CN100523210C - Method for enzyme hydrolysis of soybean isoflavone for producing genistein and daidzin aglycon - Google Patents
Method for enzyme hydrolysis of soybean isoflavone for producing genistein and daidzin aglycon Download PDFInfo
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- CN100523210C CN100523210C CNB2006100100791A CN200610010079A CN100523210C CN 100523210 C CN100523210 C CN 100523210C CN B2006100100791 A CNB2006100100791 A CN B2006100100791A CN 200610010079 A CN200610010079 A CN 200610010079A CN 100523210 C CN100523210 C CN 100523210C
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Abstract
This invention provides the method of producing genistein and daidzein by enzymatic hydrolysis of soybean isoflovone. It uses the soybean isoflovone with different purity as raw material, and hydrolyse it with Trichoderma beta- glucosidase or bean beta- glucosidase. Then solvent extraction, ultrafiltration and freeze centrifugal isolation and purification are performed to get genistein and daidzein. This invention can use hydrolysis material with different purity. The enzyme used can be purchased directly or prepared by yourself. The buffer solution can recycle. It is easy to operate and with relatively high transformation rate (>85%). The total content of soybean isoflovone daidzein is high. By using the extracted bean beta- glucosidase from soybean came from northeast region of china, the problem of poor edible security with enzyme generated by microbiological fermenting method can be resolved. This invention is suitable for the industralization production because the low cost with commercial enzyme.
Description
(1) technical field
The present invention relates to produce the method for genistein and Daidzein by the soybean isoflavones powder of different concns.More specifically, what the present invention adopted is soybean beta-glucosidase and the commercial Richter scale wood enzyme β-glucoside enzyme hydrolysis of soybean isoflavone that extracts purifying from northeast soybean voluntarily, and separation purification process such as solvent-applied extraction, ultrafiltration and frozen centrifugation, the method for producing genistein and Daidzein by the soybean isoflavones powder.
(2) background technology
Soybean isoflavones is a class secondary metabolite that forms in the soybeans they grow process.Naturally occurring soybean isoflavones always has 12 kinds in the soybean, can be divided into 3 classes, be daidzin (daidzin) class, Genistoside (genistin) class, glycitein glycosides (glycitin) class, there are (Kudou et al., 1991a with 4 kinds of forms such as free type, glucoside type, ethanoyl glucoside type, malonyl glucoside types respectively; Kudou et al., 1991b).The chemical structural formula of genistein and Daidzein is:
Genistein (genistein) Daidzein (daidzein)
Soybean isoflavones glucoside chemical structural formula is:
Table 1 listed in the chemical structural formula of all soybean isoflavones compounds:
The chemical structural formula of table 1 soybean isoflavones compound
The higher country of scientific and technological levels such as the U.S. studies show that, in 12 kinds of soybean isoflavones that in soybean, exist, the isoflavones of two kinds of free forms is that genistein (genistein) and Daidzein (daidzein) have higher biological activity, can effectively prevent and suppress the generation (Zhang Yongzhong etc. of multiple diseases such as leukemia, osteoporosis, colorectal carcinoma, lung cancer, cancer of the stomach, mammary cancer and prostate cancer, China's grain and oil journal 2004,4).But the isoflavones of these two kinds of aglycon forms its content in soybean is considerably less, 3% of not enough soybean isoflavones total amount.Have the soybean isoflavones deep processed product of higher physiological activity, market still belongs to blank at home at present.The purity that is mostly in the manufacturer production of the many production soybean isoflavones of China is the soybean isoflavones product of 20%, 40%, 60% content, and wherein aglycon content is few.The chemical structure of material is determining Substance Properties, is also determining the biological activity of material.The structure of genistein and Daidzein and female hormone structural similitude have stronger biological activitys such as weak estrogen activity.Isoflavone genin is the isoflavones s-generation product with high biological activity, has powerful growth momentum, and expectation will become the leading of following natural galenical market.External a lot of to the research of isoflavones, make progress also very soon, the preparation of isoflavone has been had patent report, and set up industrial production line, and China takes the direct solvent extraction to the production of soybean isoflavones, uses resin purification more more.The soybean isoflavones for preparing like this still is the combined form more than 95%, and wherein the genistein of free form and Daidzein content are very few, is difficult to play adjusting human body physiological function.China also someone utilizes acid hydrolysis to obtain the free form soybean isoflavones, but strong acid condition is difficult for carrying out industrialization.Because the strong acid hydrolysising by-product is many, and hydrolysate is mainly used in food or medicine, therefore also has the potential unsafe factor.External main research enzymic hydrolysis.The enzyme hydrolysis condition gentleness adopts weakly acidic buffered soln more, and isoflavone genin is volatility not, is that the protective foods of isoflavone genin or the very promising approach of medicine are rich in industrial preparation.The essence of enzyme is protein.Can there be the edible potentially dangerous of human body in enzymic hydrolysis, is a kind of ideal hydrolysis approach.External highly active soybean isoflavone glucoside hydrolase is also in the development stage at present, studying maximum soybean isoflavone glucoside hydrolases is exactly beta-glucosidase (Sun Yanmei etc., 2002), enzyme that can the hydrolyzed soy bean isoflavone glucosides also has (Shen, et al 1998 such as gluconic acid enzyme, tilactase, biological Sumylact L, fungus lactase and Sumylact L F; Bryan, BarbaraA et al 2002; Tsuruhami, Kazutakaet al 2003), they all have the ability of very strong hydrolyzed soy bean isoflavone glucosides.Purifying is extracted in use voluntarily from northeast soybean soybean beta-glucosidase can overcome microbial fermentation system enzyme, and the problem of edible safety difference can make product safer.The enzyme of commodity in useization can reduce production costs.
