsifiers and stabilizers include guar gum, acacia and other gums, starch, proteins, various water soluble polymers and homologues thereof. For example, Encyclopedia of
Chemical Technology (Encyclopedia of Chemical T
echnology), edited by Kirk Osmer, 3rd edition, Wily-Intersiene, New York, New York, 1979, Volume 8, pages 900-910.
Pages 918, 923-925 and Volume 12, Pages 55-56
See page. Gum arabic is preferred for many uses, especially in terms of its storage stability during chilled or frozen storage of the emulsion.

Gum arabic is a branched substituted heteropolysaccharide, which is highly water soluble, low viscosity and odorless, colorless or tasteless. Gum arabic is used as an emulsifier and stabilizer in foods such as confectionery, syrups, flavor oil emulsions, ice creams and beverages and inks,
Used in adhesives, textile materials and lithographic solutions.

Gum arabic is a naturally occurring gum produced in Central East Africa. Since gum arabic is obtained from this area, it is expensive and its supply and quality are unpredictable. Therefore, a long-term storage-stable, inexpensive gum arabic alternative has been sought in the industry and products derived from starch have been proposed for such use.

U.S. Pat. No. 2,661,349 issued Dec. 1, 1953 by Caldwell et al. Describes substituted dicarboxylic acid anhydride starch half ester derivatives. Some of these derivatives form stable oil-in-water emulsions suitable for use in beverage emulsions, flavor emulsions and other emulsions. For example, P. Trubiano, Chapter 9, Modified Starch: Properties and Uses, CRC Press, Boca Raton (Boc
aRaton), Florida, 1986, pp. 134-147.
Cold water soluble, low viscosity octenyl succinate starch derivatives have been successfully used to replace gum arabic in carbonated beverages. The relatively high viscosity octenyl succinate derivative is useful as a gum arabic substitute in salad dressings. Such substituted dicarboxylic acid starch half-ester derivatives are also hydrophobic substances such as flavors,
Used as an alternative to gum arabic for encapsulating vitamins, fragrances and oils. Capsule encapsulation is usually prepared by spray drying an oil-in-water emulsion. Some of this capsule encapsulation is modified to produce a composition that results in a gradual or limited release of the encapsulated flavor or oil. Other substances can dissolve in water at higher solids than gum arabic equivalents and are superior to gum arabic for certain uses.

The low viscosity (modified) starches used in beverages and flavor emulsions are usually prepared by acid-degrading starch. Methods for making low viscosity starch are well known. Richards et al., U.S. Pat. No. 4,035,235, issued Jul. 12, 1977, disclose a method for degrading lipophilic substituted starch, which is α-
Amylase digestion is used to produce low viscosity starch as an alternative to acid-degradation. The lipophilic substituted starch is suitable for flavor encapsulation and for oil-in-water emulsions. This process produces a starch product suitable for oils used in beverages as an emulsifier and capsule encapsulant. One disadvantage of using the known starch-derived products instead of gum arabic is that the known starch derivatives are not very stable during storage. This starch derivative exhibits shorter shelf life and worse chilling and freeze / thaw stability compared to gum arabic. Therefore, in certain applications, such as flavor syrup bases used in the manufacture of soft drinks and similar types of beverages,
This starch-derived substitute does not work as well as gum arabic. Maintains stability during flavor oil emulsion and storage as beverage manufacturers transport flavored syrup bases to bottlers in different districts where they are placed in refrigerated storage for a long time before being used for bottling operations. There must be. Further, the cooling temperature changes from bottler to bottler, or it changes from day to day, so
The flavor oil emulsion must be able to withstand temperature cycling, including freeze / thaw cycles.

Stability problems in beverage use are believed to occur due to the aging tendency of the starch product and do not result in temperature cycling or long term storage in flavor oil emulsions. In severe cases starch ages to form gels and flavor oils separate completely from the aqueous phase. Aging of starch is essentially a crystallization process. This occurs when the linear portions of the starch molecule, algin, which are adjacent to each other, form internal chain hydrogen bonds with the hydroxyl groups. Molecules are associated with the formation of molecular aggregates if sufficient internal chain hydrogen bonding occurs. This aggregate exhibits a reduced capacity for hydration and therefore a low water solubility. This aggregate precipitates or forms a gel in a more concentrated solution. The aging tendency is more pronounced in starch containing high levels of linear amylose molecules. The aging tendency is less pronounced in starch containing linear (amylose) and branched (amylopectin), or containing only branched molecules. At lower temperatures, starches containing amylose and amylopectin show a greater aging tendency.

Partially overcomes aging by chemically inducing starch molecules in certain uses, interfering with the association of starch molecules or between parts of the same molecule, thereby hydrating the starch during storage. Stabilize starch by reducing the tendency to lose. For example, starch is reacted with a reagent to introduce substituents, such as hydroxypropyl-, phosphate-, acetate- or succinate groups, to stabilize the starch molecule during storage. This derivatization reaction is performed on starch which is further modified by cross-linking or degradation to obtain starch for a particular application. However, this derived starch does not produce the stable emulsifying properties typical of gum arabic.

