TWI241347B - A process for producing an aglucone isoflavone enriched vegetable protein whey from a vegetable protein whey and materials produced therefrom - Google Patents

Abstract

An aglucone isoflavone enriched vegetable protein whey, whey protein material, high genistein material, high daidzein material, and aglucone isoflavone material are provided, as well as a process for producing the same from a vegetable protein whey. Isoflavone conjugates in a vegetable protein whey are converted to isoflavone glucosides by treating the whey at a temperature and a pH for a period of time sufficient to effect the conversion. The isoflavone glucosides are converted to aglucone isoflavones by enzymatic reaction to produce an aglucone isoflavone enriched vegetable protein whey. Aglucone isoflavone whey protein material is recovered from the aglucone isoflavone enriched vegetable protein whey. A high genistein content material, a high daidzein content material, and an aglucone isoflavone material are produced from an alcohol extract of the aglucone isoflavone whey protein material.

Description

1241347 A7 B7 Five 'invention description (printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economics Background of the Invention The present invention relates to a vegetable protein milk rich in aglycone isoflavones, a milk protein protein rich in aglycone isoflavones, Base isoflavone substance, high 5,7,4-dihydroxy isoflavone content substance and high '-dihydroxy isoflavone content substance, and a method for producing them from plant protein milk. , Including plant-based protein substances such as U. These compounds include 7,4, dihydroxy isoflavone glycosides, 6 " _ = Ac 7,4'_ dihydroxy isoflavone glycosides, 6, 0] y [al 74, _Dihydroxyisodione sugar #, 7,4'_dihydroxy isoflavones, 5 7 4, trihydroxy isoflavone glycosides, 6 " _Ac 5,7,4, trihydroxy isoflavone glycosides, 6, , _〇Mai 1,7,4'_trihydroxy isoflavone glycofen, 5,7,4, _trihydroxy isoflavone, 6_oxy_7,4 _ monohydroxy isoflavone sugar, 6 " _〇Ac "Methoxy_dihydroxy isoflavone saccharin, 6 ,, 〇〇Mal L methoxy_74, _dihydroxy flavonoid saccharin, 6-methoxy_7,4, _dihydroxy isoflavone, 4 , · Methoxy _ Dihydroxy isoflavones, 4, methoxy_57_ dihydroxy isoflavones methoxy isoflavones, and coumestrol (CD⑽can called. In commercial production ρα < manufacturing, such as plant protein isolates and concentrated : Chuan Lidian: remove these substances. For example, in the preparation of soy protein isolate (Chuan,-Wu Wanzhong 'where the soybeans are extracted with an aqueous vehicle solution, 2 yellow protein is dissolved in the extract together. Protein The self-extract is separated from the acidified single precipitate and separated to form an isolate, leaving a lot of solubilized liptonone (milk. The residue left in the acid-precipitated protein isolate is thoroughly washed and removed. Milk Pulp and cleaning solution are typical: discarded. Isoflavones in plant-based protein milk include isoflavone _ fine nail 4, reduced §. «Ϋ« ι ϋϋ 4—lt 1 · .—.— · —B ^ ν —B ^^ i · 1 (Please read the precautions on the back before filling in this page} μ scale is applicable to China-4- Printed by the Central Consumers Bureau of the Ministry of Economy Staff Consumer Cooperative 1241347 A7 V. Description of the invention (2 glycosides ), Isoflavone conjugates and aglycone isoflavones. Isoflavone glycosides are attached to the compound Isoflavone moiety of glucose molecule. Isoflavone co-prototypes have additional moiety attached to glucose molecule, such as 6 " -〇Ac 5,7 4, _tri-isoflavone with acetate group attached to the 6 position of glucose molecule Ligand isoflavones include isoflavone moieties without linked glucose molecules. Soymilk contains three "family," isoflavone compounds with corresponding glycans, conjugates and ligand members: 5,7,4 , Trihydroxy isoflavones, 74, dihydroxy isoflavones, and 6-methoxy-7,4, dihydroxy isoflavones. The 5 7 4, _ trihydroxy isoflavone family includes the glycocalyx 5,7,4'_trihydroxy isoflavone glycocalyx ', the conjugate 6〃-0Mal 5,7,4'_trihydroxy isoflavone glycocalyx (574 , _Trihydroxy isoflavone glycoside 6 " -malonate) and 6,0Ae 5 7,4, trihydroxy ^ flavonoid glycoside (5,7, V-trihydroxy isoflavone glycoside 6 " _B Acid ester) and ligand 5.7.4, trihydroxy isoflavones. 7,4, dihydroxy isoflavone family includes glycocalyx 7,4'_ diacyl isoflavone sugar taste, a total of 6 〃-〇Mal 7,4, _ diacyl isoflavone glycoside and 6 " -0Ac 7,4'-dihydroxy isoflavone glycocalyx and ligand 7.4, dimeryl isoflavone. 6-methoxy-7,4, dihydroxy isoflavone family includes glycoside 6-methoxy-7,4, dihydroxy isoflavone glycocalyx, conjugate OMal 6 -methoxy-7,4f -dihydroxy Isoflavones: y: and ligand 6_methoxy_7,4f-dihydroxy isoflavones. Although all isoflavones are important for medical evaluation, the ligand is the most important specific isoflavone I. 5,7,4'-trisyl isoflavones and 7,4, -dimeric isoflavones can significantly reduce cardiovascular risk factors. "Effects of plant and mammalian estrogen on blood lipids of female monkeys 'Eave Oak', Vol. 90, page 1259 (October 1994). Also -5- This paper size applies Chinese National Standard (CNS) A4 specification (210x297 mm)

1241347 Α7 Β7 Invention description (3) It is believed to reduce symptoms in women caused by a reduced or altered amount of endogenous estrogen, such as menopause or premenstrual syndromes. Furthermore, it is understood that in plant-based substances such as soybeans The contained isoflavones can inhibit the growth of human cancer cells, such as breast cancer cells and prostate cancer cells, as described in the following article: "Human breast cancer cell growth inhibition of 5, 7, 4'-trihydroxy isoflavones, independent For estrogen receptors and multidrug resistance genes ", by Bitson and Barnes, Newsletter of Biochemical and Imperial Physics Research, Volume 179, Issue 1, pages 661-667, August 30, 1991;" Day 5, "7,4'_trihydroxy isoflavones and 4, -methoxy-5,7-di-free isoflavones inhibit the growth of human prostate cancer cells, but there is no epidermal growth factor receptor glycine autophosphorylation." Pittson and Barnes, Prostate, Vol. 22, 3 3 5 _ 3 4 5 (19 9 3); and "Soybeans Inhibit Breast Tumors in Breast Cancer Model" by Barnes et al. Mutants and defamatory mice, 23 9-25 3 (1 990). As mentioned above, aglycone isoflavones include 7, 4, 2 Isoflavone, 5,7,4, genistein and 6-methoxy-7, 4, Dihydroxyisoflavone of such ligands having the formula: Ministry of Economic Affairs Bureau of Standards printed crack employees consumer cooperatives

Wherein Ri, R2, R3 and R4 can be selected from the group consisting of H, OH * OCH3. This paper size applies to China National Standard (CNS) A4 specification (210 × 297 mm)

1241347 A7

V. Description of the invention (4) 5,7,4'-trihydroxy isoflavones have the above formula, where Ri = 0h, R2 = h, R3 = OH and R4 = 0H, 7,4'_dihydroxy isoflavones With the above formula, where Ri = OH, R2 = H, R3 = H and R4 = 0H, and 6-methoxy_7 4, dihydroxy isoflavones have the above formula, where ri = 0h, r2 = 0ch3 , = R4 = 0H 0 Therefore, the present invention relates to the enrichment of ligands and vegetable protein milks and milk protein materials with these compounds, and particularly high content of 5, 7, 4 triseptyl isoflavones , High 7, V-dihydroxy content substances and aglycone isoflavone substances. The invention also relates to the production of algae-rich plant-based protein milk, algae-rich plant-based milk protein material, high 5,7,4 ^ trihydroxy isoflavone content material, high 7,4, dihydroxy Method of isoflavone substance and aglycone isoflavone substance. The common method for printing and converting plant protein isoflavone conjugates to aglycone isoflavones by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economics is known and provided in co-examined US patent application serial number 08 / 477,102 '1995 Apply on June 7 and be owned by the assignee of this specification. Methods for converting isoflavone glycosides to aglycone isoflavones are also known. The method for converting isoflavone glycosides to aglycone isoflavones in vegetable protein milk is provided in a co-trial application PCT / US / 94/10699, which is owned by the assignee of this specification. Other methods of converting isoflavone glycosides to aglycone isoflavones are also known in the art, as described in Japanese Patent Application No. 2 5 8, 6 6 9 and 0 b at a et al. This method does not provide the conversion of isoflavone conjugates to aglycone isoflavones or a substance with a high 5,7,4'-trihydroxy isoflavone content, a substance with a high 7,4f-dihydroxy isoflavone content or an aglycone isoflavone. Furthermore, these only reach the standard of converted sugar paper which is applicable to the Chinese National Standard (CNS) A4 specifications (210X297 mm), mouth 41347 A7 B7 grams, invention description (5 moderate degree of substitution with ligands, and the time required to substantiate In order to complete this intermediate process, JKmg this method is not required for a large number of commercial operations. This method is aimed at providing plant protein milks rich in aglycone isoflavones and preparation from plant protein milks Their method. A further object of the present invention is to provide a ligand isoflavone milk protein material and a method for preparing them from a plant-based protein milk. A still further object of the present invention is to provide a high 5,7,4, _II Hydroxyl isoflavone content materials and methods for preparing them from plant protein milk slurry. A still further object of the present invention is to provide high 7,4, dihydroxy isoflavone content materials and methods for preparing them from vegetable protein milk slurry. A still further object of the present invention is to provide a ligand isoflavone substance and a method for preparing them from a vegetable protein milk slurry. These and other objects are described in detail in the description below Brief description of the invention. Brief description of the invention. Brief summary of the invention. Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. The present invention is a plant protein milk slurry containing aglycone isoflavones and a plant protein milk slurry containing isoflavone conjugates. Method for phytoprotein milk based on isoflavones. The method includes treating a phytoprotein milk containing isoflavone conjugates at a temperature and a pH for a period sufficient to convert the isoflavone conjugates to isoflavone glycosides. Time. Enzymes are contacted with isoflavone sugars in vegetable protein milk at a temperature and at a pH for a period of time sufficient to convert most isoflavone sugars to aglycone isoflavones. In a specific embodiment of the present invention The isoflavone conjugate is converted into isoflavone glycocalyx, which is made from plant protein milk. Between about 2 ° C and about 1 2 1 ° C -8-This paper size applies Chinese National Standard (CNS) A4 specifications (210X297 mm) 1241347 A7 B7 V. Description of the invention (Processed at a temperature of 6 and at a temperature of about 6 to about 13.7 to 11 ° C. In another specific embodiment of the present invention, Isoflavone sugar Converted into Isoflavones are obtained by contacting isoflavone saccharin with enzymes in a vegetable protein milk slurry at a temperature of about 5. (: to about 75. (: ㈤) and at a temperature between about 3 to about 9. Iso黄 明 共 秘 至 异 黄 嶋 # and the high conversion rate of the isotope to the isoflavones are achieved. In a specific embodiment, at least _isohuang 嗣 conjugates are converted to 黄 yellow _ oath and at least 8 () % Isoflavone sugar is converted into aglycone isoflavones. In another element, the present invention is to prepare aglycone isoflavone milk for aglycone isoflavone milk protein material and a vegetable protein milk containing isoflavone co-consumers. Method for pulping proteinaceous materials. The aglycone flavonoid milky protein material containing protein and aglycone isoflavones is recovered from the agar-rich plant protein milk. In a specific embodiment of the present invention, the aglycone isatimilumin milk polyprotein material is recovered by at least one of ultrafiltration, thermal coagulation, and dehydration. In yet another element, the present invention is a high 5,7,4, _ trihydroxy isoflavone content substance and a 5,7,4'-trihydroxy isoflavone content prepared from a plant protein milk slurry containing an isoflavone conjugate. Material methods. The aglycone isoflavone serum protein material derived from the vegetable protein serum is extracted with an aqueous alcohol extractant to produce an aspirin-containing isoflavone extract. The extract is in contact with the adsorbed substance for a period of time sufficient to separate the extract from the extract with a high 5,7,4'_trihydroxy isoflavone content. u In another element, the high 7,4, _dihydroxy isoflavone-containing substance and the isoflavone conjugate-containing vegetable protein milk of the present invention are high in the content of the extract.