The chemical equation of enzymatic hydrolysis reaction is as follows:
Genistein (genistein)
Daidzein (daidzein)
The Chinese patent ZL97120435.7 of Protein Technologies International, Inc discloses and has used enzymic hydrolysis to prepare the method for isoflavone.This patent uses the enzyme of cleavable 1,4 sugared sweet key to prepare isoflavone.The enzyme source of institute's cracking 1,4 sugared sweet key is in aspergillus niger, aspergillus oryzae, newborn Crewe Wei Shi yeast and crisp wall gram aspergillus niger.Preferred enzyme is α-galactase, beta galactose enzyme, glucoamylase and polygalacturonase.
The Chinese patent of the mediate application of Jin Feng (application number CN03133637.X) discloses the method hydrolyzed soy bean isoflavone glucosides with microbial fermentation, with the preparation isoflavone genin.Wherein employed microorganism comprises bacterium, mould, yeast and basidiomycetes.The microorganism of using in embodiment comprises high temperature oxygen consumption mould Clostridium thermocopriea, aspergillus niger (Asperillus niger), aspergillus oryzae (Aspergillusoryzae), candiyeast and Agaricus bitorqui bacterium (Agaricus bitorguis).Enzymolysis product separates with ethanol sedimentation or n-butanol extraction, can obtain isoflavone genin.The transformation efficiency of this method is 50-80%.
The Chinese patent (application number CN200410058379.8) of Wang Zhe application discloses the method for producing isoflavone genin with beta-glucosidase and membrane technique.This method is a basic raw material with the soybean isoflavones powder, make inductor with soybean protein and soybean isoflavones, with cultivating the extracellular enzyme beta-glucoside enzyme hydrolysis of soybean isoflavone that mould discharges, and utilize the membrane technique that is selected from ultrafiltration and dialysis to carry out the aftertreatment of enzyme hydrolyzate, obtain isoflavone genin.
(3) summary of the invention
The purpose of this invention is to provide a kind of method that is suitable for suitability for industrialized production genistein and Daidzein.
The object of the present invention is achieved like this: the soybean isoflavones powder with different purity is a raw material, use a kind of being hydrolyzed in Richter scale wood enzyme β-glucuroide or the soybean beta-glucosidase, again hydrolyzed solution is carried out solvent extraction, ultrafiltration and frozen centrifugation separation and purification, obtain genistein and Daidzein.Its hydrolysising condition is: the enzyme-to-substrate ratio is 6000～14000U/g, adds pH and is 4.00～6.00 buffered soln and be diluted to that to make the reaction soln cumulative volume be the 800mL/g substrate, 37～60 ℃ of hydrolysis 1～12 hour; Hydrolyzed solution earlier by ultra-filtration and separation, have the organic solvent of close characteristic to extract with the ether of 1:1～1:2, methylene dichloride, ethyl acetate or with them again, boil off organic solvent, vacuum-drying must be rich in genistein and Daidzein power-product; Wherein, the genistein hydrolysis efficiency can reach more than 90%, and the Daidzein hydrolysis efficiency can reach more than 98%.