Other known methods of limiting starch aging at low temperatures also do not result in stable emulsified starch. For example, U.S. Pat. No. 3,525,672 issued August 25, 1970 by Wurzburg et al. Describes a cross-linked inhibitory starch thickener treated with an enzyme such as β-amylase and frozen. / Providing thawing stability to pie fillings, puddings, and other thickened foods that are exposed to low temperature storage. In addition to the inhibition treatments described, it is sometimes stated that it is advantageous to partially derivatize the starch base. Typical substituents include ester groups such as acetate-, succinate-, phosphate- and sulphate groups and ether groups.

U.S. Pat. No. 4,428,972 issued January 31, 1984 by Wurzberg et al. Discloses a waxy starch thickener having even better low temperature stability in aqueous dispersions. The starch is selected WXSU ₂
Obtained from genetic plants.

But in overcoming aging during low temperature storage,
This invention does not disclose any starch product useful for making shelf stable oil-in-water emulsions. Thus, there remains a need for a product that has emulsifying properties with stability during storage, cooling and freeze / thaw cycles and that can be used in place of gum arabic.

Therefore, improved and modified starches showing the stable emulsifying properties of gum arabic are required. According to the present invention, such a modified starch is provided together with a method for producing such a modified starch.

Products and beverages containing starch-based emulsions with good storage stability, in particular flavor oil emulsions, are also provided.

The present invention consists of a starch derivative containing a hydrophilic group and a hydrophobic group, up to 70% by weight of which the 1,4-α-D-glucoside bond can be cleaved from the non-reducing end of starch. Degraded by an in vitro enzyme that is unable to cleave the 1,6-α-D-glucoside bond,
Provided is a modified starch having the emulsifying properties of an emulsion that exhibits improved stability during storage, resistance to oiling and resistance to gelation.

The invention further comprises the following steps: a) a gelatinized starch derivative which is characterized by respective substituents containing hydrophilic and hydrophobic groups; b) up to 70% by weight of starch derivative. Maltose that can cleave 1,4-α-D-glucoside bonds from the non-reducing end of starch, but cannot cleave 1,6-α-D-glucoside bonds in starch And a modified starch having the emulsifying properties of an emulsion exhibiting improved stability during storage and resistance to oiling and gelling.

The enzymatic treatment used in the method of the invention is carried out after derivatizing the starch so that it contains hydrophilic and hydrophobic groups, which have emulsifying properties.

Alternatively, the enzyme treatment is carried out before producing the starch derivative. A method for producing such a derivative is disclosed in the above-mentioned US Pat. No. 2,661,349. A preferred starch derivative is a starch alkenyl succinate half ester in which the carboxyl group is present as an acid or carboxylate salt. However, it is stated that such modified starch can be produced using any method which produces the desired hydrophobic function or both hydrophobic and hydrophilic functions in the starch molecule, thereby producing emulsifying properties. There is. Suitable derivatives and methods for making them are described in US Pat. No. 4,626,288 issued Dec. 2, 1986 by Trzasko et al.

Although the 1,4-α-D-glucoside bond can be cleaved from the non-reducing end of the starch molecule,
Treatment with an in vitro enzyme that cannot cleave the 6-α-D-glucoside bond decomposes up to 70% by weight (preferably 55% by weight) of starch derivatives containing hydrophilic and hydrophobic groups into maltose. Carry out until.

In the production of the modified starch of the present invention, the desired starch is slurried in water at a rate necessary to achieve the desired enzyme-substrate concentration or at a rate calculated to be suitable for end use. If desired, starch is used in granular form, but the enzymatic degradation of granular starch proceeds slowly. The temperature and pH of the mixture is then adjusted to the optimum conditions for the particular enzyme used by the manufacturer or supplier to produce the starch product.

The enzyme is 1,4-α-D-from the non-reducing end of the starch molecule.
It must be an in vitro enzyme capable of cleaving the glucosidic bond, but not the 1,6α-D-glucosidic bond of the starch molecule. β-amylase-
This is preferred-a highly specific bioenzyme, the complex of which reacts only from the non-reducing end of a starch molecule having a maltoside group which binds to the glucose group via its 1,4-α-D-glucosidic bond. Can be formed. The enzyme thereby only attacks starch at the non-aldehyde end (non-reducing end) and separates maltose units from this outer branch until it reaches the branch point (1,6-bond). A single maltose or glucose unit remains at each branch point of the starch molecule after enzymatic treatment. This in vitro enzyme can separate 1,4-bonds of starch branching, but cannot break down 1,6-bonds, so the rest of the degradation process is a dense branched structure. It has no upper or outer branches or contains only short outer branches. The product is therefore free of long outer chains that are subject to aging and syneresis in aqueous starch dispersions that have been subjected to long-term storage and / or repeated freeze / thaw cycles.

The enzyme provides up to 70% (preferably 55% by weight) starch to decompose to maltose or to the desired end point (ie improved storage stability of special emulsions made with starch). The starch can be digested until it reaches sufficient decomposition)
The end point is determined by the change in viscosity, by the reducing sugar content or by any other known method for measuring the amount of enzymatic degradation of starch molecules. Alternatively, enzymatic degradation may be continued until substantially all of the resulting maltose units have been removed from the starch molecule. Usually decomposition is temperature,
Hours to hours depending on enzyme and substrate concentrations and other changes
It is held for a period of 24 hours or more. The enzymatic degradation is then terminated by heat, chemical addition or other known method for inactivating the enzyme or separating it from its substrate.

The resulting decomposed starch composition is spray dried, drum dried or otherwise recovered in a form suitable for the present invention.