I___-9-This paper size applies the Chinese National Standard (CNS) A4 (210 X 297 mm) 1241347 A7 B7 V. Description of the invention (7) 7,4, Dihydroxy isoflavone content method. A vegetable isoflavone-derived aglucone isoflavone serum protein material was extracted with an aqueous alcohol extractant to produce an aglucone isoflavone-containing extract. The extract is contacted with the adsorbed substance for a period of time sufficient to separate the substance with a high 7,4'-dihydroxy isoflavone content from the extract. In yet another element, the present invention is a ligand isoflavone substance and a method for preparing a ligand isoflavone substance from a vegetable protein milk slurry including an isoflavone conjugate. The aglycone isoflavone serum protein material derived from the vegetable protein milk slurry is extracted with an aqueous alcohol extractant to produce an aglycone isoflavone-rich extract. The extract is concentrated to about 5% to about 30% of its original volume, and the isoflavone substance is precipitated from the extract by adding water to the extract. In a specific embodiment, the substances with a high content of 5, 7, 4, and trihydroxy isoflavones are separated from the ligand isoflavones. The ligand isoflavone substance is dissolved in the alcohol solution, and the alcohol solution is in contact with the adsorbed substance for a period of time sufficient to separate the substances with a high content of 5,7,4, and tribasic isoflavones. Printed by the Consumers' Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs. In another specific embodiment, the substances with high 7,4, _dihydroxy isoflavone content are separated from the aglycone isoflavone substance. The ligand isoflavone substance is solvent-soluble in an aqueous alcohol solution, and the alcohol aqueous solution is brought into contact with the adsorbed substance for a period of time sufficient to separate a substance having a high content of 7,4, dilight isoflavone. Description of a preferred embodiment The starting material of the present invention is a plant-based protein milk slurry, wherein the plant-based protein milk is defined as the remaining amount after removing the plant protein-depleted mass from the plant-based protein extract; Aqueous protein, isoflavones or other water-soluble compounds. In a preferred embodiment, the starting material is soybean milk -10-

1241347 A7 B7 V. Description of the invention (8 oars' because the present invention is particularly suitable for the production of an isoflavone-containing cream from a soybean material, an isoflavone cream protein material, a high 5,7,4'-three Starting material isoflavone content, high 7,4, dihydroxy isoflavone content material and aglycone isoflavone material. The plant protein milk starter contains isoflavone conjugate, isoflavone saccharin and aglycone isoflavone. For example, soymilk contains isoflavone sugar 甞 5,7,4, _ dihydroxy isoflavone sugar: y :, 7, 4, 4-dihydroxy isoflavone sugar 甞 and 6_methoxy _ 7'4- Isoflavone sugar replacement per base; isoflavone conjugates_5,7,4'-trisyl isoflavone glycosides, 7,4'-dihydroxy isoflavone glycosides and 6_methoxy_7, 6〃-malonic acid esters of 4, _ monohydroxy isoflavone glycosides and 5,7,4,6〃-acetates of trihydroxy isoflavone glycosides and 7,4, dibenzyl isoflavone glycosides; and Ligand isoflavones_5,7,4, trihydroxy isoflavones, 7,4, dihydroxy isoflavones, and 6-methoxy 7,4, dihydroxy isoflavones. Isoflavones in soy milk starting materials Isoflavone conjugates are advantageous. Member of the Central Standards Bureau of the Ministry of Economic Affairs Printed by industrial and consumer cooperatives, plant protein milk starting materials are typically obtained as by-products of the traditional plant protein isolate production process. Plant protein sources, such as soy flakes, from which the oil has been extracted by falling agents, can be extracted by water Co-treatment with agents, = has a pH higher than the isoelectric point of the protein of vegetable protein source to produce each protein, isoflavones, and extracts of other compounds dissolved from the extract by the vegetable protein source. The extract is insoluble in the extract Liquid vegetative nature: two separate. The solubilized protein and isoflavone-containing extracts from the imprinting point to about the isoelectric point of the protein, soy protein about (4-4.6, egg :: liquid precipitates the protein. The precipitated protein is separated The production of plant-based shellfish is a separate matter, and the starting of the plant-based milk is ⑼ big _ isoflavones 1241347 A7 __________ B7 V. Description of the invention (9) Keep 'grain in the gift slurry. For the best in the slurry Isoflavone recovery, additional cleaning of precipitated protein I may be required, and each cleaning solution is added to the slurry. The vegetable protein slurry starting material can be spray-dried in water Plant & protein% gift pulp. For each treatment, the plant protein milk can be spray-dried to recover milk protein (still soluble in milk < protein, after Shendian protein isolate), isoflavones and others The compound is a solid substance. The spray-dried shellfish can be added to water to reduce the plant protein milk slurry starting material. In a more specific embodiment, the slurry contains about 2-10 grams of spray-dried material per 100 grams of water to confirm that The gift set is not too thick, but provides enough isoflavones to produce the desired isoflavone-containing isoflavone-containing milk, aglycone isoflavone milk protein material, high 5'7,4-dihydroxy isoflavone content material, High 7,4, -dihydroxy isoflavone content substances and aglycone isoflavone substances. In the first transformation step or operation, the isoflavone conjugate is converted to isoflavone glycocalyx in the plant protein milk slurry starting $ to produce isoflavone glycocalyx-rich plant protein milk Pulp. Conversion was found to depend on the pH and temperature of the slurry. Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. The pH range for converting isoflavone conjugates to isoflavone glycocalyx is from about 6 to about 13.5. The ρ η of the vegetable protein milk should be adjusted to the desired pH when needed. Huang Yu protein milk paddles typically have a pH of about 4.4-4.6 and should be adjusted to the desired pH range with an alkali or alkaline reagent. The pH can be adjusted with any suitable alkali, caustic or alkaline reagent, which will increase the pH of the system, including sodium oxychloride, potassium hydroxide and calcium hydroxide. The transformation was found to proceed more easily under extremely alkaline conditions, preferably 9-1 1. The pH should be maintained below pH 12, because isoflavone glycosides 7,4'-trihydroxy isoflavones: y :, 7, dihydroxy

