CN102960644B - 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 and take foods processing technique and the functional food additives technical field that agricultural byproducts are raw material, be specifically related to a kind of method that enzyme process is prepared 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 that edible cell, polysaccharide, lignin, colloid, modified cellulose, clayey, oligosaccharides, pectin and a small amount of constituent that can not be digested and assimilated by human body endogenous enzymes are as wax, cutin, cork matter etc.According to the dissolubility in water, can be divided into water-soluble dietary fiber (SDF) and the large class of water insoluble dietary fiber (IDF) two.Insoluble diedairy fiber refers to not by human body alimentary canal enzymic digestion and is insoluble to the part dietary fiber of hot water, it is the main component that forms 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 does not refer to and is digested by human body digestive ferment, 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 being present in apple, tangerine class, the glue in vegetable seeds, the algin in 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 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 a tortuous process 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 current 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 proposes for 2000, the suitable absorption of adult's dietary fiber 30.2g/d.
At present, it is found that and the dietary fiber developed has tens kinds more than.Conventionally by dissolubility, divide, dietary fiber divides two kinds of water-soluble and water-insolubles.The representative of water-soluble dietary fiber has glucan, pectin, guar gum and fungi polysaccharide etc.; Water insoluble dietary fiber mainly be take soybean dietary fiber, shitosan and hemicellulose as main.
Pumpkin, annual herb climbs climing plant, belongs to Curcurbitaceae, Cucurbita.The annual pumpkin output of China accounts for the more than 13% of Gross World Product, occupy No. 1 in the world, and its resource is very abundant.In < < Compendium of Materia 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 ".Current research both domestic and external shows; 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 body; especially there is remarkable efficacy preventing and treating aspect diabetes; in addition; in pumpkin starch, resistance starch content is higher, is having certain advantage, therefore aspect anti-digestibility; pumpkin has medical treatment widely and health-care effect, is a kind of desirable natural health dietotherapy product.
Current 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 due to research complex technical process, cost is too high, mostly still rests on laboratory sample and grinds the examination stage.Therefore, improve the preparation technology of dietary fiber, more kinds of resources that dietary fiber is prepared in developing, further method and the condition of the applicable suitability for industrialized production of research have important practical significance.
In pumpkin, contain abundant dietary fiber, the organic molecule of dietary fiber energy absorption of bile acid, cholesterol mutagen, after 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 excretion 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 food, eliminates postprandial hyperglycemia, diabetic is had to the effect of steady blood sugar and mitigation symptoms, to normal person, also can play prevention effect.Jin Ping etc. also studies confirm that, some soluble polysaccharides in pumpkin have hypoglycemic and effect that rise insulin.In addition, in pumpkin, abundant soluble dietary fiber can also reduce the cholesterol levels of blood plasma and liver, promotes the discharge of body inner cholesterol.The acid of pectin in 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; Pectin fiber can also reduce gastrointestinal hormone " gastric inhibitory potypeptide " (Gastric inhitory polyperide, GIP) secretion, postprandial blood sugar and serum insulin levels are declined, simultaneously pectin has fabulous absorption property, can bond and effective heavy metal and the radioactive element such as the cytotoxin in elimination body and lead, mercury.
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, preparation method is simple, easy control of process conditions, production cost is low, is easy to large-scale industrialization and produces.
No matter the pumpkin dietary fiber that the present invention provides described preparation method to prepare simultaneously, be water-soluble dietary fiber or insoluble diedairy fiber, all has advantages of that purity is high, performance good.
Above-mentioned purpose of the present invention is achieved by following technical solution:
The preparation method that a kind of pumpkin dietary fiber is provided, comprises the following steps:
(1) get fresh pumpkin, clean, remove the peel, go to smash to obtain pumpkin fruit to pieces after seed;
(2) in the pumpkin fruit after smashing to pieces, adding pH value is 5～7 phosphate buffer, and the weight ratio of pumpkin fruit and phosphate buffer is 1:6, obtains 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 to the enzyme that goes out, suction filtration, obtain filtrate and filter residue;
With clear water, filter residue is washed till to neutrality, dries, obtain insoluble diedairy fiber; By gained filtrate ethanolic solution suction filtration after precipitation process at room temperature, filter residue is dried and obtained water-soluble dietary fiber.