The present invention includes all emulsified compositions in which the emulsifier is starch, which starch is enzymatically modified to improve the storage stability of the emulsion. Therefore, an emulsion consisting of a mixture of modified starch and gum or other emulsifiers should be included.

Use of the modified starch of the present invention in products where gum arabic is used as an emulsifier, stabilizer or homologue thereof, and in products where a polymeric water-soluble emulsifier is used to form or stabilize an emulsion. Is advantageous.
It can therefore be used in beverages, confectioneries, ice creams, other beverages and other foodstuffs requiring a storage-stable emulsifier, flavored with oils such as orange or lemon oil. It can be used in beverages based on water and alcohol. The starch can also be used in the production of spray dried flavor oils which are reconstituted with water to give flavor emulsions and in inks, textile materials and other non-food end uses.

Suitable starches used in the production of enzymatically degraded starch according to the present invention include corn, potatoes, sweet potatoes, wheat, rice, sago, tapioca, vulgaris, sorghum and all plant sources including homologues. , Derived from. Also included are modified products derived from any of the above starches. This is for example oxidation, α
-Flowable or weakly boiling starches and derivatized starches produced by amylase modification, mild acid hydrolysis or thermal dextrinization, such as ethers and esters.

The starch is preferably gelatinized starch (pre-heat-treated non-granular starch), and may be fluid starch modified by mild acid decomposition or a known thermal dextrinization method. (Eg MW Rutenberg, "Starch and Its Chemicals", Water Soluble Gases and Resins-Handbook, RL Davidson, Editor, McGraw Hill, Inc., New York, New York 1980, See pages 22-36). One or several combinations of this chemical engineering technique are used. The modification is generally performed before treatment with the hydrophobic / hydrophilic (or simply hydrophobic) agent and before the β-amylase treatment. If desired, starch is modified by treatment with α-amylase enzyme to produce free-flowing starch in the manner described in US Pat. No. 4,035,235. Chemical modification with α-amylase is also generally performed before β-amylase treatment. If a high viscosity emulsion system is desired, it is not necessary to modify the starch.

The starch is derivatized by treatment with all agents which give the starch its emulsifying properties. Derivatization is performed before or after β-amylase treatment. The agent must have a hydrophobic portion and may contain a hydrophilic portion. The hydrophobic part must be an alkyl-, alkenyl-, aralkyl- or aralkenyl-group containing at least 5 C-atoms, preferably 5-24.
The hydrophilic moiety is provided by the reagent as in the preferred embodiments below or the hydroxyl group of starch itself acts as the hydrophilic moiety as in other embodiments, the agent provides only the hydrophobic moiety. .

In a preferred embodiment, starch is prepared according to US Pat.
It is derivatized by reaction with an alkenyl anhydride cyclic dicarboxylic acid according to the method described in Japanese Patent No. 349. However, using any method of derivatizing starch that produces the desired mix of hydrophobic and hydrophilic functions in the starch molecule, thereby producing stable emulsifying properties, to produce the modified starch according to the invention. You can This includes methods not previously known.

If a low viscosity emulsifier is desired, a preferred embodiment is an amylopectin-containing starch, such as the octenyl succinate half ester derivative of waxy corn. It has been engineered to a water flow rate (WF) of up to about 60. Water fluidity is an empirical test of viscosity measured on a scale of 0-90. At this time, the fluidity is the reciprocal of the viscosity. The water fluidity of starch is generally measured using a Thomas rotary shear-type viscometer (Arthur H. Thomas, Philadelphia, PA). This meter has a viscosity of 2 at 30 ° C.
Standardized with standard oil having 4.73 cps. This oil requires 100 revolutions 23.12 ± 0.05 seconds. An accurate and reproducible measure of water fluidity is obtained by determining the elapsed time of 100 revolutions at various solid levels depending on the modification rate of starch (viscosity decreases with increasing modification). In a preferred embodiment, modified starch is treated with at least 0.25% octenylsuccinic anhydride, preferably 3.0%. If desired, hydroxypropyl octenyl succinate derivatives can be used.

For other products, the degree of substitution or modification level that produces the desired viscosity and emulsifying properties is used. For example, U.S. Pat. No. 4,035,235 discloses suitable embodiments. It consists of a process for producing lipophilic derivatives of starch which are used as alternatives to gum arabic for encapsulating water insoluble substances such as volatile flavor oils and perfumes.

In the preferred embodiment, the next step after production of the starch derivative must be to gelatinize the derivatized starch. The gelatinization process causes the starch molecules to evolve from the granular structure, which allows the enzyme to more easily and uniformly break down the outer branches of the starch molecules. After gelatinizing the starch-based slurry, the solids, temperature and pH of the slurry are adjusted to produce optimal enzyme activity.

Optimal parameters for enzyme activity will vary depending on the enzyme used. Therefore, the rate of enzymatic degradation is
Depending on factors such as the type of enzyme used, enzyme concentration, substrate concentration, pH, temperature, presence or absence of inhibitor and other factors. Depending on the type of enzyme, i.e. its origin, various parameters require adjustment to achieve the optimum digestion rate. Generally preferred enzymatic digestion reactions are performed at maximum solids content. The rate is such that it can facilitate the subsequent drying of the starch composition while maintaining an optimal reaction rate. For barley β-amylase used according to the invention for producing emulsifiers which are applied, for example, to beverages, preheated starch dispersions with a solids content of up to 33% are preferred. However, the more solids are used, the more solids are more difficult or ineffective to stir, and the starch dispersion becomes more difficult to handle.