The present invention can also comprise some features like this:
1, described soybean beta-glucosidase adopts following method to obtain:
(1) extraction of soybean beta-glucosidase
Get the full beans of northeast soybean, in 20 ℃ water, soaked 20 hours, use cold rinse; In the soybean of soaking into, add 0.1molL
-1, the phosphate buffer soln of pH6.60 grinds homogenate; Soya-bean milk is with 4000 rev/mins of centrifugal 3min; Supernatant liquor 0.1molL
-1HCl be acidified to pH5.0, the back is with automatic refrigerated centrifuge 8000r/min, 4 ℃, centrifugal 10min; Supernatant liquor is slightly carries enzyme;
(2) purifying of soybean beta-glucosidase
Add ammonium sulfate and make saturation ratio reach 65～75% above-mentioned slightly carry in the enzyme, left standstill 15～20 hours; 8000 * g, 4 ℃, frozen centrifugation 10～15min, collecting precipitation protein is dissolved in 0.05molL
-1, in Sodium phosphate dibasic-citric acid solution of pH5.0; This enzyme solution is poured dialysis tubing into, and dialysis is two days in identical buffered soln, and the buffered soln that changes dialysis usefulness is more than three times; Precipitation appears in enzyme liquid, 4000 rev/mins of centrifugal 10min again, and clear liquid is partially purified enzyme liquid, and said process is 4～10 ℃ of operations down.The purifying of soybean beta-glucosidase also can adopt ultrafiltration process.
Compared with the prior art, method of the present invention can be used the soybean isoflavones powder of different purity.The enzyme that the present invention uses is the commercialization enzyme that can directly buy, can be applied to food or medicine; Also can prepare voluntarily.The commercial enzyme that is adopted is Richter scale wood enzyme β-glucuroide.Zhi Bei enzyme is the soybean beta-glucosidase that extracts purifying from northeast soybean voluntarily voluntarily.This soybean beta-glucosidase liquid is when pH5.00, and the optimum temps of reaction is 45 ℃.This soybean beta-glucosidase enzyme when being lower than 40 ℃ is more stable, and enzyme activity had only remained about 10% after enzyme activity reduced by 23%, 60 ℃ of heating 5min behind 50 ℃ of heating 5min, to 70 ℃ of enzymic activity total losses.This soybean beta-glucosidase less stable under acidic conditions, pH value are 3.00 and 4.00 o'clock, place 24 hours in 5 ℃, and enzyme activity remains 10% and 46% respectively, the pH value be 5.00～7.00 o'clock more stable.
This patent adopts the soybean beta-glucosidase that extracts purifying from northeast soybean voluntarily.The optimum temps of the soybean beta-glucosidase enzyme reaction of purifying is 45 ℃, and the best pH of enzyme reaction is 5.00.The pH value be 5.00～7.00 o'clock more stable.More stable when being lower than 40 ℃, 70 ℃ of basic total losses of enzymic activity.MMatsuura (Matsuura M.; Obata A., β-Glucosidases from soybeans hydrolyze daidzinand genistin.J.Food Sci., 1993.58 (1), 144-147.Matsuura M.; Sasaki J.; Murao S., Stuies on β-Glucosidases from soybeans that hydrolyze daidzin and genist in:Isolation and characterization of an isozyme.Biosci.Biochem Biochem., 1995.59 (9), 1623-1627.) etc. studied the character of beta-glucosidase in the commercially available soybean of the U.S., they have been purified 456 times by the soybean beta-glucosidase of research, 45 ℃ of optimal reaction temperatures, identical with No. 42 soybean beta-glucosidases of east farming.Optimal ph is 4.50, and is close with the optimal ph of No. 42 soybean beta-glucosidases of east farming.Thermostability (this enzyme is behind 50 ℃ of heating 5min, and enzyme activity reduces 59%) is more very different than east No. 42 soybean beta-glucosidases of farming, and pH stability (the pH value is 4.00 o'clock, and enzyme activity reduces hardly) is better than east No. 42 soybean beta-glucosidases of farming.The beta-glucosidase that exists in the commercially available soybean of this U.S., in the time of 45 ℃, hydrolyzed soy bean isoflavone glucosides 3 hours, hydrolysis efficiency are 22-29%.No. 42 soybean beta-glucosidases of east farming in the time of 45 ℃, hydrolyzed soy bean isoflavone glucosides 3 hours, hydrolysis reaction efficient is 72%.Although concrete experiment condition difference because hydrolysis efficiency differs greatly, has higher hydrolyzed soy bean isoflavone glucosides ability can east No. 42 soybean beta-glucosidases of farming are described, in processing high-content soybean isoflavone product, higher utility value is arranged.Use the soybean beta-glucosidase and carry out enzymic hydrolysis, mild condition, save energy, isoflavone genin is volatility not, and is safe and reliable, is the very promising approach of industrial preparation isoflavone genin.
