CN102960644A - Preparation method of pumpkin dietary fiber and pumpkin dietary fiber - Google Patents
Preparation method of pumpkin dietary fiber and pumpkin dietary fiber Download PDFInfo
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Abstract
The invention discloses a preparation method of a pumpkin dietary fiber and the prepared pumpkin dietary fiber. According to the invention, pumpkin pulp is used as a raw material, which is treated with appropriate technology to remove starch and protein, so as to obtain insoluble dietary fiber and water-soluble dietary fiber; and purity of insoluble dietary fiber reaches higher than 85%, and purity of water-soluble dietary fiber reaches 90.5%. The dietary fiber provided by the invention has excellent quality, good water-holding capacity, swelling capacity and oil holding capacity; in addition, the dietary fiber has on sodium nitrite adsorption capacity, cholesterol adsorption capacity and cationic exchange capacity better than soybean dietary fiber on the market, has effects of lowering lipid, reducing weight and preventing cancer of pumpkin effects, and can be used as a functional food ingredient.
Description
Technical field
The invention belongs to foods processing technique and functional food additives technical field take agricultural byproducts as raw material, be specifically related to the method that a kind of enzyme process prepares pumpkin dietary fiber.
Background technology
Dietary fiber (Dietary Fiber, DF) word is proposed in nineteen fifty-three by Hipsley the earliest, and it refers to edible cell, polysaccharide, lignin, colloid, modified cellulose, clayey, oligosaccharides, pectin and a small amount of constituent that can not be digested and assimilated by the human body endogenous enzymes such as wax, cutin, cork matter etc.Can be divided into water-soluble dietary fiber (SDF) and water insoluble dietary fiber (IDF) two large classes according to the dissolubility in water.Insoluble diedairy fiber refers to not by the human body alimentary canal enzymic digestion and is insoluble to the part dietary fiber of hot water, it is the main component that consists of cell membrane, the chitin and chitosan that comprises cellulose, hemicellulose, lignin, protopectin and animality, its lignin does not belong to polysaccharide, is to make cell membrane keep the aromatic hydrocarbons of certain toughness.Water-soluble dietary fiber refers to not by the human consumption enzymic digestion, but it is water-soluble again by the reppd part dietary fiber of ethanol to be dissolved in warm water or hot water and its.Mainly comprise the pectin that is present in apple, the tangerine class, the glue in the vegetable seeds, the algin in the marine alga, carragheen, agar and microbial fermentation product xanthans etc.Source by dietary fiber is divided, and can be divided into the different dietary fibers in source such as plant origin dietary fiber, animality source dietary fiber, seaweeds polysaccharide dietary fiber, microbial polysaccharide, synthetic semi-synthetic class dietary fiber.Its chemical nature is widely different, but ultimate constituent is then more similar, and the mutual main distinction shows the relative amount of each constituent, the difference of the glycosidic linkage of molecule, the degree of polymerization and branched structure aspect.
People have experienced the process of complications to the understanding of dietary fiber.Think at first the dietary fiber utilization that can not be absorbed by the body, useless to human body.Along with human knowledge's development, find that dietary fiber has very important physiological function to human body.Insoluble diedairy fiber has the effects such as prevention of obesity disease, constipation, colon cancer, and soluble dietary fiber can reduce serum cholesterol, suppresses postprandial blood sugar, improve gut flora.Relevant regulations has also been done to its intake by present a plurality of country.The intake of the total dietary fiber that U.S. FDA is recommended is adult 20～35g/d, and it is 22～24g/d that the intake of everyone dietary fiber is recommended by Canada.Chinese Soclety of Nutrition proposed in 2000, the suitable 30.2g/d that takes in of adult's dietary fiber.
At present, it is found that and the dietary fiber developed has tens kinds more than.Usually divide by dissolubility, dietary fiber divides water-soluble and two kinds of water-insolubles.The representative of water-soluble dietary fiber has glucan, pectin, guar gum and fungi polysaccharide etc.; Water insoluble dietary fiber is mainly take soybean dietary fiber, shitosan and hemicellulose as main.
Pumpkin, annual herb are climbed climing plant, belong to Curcurbitaceae, Cucurbita.The annual pumpkin output of China accounts for more than 13% of Gross World Product, occupy the No. 1 in the world, and its resource is very abundant.In Compendium of Material Medica, Li Shizhen (1518-1593 A.D.) is put pumpkin and glossy ganoderma together, says that it has the effect of " bowl spares, tonifying liver gas, the beneficial motive, beneficial lung qi, beneficial tangerine gas ".Present both domestic and external studies show that; pumpkin all has good medical value at expelling parasite, vision protection, reduction cholesterol, anti-cancer, the aspects such as heavy metal anticancer, that remove in the body; especially remarkable efficacy is arranged aspect the diabetes preventing and treating; in addition; resistance starch content is higher in the pumpkin starch, certain advantage is being arranged, therefore aspect the anti-digestibility; pumpkin has widely medical treatment and health-care effect, is a kind of desirable natural health dietotherapy product.
Present existing commercial diets kinds of fibers mainly contains Testa Tritici fiber, soybean dietary fiber, banana fiber, apple fiber, carrot fiber etc., but the activity that commercially available dietary fiber has is not fine, have since research complex technical process, cost is too high, mostly still rests on the laboratory sample and grinds the examination stage.Therefore, improve the preparation technology of dietary fiber, more kinds of resources of developing preparation dietary fiber, further study the method and the condition that are fit to suitability for industrialized production and have important practical significance.