When the method of the present invention is described using β-amylase as the enzyme component, other enzymes such as in vitro-α-1,
Select 1,4-linkage of starch molecule from 4-glucosidase, in vitro-1,4-α-D-glucan maltotetrahydrolase, in vitro-1,4-α-D-glucan maltohexahydrolase or non-reducing end Other bioenzymes, which are cleaved selectively but which retain the 1,6-linkage, are used to produce the modified starches according to the invention.

Such in vitro enzymes separate glucose, maltose, or relatively large saccharidic units only from the outer branches of the amylopectin molecule, and do not debranch the branches, so the enzymatic degradation method leaves short-chain branch points intact. Give rise to molecules. In amylopectin, branch points occur in about 4-5% of the monosaccharide units of the molecule. If the enzymatic degradation of amylopectin is continued, the outer branches of the molecule are shortened to the point of reducing or eliminating association with other branches, and the remaining 1,6-branch points associate the inner branches. Protect from. Therefore, the degraded molecules exhibit improved aging resistance during storage and during freeze / thaw cycles. For this reason, an in vitro enzyme capable of cleaving the 1,4 bond from the non-reducing end of the starch molecule but not the 1,6 bond is an essential requirement of the present invention.

Although the method of the present invention uses the enzyme in solution, the method of using the enzyme supported on a solid support is suitable for use in the present invention.

The optimal concentration of enzyme and substrate is limited by the level of enzyme activity. Enzyme activity is expressed in diastase potency (DP °) per ml of aqueous enzyme solution. DP ° is 5% of sample enzyme preparation
The amount of enzyme contained in 0.1 ml of solution. This preparation produces sufficient reducing sugars to reduce 5 ml of the Fehling's solution when the sample is incubated with 100 ml of substrate for 1 hour at 20 ° C. The procedure for determining DP ° is described in Food Chemicals Codex, Third Edition, National Academia Press, Washington, DC, 1981, p.484.

An excess of 334 DP ° per 100 g starch per dry weight must decompose a gelatinized starch solution containing up to 33% solids within 8 hours under optimal conditions of temperature and pH.
DP ° 840-1,110 (pH of 100 g of starch per dry weight)
5.3 and a temperature of 57 ° C) is preferred.

This reaction proceeds in the presence of buffer, ensuring that the pH is at optimal levels during degradation. Buffers such as acetates, citrates or salts of other weak acids are preferred. Other agents can be used for optimal oxygen activity. Reaction is pH
Range about 3-10, preferably 5-7, most preferably 5.7
At 55-60 ° C.

Aqueous starch dispersion at 20-100 ° C, preferably 55-60
It must be maintained during enzymatic digestion at a temperature of ° C, most preferably 57 ° C. However, if a relatively short reaction time is desired, temperatures are used in the range 60-63 ° C or even higher enzyme concentrations. If other parameters of the enzymatic reaction are used, the preferred and optimum temperatures will vary with other parameters, such as substrate concentration, pH and other factors affecting the enzyme activity, and can be determined by the practitioner.

The enzymatic reaction can be continued until the desired level of degradation is reached or until all of the resulting maltose has been removed from the starch molecule. The progress of the enzymatic reaction is measured by various methods. If all critical parameters are to be demonstrated for the achievement of a particular starch composition, then the reaction must be advanced in time (ie 8 hours in Example 1) to a predetermined relative end point. Furthermore, the final point is traced, and the concentration of reducing sugar is measured and limited. 1,4-α-
Maltose produced by D-glucosidase activity is a reducing sugar and is quantified by a known method. Other techniques such as tracking changes in viscosity or changes in molecular weight are used to limit the reaction endpoint.

In a preferred embodiment, the end point of decomposition is measured by determining the percentage of reducing sugars contained in the reaction medium. Each maltose unit contains two glucose units but only one reducing sugar that can be detected by the reducing sugar assay. Therefore, the weight percent of reducing sugars (calculated as glucose) is approximately equal to 1/2 of the weight percent of maltose units formed by the breakdown of starch molecules. When β-amylase is contaminated with high levels of α-amylase, the reaction medium contains even higher levels of reducing groups, and reducing sugar measurements take into account the effects of such contamination in measuring the progress of enzymatic reactions. Must be standardized in. Alternatively, maltose may be measured directly by other known types of analytical methods.

The degree of starch degradation required to substantially improve the low temperature stability of the starch composition is subject to change.
It is based on the type of starch used, the presence and nature of all substituents and the degree of chemical modification, if any. Starch degradation varying from 13-55% by weight is known to result in improved low temperature stability in thickeners (see US Pat. No. 3,525,672). For low viscosity emulsified applications (i.e. starch modified to WF40-60), degradation is continued until up to 70% by weight starch (as measured by the reducing sugar content of the starch dispersion) is hydrolyzed to maltose.

The maximum amount of enzymatic degradation to maltose that can theoretically be achieved using pure β-amylase and modified starch is about 55% by weight starch. Pure β-amylase is not readily available on the market. Β available on the market
-Amylases contain small amounts of α-amylase, and in vitro enzymes that randomly cleave internal starch branches separate these branches of β-amylase activity. Starch modification by acid, heat or oxidation has a similar effect, but not as markedly as the β-amylase degradation of starch. Degradation of up to 70% by weight starch can therefore be achieved using commercially available β-amylase contaminated with small amounts of α-amylase.