1241347 A7 B7 V. Description of the invention (10-based isoflavone glycoside and 6-methoxy ^ H 7,4 _ dihydroxy isoflavone glycocalyx, in particular 7,4'-dihydroxy isoflavone glycoside -1 m ^ Degradation tends to be at pH 12 and above. The reaction is less likely at lower pH conditions and easier to carry out, such as about pH 6, however, the reaction will be performed at higher temperatures and / or under increased pressure. The temperature range for the conversion of isoflavone conjugates or isoflavone glycocalyx is from about 2X: to about 12 1. The temperature range in which conversion is liable to occur depends on the pH of the slurry. The inventors have found that the conversion occurs at extremely high pH It is easy to carry out at lower temperatures. For example, at a pH of the slurry of about u, the conversion is easily and efficiently performed at a temperature range of about 5x: to about 50x: at a slurry of about 9? 11, The conversion occurs efficiently at a temperature range of about 4 5 C to about 7 3 C. When the pH of the slurry is extremely low, the conversion occurs at a higher temperature. For example, at a pH of 6 of the slurry, the conversion occurs at about 8 〇. (: To a temperature range of about i 2 丨 t. In a preferred embodiment, the pH of the slurry is about 丨 丨 at about 35 Completed. In another preferred embodiment, the transformation is completed at a pH of about 7 3 at a pH of about 9. The time period required to substantially complete the conversion of the isoflavone conjugate to the isoxone glycocalyx is plant-dependent The temperature and temperature of the protein emulsion depend on the temperature. The time period ranges from 15 minutes to 24 hours. The transformation occurs more easily at higher pH and at higher temperature. At a pH of about 9-10, the transformation is at 73 ° Substantially complete in about 4 to about 6 hours at C. At a pH of about 10-11, conversion is substantially complete in about 30 minutes to about 30 hours at 35 ° C. In the best specific embodiment The isoflavone conjugate is converted to isoflavone sugars at a pH of about 11 and at a temperature of about 35 ° C in about 45 minutes: y: The first conversion step is quite effective. From about 80% to about -13- This paper size applies to the Chinese national standard (Cns) A4 specification (21〇 × 297 mm) Please read the note first Η if printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 1241347 A7 B7 V. Description of the invention (12 ^ 佳 <In the specific embodiment, supplementary enzymes are added to human milk, and $ 疋 α 'is sufficient Retaining enzymes are present in the milk because supplementation and dramatic reductions are required to substantially complete the conversion of sugar to phytophysin :: plus: Enzymes are based on optimal activity under selected pH and temperature conditions. Supplementing enzymes are capable of cleavage Isoflavones and isoflavones. The enzymes of the bond between Shaofen, such as sugar enzymes that can cleave sugar vow bonds. The supplementary enzymes are commercial sources. Β-galactosidase enzyme, glucoamylase enzyme and pectinase enzyme. Particularly preferred are enzymes such as biological pectinase 100L (which is preferably used in a pH range from about 3 to about 6) ), Biological pectinase 300L (optimal pH range from about 3 to about 6), biological pectinase OK 70L (optimal pH range from about 3 to about 6), biological lactase 3,0. 〇〇 (optimum 1) 11 ranges from about 3 to about 6) and neutral lactase (optimal pH Fanyuan from about 6 to about 8), all of which are obtained from Discovery International, 1 8 3 3 5 7th Street, mailbox 3 9 1 7, Sarasota, Florida 3 4243. Also particularly preferred are lactose F (optimal pH range from about 4 to about 6) and lactase 5 0,000 (optimal pH range from about 4 to about 6), which are obtained from Oman International Enzyme Company, mailbox 1000, TOY, Virginia 22974. Other particularly good supplementary enzymes include G_Zyme G990 (optimum p 约 from about 4 to about 6) and Enzec 〇 眞 bacteria lactase enzyme concentrate (optimal p Η range from about 4 to about 6), obtained from the enzyme development company, No. 2 Bin Plaza, Room 2 4 3 9, New York City, New York 1 〇 1 2 1 ϊ Lactozyme 3 0 0 0 L (optimal pH range from about 6 to about 8) and a-Gal 600L (optimal pH from about 4 to about 6.5) Obtained from Novo Nordisk Bio-Industrial Corporation, No. 33, Janna Road, Danbourg, Connecticut 06 8 1 3; Neutral lactase (best pH range Fan-15-This paper size applies to Chinese national standards (CNS ) A4 size (21 × 297 mm) 1241347 A7 B7 V. Description of the invention (13) from about 6 to about 8), obtained from Pfizer Food Science Group, 205 East 42nd Street, Wuyue City, New York 10017; and Maxilact L2000 (the optimal pH range is from about 4 to about 6), obtained from Gist Br cades component company, King of Prussia, Pennsylvania 1940. Once a sufficient concentration of enzyme is present, or from residual enzymes, supplementary enzymes, or both, the enzyme and the isoflavone core are contacted in the milk at a pH and temperature for a period sufficient to convert at least most and preferably substantially heterozygous Time of flavonoid saccharin to aglycone isoflavone. When needed, the pH of the isoflavone glycocalyx-rich milk should be adjusted to a pH range where the enzyme reacts with the isoflavone glycocalyx. Within this pH range, the combined residual and supplementary enzymes react with the isoflavone glycocalyx from about 3 to about 9. The inventors have found that the enzymes remaining in the milk are effective in a pH range of about 7 to about 9, because of course, the pH of the milk is low during the reaction process. Supplementary enzymes are effective in the optimal pH range specified by enzyme manufacturers, as mentioned above for some specific enzymes. Typically, supplemental enzymes are effective in the neutral pH range from about 6 to about 8, or in the acidic pH range from about 4 to about 6. Acid enzymes have also been shown to be effective at a pH of about 3. The pH of the cream produced by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs can be adjusted. In most cases, the pH from the first step is extremely high or the pH is reduced. Acid, hydrochloric acid or any suitable reagent. Preferably, the reagents used will be food grade reagents or acids. The temperature range for conversion of isoflavone glycosides to aglycone isoflavones ranges from about 5 t to about 7 5 C. Temperature significantly affects enzyme activity and therefore conversion. Supplement enzymes can be effective above 72.5 ° C, for example, a-Gal 600L is effective at 75 -16-

1241347 Α7 Β7 V. Description of the invention (14) c 'However, it is preferable to perform transformation at a lower temperature to prevent deactivation of the enzyme. In a preferred embodiment, the conversion is between about 35 and about 45 π. Preferably, the 'enzyme reaction is performed at the same temperature as in the first conversion step, thereby eliminating the need to change the temperature of the slurry after the first conversion step. Optimally, both the second transformation step and the first transformation step are performed at 3 5c. Also preferably, the fixed temperature is maintained throughout the conversion of the isoflavone saccharin to aglycone isoflavone. However, in some cases, it may be necessary to raise, lower, or change the temperature during the reaction. The time required for the two transformation steps depends on the factors related to the enzyme, in particular; the temperature and the temperature and pΗ of the milk. In most cases, complete conversion is likely to be achieved within 24 hours, however, it is preferred that supplementary enzymes be added to dramatically increase the reaction rate. The selected supplementary enzyme, enzyme concentration, pH, and temperature preferably result in substantially complete conversion within 2 hours, and optimally within 1 hour. Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs The degree of conversion of isoflavone glycosides to aglycone isoflavones in the second conversion step is significant, typically from at least about 80% to 100%. Conversion of at least 95% isoglucanose to aglycone isoflavones is usually achieved. After the isoglucan sugar is converted to aglycone isoflavones, the aglycone isoflavone-rich milk can be used as desired 'without drying or removing proteins in the milk' or alternatively aglycone isoflavone milk proteins The material may be recovered to concentrate aglycone isoflavones in the protein material. Aglycone isoflavone milk protein material, as used herein, is defined as protein-containing, aglycone isoflavones. ·· 17_ This paper size applies the Chinese National Standard (CNS) A4 specification (21 〇 × 297 mm) 1241347 A7 V. Description of the invention (15) Ketones and residual plant compound I substances that can be precipitated and separated from vegetable protein milk slurry.配 Protein materials containing aglycone isoflavones can be recovered by conventional procedures such as ultrafiltration, thermal coagulation, and dehydration. The resulting aglycone isoflavone serum protein material can be dewatered and dried by conventional methods. The aglycone isoflavone serum protein material can also be recovered from the cooling slurry from the slurry. The aglycone isoflavone milk protein material is insoluble in the cooled milk and the precipitate can be separated from the milk by centrifugal cooling. Preferably, the slurry is cooled to about 4 ° C to precipitate proteinaceous matter. In a preferred embodiment, the slurry is concentrated to enhance the recovery of proteinaceous material of the isoflavone slurry. It was found that increasing the concentration of solids to liquid ratio in the milk by concentrated milk increased the capture of aglycone isoflavone milk protein protein from the milk. The slurry can be heated and concentrated under reduced pressure or both. Preferably, the slurry is concentrated to a solid to liquid ratio of about 1: 3 to about 1: 6, most preferably about 1: 3. Substances with a high 5,7,4, trihydroxy isoflavone content and substances with a high 74, dihydroxy isoflavone content can be produced from the recovered isoflavone serum protein material. As used herein, substances with a high 5,7,4, trihydroxy isoflavone content are defined as containing up to 40% 5,7,4, trisyl isoflavones and optimally at least 90% 5,7 4, _ The plant-based substance of Sanjingji Isoflavones and residual plant substances printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs. , Residual soy material. High 7,4, dihydroxy isoflavone content substances contain at least 40% 7,4, dihydroxy isoflavones and residual plant matter. In order to produce high 5,7,4'-trihydroxy isoflavones and high 7,4, dihydroxy isoflavin content substances, the aglycone isoflavone milk protein material was initially washed to remove ______18_ This paper is again suitable for China Standard (CNS) A4 specification (2 丨 0 '乂 297 mm)' — 1241347 A7 B7 V. Description of the invention (16 Employees' cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs print undesired salts and sugars and then dry them. For cleaning Base isoflavone serum protein material, the material is diluted with water, preferably to about 10/0 solids to about 60/0 solids, and most preferably to about 2% solids. The washing water can be at any temperature However, it is preferably between about 25 ° F. and about 75 ° C., and most preferably about 60 ° C. After washing the isoflavone_milk protein material, the material is separated from the cleaning solution and dried. In a preferred embodiment, the substance is separated by centrifuging the substance and the supernatant from which the substance is poured. The aglycone isoflavone serum protein material can then be extracted with an aqueous alcohol extractant to remove the aglycone isoflavones from the milk protein and produce a rich formula Isoflavone extracts. Low molecular weight alcohols such as methanol In particular, ethanol is preferred as the alcohol component of the extractant. Aglycone isoflavones have been found to dissolve in almost all alcohol concentration extractants. Aglycone isoflavones are particularly soluble in the extractant and contain about 30% alcohol to about 90 % Alcohol, most preferably between about 600% methanol and about 80% alcohol. Although aqueous alcohols are the preferred solvents, other solvents include water, acetonitrile, dichloromethane, acetone, and ethyl acetate and these The solvent mixture can be used to complete the extraction of aglycone isoflavones from the serum protein material. The extraction is performed using a minimum amount of extractant. Preferably, the weight ratio of the extractant to the aglycone isoflavones protein material does not exceed u : 丨. In a specific embodiment, the substance is extracted using a countercurrent extraction process, wherein the weight ratio of the extractant to the substance moxibustion is between about 6: 1 to about 8: 丨. In another embodiment, the substance can be extracted in 2 parts Extraction, where the combined weight ratio of extractant to substance 値 does not exceed 1 i: i. Although the extraction can be performed at any pH, it is preferred that the extractant has about the isoelectric point of the protein in the isoflavone slurry protein material ipH. with minimal (Please read the notes on the back before filling this page)