Contain abundant dietary fiber in the pumpkin, the organic molecule of dietary fiber energy absorption of bile acid, cholesterol mutagen behind the edible dietary fiber, can significantly suppress total cholesterol concentration and raise.Dietary fiber can also reduce the synthetic of cholic acid and its esters and absorb, thereby obstruction neutral fat and cholesterol absorb again, limit the circulation of cholic acid liver sausage, and then has accelerated the drainage of fat composition, thereby can prevent cholelithiasis.Thereby dietary fiber can also slow down the generation that the rising of blood sugar reduces blood fat.The effect of soluble dietary fiber is the absorption that delays glucose in the food, eliminates postprandial hyperglycemia, to the effect that the diabetic has steady blood sugar and mitigation symptoms, also can play prevention effect to the normal person.Jin Ping etc. studies confirm that also some soluble polysaccharides in the pumpkin have hypoglycemic and effect that rise insulin.In addition, abundant soluble dietary fiber can also reduce the cholesterol levels of blood plasma and liver in the pumpkin, promotes the discharge of body inner cholesterol.The acid of pectin in the pumpkin, cyclopropylamino (CTY) and some trace elements zn, Cr etc. play an important role to prevention and treatment diabetes and hyperglycemia.Pectin can reduce cholesterol in human body content in conjunction with cholesterol, prevention and treatment atherosclerotic; The pectin fiber can also reduce gastrointestinal hormone " gastric inhibitory potypeptide " (Gastric inhitory polyperide, GIP) secretion, postprandial blood sugar and serum insulin level are descended, pectin has fabulous absorption property simultaneously, can bond and effective cytotoxin and heavy metal and the radioactive elements such as lead, mercury of eliminating in the body.
But have no at present the correlation technique report about pumpkin dietary fiber.
Summary of the invention
An object of the present invention is the deficiency for existing dietary fiber technology of preparing, especially the technical deficiency of pumpkin dietary fiber, a kind of preparation method of pumpkin dietary fiber is provided, successfully prepare pumpkin water-soluble dietary fiber and insoluble diedairy fiber, the preparation method is simple, easy control of process conditions, production cost is low, is easy to large-scale industrialization production.
No matter the pumpkin dietary fiber that the present invention provides described preparation method to prepare simultaneously is water-soluble dietary fiber or insoluble diedairy fiber, all has purity height, advantage that performance is good.
Above-mentioned purpose of the present invention is achieved by following technical solution:
A kind of preparation method of pumpkin dietary fiber is provided, may further comprise the steps:
(1) gets fresh pumpkin, clean, remove the peel, go to smash to get pumpkin fruit to pieces behind the seed;
(2) adding the pH value in the pumpkin fruit after smashing to pieces is 5～7 phosphate buffer, and the weight ratio of pumpkin fruit and phosphate buffer is 1:6, gets the mixture of pumpkin fruit and phosphate buffer;
(3) in the mixture of step (2) gained pumpkin fruit and phosphate buffer, add AMS and carbohydrase enzymolysis;
(4) in step (3) enzymolysis gained enzymolysis liquid, add protease hydrolyzed;
(5) step (4) enzymolysis gained enzymolysis liquid is boiled the enzyme that goes out, suction filtration, obtain filtrate and filter residue;
With clear water filter residue is washed till neutrality, oven dry obtains insoluble diedairy fiber; With gained filtrate ethanolic solution suction filtration after the precipitation process at room temperature, the filter residue oven dry is obtained water-soluble dietary fiber.
Consisting of of step (2) described phosphate buffer: contain 0.2M sodium hydrogen phosphate 123ml in the 1L phosphate buffer, 0.2M sodium dihydrogen phosphate 877ml;
Preferably, the pH value of the described phosphate buffer of step (2) is 6.Under preferred condition, can provide for the enzymolysis processing of subsequent step of the present invention best enzymolysis environment.
Preferably, the addition of AMS and carbohydrase is respectively 55.335～719.355U/g and 412.5～5362.5U/g in the step (3).The time of the described enzymolysis of step (3) is 15～105min;
More preferably, the addition of AMS and carbohydrase is respectively 608.685U/g and 4537.5U/g in the step (3).The time of the described enzymolysis of step (3) is 90min.
Preferably, the time of the described enzymolysis of step (4) is 15～105min, and the more preferred time of enzymolysis is 75min.
Preferably, the addition of the described protease of step (4) is 2500～12500 U/g.
The most preferably, the addition of protease is 7500U/g in the step (4), and enzymolysis time is 75min.
Further, the total addition level of described AMS and carbohydrase can be determined according to 0.5～1.1% of pumpkin fruit weight; The part by weight of described AMS and carbohydrase is preferably 1:1～1:9, most preferably is 1:7.
The temperature of the described enzymolysis of step (3) is 40～80 ℃, preferred 60 ℃
For step (3), the invention provides a highly preferred technical scheme, the addition of described AMS and carbohydrase is respectively 608.685U/g and 4537.5U/g, and the ratio of the two is 1:7, and the temperature of enzymolysis is 60 ℃, and the time is 90min.
The preferred flavor protease of the described protease of step (4), alkali protease or papain, more preferably alkali protease.