After obtaining the desired degree of starch degradation, the enzyme is inactivated. It is expedient to inactivate the β-amylase rapidly at a temperature of about 100 ° C. and therefore terminate the reaction by raising the temperature of the starch dispersion to at least 75 ° C. for at least 15 minutes.

It will be appreciated that the order of the steps in the method of the present invention can be carried out in any order and is not limited to the preferred embodiments described above. Therefore, the order is reversed in the second preferred embodiment to complete the enzymatic degradation prior to the derivatization step.

Where the end use application requires purification of the starch composition, maltose and other reaction impurities and by-products are dialyzed, filtered, centrifuged or any other known known for isolation and concentration of starch compositions. It can be removed by a method.

If a dried starch composition is desired for the end use application, the starch composition is dehydrated in all known ways.

In the beverage flavor emulsion, waxy corn starch with a water fluidity of about 40-60 has been preferred.

For low viscosity emulsifiers, viscosity reduction during the enzymatic reaction is used to determine when the desired level of degradation is achieved. A method for tracking the viscosity of the reaction solution is described in Example 1. Any of the many known methods for measuring viscosity can be used. However, the viscosity is modified by factors other than the desired enzyme activity. For example, if the β-amylase contains large amounts of α-amylase, the decrease in viscosity is β-amylase.
It cannot be directly linked to amylase activity. Therefore, the level of α-amylase contamination must be carefully followed and adjusted to see if viscosity changes are used to determine the level of enzymatic degradation. If the β-amylase or any other 1,4-α-D-glucosidase used according to the invention contains high levels of α-amylase contaminants, the β-amylase should be purified prior to use or the α-amylase Inhibitors can be added to the reaction dispersion.

The following examples detail specific examples of the present invention. All parts and percentages in this example are stated on a dry weight basis and all temperatures are degrees Celsius unless otherwise noted.
Storage stability is measured by promoting aging at low temperature and shortening the test period.

Example I This example illustrates the preparation of a modified starch of the present invention for use in beverage flavor emulsification.

An octenyl succinate derivative (OSA) of waxy maize starch is prepared by the method disclosed in Example II of US Pat. No. 2,661,349, except that cornstarch is replaced with waxy corn. 3 more starch
% 3% rather than 0.5% as disclosed in the literature
React with octenyl succinic anhydride. A 28% aqueous slurry of OSA waxy corn is jetcooked at about 149 ° C (300 ° F). OSA then heat treated
Place waxy corn in a constant temperature bath for 55-60
Keep at constant stirring at ℃. Adjust the pH to 5.3 with 3% hydrochloric acid.

The heat-treated OSA waxy corn dispersion was divided into four batches with varying levels of barley β-amylase (1,4-α-D-glucan maltohydrolase (EC3.
2.1.2), from Huelmcovaiochemics, Inc., Elk Grove Village, Illinois. ) To each batch. The amount of enzyme added was 168,334,840 and 1,110 per 100 g of OSA waxy maize dry matter.
It is DP °. Preliminary tests determine that this relative range of enzyme concentrations produces the desired starch degradation in about 8 hours.
Three batches containing at least 334 DP ° per 100 g starch achieve the desired degree of decomposition in 3-8 hours.

The degree of degradation is determined by following the final viscosity of the dispersion.
Therefore the amount of α-amylose present in barley β-amylose was traced and limited to only 0.4 DU / ml of enzyme solution, but the viscosity is not affected by this change. DU (dextrinization unit) dextrinizes soluble starch in the presence of excess β-amylase at 20 ° C at a rate of 1 gm / h α
The amount of amylase.

To measure the viscosity through the funnel, weigh 38 g of Kako starch (anhydrous base) in a tared 250 ml beaker (stainless steel) equipped with a thermometer, add distilled water to make a total of 20 g.
0g. The sample is mixed to dissolve all mass and heated or cooled to 22 ° C (72 ° F). A total of 100 ml of the heat-treated starch dispersion liquid is put into a graduated cylinder for measurement. Then pour into the graduated funnel while closing the hole with your finger. Pour a small amount into a metric glass to remove any trapped air and pour back any remaining residue in the metric glass back into the funnel. A timer is used to record the time required for 100 ml of sample to flow through the top of the funnel.

The funnel is a standard 58 °, thick wall, resistant glass funnel. The diameter of the apex is 9-10 cm and the inside diameter of the barrel is about 0.381 cm. The funnel is calibrated by the above procedure so that 100 ml of water will pass in 6 seconds.

The parameters of the funnel passage viscosity test are carefully adjusted to correlate the degree of starch degradation by β-amylase with viscosity loss by limiting α-amylase contamination. The reducing sugar is measured by the Fering method to confirm the degree of decomposition.

In this example, the desired end point of the enzymatic reaction is obtained within the funnel passage viscosity range of 9-50 seconds. The reducing sugar content in this example is 29-
35%. Corresponding decomposition of starch is 58-
70%. When the desired viscosity is obtained, the enzyme is inactivated by pouring fresh steam into the reaction solution until a temperature of at least 75 ° C is reached and maintained for at least 15 minutes. Then standard # 22 1 1 available from Spraying Systems, Inc. with inlet temperature 200-210 ° C and outlet temperature 85-90 ° C.
Spray dry the batch using a / 4 J nozzle. The spray dried starch product is screened through a # 40 mesh screen.