Trichodermareesei beta-glucosidase (buying in Shanghai) is a faint yellow solid, and it is dissolved into the aqueous solution, studies its zymologic property.At first determine to measure the test conditions of Trichodermareesei beta-glucoside enzyme activity: concentration of substrate 3mmolL
-1, 50 ℃ of temperature of reaction, pH value are 5.0,10 minutes reaction times.Discover the better heat stability of this enzyme, 60 ℃ of heating are after 60 minutes, and enzyme activity descends 29%; Enzyme is more stable when pH the about 4.5-5.LmmolL
-1Ba
2+, Ca
2+, Al
3+, Mg
2+And Fe
2+To enzyme no obvious suppression effect alive, 1mmolL
-1Ag
+Work has the obvious suppression effect to enzyme, and enzyme activity has reduced 64%.The top condition of Trichodermareesei beta-glucosidase hydrolysis soybean isoflavone glycoside is: enzyme concentration 20 units, 50 ℃ of temperature, pH4.5, hydrolysis time 90 minutes.To Genistoside percent hydrolysis 99.03%, daidzin percent hydrolysis 98.06%.
Easy handling of the present invention has higher transformation efficiency (〉 85%), product isoflavone genin total content is higher.The enzyme that the present invention uses is the commercialization enzyme that can directly buy, also can be the enzyme for preparing voluntarily.The buffered soln of used enzyme can recycle.Purifying is extracted in use voluntarily from northeast soybean soybean beta-glucosidase can overcome microbial fermentation system enzyme, and the problem of edible safety difference can make product safer.The enzyme of commodity in useization can reduce production costs.Therefore, method of the present invention is more suitable in suitability for industrialized production.
(4) specific embodiments
For a more detailed description to the present invention for example below:
Below being defined as of enzymic hydrolysis aglycon yield among each embodiment:
Isoflavones total amount=glucosides type isoflavone content+malonyl-fundamental mode isoflavone content+ethanoyl isoflavone content
Isoflavone content is measured and is adopted high performance liquid chromatography.
The preparation of embodiment 1 soybean beta-glucosidase and hydrolyzed soy soybean isoflavones
The full beans 500.0g of soybean (No. 42, east farming is by providing of Northeast Agricultural University's soybean research institute), immersion is 20 hours in 20 ℃ water, uses cold rinse.Add 0.1molL in the soybean of soaking into
-1, the phosphate buffer soln 5000mL of pH6.60, electronic refiner homogenate.Soya-bean milk is with 4000 rev/mins of centrifugal 3min.Supernatant liquor 0.1molL
-1HCl be acidified to pH5.0, the back is with automatic refrigerated centrifuge 8000r/min, 4 ℃, centrifugal 10min.Supernatant liquor is and slightly carries enzyme.Add ammonium sulfate and make saturation ratio reach 75% slightly carrying in the enzyme, left standstill 20 hours.8000r/min, 4 ℃, frozen centrifugation 10min, collecting precipitation protein is dissolved in 0.05molL
-1, in Sodium phosphate dibasic-citric acid solution of pH5.0.This enzyme solution is poured dialysis tubing into, and dialysis is two days in identical buffered soln, and the buffered soln that changes dialysis usefulness is more than three times.Precipitation appears in enzyme liquid.4000 rev/mins of centrifugal 10min again, clear liquid is partially purified enzyme liquid.
The soybean isoflavones powder that takes by weighing 100 grams 40% is pressed the 6000U/g enzyme substrates than adding soybean beta-glucosidase enzyme liquid as hydrolysis substrate, and adding buffered soln adjustment pH is 5.00, at 45 ℃ of following hydrolysis 6hr.After the hydrolysis, reaction solution makes soybean beta-glucoside enzyme-deactivating at 80-90 ℃ of heating 5min.Then, use ethyl acetate extraction three times, steam ethyl acetate, get isoflavone genin powder 20.3 grams after the vacuum-drying, wherein genistein content is 19.6%, and Daidzein content is 17.2%.
Embodiment 2 Richter scales wood enzyme beta-glucosidase (commercial enzyme gets cellulase by Richter scale wood enzymic fermentation, extracts purifying and get beta-glucosidase from cellulase) hydrolyzed soy bean isoflavone powder prepares genistein and Daidzein
Get 100g 20% soybean isoflavones powder (grain institute in Heilongjiang Province's provides), add phosphoric acid disodium hydrogen-citric acid solution and transfer pH to 4.5, by the enzyme-substrate concentration ratio be 5% add beta-glucosidase (the ratio vigor of beta-glucosidase is: 230,000 U/g), at 50 ℃ of following water-baths vibration hydrolysis 90min.Centrifuging separates then, and the isoflavone genin after the separation is deposited in 70 ℃ of following waters (volume of water be original liquid 4/5) and washes 30min, and placement is spent the night.Centrifuging separates, the precipitation that obtains uses 70 ℃ of following waters (volume of water be original liquid 4/5) to wash 15min again, and centrifuging separates, and the precipitation that obtains is with acetone solution (amounts of acetone is sedimentary 5 times), centrifugation, the precipitation (being mainly soybean saponin) that is insoluble to acetone reclaims.Acetone soln rotation reduction vaporization is removed acetone, and what obtain is deposited in 50 ℃ of following vacuum-dryings, product 5g, yield is 5%.All aglucone content is 10% in the product, and wherein Daidzein 2.5%, and genistein content is 7.5%.