Adopt alkali protease, the enzyme that goes out behind step (3) enzymolysis makes AMS and carbohydrase inactivation, then be cooled to 40～50 ℃, add again alkali protease after regulating pH value and be 9～10, regulate preferably employing NaOH of pH value, the concentration of NaOH is not done strict restriction, preferred 2M.
Described AMS, carbohydrase and protease all can adopt commercial conventional products.
Preferably, the consumption of the described ethanol of step (5) is preferably 4 times of volumes of gained filtrate, and the volume by volume concentration of ethanol is 95%, and the time of described precipitation process is 1 hour;
Preparation method of the present invention can also comprise step (6), and step (5) gained insoluble diedairy fiber and soluble dietary fiber are pulverized, and namely obtains pulverous dietary fiber.
The invention provides the pumpkin dietary fiber that described preparation method prepares, comprise insoluble pumpkin dietary fiber and water-soluble pumpkin dietary fiber.The dissolubility of water-soluble pumpkin dietary fiber is 83.19%, and holding oily power is 0.7221 ± 0.0252g/g, and expansive force is 5.9919 ± 0.0003mL/g, and retentiveness is 1.5524 ± 0.3288g/g; It is 2.644 ± 0.048g/g that insoluble pumpkin dietary fiber is held oily power, and expansive force is 8.4587 ± 0.0080mL/g, and retentiveness is 8.2642 ± 0.3196g/g.
Beneficial effect of the present invention:
The present invention successfully adopts stock number pumpkin large, of high quality and at a reasonable price as raw material, comprehensive process is extracted and is obtained high added value, highly active dietary fiber, it is costly a bit to give full play to functional mass rich content in the pumpkin, comprehensive nutrient, for the dietary fiber of deep exploitation pumpkin provides important technical foundation, increase a kind of good dietary fiber new varieties, had huge consumption market and very high economic worth.
The present invention is based on the long-term a large amount of research and analysis experiments to the pumpkin fiber, targetedly enzymolysis process is provided, at first utilize the starch in AMS and the carbohydrase enzymolysis removal pumpkin, then adopt the protein in the protease removal pumpkin, the selectivity of enzyme is high, starch, protein in the degraded pumpkin that can be single-minded, and do not destroy other compositions, so that soluble cellulose, hemicellulose isoreactivity composition can be preserved effectively, the physiologically active of dietary fiber is improved greatly like this.
The pumpkin dietary fiber purity that the present invention prepares is high, and the purity of insoluble diedairy fiber reaches more than 85%, and the purity of water-soluble dietary fiber reaches 90.5%.The expansive force of dietary fiber reaches every gram more than 5.99 milliliters; In addition, pumpkin dietary fiber has ability and the cation exchange capacity (CEC) of good absorption cholesterol, natrium nitrosum, has the effect such as lipopenicillinase, fat-reducing, anti-cancer of pumpkin.
Operating procedure of the present invention is simple, and process conditions are gentle, and equipment is simple, and is with low cost, is fit to industrialness and promotes and large-scale production, provides powerful support for for the utilization of a large amount of pumpkin resource of China provides.
Description of drawings
The addition of Fig. 1 AMS and carbohydrase is on the impact of TDF yield
The ratio of Fig. 2 AMS and carbohydrase is on the impact of TDF yield
The hydrolysis temperature of Fig. 3 AMS and carbohydrase is on the impact of TDF yield
The enzymolysis time of Fig. 4 AMS and carbohydrase is on the impact of TDF yield
Fig. 5 protease addition is on the impact of TDF yield
Fig. 6 protease hydrolyzed time is on the impact of TDF yield
Fig. 7 cholesterol calibration curve
Fig. 8 pumpkin dietary fiber is to the absorption of cholesterol
Fig. 9 natrium nitrosum calibration curve
Figure 10 pumpkin dietary fiber (IDF) is to the absorption (pH=2) of natrium nitrosum
Figure 11 pumpkin dietary fiber (SDF) is to the absorption (pH=2) of natrium nitrosum
Figure 12 pumpkin dietary fiber (IDF) is to the absorption (pH=7) of natrium nitrosum
Figure 13 pumpkin dietary fiber (SDF) is to the absorption (pH=7) of natrium nitrosum
Figure 14 pumpkin dietary fiber cation exchange capacity (CEC)
Figure 15 pumpkin dietary fiber cation exchange capacity (CEC).
The specific embodiment
Describe the present invention in detail below in conjunction with specific embodiment.Unless stated otherwise, the raw material of the present invention's employing and equipment are conventional commercial raw material and equipment of the art.
Embodiment 1Pumpkin fiber raw materials composition and engineering list factor research experiment
AMS can become the Starch Hydrolysis in the pumpkin raw material dextrin and compound sugar, and carbohydrase can be hydrolyzed dextrin single-mindedly, compound sugar becomes glucose and is dissolved in organic solvent.So the present invention intends adopting AMS and Glucoamylase hydrolysis pumpkin.But enzymolysis is removed the effect of starch except closely related with the kind of enzyme, and the factors such as the mutual proportioning in the interpolation total amount of the addition of each enzyme, mixed enzyme, the mixed enzyme between the various enzymes, enzymolysis time, hydrolysis temperature are closely related, also there is mutual influence between these factors, the mechanism of action more complicated.Simultaneously, enzymolysis starch step operating result also exists important impact to the follow-up preparation process of pumpkin dietary fiber, need to scientifically consider.