Example II This example illustrates that the aqueous dispersion of the product prepared in Example I is stable for multiple freeze / thaw cycles, in other words stable for longer storage. This example further illustrates that the β-amylase of the present invention is more stable than non-degraded control starch.

Using the method described in Example I, OSA waxy corn and acid degraded 50WF OSA waxy corn are digested with β-amylase. The β-amylase degraded starch is dispersed at 20% solids in water, along with a control consisting of oxidized OSA waxy corn currently used in beverage flavor emulsions, and placed in a freezer. Freeze dispersion 6 times in one set of tests
Subject to thawing cycle. In this case the frozen part of the cycle
1 to 5 days in 22 days. In another set of tests, the dispersion is exposed to a series of 6 freeze / thaw cycles.
In this case, the frozen portion of the cycle is 1-5 days over a period of 18 days. After each freeze / thaw cycle, use a Bookfield viscometer to measure the viscosity at room temperature (about 22 ° C (72 ° F)).
Measure with 3 spindles at 10 rpms.

Viscosity of control increases and visible floc (floc)
Are formed and then the viscosity decreases. The viscosity of the enzyme-treated sample is low and shows almost no change other than the change inherent in the Hookfield viscometer reading. The flocs formed in the control after one more freeze / thaw cycle are not present in the enzyme treated sample even after six freeze / thaw cycles. Thus, the enzymatically degraded starch composition exhibits greater resistance to aging during temperature changes, i.e. during storage and storage, as compared to control compositions currently used in place of gum arabic in beverages.

Example III This example illustrates that modified starch purified by removal of β-amylase liberated maltose still retains aging resistance during storage.

The β-amylase degraded OSA waxy corn produced by the method described in Example I above is dialyzed to remove maltose. Starch is dispersed in distilled water at 15-20% solids and placed in dialysis tubing (obtained from a Spectropor membrane) that maintains molecules with an excess of 6,000-8,000. The dispersion is dialyzed against distilled water until the dialysate is free of all organic material. The starch is then precipitated with ethanol and collected.

In addition to the maltose-containing sample prepared as in Example I, the maltose-free sample is cooled at 5-7 ° C for 83 days. Samples are taken out periodically, and differential scanning calorimetry (DS
C) Subject to analysis to determine the minimum number of days required for the sample to age. DSC analysis is AC Eliasson
"Aging of Starches Measured by Differential Scanning Calorimetry", Prog, Biotechnol, 1: 93-
8 (1985).

The maltose-free sample shows no aging after storage for 83 days under cooling. Thus, like maltose-containing β-amylase-degraded starch, maltose-free starch exhibits increased resistance to aging compared to the recently used OSA waxy corn emulsifiers. (See Table I). Therefore the presence of maltose is not essential for improving the stability of degraded starch.

Example IV This example illustrates that β-amylase degraded starch exhibits improved aging resistance during storage.

A 50% solids aqueous dispersion is prepared using the following starches: 1) β-amylase degraded OSA waxy corn (Example I above), 2) β-amylase degraded 50WF oxidized waxy corn ( Example II) above, 3) β-amylase degradation produced by the following method,
50WF oxidised waxy corn myristate ester, 4) 50WF oxidised waxy maize control myristart ester, 5) α-amylase modified OSA waxy corn Control, 6) OSA waxy corn control that has been oxidized, and 7) OSA waxy corn control.

All β-amylase degraded samples are prepared by the method described in Example I. All OSA samples are prepared as described in Example I. Millistate ester sample 300 m of water
Slurry 200g of waxy corn which was 50WF oxidized in 1 l, adjust the pH to 8.0 by adding 3% NaOH, N-
Myristyl-N-methylimidazolium chloride 20
Prepared by adding g over 10-15 minutes. The reaction is continued for 2 hours and more water is added so that it can be stirred when the reaction mixture becomes viscous. The reaction mixture is filtered, the starch resuspended, the pH adjusted to 5.5, then filtered, washed with methanol and dried. The starch product contains about 5% by weight of myristart ester.

In addition to the control and the maltose-free sample of Example III above, this starch dispersion is stored under cooling at 5-7 ° C. Periodically samples are evaluated for aging by the DSC analysis method described in Example III.

Table I shows the points that can be detected by DSC analysis.
The number of days required for each starch composition to age is summarized.

The results show that the controls not treated with β-amylase age rapidly, while the treated samples do not age to the point detectable by the 83 day DSC analysis under study.

Example V This example illustrates that various starches and starch derivatives show improved storage stability and, in some cases, improved emulsion forming ability after β-amylase degradation.

A group of starches containing oxidized tapioca, 40WF flow rate corn and hydroxypropyl 80WF oxidized waxy corn derivative with 19% solids and a funnel passage viscosity of 33.6 seconds at 22 ° C (72 ° F) is given in Example I. It is reacted with octenylsuccinic anhydride by the method described to give an OSA derivative. A 5% myristate ester derivative of 50WF waxy corn is prepared by the method described in Example IV.