, 1T • i ^ n In m n-I—- 1 ------ -19- 1241347 A7 A ~ — ---- ~ — B7 5. Description of the invention (17) The solubility of the protein in the extractant. Preferably, the extractant has between about 3 to about 6 &lt; PH &gt; and most preferably about 45, in the milk protein protein. % Extraction can be performed at any temperature up to the boiling point of the extractant, and preferably between about 25 ° C and about 70 ° C. In order to reduce the time required for the extraction of aglycone isoflavones from the protein substance of the aglycone isoflavone slurry, it is preferable to perform the extraction at a temperature of about 60 ° C, preferably at about 60 ° C. After extraction, substances with high content of 5,7,4'-trihydroxy isoflavones and substances with high content of 7,4 and dihydroxy isoflavones can be separated from the extraction solution rich in aglycone isoflavones. Time sufficient to separate 5,7, V-trihydroxy isoflavones and 74, dihydroxy isoflavones from the extract. In a preferred embodiment, the substances with high 5,7,4, trihydroxy isoflavones and high 7, V-dihydroxy isoflavones are self-extracted and separated by reverse phase high performance liquid chromatography ("HPLC"). . 5,7,4, trihydroxy isoflavones and 7,4, dihydroxy isoflavones are separated from other isoflavones and impurities in the extraction solution, and the particles of the adsorbed substance are dissolved by the extraction solution, which are releasably specified by a compound The method combines 5,7,4, trihydroxy isoflavones, 74, -dityl isoflavones, other isoflavones and impurities, so that the compounds can be separated. Printed by the Consumers' Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs, the aspirin-rich isoflavone-rich extract was filtered to remove insoluble materials and connected to HPLC columns. HPLC columns are prepared by filling traditional commercial-obtained HPL C columns with granular adsorbed substances, which releasably bind 5,7,4, trihydroxy isoflavones, 74 \ dionyl isoflavones, and other isomeric compounds in a specific way. Flavonoids and impurities. The adsorbent can be any reverse-phase HPLC filling material. However, the preferred filling material can be loaded with capacity and separation efficiency. -20- This paper size applies to China National Standard (CNS) A4 (210X 297 mm). 1241347 Central Bureau of Standards, Ministry of Economic Affairs Printed by employee consumer cooperative A7 B7 V. Description of invention (18 Force and cost criteria selected.-A better filling material of this kind is Kromasil C18 16 micron 100 Angstrom beads, obtained from Ika. Nobel, Nobel Industries, Sweden The filtered extract was passed through a packed HPLC column until the binding sites of all the columns were completely saturated with isoflavones, which was detected by the appearance of flavonoids in the effluent from the column. The HPLC column can then be eluted with a polar eluent to The separation is completed. In a preferred embodiment, the eluate is an aqueous alcohol. The aqueous alcohol eluate may have an alcohol content between about 30% and about 90% alcohol, and preferably an alcohol content of about 50% alcohol In order to provide good separation and good solubility of isoflavones, the alcohol is preferably methanol or ethanol, of which ethanol is preferred, when high 5,7,4, -trihydroxy isoflavones or high 7 4, · dihydroxy Isoflavones Content product substances are to be used in food or pharmaceutical applications. High 5,7,4'-trihydroxy isoflavones and high 74, dihydroxy isoflavone content substances are collected from the column effluent. Outflow containing 7, dihydroxy isoflavones The liquid separation was first separated from the column, followed by the 6-methoxy-7,4, _dihydroxy isoflavone separation solution, followed by the more polar 5,7,4, and trihydroxy isoflavone separation solution. 7,4 '-Dihydroxy isoflavones and 5,7,4, _trihydroxy isoflavone fractions are collected as they elute from the column. 6-methoxy-7,4, dihydroxy isoflavone fractions are also available when desired. Can be collected. The alcohol in the separation liquid can be removed by evaporation, and then high 5,7,4 乂 trihydroxy isoflavone 1 and high 7,4'-dihydroxy isoflavone content and high 6_methoxy-7, 4. A substance with an isoflavone content can be recovered by traditional separation methods such as centrifugation or filtration. The recovered content of 5,7,4 trihydroxy isoflavones contains at least 40./05,7,4, trihydroxy isoflavones. And preferably at least 90% 5 7 4, 3-21-μ's scale is applicable to Chinese National Standard (CNS) a4 specifications (210X297 mm)

1241347 A7

V. Description of the invention (19) Hydroxyl isoflavones, together with residual plant-based substances, are residual soy substances when 5,7,4, and trihydroxy isoflavones are recovered from soybean milk. Recovered &lt; high 7,4 '-dilight isoflavone content material contains at least 40% 7,4, dihydroxyisogonone, which, along with residual plant matter, typically includes a significant amount of methoxy-7 , 4, Erweiji different yellow _. In another specific embodiment of the present invention, the aglycone isoflavone substance is produced from the S-containing aglycone isoflavone extract. As used herein, aglycone isoflavone substances are defined as those containing at least 10% 5,7,4 trihydroxy isoflavones and at least 5% 7,4,2,2-derived isoflavones and other isoflavones and residual plant compounds substance. After the extraction of aglycone isoflavone proteinaceous material, the aglycone isoflavone-rich extract can be concentrated to promote the precipitation of aglycone isoflavone from the extract. The extract can be concentrated by heating the extract, placing the extract under reduced pressure, or both. In a preferred embodiment, the extract is concentrated to between about 15% and about 30% of its original volume. Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs. The aspirin isoflavones are extracted from the extraction solution by adding water to the extraction solution. In a preferred embodiment, between about 6 to about 8 parts of water is added to each concentrated extract. When adding water to the extract, some aglycone isoflavone substances passed through Shen Dian. To maximize the recovery of the isoflavone material from the extract, the extract and water are thoroughly mixed and then cooled. The extract and water are mixed together for a period of time, preferably from about 30 minutes to about 1 hour. In a preferred embodiment, the extract and water are mixed at a temperature between about 50 ° C and about 75 ° C, and most preferably about 70 ° C. After the water and the extraction liquid are thoroughly mixed, the mixed liquid is cooled to -22- This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) 1241347 Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs A7 B7 V. Description of the invention (20) Ligand isoflavone substance is precipitated. Preferably, the extract / water mixture is cooled to about 5 ° C to about 20 ° C, and most preferably to about 10 ° C, for a period of time that is sufficient for Shen Dian to have all the isoflavone substances. The precipitated isoflavones can then be separated from the extract / water mixture in a conventional manner such as by centrifugation or filtration. The separated aglycone isoflavone material can then be washed with water. In a preferred embodiment, the isoflavone material is washed with water at a temperature of about 70 ° C. for about 5 minutes, wherein the weight ratio of the water cleaning solution to the material is about 0: 8 to about 2: 1 between. The aglycone isoflavone material is separated and dried from the cleaning solution by conventional methods such as filtration or centrifugation. The recovered isoflavone material typically contains at least 20% 5,7,4'-trisyl isoflavones and at least ι0% 7,4'-dimethy isoflavones, and the remaining content of the material is determined by the residual Plant-based substances are formed, including other aglycone isoflavones. Residual plant matter is a soy substance when the aglucone isoflavone substance is separated from the soybean milk. The recovered aglycone isoflavone material may be further purified to produce at least 40% 5,7,4'-tri # isoflavones and preferably at least 900/05, 7, 4, 4 The base isoflavones have a high content of 5,7,4 '· trihydroxy isoflavones and a content of 7,4f-dihydroxy isoflavones with a high content of 7,4 and dihydroxy isoflavones. The ligand isoflavone substance is solvent-soluble in an aqueous alcohol solvent. Low molecular weight alcohols are preferred as the alcohol component of the solvent, with ethanol being the most preferred for food and pharmaceutical applications because of its low toxicity. The alcohol content of the solvent is preferably between about 30% and about 90%, with an alcohol content of about 80% being optimally about to provide good solvent fusion of the isoflavone material. The aqueous alcohol solution containing the falling agent isopropyl alcohol isoflavone substance can be in contact with the adsorbing substance for a period of time sufficient to separate 5, 7, 4, and trihydroxy isoflavones from the aqueous alcohol solution (please read the precautions on the back before filling this page} 'Booklet-23-

1241347 A7--'~ -__ B7 V. The description of the invention (21) and the time of 7, 4, and ditriflavone content. In a preferred embodiment, the 5,7,4, trihydroxy isoflavones and the high 7,4, dihydroxy isoflavones are separated from the aqueous alcohol solution by the reverse phase H p lc. Η The PLC column system was prepared as before, and the aqueous alcohol solution containing the ligand isoflavone substance was loaded into the column and the horse's 5,7,4 '· tri-light isoflavone content substance and high 7,4, 2-side isoflavone The content material was eluted from the column in the manner described above. A high 5,7,4, trihydroxy isoflavone content substance contains at least 40% trihydroxy isoflavones, preferably at least 90% trihydroxy isoflavones, together with the remaining plant-derived substances, which are at 5.7.4, When trihydroxy isoflavones are recovered from soy milk, they are deficient in the residual xanthate.咼 7,4’_dijingyl isoflavone content substance contains at least 40% 7,4, _diweiyl isoflavone g, same as the remaining plant-based substances. Experiments The present invention is explained in more detail by the following examples, using soybean milk as a vegetable protein milk. The examples are intended to be illustrative and should not be construed to limit or obstruct the scope of the invention in any way. Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs as mentioned above. Soymilk includes 5,7,4, trihydroxy isoflavones, 7, 4, dihydroxy isoflavones, and 6-methoxy-7,4, dihydroxy groups. Isoflavones ", the isoflavones' have corresponding sugar tastes, conjugates and ligand members, of which the 5,7,4, _ trihydroxy isoflavone family includes the conjugate 6 &quot; -〇Mal 5,7 , 4, · tritrisyl isoflavone glycosides and 6〃-0 Ac 5,7,4, trihydroxy isoflavone glycosides, glycosides 5.7.4, trihydroxy isoflavone glycosides and ligands 5,7,4 ' -Trihydroxy isoflavones; 7.4, Dihydroxy isoflavone family conjugates 6 &quot; -01 ^ &amp; 17,4, diprotection isoflavone glycosides and 6 &quot; -OAc 7,4 ^ dihydroxy isoflavone glycosides , Sugar: y: 7,4, dihydroxy isoflavone glycosides and ligands 7,4 ^ dihydroxy isoflavones; and 6_methoxy-24. This paper size applies Chinese National Standard (CNS) A4 specification (210X297 mm) ) 1241347 A7 B7 V. Explanation of the invention (22) ^^^-Dihydroxy isoflavone family conjugates ^^ Laos "monomethoxy-7,4'-dihydroxy isoflavone glycocalyx, glycocalyx 6- Methoxy_7,4, _ diamyl isoflavone glycosides and ligand 6-methoxy -7,4'-dihydroxy isoflavone. In the following table, the relative concentrations of isoflavones are determined as a percentage of the isoflavone family. For example, in the 5,7,4'_trijingyl isoflavones | family: 〇 / 〇5,7,4, _trijingyl isoflavone sugar 甞 +% 6〃-0Mal 5,7,4'_ Trihydroxy isoflavone sugar taste +% 6 ,, _ OAc 5,7,4'-trihydroxy isoflavone glycocalyx +% 5,7,4, _trilight isoflavone = 100%. The degree to which the conjugate is converted to glycocalyx and glycoside to formulate a compound can be determined by comparing the percentage of each type of compound in the isoflavone family. Example 1 In the first experiment, the conversion of isoflavone conjugates to isoflavone glycofen was examined. The degree of conversion is determined by a quantitative decrease in the percentage of malonate and acetate acetate of the isoflavone family and a corresponding increase in the corresponding sugar percentage of the same isoflavone family. Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. In the first step, the effects of different conditions in the conversion of isoflavone conjugates to isoflavone glycocalyxes were measured at two different temperatures. The spray-dried soymilk slurry was slurried in water to form a 2% heavy solids soymilk suspension. Soymilk was divided into 2 groups of 4 samples. The samples of each group were adjusted to p H 6 · 0, 7 · 0, 9.0, and 1 1 · 0. The sample group was incubated for 24 hours, while the group sample was incubated at 45 ° C and the other group was at 7 2.5. (: Subculture. Weekly analysis was performed on each sample at 0.2, 4, 6, 8, and 24 hours to determine the isoflavone content of the sample. Table 1 below illustrates the changes and Distribution. -25 This paper size applies to Chinese National Standard (CNS) A4 (210X297 mm) 1241347 A7 B7 Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs

This paper size applies to China National Standard (CNS) A4 (210X297 mm) 1241347 A7 B7 V. Description of the invention (24) e-cm β »Mevacyl πανϋ. XK4f-Three 5J 糸 Three Articles Three and 1 XI Articles m ΊΑ 二 ... formyl