The applicant determines to adopt AMS and carbohydrase as the starch in the mixed enzyme hydrolysis pumpkin at first based on the specific aim research to the pumpkin constituent, obtains desirable effect.The addition of determining mixed enzyme (AMS and carbohydrase) is 0.1%, 0.3%, 0.5%, 0.7%, 0.9%, 1.1%.Pressing enzyme work calculates, the addition of AMS then is 55.335U/g, 166.005U/g, 276.675U/g, 387.345U/g, 498.015U/g, 608.685U/g and 719.355U/g, and the addition of carbohydrase is 412.5U/g, 1237.5U/g, 2062.5U/g, 2887.5U/g, 3712.5U/g, 4537.5U/g and 5362.5U/g.Accompanying drawing 1 provides mixed enzyme Different adding amount of the present invention to remove the result of pumpkin starch, and (total dietary fiber, TDF=IDF+SDF) yield is index with TDF.Increase along with the mixed enzyme consumption, TDF increases first rear minimizing, this is because the adding of mixed enzyme agent, enzyme promptly decomposes the partial starch in the pumpkin fruit, and the dietary fiber of extraction contains certain starch, so yield is high, carrying out along with increase and the reaction of enzyme dosage, starch in the pumpkin is fully degraded, and yield descends, and the present invention has selected these two kinds of enzymes to form mixed enzyme and removed starch.
More than experiment, analyze through SPSS, selected the addition of AMS and carbohydrase to be respectively 608.685U/g and 4537.5U/g, be 1.1%, the ratio selectable of the two is selected 1:1,1:3,1:5,1:7,1:9, both can effectively remove desizing, and can remove again dextrin and compound sugar in the Soluble Fiber.Accompanying drawing 2 provides the affect result of the ratio of mixed enzyme on the TDF yield.Experiment shows that 1:3 is optimum ratio.
Among the present invention, adopt different enzymolysis times to test.Here chosen 15,30,45,60,75,90,105min is that example describes.Accompanying drawing 4 provides the experimental result of enzymolysis time.Along with the increase of time, the yield of TDF increases, and is later on downward trend to 90min, analyzes through SPSS, and 75min and 90min are without significant difference, and all there were significant differences with other for 75min, and for saving time, Decrease production cost is selected 75min.
Behind the Starch Hydrolysis, for the removal of pumpkin fruit protein, present embodiment has carried out the research experiment of hydrolysis result.Present embodiment provides the experimental result of preferred three kinds of protease, is shown in Table 1.Be flavor protease, papain and alkali protease.The experimental result demonstration, in described three kinds of protease, the better effects if of alkali protease, protein place to go effect is more thorough.
Get alkali protease and carry out the experiment of Different adding amount, addition take alkali protease is respectively 0.2%, 0.4%, 0.6%, 0.8%, 1.0% as example, press enzyme work calculation addition and then be 2500U/g, 5000U/g, 7500U/g, 10000U/g, 12500U/g, accompanying drawing 5 is that the alkali protease addition is on the result that affects of TDF yield.Experimental result shows: when the protease consumption was between 2500～7500 U/g, along with the increase of protease consumption, yield increased sharply; When the protease consumption was between 7500～12500 U/g, yield reduced along with the increase of enzyme amount.Analyze through SPSS, determine that addition take alkali protease is as 0.6%(7500 U/g) as optimum addition.
Present embodiment has carried out the experiment of different enzymolysis time, chooses 15,30,45,60,75,90,105min is example, and accompanying drawing 6 has provided the enzymolysis time result.Experimental result as can be known, in the time of 15～45 min, yield is in rising trend, during 45～105 min, yield descends.Analyze former because: the reaction time prolongation can make enzyme and protein molecule fully act on, thereby form more covalent cross-linking, yield is increased, reach certain hour, protease is fully degraded, and because selectivity and the high efficiency of enzyme, and the residual etc. of the mixed enzyme used of hydrolyzed starch considers, continue overtime at enzymolysis time, the dietary fiber yield descends again to some extent, and the present invention determines that protease hydrolyzed time 45min is for well.
Embodiment 2The pumpkin dietary fiber optimum preparation condition
On the basis of single factor experiment, for obtaining the optimised process of preparation dietary fiber, the present invention has chosen mixed enzyme addition, mixed enzyme enzymolysis time, mixed enzyme ratio, protease addition, protease hydrolyzed time and has carried out further orthogonal test, these five factors of integrated survey are on the impact of pumpkin dietary fiber preparation, and evaluation result represents with pumpkin total dietary fiber (TDF) yield.Adopt L
16(4
5) orthogonal test table (being shown in Table 2) is tested, and definite best of breed.
Pumpkin dietary fiber optimum preparation condition Orthogonal experiment results
Experimental result adopts SPSS to analyze, can find out from the result of analysis of variance table shown in the table 3, the addition of mixed enzyme has larger impact (P=0.000) with the mixed enzyme enzymolysis time to pumpkin total dietary fiber (TDF) yield, next is protease hydrolyzed time (P=0.001), mixed enzyme ratio affects not quite (P=0.072) to the result, and the protease addition affects also little (P=0.128) to the result.
Can find A in conjunction with mean estimated statement 4 and paired comparisons table table 5 in single factor statistics scale
1, A
2Mean large (1.519,1.510), and all with A
3, A
4There were significant differences (P＜0.05).From economic angle, save enzyme dosage, select A
1Be the best.