A mixture of C _16- C ₂₄ hydrocarbon chain ethers of waxy corn starch, 100 g of starch and 3-chloro-2.
It is prepared by mixing 3 g of hydroxypropyl-cocoalkyl-dimethylammonium chloride (Quab 360 [C _16-24 H _25-33 NOCl ₂ ], obtained from Degussa) at pH 11.5-12. The starch mixture is heated to 40 ° C., reacted for 16 hours, then neutralized to pH 5.5-6, washed 3 times with water and dried. This starch ester derivative is subjected to β-amylase degradation in the manner described in Example I and then (except one) spray dried. The exception is that the viscosity of the reaction medium through the funnel is 50.
When the viscosity of a control mixture of C _16- C ₂₄ hydrocarbon chain ethers of WF waxy corn starch was reached, β
-To end the amylase degradation. A control starch ether is prepared by the method described above, where the starting compound is 50WF waxy corn starch which has been oxidized.

All other samples of the respective starch derivative are subjected to oxygenolysis in the manner described in Example I and then spray dried.

A sample of the spray-dried starch derivative (20 g) is mixed with 180 g of distilled water. The dispersion is stirred in a Waring blender for 2 minutes at low speed with a power stat set at 25-30. Add citrus oil mixture (80 g) at slow speed to the center of the vortex. The citrus oil mixture is from the Fritzsche-Dodge
-Olcott) is a mixture of 12 parts of a single fold orange oil obtained from Hercules and 3.4 parts of ester gum # 8 BE obtained from Hercules. Emulsify the starch dispersion and oil for 2 minutes at high speed in a Waring blender. The resulting emulsion is placed in a glass jar, stoppered, placed in a freezer, removed periodically,
Warm up to room temperature. The evaluation consists of visually examining the emulsification via separation, oilification and gelation. All samples are also evaluated after chilled storage except for those that do not form stable emulsions at room temperature.

The control 40WF oxidized OSA corn starch gels at room temperature and does not form an emulsion. Control hydroxypropyl 84 WF oxidised waxy corn and 50 WF oxidised waxy corn control myristate ester immediately form an emulsion that separates at room temperature. Similarly, control oxidised tapioca does not form an emulsion.
50WF Oxidized waxy maize control ethers form inadequate emulsions. It exhibits a pale yellow cream at room temperature and becomes oily after cooling for 1 day.

In contrast, β-amylase-degraded 40WF oxidizer OSA corn starch forms an emulsion that separates only after two freeze / thaw cycles, while β-amylase-degraded oxidizer OSA tapioca starch does not. Form an emulsion that separates after the freeze / thaw cycle. β-amylase degraded hydroxypropyl-OS
A is stable during 7 freeze / thaw cycles. The β-amylase-degraded 5% myristate ester of waxy maize forms an emulsion that is stable during one freeze / thaw cycle and after 3 weeks of cold storage. The waxy maize β-amylase degrading ether forms an emulsion that is stable at room temperature and remains stable after two freeze / thaw cycles for 12 days. All other β-amylase treated samples form emulsions that are stable after 6 months of cold storage. In all cases, the β-amylase degraded sample performed better than its respective control.

The results show that β-amylase degradation of OSA starches such as waxy corn, corn and tapioca derivatives improves the storage stability of these starch-formed oil-in-water emulsions. Also the results obtained are that starch is a hydrophobic / hydrophilic substituent, eg OSA,
Or it is the same only with a hydrophobic substituent, whether derivatized with, for example, myristart. Furthermore, the results show that specific applications require derivatives such as hydroxypropyl OSA waxy maize, and enzymatic degradation of such derivatives results in low temperature stable oil-in-water emulsions.

Example VI This example illustrates that the modified starch of the present invention, like gum arabic, resists changes during storage and is more resistant than the starch-based emulsifiers currently used to emulsify flavor oils in beverages. .

Oxygen Degraded OSA Derivatives of Waxy Maize and 50 WF Oxidized OSA Waxy Maize are prepared by the method described in Example I. The starch composition and control-which contains gum arabic and the oxidized OSA waxy corn recently used in beverage flavor oils-is emulsified in the manner described in Example V.

These emulsions are stored refrigerated for 3 months. Bookfield viscosity is measured at the beginning and then after 3 months. The measurement is to use a # 5 spindle with a viscosity of 1000-50.
Except for the use of 00cps, the procedure is as described in Example II. Good emulsion stability is observed when the enzymatically degraded sample is subsequently visually observed under cooling for 3 months. The viscosity measurements are summarized in Table II.

The results show that the emulsion prepared from the β-amylase-degraded starch composition is as resistant as gum arabic emulsion to viscosity changes on storage, gelling and oiling.

The modified starches according to the invention in aqueous dispersions and in emulsions exhibit improved aging resistance, which is an improvement over the storage stability of currently known starch emulsifiers.

Example VII This example demonstrates that the modified starch of the present invention can be prepared by reversing the sequence of steps described in Example I.

A 28% solids aqueous slurry of waxy corn starch is prepared, the pH is adjusted to 6.0-6.3 by the addition of 3% NaOH and the slurry is jet heat treated at about 149 ° C (300 ° F). Place the heat-treated starch in a constant temperature water bath,
Keep at 55-60 ° C with constant stirring. The barley β-amylase used in Example I is added to the heat-treated starch at a concentration of 1.650 DP ° per 100 g dry weight of starch. The batch of β-amylase used in this example has an α-amylase activity of 0.9 DU / ml. The degree of degradation is followed by the funnel passage viscosity procedure described in Example I.