, Super-based iso-rich base, real-based base, iso-based base, iso-based base, iso-based base, iso-based base, iso-based base, iso-based base, iso-based base, iso-based base, iso-based base, etc. -7甞 《I 甞 S ^ l # Huang Hu's job title is different ★ Hu quilt base is different from W page fraction '(Please read the precautions on the back before filling this page) Lu t 711 * C t-0 16 6S 0 19 15 65 2 18 32 33 36 t-2 hours 33 41 0 19 33 48 2 17 39 27 34 hours 43 39 0 17 42 39 2 17 A6 2! 33 iH, hours 51 33 0 17 49 32 3 17 50 19 31 Hours Η 21 0 16 54 27 3 16 57 14 29 hours ao 5 0 15 77 5 3 16 66 0 34 'order t-0 16 65 0 19 15 65 2 18 32 32 30 ** 2 hours 50 X 0 16 50 ^ 4 0 15 50 20 30 μh 57 27 0 15 57 27 0 16 A9 17 34 t-6 hours 62 23 0 15 62 23 0 IS 54 13 34 hours 67 19 0! 4 67 11 0 15 57 10 34 24 hours 70! 7 0 13 63 17 0 20 50 10 39 t-0 16 65 0 19 13 65 2 18 32 33 36 r-2 hours 15 0 0 15 η 0 0 It 62 0 3t t—small 4 0 0 15 SI 0 0 19 63 0 37 t- «hour 16 0 0 Μ 79 0 0 21 61 0 39 tl hour t7 0 0 13 77 0 0 23 60 0 40 t-24 hours 90 0 0 10 53 0 0 47 46 0 54 pH 7 η 1-0 16 65 0 19 15 65 2 II 32 33 36 * • 2 hours 41 43 0 17 41 39 2 17 47 25 29 hours 52 • 32 0 1 $ 50 32 2 16 49 IS 33 hours 5t 27 0 IS 56 26 2 16 3. 13 35 Bu 8 small 4 64 21 0 IS H 20 2 16 55 12 32 1-24 small "59 4 0 31 61 3 0 36 50 0 50 -27- This paper size applies to Chinese national standards (CNS) Α4 specification (210 × 297 mm) Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs mLmx 1-0 16 65 0 19 15 65 2 It 32 33 36 1.2 hours 13 4 0 13 12 4 0 14 64 0 36 H hours II 2 0 1! Μ 2 0 13 65 0 35 * · M, hours 90 0 0 10 IS 0 0 15 65 0 33 1 hour-91 0 0 9 hemp 5 0 0 IS 65 0 35 * 24 small 4 100 0 0 0 IS 0 0 13 tuo 0 0 1241347 A7 B7 V. Description of the invention (25) 5,7,4-two 5,7, winter two 5,7, year-two super 5,7,4 · 7, 4- ^ ¾ 7,-^ 0-¾ 74r ~ ji. -74f ^ zJ »'6- ^ 5 A / cm» λ Dakou Super i-Iso- ▲ Iso-huang dimedi' Iso-qi 'Iso- yellow & Different: Huang / ΪBased Isoisocyanate-7 ， · ^ · 二 1¾ ^ -7 Surgery 2 ^ 00 黄 9 ^ | # X 明 Ϋ # 明 阴 Ϋ: 异 黄 W Ming Shao Ming Yan隹 Huang Hu Snail ^: Yi 'Huang said to be poor ^ Different page fraction, t-2 hours 8 6 r »4 hours 87 t» 6 hours Π t-β hours 88 t-24 hours 78 65 0 0 0 0 19 14 13 13 12 22 76 72 67 65 0 0 0 0 24 28 33 39 76 32 57 54 48 33 0 0 0 0 0 36 43 46 49 52 Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs as described by 6 &quot;-〇 M al and 6 &quot;-0 A c The relative concentration of isoflavone conjugate compounds is reduced and The corresponding increase in the glycosides 5,7,4, trihydroxy isoflavone glycocalyx, 7,4f-dihydroxy isoflavone saccharide and 6-methoxy 7, V-dihydroxy isoflavone glycoside are shown in the first This transformation step is the fastest and most complete at higher, more experimental ρΗ conditions and higher temperatures. Substantially complete conversion of the isoflavone conjugate into isoflavone sugars occurs at pH 9 and 11 samples at both 45 ° C and 72.5 ° C. Conversions were also performed to near complete at pH 6 and 7 samples at 72.5 ° C. Example 2 In a second experiment, the isoflavone glycosides were converted into aglycone isoflavones and examined. The isoflavone saccharin rich milk produced from the first transformation step was used to examine the second transformation step. The degree of conversion is determined by the quantitative decrease of the percentage of glycosides of the isoflavone family, and the corresponding quantitative increase of the percentage of ligands coupled to the same isoflavone family. Soybean milk is transformed into isoflavone glycoside-rich milk. The pH of the milk is adjusted to 1 1.0 and incubated at 35 ° C for 30 minutes. A sugar-rich milk sample was cultured at 45 ° C for 24 hours to determine the isoflavone glycofen that was converted to aglycone isoflavones and the enzymes remaining in the milk. Other syrup rich in sugar Ru-28 This paper size applies to Chinese National Standard (CNS) A4 specification (210X 297 mm) —Ini #! (Please read the notes on the back before filling this page) Order 1241347 A7 B7 5 2. Description of the invention (26 samples were inoculated with supplemental enzymes from the following commercial sources: biological pectinase 100L, biological pectinase 300L, biological pectinase 0K70L, lactase F, α-Gal 600L, G-Zyme G990, exploring organisms Lactase 3 0,000 &gt; Novo Lactozyme 3 000L &gt; Maxilact L2000 E nzec 〇 lactase, P fizer neutral lactase and explore neutral lactase. With oc-Gal 600L, G-Zyme G990, bio pectinase 100L, biological pectinase 300L, biological pectinase OK70L, lactase ρ and Enzeco bacterial lactase inoculated samples pH adjusted to pH 4.5 before inoculation. Novo Lactozyme 3 000L, Maxilact L2000, Pfizer neutral lactase, Explore biological lactase 30,000 and neutral lactase inoculated samples, adjust pH to pH 4.5 and 7.0 before inoculation. Supplementary enzyme samples are then cultured at 50% (except lactase F Sample at 35 ° C Samples of nutrient and biological pectinase 300L and biological pectinase OK70L were cultured at 40 ° C. The samples were taken out after a period of time and the isoflavone content was measured. The following Table 2 shows the distribution of isoflavones during the experiment. (Please read the notes on the back before filling this page) * Order——Γ. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs -J— · I =--I ϋ— -29 (CNS) A4 specification G χ 297 Male #) 1241347 Α7Β7 V. Description of the invention (27 Table 2 e-om 冰-冰 ία'- 二 6 * methylamino group &amp; methyl group &amp; methyl group page fraction残 # 酶: 〇Η90. T-0 86 t-2 hours 89 44 small b 92 t »6 hours W 1.24 hours 0 9 9 S 7 100 79 SI 82 12 Ο 18 U IS IS 100 100 100 100 100ο Ο Ο Ο Ο 100 biological jMfg »3QCL | JM5 5CTC QIAIOO grams of sugar-rich IL pulp 1-0 74 〇t-0.5 hours 46 〇t-1 Xiaosheng · 22 〇-1.5ψϋ II 〇1 * 2. , Ί, hinder 6 〇11 0οοο IS 54 71 89 94 100 Α6 24 Μ 7 Ο 51 73 52 90 77 75 66 73 70 Ο Ο 10 Ο Ο 23 25 24 27 30

Id— (Please read the notes on the back before filling in this page) • pLi- HB1 ·· —ten-Bookmarked by the Consumers' Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs-30- This paper size applies to China National Standard (CNS) A4 Specifications (210X297 mm) 1241347 A7 B7 ^ jm, V. Description of the invention (28) (^ -ΟΜί, 6 &gt; Methanyl e〇Mii 5,7,4'-three 5,7 half-three 5,7, &amp; Sanqing 7 + 1 7 Shao-two 1¾ 7, 々-two ^ 7,4'-di-7,4 '· ^ $' 6 · methyl II group 6 »methylamino group ^ search base isomeryl radical Isoflavoryl, isomeryl, isoflavyl, isoflavogenous, isobyl, isoflavage-7, Benji ^ -7 · 4Ί Sample yellow g 甞 甞 明 * | 甞 Sun eaves 甞: Isomerized yellow Hu Ming 甞 甞 明 甞: Huang 嗣 嗣 站Bizarre suspicion that the base is different from the yellow page, although the biological early release is OKTOL rf! 4 &lt; 4 &lt; rg (Η_gram rich in nuclear poverty ^ feeΗ) 74 0 11 15 100 0 0 0 77 0 23 HJ. 5 hours 69 0 0 31 70 0 0 30 76 0 24 1 hour 54 0 0 46 53 3 0 44 76 10 24 M.5! Hours AA 0 0 56 43 0 4 52 75 0 23 t * 2 37 0 0 63 35 3 0 62 74 0 26

Bio coffee LpH45 4Q ° C 〇14 such as 00 grams of rich silk pulp 1-0 50 2 0 47 61 2 0 37 60 0 40 t-1 hours 25 2 0 73 31 2 0 67 54 0 55 t-2 hours 12 2 0 t6 15 1 0 S3 51 0 50 1-3! hours 7 2 0 92 9 1 0 90 0 0 100 qi grams αοο grams of sugar-rich milk ϋ —ϋ ϋ (Please read the notes on the back before filling in this (Page) t-0 47 I 0 45 45 7 0 4t 74 0 26 Ι · 0.5 hours 9 9 0 η S 9 0 S3 55 0 39 Μ small broken 3 S 0 19 2 t 0 90 46 0 Μ 2 hours 0 9 0 91 0 t 0 92 32 0 (Λ 5 vermiculite 100 g is rich like L slurry H) S3 〇tl hours 4 0 bu 2 small 4 1 〇 3 small poem 0 〇m g αοοg 1 H) 13 H). 5 hours 17 bu 1; hours 6 bu 2 small 4 〇17 S3 0 0 17 SO 96 2 0 0 91 23 99 0 0 0 100 10 100 0 0 0 100 1 16 79 3 1 17 15 η 16 4 3 77 39 93 5 4 3 17 26 99 0 4 3 92 4 20 77 76 7f 15 55 'Booklet. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs ny〇9w pH 4 &lt; «rc Q1g / 100g is rich in scent ^ S3 0 0 17 S3 0 0 17 to 0 20 1-1 hours 49 1 0 31 41 0 0 59 t2 0 IS 2 hours 30 1 0 69 21 0 0 79 79 0 21 1-3 hours II 0 0 12 11 0 0 t9 69 11 19 -31 This paper size applies Chinese National Standard (CNS) A4 specification (210 X 297 mm) 1241347 A7 B7 V. Description of the invention (29 5,7,4'-three 5,7, Dongzhu 5,7 , Dongsanqing 5,7,4,-, Leptyl isoryl, inflammatory base, isoflavone, triseptyl sample, Huang Huangming, 甞 甞 甞 甞 甞: Isocyanine 7, 4, Ί 7, 4 Ί ~ methylamino -7, ... 6 &gt; Methyl isoflavyl isoflavyl isoflavyl isoflavone isotyl isoflavone-7, wood Ⅱ-7 this two s Q2 ^ g〇Q,