Can find B in conjunction with mean estimated statement 6 and paired comparisons table table 7 in single factor statistics scale
3Mean is large (1.513), and and B
2, B
4There were significant differences (P＜0.05), with B
1Without significant difference (P〉0.05).
Can find E in conjunction with mean estimated statement 8 and paired comparisons table table 9 in single factor statistics scale
4Mean is large (1.513), and and E
1, E
3There were significant differences (P＜0.05), with E
2Without significant difference (P〉0.05).
Conclusion: by the actual conditions of above orthogonal experiments in conjunction with the pumpkin dietary fiber experiment, summing up the optimum extraction process that obtains pumpkin dietary fiber is A
1B
3C
4D
4E
4Be that the mixed enzyme addition is that the addition of 0.5%(AMS and carbohydrase is respectively 276.675U/g and 2062.5U/g), the mixed enzyme enzymolysis time is 90min, ratio is 1:7, the protease addition is 0.8%(10000U/g), the protease hydrolyzed time is 75min.With this understanding, pumpkin total dietary fiber yield (TDF) is 39.1%.
Pumpkin dietary fiber prepares by following preparation method in the present embodiment:
(1) gets fresh pumpkin, clean, go seed, adopt tissue mashing machine to smash to get pumpkin fruit to pieces;
(2) the pH value that adds 0.2M in the pumpkin fruit after smashing to pieces is 6 phosphate buffer, and the mass ratio of pumpkin fruit and phosphate buffer is 1:6;
Consisting of of phosphate buffer: contain 0.2M sodium hydrogen phosphate 123ml in the 1L phosphate buffer, 0.2M sodium dihydrogen phosphate 877ml;
(3) mixed in step (2) in the pumpkin fruit of phosphate buffer and add mixed enzyme, the amount that mixed enzyme adds is determined according to 0.5% of phosphate buffer and pumpkin fruit gross weight; AMS and carbohydrase weight ratio are that 1:7(is that AMS and carbohydrase are respectively 216 U/g and 2398 U/g in the mixed enzyme), in 60 ℃ of enzymolysis 90min, be cooled to room temperature after boiling the enzyme that goes out, the pH value to 10 of regulating enzymolysis liquid with the NaOH of 2M gets enzymolysis liquid A;
(4) add alkali protease in step (3) gained enzymolysis liquid A, the addition of alkali protease is 0.8% of enzymolysis liquid A weight, i.e. 10000 U/g.In 50 ℃ of enzymolysis 75min, boil the enzyme that goes out, be cooled to room temperature and get enzymolysis liquid B;
(5) enzymolysis liquid B is filtered, filter the filter residue that obtains and be washed till neutrality with clear water, freeze drying (subzero 45 ℃ of freeze-drying, freeze drier) is pulverized, and gets insoluble diedairy fiber (IDF);
Enzymolysis liquid B filtered the filtrate obtain with 4 times of filtrate volume ethanol precipitation process under room temperature, the volume by volume concentration of described ethanol is preferably 95%, and the sedimentation time is about 1 hour, filter after the precipitation process, with filter residue freezing (with aforementioned) drying, pulverize, obtain soluble dietary fiber (SDF).
Present embodiment pumpkin total dietary fiber yield (TDF) is 39.1%.The purity of insoluble diedairy fiber reaches more than 85%; The purity of water-soluble dietary fiber reaches 90.5%.Detection method is with reference to GB GB/T5009.88-2008 and GB/T5009.88-2003).
(2) the functional characteristic analysis of pumpkin dietary fiber
(1) expansive force: take by weighing 0.2 g water-soluble dietary fiber powder (SDF) and put into graduated cylinder, write down the volume of water-soluble dietary fiber dry product, accurately pipetting 10 mL distilled water with pipette adds wherein, under room temperature, place 24 h after shaking up, then the absolute ethyl alcohol that adds 5 times of volumes reads the volume of the water-soluble dietary fiber (SDF) that precipitates in the liquid.
Accurately take by weighing 0.1g insoluble diedairy fiber powder (IDF) and place tool plug scale test tube, add 10 mL distilled water, leave standstill 18h under the room temperature behind the mixing, read volume after the sample sedimentation.
Expansive force=[volume after the swelling (mL)-dry product volume (mL)]/sample dry weight (g)
(2) hold oily power: take by weighing 1.0 g dietary fibers in 50 mL centrifuge tubes, add peanut oil 10 mL, 37 ℃ leave standstill l h, and the centrifugal 20min of 4000 r/min discards oil and the residue on upper strata, blots free oil with filter paper, weighs.
Hold oily power (g/g)=[heavy (the g)-sample dry weight (g) of sample oil]/sample dry weight (g)
(3) retention ability: take by weighing 1 g water-soluble dietary fiber powder (SDF) and put into beaker, then add entry 100 mL, at room temperature, behind electromagnetic agitation 24 h, add the absolute ethyl alcohol precipitation of 5 times of volumes, suction filtration claims quality.Calculate retention ability.
Accurately take by weighing 0.8g sample (IDF), place test tube, add 40mL distilled water, leave standstill 20h under the room temperature behind the mixing, the centrifugal 20min of 4000 r/min claims quality after removing supernatant.