When the starch reaches a funnel viscosity of 30 seconds, the enzyme is added to 10
The pH is lowered to 3.5-4.0 by the addition of% HCl to inactivate it and the starch is kept at this pH for 30-60 minutes. PH is 3% after deactivation
Adjust to 7.0 by adding NaOH.

The OSA derivative is prepared by thorough mixing with a debranched starch dispersion neutralized with 3 g of octenylsuccinic anhydride per 100 g of dry weight of starch. The reaction can be continued for 4 hours at room temperature with good stirring.

A portion of this starch dispersion is spray dried as described in Example I.

Spray-dried aqueous dispersions of debranched starch from commercially available control starch (oxidized OSA waxy corn derivative) for use as an emulsifier in orange oil emulsions as in Example V It is manufactured from a sample. The emulsion is exposed to at least 8 freeze / thaw cycles for 1-10 days and evaluated as in Example V.

The control starch forms an emulsion covered by a thick orange oil film after one freeze / thaw cycle. In contrast, debranched starch forms an emulsion that is stable for at least 5 freeze / thaw cycles. The spray dried starch sample is stable for 6 cycles. The aqueous starch dispersion sample shows a thin orange oil film on the water surface at the beginning of the sixth cycle.

The emulsions using the modified starches of the present invention-which are produced by enzymatic degradation of starch either before or after the production of starch derivatives-use one of the improved aging resistance and commercially available starch emulsifiers. The storage stability of the prepared emulsion is improved.

Claims (10)

(57) [Claims] 1. A starch derivative containing a hydrophilic group and a hydrophobic group, up to 70% by weight of which the 1,4-α-D-glucochid bond can be cleaved from the non-reducing end of starch. Emulsifying properties of emulsions showing improved stability during storage, resistance to oiling and gelation, which are degraded by in vitro enzymes that are unable to cleave the 1,6-α-D-glucoside bond of starch. Kako starch with. 2. Starch is acid or heat-modified or converted to a water fluidity (WF) of up to about 60 by α-amylase, the in vitro enzyme is β-amylase, and the starch is gelatinized waxy corn. Starch and starch at least octelsuccinic anhydride per dry weight of starch
The chemically modified starch according to claim 1, which is treated with 0.25% to induce. 3. The hydrophobic group comprises an alkyl-, alkenyl-, aralkyl- or aralkenyl group having at least 5 C atoms, and the starch derivative having a hydrophobic group and a hydrophilic group is represented by the following formula: (Wherein R is a group selected from the group of dimethylene- and trimethylene groups, R'is a hydrophobic group consisting of an alkyl-, alkenyl-, aralkyl or aralkenyl group having at least 5 C atoms, and the carboxyl group COOH is The modified starch derivative according to claim 1, which is a hydrophilic group. 4. The following steps: a) yielding a gelatinized starch derivative, which derivative is characterized by respective substituents containing hydrophilic and hydrophobic groups; b) up to 70% by weight of starch derivative. Is capable of cleaving the 1,4-α-D-glucoside bond from the non-reducing end of starch, but is degraded by an in vitro enzyme that cannot cleave the 1,6-α-D-glucoside bond of starch. A method of producing a modified starch having the emulsifying properties of an emulsion which exhibits improved stability during storage, resistance to oiling and resistance to gelation, consisting of maltose. 5. Starch is waxy corn starch, which is acid or heat-modified or converted to a WF of up to about 60 before being treated with an in vitro enzyme by α-amylase, and the starch is dry weight of starch. 5. The method of claim 4, which is induced by treatment with at least 0.25% octenyl succinic anhydride per unit. 6. The in vitro enzyme is β-amylase, which has an enzymatic concentration of 3-10 at an enzyme concentration of 334-1110 degrees of diastase force per 100 g of starch per dry weight in an aqueous dispersion containing up to 33% solids of enzymatic degradation. A process according to claim 4, which is carried out in the pH range and in the temperature range 20-75 ° C. 7. The hydrophobic group of the starch derivative is an alkyl-, alkenyl-, aralkyl- having at least 5 C-atoms.
Alternatively, a starch derivative consisting of an aralkenyl group and containing a hydrophilic group and a hydrophobic group is represented by the following formula (Wherein R is a group selected from the group of dimethylene- and trimethylene groups, R'is a hydrophobic group consisting of an alkyl-, alkenyl-, aralkyl- or aralkenyl group having at least 5 C-atoms, and the carboxyl group COOH is hydrophilic. The method according to claim 4, wherein the substituent is a cation group. 8. The following steps: a) Up to 70% by weight of gelatinized starch can be cleaved from the non-reducing end of starch to the 1,4-α-D-glucoside bond, but to give 1,6 of starch. -Decomposing into maltose by decomposing with an in vitro enzyme that cannot cleave the α-D-glucoside bond, and b) reacting enzymatically-degraded starch with an agent that produces a starch derivative containing a hydrophilic group and a hydrophobic group. A method of producing a modified starch having the emulsifying properties of an emulsion, which exhibits improved stability during storage, resistance to oiling and resistance to gelation, comprising: 9. An industrial or food emulsion containing the modified starch according to claim 1. 10. An industrial or food emulsion containing the modified starch produced according to claim 4.