JMCE 丨 Glycol-rich milk paste Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs

liULi Η) 7 Li 7 0 15 77 7 0 16 75 6 19 1 hour 71 f 0 M 10 7 0 13 0 14 t-4 hours 77 t 0 15 to 7 0 &quot; M 0 16 iHTft t-0 71 7 0 15 n 7 0 16 75 6 19 t-ι hours η plug 0 20 77 7 0 16 72 0 2t tM small 4 61 1 0 24 74 7 0 19 61 0 39 Um; I ^ V -ww »vrr 02富 练 # 浆 liLU t-0 7 7 7 15 15 7 7 0 16 75 6 IV t-1 hours 71 7 0 13 n 7 0 16 75 6 19 l * -4 hours 76 7 0 17 76 7 0! 7 73 6 21 nH7tt H) 71 7 0 15 77 7 0 16 75 6 19 t-1 hour 71 7 0 22 73 6 0 21 5 «7 31 bu 4 hours 65 7 0 21 69 5 0 26 32 0 AM ffisr neutral milk _ ^ r 〇2 grams plus 0 grams of sugar-rich syrup • ULii t-0 7S 7 0 IS 77 7 0 16 75 6 19 lp hours 77 7 0 16 77 6 0 17 73 7 20 M small 蛉 77 0 7 16 77 6 0 17 76 0 24 Η 70 Η) 71 7 0 15 77 7 0 16 75 6 19 1-1 hours 70 7 0 23 72 5 0 23 70 6 24 hours 55 7 0 3t 60 6 0 34 66 0 W -32- This paper size is in accordance with Chinese National Standard (CNS) A4 (210X297mm) 1 ^ 1- n ^ i * »-—-ϋϋ · 1 ^ 1 I (Please read the precautions on the back before filling this page ) "Order_ A7 1241347 B7 V. Description of the invention (30) ^ ava, W · 3 5,7, winter master ^ • OAq Sanqing 5Z winter 7 ^ 4 k 6» A gas group 6. A n group. 6 · A # 1 基 样 黑 &amp; kiyipu three hides: yihuan IK Huanji Yizhoupuyi yihuangming H station base yihuang 《、 Percentage exploration creature ## 1ug 30J0050 ° C Q2j? / 100 Kefu training pulp «LU t-0 71 7 0 15 77 7 0 16 75 6 19 1-1 hours 0 6 0 94 0 6 0 94 0 0 100 M hours 0 4 0 96 0 5 0 95 0 0 100 Μ ) 71 7 0 15 77 7 0 16 75 6 f9 t * l hours 2 7 0 91 3 7 0 90 29 0 71 4 hours 0 7 0 93 0 6 0 94 0 0 100 Explore creatures, hidden 50eC 〇2ίΑ〇〇 ίϊ Contains H3JLX Η) 78 7 0 15 77 7 0 16 75 6 19 t-bu], hour 73 6 0 21 76 5 0 19 79 0 21 Μ small broken 73 6 0 21 76 5 0 19 76 0 24 atLlfl t-0 71 7 0 15 77 7 0 16 75 6 19 1 hour 2 7 0 91 7 4 0 19 15 0 15 μh 0 7 0 93 0 4 0 96 0 0 100 If converted by 5, 7, respectively , 4 f -trihydroxy isoflavone sugar taste, 7,4, _dihydroxy isoflavone sugar taste and 6-methoxy-7,4, -dihydroxy isoflavone sugar ^ cheng (please read first (Please read the notes on the reverse side and fill in this page again) Print the 5,7,4 \ trihydroxy isoflavones, 7,4 \ dihydroxy isoflavones and 6-methoxy-7, V printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs -As shown by the diacid isoflavones, substantially complete conversion of the isoflavone sugars to the aglucone isoflavones was achieved. Supplemental enzymes significantly increase the conversion rate ’and some supplementary enzymes complete substantially complete conversion within 1 hour. The most effective supplemental enzymes found at pΗ4.5 are biopectinase 1000L, biopectinase 3 00L, lactase F, α-Gal 600L, G-Zyme G990, and exploration of biological lactase 30,000 and Enzeco P. lactase. Found at pH 7.0_-33- This paper size applies to Chinese National Standard (CNS) A4 (210 X 297 mm) 1241347 A7 B7 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (31) The effective supplement enzymes are exploring biological lactase 30,000 and neutral lactase. Example 3 In another experiment, the aglycone isoflavone milk protein material was recovered from a soybean milk rich in aglycone isoflavones. The first soymilk sample rich in aglycone isoflavones weighed 1,000 grams and contained 30 mg of 5,7,4, _trihydroxy isoflavone, 37 mg of 7,4, dihydroxy isoflavone, and 7 mg of 6-methyl The oxy_7,4, _dilightyl isoflavone is evaporated from low heat to 63 grams of concentrated (concentration ratio 値 -1: 6 · 1). The concentrated milk is heated to the proteinaceous matter coagulated in the milky slurry and centrifuged to further concentrate the milky proteinaceous matter. 21 grams of serum protein containing 25 mg of 5,7,4'-trihydroxy isoflavones, 32 mg of 7,4f-dihydroxy isoflavones and 6 mg of 6-methoxy-7,4'-dihydroxy isoflavones The substance is separated from the milk. Recovered milk protein material contains 82% 5,7,4'_trihydroxy isoflavones, 88% 7,4'-disuperyl isoflavones and 77% 6-methoxyl in the combined milk and milk protein materials Radicals 7,4, dihydroxy isoflavones. The second soymilk sample rich in aglycone isoflavones weighed 400 grams and contained 12 mg of 5,7,4, trihydroxy isoflavones, 15 mg of 7,4, dihydroxy isoflavones, and 3 g of 6- Methoxy-7,4'-diazonyl isoflavones are heated to the proteinaceous material that coagulates in the milk 'without concentrating the milk. The coagulated milk protein material and the milk are centrifuged to further concentrate the milk protein material. 8 7 grams of milk protein material was recovered and contained 5 mg of 5,7,4, trihydroxy isoflavones, 7 g of 7,4, diacryl isoflavones, and 1 mg of 6-methoxydiboryl isoflavones. Recovered milk protein material contained 44% 5,7,4'_trisyl isoflavone, 47% 7,4, -dihydroxy isoflavone and 34% 6-methyl in the combined milk protein material and milk Oxy-7,4 'dihydroxy isoflavone. -34 This paper size is applicable to China National Dejin (CNS) Α4 specifications (辂,-(Please read the precautions on the back before filling this page)