Retention ability=[(fiber wet weight g-fiber dry weight g)/fiber dry weight g] * 100%
(4) pumpkin dietary fiber is to the absorption of cholesterol:
Get the yolk of commercially available fresh hen egg, fully beat into emulsion with 9 times of amount distilled water.Get respectively present embodiment 2.0 g pumpkin insoluble diedairy fibers (IDF), pumpkin soluble dietary fiber (SDF) in the triangular flask of 200 ml, add 50g dilution egg yolk liquid, stir, regulation system pH value to 2.0 and 7.0, put in the shaking table 37 ℃ of vibration 3 h, centrifugal 20min(precipitation dietary fiber under the 4000r/min), draw the 0.04ml supernatant, adopt o-phthalaldehyde method colorimetric estimation cholesterol level under 550nm.Take cholesterol as standard substance production standard curve, y=0.0012x+0.0039, R
2=0.9976, see shown in the accompanying drawing 7.
Dietary fiber is to the adsorbance (A) of cholesterol=(before the absorption in the egg yolk liquid cholesterol amount-absorption afterwards cholesterol amount in the supernatant)/dietary fiber weight.Pumpkin dietary fiber of the present invention the results are shown in shown in the accompanying drawing 8 adsorption experiment of cholesterol.As seen from the figure, pumpkin dietary fiber is better than soybean dietary fiber far away to the cholesterol adsorption capacity, and wherein the adsorption effect of insoluble diedairy fiber is best, reaches 18.365mg/g, and the adsorption effect in the pH=7 environment is better than pH=2's.
(5) pumpkin dietary fiber is to the absorption of natrium nitrosum:
A. NO
2 -The formulation of calibration curve
Press the GB5009.33-2008 method, accurately pipette 0.00mL, 1.0mL, 2.0mL, 3.0mL, 4.0mL, 5.0mL, 7.5mL, 10mL, 12.5mL NO
2 -Solution (every milliliter is equivalent to 5 μ g) adds respectively 2 mL, 0.4% sulfanilic acid solution, mixing in 9 50 mL colorimetric cylinders, after placing 3 ~ 5min, add respectively l mL 0.2% hydrochloride naphthodiamide solution, add water to scale, mixing, leave standstill 15 min, use the 1cm cuvette, take zero pipe as reference, measure 538nm wavelength place absorbance at spectrophotometer, the drawing standard curve is seen shown in the accompanying drawing 9.
B. adsorption effect is measured
It is that the pH value is respectively 7.0 and 2.0 (regulating the pH value with hydrochloric acid and NaOH respectively, simulation small intestine and gastric environment) that the absorption environment is set, and reaction is carried out in the 250mL conical flask, and reaction volume is 100mL, NO
2 -Concentration is 220mol/L, adds 0.5g pumpkin dietary fiber of the present invention, and the concentration that lmL sample liquid is measured NO2-is respectively got in the electromagnetic agitation reaction at set intervals under 37 ℃ of constant temperature, respectively does the blank assay contrast simultaneously, and experimental result is seen shown in accompanying drawing 10 ~ accompanying drawing 13.As seen from the figure, pumpkin dietary fiber has suction-operated to nitrite in pH=2, does not have suction-operated in pH=7.The suction-operated of insoluble diedairy fiber (IDF) be greater than (adsorption time of solubility will be grown) of soluble dietary fiber (SDF), and the adsorption capacity of two kinds of pumpkin dietary fibers is all greater than soybean dietary fiber.
(6) cation exchange capacity (CEC)
Take by weighing 0.25 g dietary fiber sample of the present invention, add HCl 10 mL of 0.1 mol/L, shake up, room temperature is placed 24 h.Filter paper filtering repeatedly cleans with distilled water and to remove unnecessary acid, and (the AgNO3 solution with 10% is identified and no longer contained till the chlorion in triangular flask to shift residue, vacuum drying), add the NaCl solution of 100 mL 15%, use magnetic stirrer, make indicator with 0.5% phenolphthalein, with the slow titration of 0.01 mol/L NaOH solution, stop titration when solution is little when red, jolting is dripped after fading again, until jolting 5min still colour-fast be terminal point, the record titration process in the pH value.Make V
NaOHWith pH value graph of a relation.Replace HCl with distilled water, measure the amount of the blank NaOH that consumes.Pumpkin dietary fiber cation exchange capacity (CEC) experimental result is seen shown in accompanying drawing 14 and the accompanying drawing 15, as seen from the figure, the dietary fiber curvilinear motion is relatively slow in the titration curve, cation exchange capacity (CEC) is relatively poor, this may be because leaching process has to a certain degree destruction to the side-chain radical structure in the dietary fiber chemical constitution, thereby causes reducing of its cation exchange capacity (CEC).But the cation exchange capacity (CEC) of pumpkin insoluble diedairy fiber (IDF) is better than the soybean insoluble diedairy fiber.