Embodiment 3 preparation high-content genistein and Daidzein methods
Prepare high-content genistein and Daidzein if desired, can use the high-purity soybean isoflavone powder is that substrate is hydrolyzed.Before the hydrolysis soybean isoflavones powder is further carried out purifying, be hydrolyzed again.Concrete grammar is as follows:
Accurately take by weighing the soybean isoflavones powder, press 1:20 and add leaching agent acetone, at 60 ℃ of following heated and stirred reflux extraction 3h, extracting solution is centrifugal 20min under the 4000r/min condition, gets supernatant liquid.Precipitation with 1:10 acetone once more under 60 ℃ in magnetic force heated and stirred reflux extraction 1h, centrifugation merges extracted twice liquid.The decompression rotary evaporation is removed acetone.Soybean isoflavones behind the purifying is used for the enzymic hydrolysis substrate, by after the hydrolysis of above-mentioned enzyme optimum hydrolysising condition, hydrolyzed solution is placed 4 ℃ the time spend the night.Centrifugal 30min under the condition of 4000r/min on the 2nd is dissolved in 50-60 ℃ of warm water with precipitation, places 4 ℃ the time and spends the night.Next day is centrifugal under the condition of 4000r/min again, and it is dry that precipitation is put into vacuum drying oven.Dried solid precipitation is ground into powder, is prepared into corresponding purity product.
Claims (1)
1. an enzyme hydrolysis of soybean isoflavone is produced the method for genistein and Daidzein, gets purity and be 20% soybean isoflavones powder, adds phosphoric acid disodium hydrogen-citric acid solution and transfers pH to 4.5; The enzyme substrates concentration ratio is 5% adding Richter scale wood enzyme beta-glucosidase, and the ratio vigor of Richter scale wood enzyme beta-glucosidase is: 230,000 U/g; At 50 ℃ of following water-bath vibration hydrolysis 90min; Centrifuging separates then; Isoflavone genin after the separation is deposited in 70 ℃ of following waters, and volume of water is 4/5 of an original liquid, washes 30min, and placement is spent the night; Centrifuging separates, and the precipitation that obtains is used 70 ℃ of following waters again, and volume of water is 4/5 of an original liquid, washes 15min, and centrifuging separates; The precipitation acetone solution that obtains, amounts of acetone are sedimentary 5 times, centrifugation, the precipitation that is insoluble to acetone reclaims, acetone soln rotation reduction vaporization is removed acetone, what obtain is deposited in 50 ℃ of following vacuum-dryings, product.
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| CN101086002B (en) * | 2007-06-22 | 2010-06-16 | 清华大学 | Method for hydrolyzing soybean isoflavone by enzyme |
| CN101701232B (en) * | 2009-10-27 | 2012-03-14 | 武汉工业学院 | Method for preparing high-purity soybean isoflavone aglycones |
| CN106244643B (en) * | 2016-07-31 | 2019-06-28 | 山西大学 | A kind of method that enzymatic hydrolysis prepares daidzein |
| CN106820142A (en) * | 2017-01-10 | 2017-06-13 | 江苏农林职业技术学院 | A kind of method for improving kudzu root flavone antioxidation activity |
| CN116200435B (en) * | 2023-02-24 | 2024-10-11 | 安徽大学绿色产业创新研究院 | Method for preparing isoflavone aglycone from yellow serofluid |
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| Title |
|---|
| 大豆β-葡萄糖苷酶水解大豆异黄酮糖苷的研究. 孙艳梅等.中国粮油学报，第21卷第2期. 2006 |
| 大豆β-葡萄糖苷酶水解大豆异黄酮糖苷的研究. 孙艳梅等.中国粮油学报，第21卷第2期. 2006 * |
| 大豆异黄酮提取工艺的研究. 关海君，朱冬梅，于殿宇，石彦国，赵焕理.中国油脂，第8期. 2003 |
| 大豆异黄酮提取工艺的研究. 关海君，朱冬梅，于殿宇，石彦国，赵焕理.中国油脂，第8期. 2003 * |
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