1T J-- 1241347 A7 --------- B7 V. Description of the invention (32) &gt; Compare 罘 1 and 2 samples of the milk protein material, and clearly concentrate the rich protein formula before separating the milk protein material. The milk aggregation of the isoflavones resulted in increased capture of aglycone isoflavones in milk protein shellfish materials. Example 4 In another experiment, the aglycone isoflavone substance was recovered, and the aglycone isoflavone serum protein substance and the self-extracting solution Shendian aglycone isoflavone substance were extracted from an aqueous alcohol extract. 821 grams of aglycone isoflavone serum protein material contains 86% dry weight protein, 4.7 grams of 5,7,4'_trihydroxy isoflavones, 22 grams of 7,4, dihydroxy isoflavones. Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. And 0.36 g of 6-methoxy_7,4 and dihydroxy isoflavones are converted from isogone and isoflavone glycosides into aglucone isoflavones and isoflavones are recovered from the slurry. Provided with milk protein material. Aglycone isoflavone serum protein The rabbit product was extracted with 6360 g of 80:20 wt% ethanol / water solution (77: j solution / aglycone isoflavone serum protein material) at 60% for 4 5 minutes. After extraction, the generated table was cooled to 25 C and filtered on Whatman No. 4 filter and paper under empty space. The wet cake weighed 1584 grams and contained 798 grams of solids, 0.8 grams of 5,7,4'-trihydroxy isoflavones, 0.4 grams of 7,4, dihydroxy isoflavones, and 0.02 grams of 6-methoxy. -7,4. Dioxo isoflavones were recovered, and together with 3397 g of clarified extract, it contained 23 g of solids, 3.9 g of 5,7,4 \ trilateral isoflavones, 1.8 g of 7,4, _ di Hydroxy isoflavones and 0.34 g of 6-methoxy-7,4, dihydroxy isoflavones. The filter cake was extracted with 2000 g of 80:20 wt% ethanol / water solution (23: 1 solution / original isoflavone serum protein material) for the second time at 25 ° C for 5 minutes. After the second extraction, the resulting pulp was then applied to Whatman No. 4 Filter Paper-35- This paper is sized to Chinese National Standard (CNS) A4 (210X297 mm) '&quot; 1241347 A7 B7 V. Description of Invention (33) filter. The wet cake weighed 1542 grams and contained 794 grams of solids, 0.3 grams of 5,7,4'-trisyl isoflavones, 0.1 grams of 7,4'-dioxane isoflavones, and 0.011 grams of 6-methoxyl. -7,4, dihydroxy isoflavones were recovered together with the second extraction solution, weighing 2042 g, containing 4.0 g of solids, 0.5 g of 5,7,4'_trihydroxy isoflavone, 0.3 g of 7,4, 2 Hydroxyl isoflavones and 0.011 g of 6-methoxy-7,4, diquinone isoflavone. The extracts were combined and contained in the first 94% of the 5,7,4, -trihydroxy isoflavones and 9 5% of the 7,4'-dihydroxy isoflavones in the isoflavone I homogenate protein material. The extract was then concentrated by evaporation in a Buchi evaporator under vacuum at 70 C to 1528 grams (20% of the original combined extract volume). Add 6000 grams of deionized water to the concentrated extract (4 ·· 1 water / extract). When water is added, a self-colored isoflavone precipitate is formed. The precipitation slurry was heated to 70 ° C for 45 minutes. The slurry is then refrigerated at 4 ° C for 24 hours to allow the isoflavones to form and stand. 7 300 grams of supernatant was decanted from Shen Dianwu, and the remaining slurry was centrifuged to recover the precipitate. The recovered precipitate was washed with 600 g of deionized water at 70 ° C for 15 minutes. The precipitate was recovered by centrifugation and dried under vacuum at 50 ° C. Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs, the dry aglycone isoflavone substance weighs 7.3 grams and contains 49% 5,7,4, triweilyl isoflavones, 19% 7, V-dihydroxy isoflavones, and 4% 6- Methoxy-7,4 \ dihydroxy isoflavone was obtained. Example 5 In another experiment, a substance having a high content of 5,7, V-trihydroxy isoflavone and a substance having a high content of 7,4'-dihydroxy isoflavone were self-ligand isoflavone materials separated by reverse-phase HPLC. 2 grams of aglycone isoflavone substance contains 55% 5,7,4, trichizyl • 36- This paper size applies Chinese National Standard (CNS) A4 specification (210X297 mm Γ 1241347 A7 B7 V. Description of the invention (34) 1 ketone, 21% 7,4'-dihydroxyisoflavones, and 4% 6-methoxy-7,4'-dihydroxy isoflavones were added in dry weight by adding 1 liter of 50:50 wt% methanol / water solution. The solution was filtered through Whatman No. 5 filter paper and then through 0 45 micron filter paper. The solution was then loaded into a 2-inch diameter 25 cm long PLC column and filled with Kromsil filling material (Kromasil C18 16 micron, 100 Angstrom beads). ). Mobile phase containing 50:50 wt% methanol / water solution then passed through the column at a rate of 64 ml / min. 7,4'-dihydroxy isoflavone, 6_methoxy_7, 4, 2 The appearance of hydroxy isoflavones, 5,7,4, and trihydroxy isoflavones from the column effluent was detected by UV absorption. 7,4'-di- light isoflavones were collected in the first differentiation solution and 5,7 , 4'-Tri # presenting isoflavones were collected in the second separation solution. The 7,4'-dihydroxy isoflavone and 5,7,4, -trihydroxy isoflavone separation solution were evaporated to remove alcohol, resulting in the Individual differentiation solution Precipitation of high 5,7,4, -trisyl isoflavones and high 7,4'-dihydroxy isoflavones. Precipitation of high 5,7,4'-trihydroxy isoflavones and high 7,4 '-di The hydroxy isoflavone content material is recovered by centrifugation and dried in an air oven. The recovered high content of 5,7,4, -trisyl isoflavones contains about 95% 5,7,4, trihydroxy isoflavones and recovered high content. 7,4 、 二 # The basic isoflavone content substance contains about 4.5% 7,4,2,2ndyl isoflavones. Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs in the above experiment, it is 6 &quot; -〇Mal_5, 7,4, · trisyl isoflavone glycocalyx, 6〃_〇Ac-5,7,4 '· trihydroxy isoflavone glycoside, 6〇-〇Mal-7,4'-diazonyl isoflavone glycoside , 6〃-〇Ac-7,4, · bis-isoflavone glycoside, 6 &quot; -OMal-6_methoxy-7,4, diside isoflavone glycoside and 6_methoxy-7, 4. All the percentages shown in dihydroxy isoflavones are calculated. The percentages of enzyme concentration shown are from every sample in the sample. -37- This paper size applies to China National Standard (CNS) A4 specifications. (2 丨 〇 &gt; &lt; 297 mm 1 1241347 A7 &quot; ------_ _ V. Description of the invention (35) Shaped matter or per 1 GG grams of milk slurry (grams of commercial enzyme preparation). The following is a description of the method for quantifying isoflavones in soy products. Isoflavones are extracted from soy products and mixed with 75 G of sample (spray-dried finely ground powder) and 50 ml of 80/20 methanol / water solvent. The mixture was allowed to incubate in an orbital vibrator at room temperature for 2 hours. After 2 hours, the remaining undissolved material was removed by filtration through Whatman No. 42 filter paper. 5 ml of the filtrate was diluted with 4 ml of water and 1 ml of methanol. Printed by the Consumer Standards Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs The extracted isoflavones were separated by HPLC (High Performance Liquid Chromatography) using HP packed C18 Hypersil reverse phase columns. The isoflavones were injected into a column and eluted with a gradient of gradient, starting with 88% methanol, 100% water, and 2% glacial acetic acid. At a flow rate of 0.4¾ liters / minute, all isoflavones-5,74, · trihydroxy isoflavone glycosides, 6 &quot; -〇_ethylenyl 5,7,4'_trihydroxy isoflavone glycosides, 6, 〇_ Malonyl 5,7,4'-trihydroxy isoflavone saccharin, 5,7,4, _trihydroxy isoflavone, 7,4 '· dihydroxy isoflavone sugar: y :, 6 &quot; _〇 _Ethyl 7,4, dihydroxy isoflavone glycocalyx, 6 &quot; -0-propanedifluorinyl 7, dihydroxy isoflavone glycoside, 7, 4, dihydroxy isoflavone, 6_methoxy-7, 4. Dihydroxy isoflavone glycosides, 6〃-0_propanedifluorenyl 6-methoxy-7,4, dihydroxy isoflavone glycosides and 6-methoxy-7,4f · dionyl isoflavones · clear Ground resolution. Spikes were detected by UV absorption at 260 μm. Identification of spikes was performed by HPLC-mass spectrometer. Quantitative use of pure standards (5,7,4, trihydroxy isoflavones, 5,7,4f-trihydroxy isoflavones, 7,4, _dihydroxy isoflavones, and 7,4'-di # to (Isoflavone), Indofine Chemical Company, Sommervill e, New Jersey. The response factor (integrated area / concentration) was calculated for each of the above compounds and used to quantify the unknown sample. No pure standard -38-This paper size applies the Chinese National Standard (CNS) A4 specification (2 丨 〇 &lt; 297 mm) 1241347 A7 ~~ ----- B7 V. Description of the invention (36) Yes The obtained co-consumption form, the reaction factor is false as the parent molecule, but the correction factor is 6_methoxy_7,4, the reaction factor of the dimer isoflavone sugar is tasted 5 * 7,4 ' A few bases of isoflavone_saccharide #, and correct the molecular weight difference. This method provides the amount of each individual yellow. For convenience, total 57,4, -trihydroxy isoflavones and total 7,4, _dihydroxy isoflavones can be calculated and represent the aggregate weight f 'of these compounds if all conjugated forms are converted to their individual Conjugate form. These totals can also be determined directly by a method using acid decomposition to convert the unconjugated form. This is only a preferred embodiment of the present invention. Different changes and modifications can be made without deviating from its spirit and broader elements, as explained in the scope of the accompanying patent application, which is interpreted in accordance with the principles of patent law including considerable doctrine. (Please read the notes on the back before filling this page)

Jn .-Order_

Printed by the Consumers' Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs -39- This paper size applies to the Chinese National Standard (CNS) A4 (210X297 mm)

£ 1 ^%, (The fields above ^ are filled by this bureau) A4 C4 1241347 ¥ Ming Chinese manual replacement page (October 1993) manual Patent

Zhongxi I Li flllf Suspension II | JJ ^ esstdttprdducinuwageucdne- i〇9SAAV.0NE ENRICHED vegetable protein whey SSi? T ^ yEGETABLE PROTEIN whey and materials -ERQDIICED THFPF 卩 ROM "___ Invented a new type of stone British surnames and national names 1. Jeremy Sienke Airoser O. Barra Asherien 4. Maria Anchor

Invention &gt; Residence, residence? Name and nationality are all in the U.S. 1. Wu Kwong, Missouri, 5937 Case Square, St. Louis, Wu Kwong Missouri, Edrew Lane, 5523, St. Louis, Posing No. 7039 Avenue • Wu Guo Collinsville, Illinois 16 8 Burnie Still Road, American Protein Technology International Corporation, USA

Line Applicant St. Louis, Missouri_Pan Guangchang Larry Rui J.Horret

i i: less!:.: ί 1241347 bu patent i! please file 〃

It was found that TT 100% Jinghuang_ conjugate formed isoflavone glycocalyx. Typically, a conversion of at least 95%. These high conversion rates are particularly attractive for use in a large number of commercial operations. "In the two conversion steps or operations, the isoflavones produced in the first step :: and the isoflavone glycosides left in the milk are converted by enzyme reactions into f /, ketones. Transformation from isoflavone glycoside-rich milks produces aglycone isoflavone-rich plant proteins. The second transformation step was found to be dependent on the concentration of enzymes present in the milk mass and its concentration. The enzymes required for the conversion are enzymes that can cleave the isoflavone moiety and the diketoglycan and glucose bonds between glucose molecules. In a preferred embodiment, the enzyme is a carbohydrase enzyme capable of cleaving 4,4-gold money. The enzyme concentration required for the transformation of the same sugar # 成 基基 黄黄 _ depends on a variety of factors = including the types of enzymes present in milk, the distribution of enzyme concentration, the concentration of enzyme isoflavones, and the milk temperature and temperature during the conversion. : Human can exist naturally in plant-based protein milk, which is grown from microorganisms :: L pulp or can be added to the milk. Naturally occurring or derived from microorganisms: Water exists (enzymes are referred to as "residues, Enzymes and enzymes added to the milk are referred to herein as "supplements, enzymes. Enriched Γ f exists in the reading to transform at least most and preferably the flavonoid sugar of Wang Shaoxing. Soil-based ketone. Generally, if there is residual enzyme in the milk, you should add it to the milk. As mentioned above, a variety of factors determine whether the yeast sound # 丄, ... Exist to carry out the transformation. When adding supplementary enzymes, supplement _ h, to the non-enzyme should be added, so that the total trace of the existing enzymes is about 0 1/0 soil about 10% of the weight of milk solids on a dry weight basis. • 14-

Claims (1)