Pumpkin dietary fiber prepares by following preparation method in the present embodiment:
(1) gets fresh pumpkin, clean, go seed, adopt tissue mashing machine to smash to get pumpkin fruit to pieces;
(2) the pH value that adds 0.2M in the pumpkin fruit after smashing to pieces is 6 phosphate buffer, and the mass ratio of pumpkin fruit and phosphate buffer is 1:6;
(3) mixed in step (2) in the pumpkin fruit of phosphate buffer and add mixed enzyme, the amount that mixed enzyme adds is determined according to 1.1% of phosphate buffer and pumpkin fruit gross weight; AMS and carbohydrase weight ratio are that 1:1(is that AMS and carbohydrase are respectively 1221.813U/g and 3021.334U/g in the mixed enzyme), in 80 ℃ of enzymolysis 15min, be cooled to room temperature after boiling the enzyme that goes out, the pH value to 10 of regulating enzymolysis liquid with the NaOH of 2M gets enzymolysis liquid A;
(4) add alkali protease in step (3) gained enzymolysis liquid A, the addition of alkali protease is 0.2% of enzymolysis liquid A weight, and namely 2500U/g in 50 ℃ of enzymolysis 105min, boils the enzyme that goes out, and is cooled to room temperature and gets enzymolysis liquid B;
(5) enzymolysis liquid B is filtered, filter the filter residue that obtains and be washed till neutrality with clear water, freeze drying (reference is conventional or adopt freeze drier-45 ℃ of lower freeze-drying) is pulverized, and gets insoluble diedairy fiber (IDF);
Enzymolysis liquid B filtered the filtrate obtain with 4 times of filtrate volume ethanol precipitation process under room temperature, the volume by volume concentration of described ethanol is preferably 95%, and the sedimentation time is about 1 hour, filter after the precipitation process, with the filter residue freeze drying, pulverize, obtain soluble dietary fiber (SDF).
The functional characteristic analysis of pumpkin dietary fiber is with reference to embodiment 3, and present embodiment pumpkin total dietary fiber yield (TDF) is 36.8%.The dissolubility of water-soluble pumpkin dietary fiber is 83%, and holding oily power is 0.7001 ± 0.0131g/g, and expansive force is 5.329 ± 0.0324mL/g, and retentiveness is 1.329 ± 0.0385g/g; It is 2.086 ± 0.0276g/g that insoluble pumpkin dietary fiber is held oily power, and expansive force is 7.376 ± 0.112mL/g, and retentiveness is 7.863 ± 0.201g/g.
Pumpkin dietary fiber prepares by following preparation method in the present embodiment:
(1) gets fresh pumpkin, clean, go seed, adopt tissue mashing machine to smash to get pumpkin fruit to pieces;
(2) the pH value that adds 0.2M in the pumpkin fruit after smashing to pieces is 5 phosphate buffer, and the mass ratio of pumpkin fruit and phosphate buffer is 1:6;
(3) mixed in step (2) in the pumpkin fruit of phosphate buffer and add mixed enzyme, the amount that mixed enzyme adds is determined according to 1.0% of phosphate buffer and pumpkin fruit gross weight; AMS and carbohydrase weight ratio are that 1:9(is that AMS and carbohydrase are respectively 222.148U/g and 4944.007U/g in the mixed enzyme), in 40 ℃ of enzymolysis 105min, be cooled to room temperature after boiling the enzyme that goes out, the pH value to 9 with the NaOH of 2M is regulated enzymolysis liquid gets enzymolysis liquid A;
(4) add alkali protease in step (3) gained enzymolysis liquid A, the addition of alkali protease is 1.0% of enzymolysis liquid A weight, and namely 12500U/g in 40 ℃ of enzymolysis 15min, boils the enzyme that goes out, and is cooled to room temperature and gets enzymolysis liquid B;
(5) enzymolysis liquid B is filtered, filter the filter residue that obtains and be washed till neutrality with clear water, freeze-drying is pulverized, and gets insoluble diedairy fiber (IDF);
Enzymolysis liquid B filtered the filtrate obtain with 4 times of filtrate volume ethanol precipitation process under room temperature, the volume by volume concentration of described ethanol is preferably 95%, and the sedimentation time is about 1 hour, filter after the precipitation process, with the filter residue freeze-drying, pulverize, obtain soluble dietary fiber (SDF).
The functional characteristic analysis of pumpkin dietary fiber is with reference to embodiment 3, and present embodiment pumpkin total dietary fiber yield (TDF) is 35.6 %.The dissolubility of water-soluble pumpkin dietary fiber is 82.1%, and holding oily power is 0.623 ± 0.0121g/g, and expansive force is 5.001 ± 0.0234mL/g, and retentiveness is 1.432 ± 0.0324g/g; It is 1.982 ± 0.0321g/g that insoluble pumpkin dietary fiber is held oily power, and expansive force is 6.985 ± 0.124mL/g, and retentiveness is 7.123 ± 0.0423g/g.
Pumpkin dietary fiber prepares by following preparation method in the present embodiment:
(1) gets fresh pumpkin, clean, go seed, adopt tissue mashing machine to smash to get pumpkin fruit to pieces;
(2) the pH value that adds 0.2M in the pumpkin fruit after smashing to pieces is 6 phosphate buffer, and the mass ratio of pumpkin fruit and phosphate buffer is 1:6;
(3) mixed in step (2) in the pumpkin fruit of phosphate buffer and add mixed enzyme, the amount that mixed enzyme adds is determined according to 0.5% of phosphate buffer and pumpkin fruit gross weight; AMS and carbohydrase weight ratio are that 1:7(is that AMS and carbohydrase are respectively 216 U/g and 2398 U/g in the mixed enzyme), in 60 ℃ of enzymolysis 90min, boil and be cooled to room temperature behind the enzyme that goes out and get enzymolysis liquid A;
(4) add flavor protease in step (3) gained enzymolysis liquid A, the addition of flavor protease is 0.8% of enzymolysis liquid A weight, i.e. 10000 U/g.In 50 ℃ of enzymolysis 75min, boil the enzyme that goes out, be cooled to room temperature and get enzymolysis liquid B;
(5) enzymolysis liquid B is filtered, filter the filter residue that obtains and be washed till neutrality with clear water, freeze-drying is pulverized, and gets insoluble diedairy fiber (IDF);
Enzymolysis liquid B filtered the filtrate obtain with 4 times of filtrate volume ethanol precipitation process under room temperature, the volume by volume concentration of described ethanol is preferably 95%, and the sedimentation time is about 1 hour, filter after the precipitation process, with the filter residue freeze-drying, pulverize, obtain soluble dietary fiber (SDF).