^ 1 ^ 5 ^ 77 patent application A8. Replacement text for patent application scope (July 1994) C8! 丨 _ 丨 丨 _ 丨 丨 ———— ^ D8 hole, application scope for patent application 1. A self-contained Method for making isoflavone conjugate-based plant protein milks A method for producing plant-based protein milks rich in aglycone isoflavones, including: a) at a temperature between 2.0 and 121 and a pH of 6 to 13 7 The reason is that the plant-based] protein milk is for a time sufficient to convert at least 80% of the isoflavone conjugates into isoflavone glycocalyxes; and 'b) the enzyme and the isoflavone saccharin are added to the plant-based protein milk The pulp is contacted for a period of time sufficient to convert at least 80% of the isoflavone material to the ligand isoflavone at a temperature between 50 ° C and 7 5 t and a pΗ value between 3 and 9. 2. According to the scope of the patent claimed In the method of item i, the time period for converting the isoflavones to isoflavone saccharin in # is from about 4 hours to about 6 hours. 3. According to the method of patent application β, the time period in which Zhongzhong converts isoflavones into isoflavone sugar is about 0.5 hours to about 1 hour. 4. According to the patent application Method, wherein contacting an enzyme with the isoflavone sugar in the plant protein milk slurry includes adding an effective amount of supplementary enzyme to the plant protein milk slurry. 5_ According to the method of claim 4 in the scope of the patent application, the enzyme is supplemented in the lotus root. Including glycosidase enzymes that can cleave 1,4-sugar: y: bonds. 6. The method according to item 5 of the scope of the patent application, wherein the supplementary enzyme is selected from the group consisting of disaccharides # enzyme enzymes, β_sugar enzymes, Galactose _ enzymes, glucoamylase enzymes, pectinase enzymes and their group members 7. The method according to item 1 of the scope of patent application, wherein the plant = protein milk slurry includes soybean milk. 8. According to the scope of the patent application Method 1 of 'Procedure-further includes self-nucleating phytoproteins of the isoflavone flavonoids «recycling protein and paper size applicable to China Standards (CNS) A4 specifications (210X297 mm 8 8 8 8 A BCD 1241347 Application for patent The flavonoid isoflavone serum protein material. 9. The method according to item 8 of the patent application scope, wherein the aglycone isoflavone milk protein overflow material is recovered by at least one of ultrafiltration, thermal coagulation, and dewatering. 10. The method according to item 8 of the scope of patent application, wherein the aglycone isoflavone lactoprotein shellfish is recovered by cooling the plant protein milk slurry and separating the aglycone isoflavone milk protein material from the cooled milk slurry. 11. The method according to item 8 of the scope of patent application, wherein the aglycone isoflavone milk protein material is recovered from the concentrated plant protein material. 12. The method according to item § of the scope of patent application, further comprising: a) Extracting the aglycone isoflavone proteinaceous material with an aqueous alcohol extractant to produce an aglycone isoflavone-rich extract; and b) contacting the extract with an adsorbent for a period of time sufficient to separate from the extract 咼 5, 7, 4 The time of the isoflavone-containing substance. 13. The method according to item 12 of the patent application scope, wherein the aqueous alcohol extractant is included between about 30% alcohol and about 90% alcohol. 14 · root The method according to item 12 of the patent application, wherein the aqueous alcohol extractant has a pH value of the isoelectric point of the protein in the protein substance of the isoflavone emulsion. 15. The method according to item 14 of the patent application Wherein the aqueous alcohol extractant has a ρ ρ value between about 3 and about 6. 16. The method according to item 12 of the scope of patent application, wherein the aglycone isoflavone slurry protein substance is extracted with the extractant, The weight ratio of the extractant to the protein content of the syrup does not exceed about n: i. 17. The method according to item 12 of the scope of the patent application, wherein the ligand isoflavone milk-2- This paper size applies to Chinese national standards (CNS ) A4 specification (210 &lt; 297 engine) 8 8 8 8 AB c D 1241347 VI. Patent application scope Pulp protein material is extracted with 2 parts of the aqueous alcohol extractant, where the extractant of these parts is used for the milk The combined weight ratio of the pulp protein material does not exceed a total weight ratio of about 1 1: 1. 18. The method according to item 12 of the scope of the patent application, wherein the adsorbent is a particulate matter. 19. The method according to item 12 of the scope of patent application, wherein contacting the extraction solution with an adsorbent further comprises releasably binding 5,7,4'-trisyl isoflavone in the extract with the adsorbent. 20. The method according to item 12 of the scope of application for a patent, wherein the extract is eluted by the eluent through the adsorbent, and the extract is separated from the extract with a high content of 5, 7, 4, and three warp-based iso yellow S. 21. The method according to item 12 of the scope of patent application, wherein the substance having a high content of 5,7,4, and trihydroxy isoflavones contains at least 40% of 5,7,4, and trihydroxy isoflavones. 22. The method according to item 21 of the scope of patent application, wherein the substance having a high content of 5,7,4 trihydroxy isoflavones contains at least 90% of 5,7,4, trihydroxy isoflavones. 23. The method according to item 8 of the scope of patent application, further comprising: a) extracting the aglycone isoflavone proteinaceous material with an aqueous alcohol extractant to produce an aglucone isoflavone-rich extract; and b) adsorbing the substance The extract is contacted for a time sufficient to separate from the extract a substance having a high 5,7,4, trihydroxy isoflavone content. 24. The method according to item 23 of the scope of patent application, wherein the aqueous alcohol extractant is included between about 30% alcohol and about 90% alcohol. 25. The method according to item 23 of the scope of patent application, wherein the water-based alcohol extractant has an isoelectric point of the protein in the protein substance of the isoflavone emulsion -3-This paper applies Chinese National Standard (CNS) A4 specification (210X297 mm) 1241347 A8 B8 C8 D8 6. pH value in the scope of patent application. 26. According to the method of item 25 of the scope of application for patents, &lt; 10,000 / to, wherein the aqueous alcohol extractant has a p Η value between about 3 and about 6. The household according to the method of the scope of application for the patent No. 23, wherein the ligand isopuertomilk polyprotein substance is extracted with the extractant, and the weight ratio of the extractant to the milk protein substances does not exceed about 丨: j. 28. According to the method of item 23 of the scope of the applied patent, the aglycone isoflavone lactate protein * in the mash is extracted with 2 parts of the aqueous alcohol extractant, and the extractant of these parts of the extract protein has The combined weight ratio of the substances does not exceed a total weight ratio of about 1 1: 1. 29. The method according to item 23 of the scope of patent application, wherein the adsorbent is a particulate matter. 30. The method according to item 23 of the scope of patent application, wherein contacting the adsorbent with the extract further comprises releasably binding 7, V-dihydroxy isoflavone with the adsorbent in the extract. 31. The method according to item 23 of the scope of application for a patent, wherein the extract is eluted with an eluent through the adsorption substance, and a substance having a high content of 7,4, dihydroxy isoflavone is separated from the extract. 32. The method according to claim 23 of the scope of patent application, wherein the content of the 7,4'-dianyl isoflavone in the south contains at least 40% of 7,4'-dianyl isoflavone. 33 · According to the method in the scope of the patent application No. 8, further comprising: a) extracting the aglycone isoflavone proteinaceous material with an aqueous alcohol extractant to produce an aglycone isoflavone-rich extract; and b) The adsorbed substance is in contact with the extract for a period of time. This paper size is applicable to the Chinese National Standard (CNS) A4 specification (210X 297 mm). 1241347 A8 B8 C8 D8. The scope of patent application includes 6-methoxy group. -7,4'-Dihydroxy isoflavone content substance time. 34. The method according to item 8 of the scope of patent application, further comprising: a) extracting the aglycone isoflavone slurry protein material with an aqueous alcohol extractant to produce an aglycone isoflavone-rich extract; b) concentrating the rich Aqueous isoflavone-containing extract to its original volume between about 5% and about 30%; and c) adding water to the extract to precipitate aglycone isoflavone material. 35. The method according to item 34 of the scope of the patent application, wherein the aqueous alcohol extractant contains an alcohol between about 30% and about 90%. 36. The method according to item 34 of the patent application, wherein the aglycone isoflavone milk protein substance is extracted with the extractant, and the weight ratio of the extractant to the milk protein substance does not exceed about 1 1: i. • The method according to item 34 of the patent scope of the patent, wherein the aglycone isoflavone milk protein material is extracted with 2 parts of the aqueous alcohol extractant, and the child part of the extractant is The combined weight ratio does not exceed a total weight ratio of 1 1: 1. 38 'The method according to item 34 of the scope of the patent application, wherein the aqueous alcohol extractant has a pH value about the isoelectric point of the protein in the protein substance of the isoflavone slurry. 39. The method according to claim 38, wherein the aqueous alcohol extractant has a ρ η value between about 3 and about 6. 40. The method according to item 34 of the scope of the patent application, wherein water is added to the extraction liquid ', and the weight ratio of water to the extraction liquid is between about 6 ·· 1 to about 8 ·· 1. 41. The method according to item 34 of the scope of patent application, further comprising washing with water-5-paper-scale paper towels _ Jia Pi (cns) ^ Grid (Others) <Tear Public Corruption] A8 B8 1241347 A ligand isoflavone substance, wherein the weight ratio of water to the ligand isoflavone substance is between about 0.8: 1 to about 2.1. 42. The method according to item 34 of the Chinese Patent Application, further comprising cooling the extraction liquid and water to maximize the Shen Dian of the same compound of isoxanthin. 43. The method according to item 34 of the scope of patent application, further comprising: a) dissolving the ligand isoflavone substance in a solvent in an aqueous alcohol solution; and b) contacting the adsorbent substance containing the ligand isoflavone substance with the solvent Zhishui Aqueous Alcohol Solution is a period of time sufficient to separate from the aqueous alcohol solution a high 5,7,4, _ di # to a base isoflavone content substance. 44. The method according to item 34 of the scope of patent application, further comprising: a) solvent dissolving the ligand isoflavone substance in an aqueous alcohol solution; and b) contacting the adsorbent substance with the ligand isoflavone substance dissolved in the solvent The substance is then held in the aqueous alcohol solution for a time sufficient to separate from the aqueous alcohol solution a substance having a high 7,4,2% isoflavone content. 45. An aglycone isoflavone-rich plant protein milk made according to the method in the scope of patent application.配 The aglycone isoflavone-rich plant protein milk according to the scope of the patent application No. 45, which comprises the aglycone isoflavone proteinaceous material manufactured according to the method of the patent scope No. 8. According to the scope of the scope of the patent application, the plant protein milk slurry rich in aglycone isoflavones includes the content of 5,7,4 -2 per base isoflavone produced according to the method of the scope of the scope of patent application]. substance. 48. The isoflavone-rich plant protein milk slurry according to item 45 of the scope of patent application, which includes manufacturing according to the method of item 23 of the scope of patent application: -6- AB c D 1241347 6. The scope of patent application is high 7 , 4'-Dihydroxy isoflavone content substance. 49. The aglycone isoflavone-rich plant protein milk according to item 45 of the patent application scope, which comprises a high 5,7,4'-trihydroxy isoflavone manufactured according to the method of item 43 of the patent application scope. Content substance. 50. The aglycone isoflavone-rich plant protein milk according to item 45 of the scope of the patent application, which includes a substance having a high content of 7,4'-dihydroxy isoflavone manufactured according to the method of the scope of the patent application 44. 51. The aglycone isoflavone-rich plant protein milk according to item 45 of the scope of the patent application, which comprises an isoflavone substance manufactured according to the method of the scope of the patent application scope 34. 52. An aglycone isoflavone substance prepared by the method of claim 1 in the scope of patent application, including soy substance, at least 10% 5,7,4'-trihydroxy isoflavone and at least 5% 7,4'- Dihydroxy isoflavones. 53. Ligand isoflavone substances according to item 52 of the scope of patent application, which include substances with a high content of 5,7,4'-trihydroxy isoflavone prepared by the method of item 12 of the scope of patent application, including soy substance and At least 40% 5,7,4'-trihydroxy isoflavones. 54. The aglycone isoflavone substance according to item 53 of the scope of the patent application, wherein the substance with a high 5,7,4'-trihydroxy isoflavone content contains at least 90% 5,7,4 trihydroxy isoflavone. 55. The aglycone isoflavone substance according to item 52 of the scope of patent application, which includes a substance having a high content of 7,4′-dihydroxy isoflavone prepared by the method of item 32 of the scope of patent application, including soy substance and at least 40% 7,4'-dihydroxy isoflavones. This paper size applies to China National Standard (CNS) A4 (210 X 297 mm)