Present embodiment pumpkin total dietary fiber yield (TDF) is 35.9%.The functional characteristic analysis of pumpkin dietary fiber is with reference to embodiment 3, and the dissolubility of water-soluble pumpkin dietary fiber is 82.5%, and holding oily power is 0.5183 ± 0.0212g/g, and expansive force is 4.312 ± 0.0223mL/g, and retentiveness is 1.043 ± 0.0121g/g; It is 1.346 ± 0.0253g/g that insoluble pumpkin dietary fiber is held oily power, and expansive force is 6.783 ± 0.213mL/g, and retentiveness is 5.428 ± 0.125g/g.
Claims (10)
1. the preparation method of a pumpkin dietary fiber is characterized in that may further comprise the steps:
(1) gets fresh pumpkin, clean, remove the peel, go to smash to get pumpkin fruit to pieces behind the seed;
(2) adding the pH value in the pumpkin fruit after step (1) is smashed to pieces is 5～7 phosphate buffer, and the weight ratio of pumpkin fruit and phosphate buffer is 1:6, gets the mixture of pumpkin fruit and phosphate buffer;
(3) in the mixture of step (2) gained pumpkin fruit and phosphate buffer, add AMS and carbohydrase enzymolysis;
(4) in step (3) enzymolysis gained enzymolysis liquid, add protease hydrolyzed;
(5) step (4) enzymolysis gained enzymolysis liquid is boiled the enzyme that goes out, suction filtration, obtain filtrate and filter residue;
With clear water filter residue is washed till neutrality, oven dry obtains insoluble diedairy fiber;
With gained filtrate ethanolic solution suction filtration after the precipitation process at room temperature, the filter residue oven dry is obtained water-soluble dietary fiber;
Wherein, the addition of the described AMS of step (3) and carbohydrase is respectively 55.335～719.355U/g and 412.5～5362.5U/g; The time of described enzymolysis is 15～105min;
The time of the described enzymolysis of step (4) is 15～105min.
2. the preparation method of described pumpkin dietary fiber according to claim 1 is characterized in that the addition of the described AMS of step (3) and carbohydrase is respectively 608.685U/g and 4537.5U/g.
3. the preparation method of described pumpkin dietary fiber according to claim 1 is characterized in that the time of the described enzymolysis of step (3) is 90min.
4. the preparation method of described pumpkin dietary fiber according to claim 1 is characterized in that the time of the described enzymolysis of step (4) is 75min.
5. the preparation method of described pumpkin dietary fiber according to claim 1 is characterized in that the addition of the described protease of step (4) is 2500～12500 U/g.
6. the preparation method of described pumpkin dietary fiber according to claim 1 is characterized in that the total addition level of described AMS and carbohydrase can be determined according to 0.5～1.1% of pumpkin fruit weight; The part by weight of described AMS and carbohydrase is 1:1～1:9, preferred 1:7.
7. the preparation method of described pumpkin dietary fiber according to claim 1 is characterized in that the addition of the described protease of step (4) is 7500U/g, and enzymolysis time is 75min.
8. the preparation method of described pumpkin dietary fiber according to claim 1 is characterized in that the temperature of the described enzymolysis of step (3) is 40～80 ℃.
9. the pumpkin dietary fiber for preparing of each described preparation method of claim 1～8.
10. the dietary fiber for preparing of preparation method claimed in claim 9 is characterized in that comprising insoluble pumpkin dietary fiber and/or water-soluble pumpkin dietary fiber; The dissolubility of described water-soluble pumpkin dietary fiber is 83.19%, and holding oily power is 0.7221 ± 0.0252g/g, and expansive force is 5.9919 ± 0.0003mL/g, and retentiveness is 1.5524 ± 0.3288g/g;
It is 2.644 ± 0.048g/g that described insoluble pumpkin dietary fiber is held oily power, and expansive force is 8.4587 ± 0.0080mL/g, and retentiveness is 8.2642 ± 0.3196g/g.
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| CN103719880A (en) * | 2014-01-17 | 2014-04-16 | 昆明医科大学 | Preparation method of high-activity purple sweet potato dietary fiber |
| CN103976371A (en) * | 2014-06-03 | 2014-08-13 | 中国农业大学 | Method for extracting ponkan dietary fibers through extrusion modification and enzymolysis coupling |
| CN104957640A (en) * | 2015-06-26 | 2015-10-07 | 北京京隆卓尚投资有限公司 | Cistanche deserticola dietary fiber and preparation method thereof |
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Effective date of registration: 20200930 Address after: Room 3, office building, No.60, Weiliu Road, Airport Economic Zone, Dongli District, Tianjin Patentee after: Zhongke Aijia (Tianjin) Medical Food Co., Ltd Address before: 510642 No. five, 483 mountain road, Guangzhou, Guangdong, Tianhe District Patentee before: SOUTH CHINA AGRICULTURAL University |