Consisting of of step (2) described phosphate buffer: contain 0.2M sodium hydrogen phosphate 123ml, 0.2M sodium dihydrogen phosphate 877ml in 1L phosphate buffer;
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, in step (3), the addition of AMS and carbohydrase is respectively 55.335～719.355U/g and 412.5～5362.5U/g.The time of the described enzymolysis of step (3) is 15～105min;
More preferably, in step (3), the addition of AMS and carbohydrase is respectively 608.685U/g and 4537.5U/g.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, in step (4), the addition of protease is 7500U/g, 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 ℃, preferably 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 after step (3) enzymolysis, makes AMS and carbohydrase inactivation, then be cooled to 40～50 ℃, regulating pH value is to add alkali protease after 9～10 again, regulates the preferred employing of pH value NaOH, the concentration of NaOH is not done strict restriction, preferably 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 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, give full play to functional mass rich content in pumpkin, comprehensive nutrient is a little costly, for the dietary fiber of deep exploitation pumpkin provides important technical foundation, increase a kind of good dietary fiber new varieties, there is 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 pumpkin fiber, enzymolysis process is targetedly provided, first utilize AMS and carbohydrase enzymolysis to remove the starch in pumpkin, then adopt protease to remove the protein in pumpkin, the selectivity of enzyme is high, starch, protein in degraded pumpkin that can be single-minded, and do not destroy other compositions, make like this soluble cellulose, hemicellulose isoreactivity composition effectively to preserve, the physiologically active of dietary fiber is improved greatly.
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, with low cost, and the utilization that is applicable to a large amount of pumpkin resource of industrialness popularization and large-scale production ，Wei China provides and provides powerful support for.
Accompanying drawing explanation
The impact of the addition of Fig. 1 AMS and carbohydrase on TDF yield
The impact of the ratio of Fig. 2 AMS and carbohydrase on TDF yield
The impact of the hydrolysis temperature of Fig. 3 AMS and carbohydrase on TDF yield
The impact of the enzymolysis time of Fig. 4 AMS and carbohydrase on TDF yield
The impact of Fig. 5 protease addition on TDF yield
The impact of Fig. 6 protease hydrolyzed time on TDF yield
Fig. 7 cholesterol calibration curve
The absorption of Fig. 8 pumpkin dietary fiber to cholesterol
Fig. 9 natrium nitrosum calibration curve
Figure 10 pumpkin dietary fiber (IDF) is to the absorption of natrium nitrosum (pH=2)
Figure 11 pumpkin dietary fiber (SDF) is to the absorption of natrium nitrosum (pH=2)
Figure 12 pumpkin dietary fiber (IDF) is to the absorption of natrium nitrosum (pH=7)
Figure 13 pumpkin dietary fiber (SDF) is to the absorption of natrium nitrosum (pH=7)
Figure 14 pumpkin dietary fiber cation exchange capacity (CEC)
Figure 15 pumpkin dietary fiber cation exchange capacity (CEC).
The specific embodiment
Below in conjunction with specific embodiment, describe the present invention in detail.Unless stated otherwise, the raw material that the present invention adopts 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 dextrin and compound sugar by the Starch Hydrolysis in pumpkin raw material, 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 the kind with enzyme is closely related, and the factors such as the mutual proportioning in the interpolation total amount of the addition of each enzyme, mixed enzyme, mixed enzyme between various enzymes, enzymolysis time, hydrolysis temperature are closely related, between these factors, also there is mutual influence, mechanism of action more complicated.Meanwhile, 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 is the specific aim research based on to pumpkin constituent first, determines and adopt AMS and carbohydrase as the starch in mixed enzyme hydrolysis pumpkin, 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 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 with TDF, (total dietary fiber, TDF=IDF+SDF) yield is index.Increase along with mixed enzyme consumption, TDF first increases rear minimizing, this is adding because of mixed enzyme agent, enzyme promptly decomposes the partial starch in pumpkin fruit, and the dietary fiber of extraction is containing certain starch, so yield is high, carrying out along with increase and the reaction of enzyme dosage, starch in pumpkin is fully degraded, and yield declines, and the present invention has selected these two kinds of enzymes to form mixed enzyme and removed starch.
More than experiment, through SPSS, analyze, 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 Soluble Fiber.Accompanying drawing 2 provides the affect result of the ratio of mixed enzyme on TDF yield.Experiment shows, 1:3 is optimum ratio.
In 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 to 90min, is downward trend later, through SPSS, analyzes, and 75min and 90min are without significant difference, and all there were significant differences with other for 75min, for saving time, reduces production cost, selects 75min.
After Starch Hydrolysis, for the removal of pumpkin fruit protein, the present embodiment has carried out the research experiment of hydrolysis result.The present embodiment provides the experimental result of preferred three kinds of protease, is shown in Table 1.Be flavor protease, papain and alkali protease.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, the addition of alkali protease of take is respectively 0.2%, 0.4%, 0.6%, 0.8%, 1.0% as example, pressing enzyme work calculation addition is 2500U/g, 5000U/g, 7500U/g, 10000U/g, 12500U/g, and accompanying drawing 5 is the affect result of alkali protease addition on TDF yield.Experimental result shows: when protease consumption is between 2500～7500 U/g, along with the increase of protease consumption, yield increases sharply; When protease consumption is between 7500～12500 U/g, yield reduces along with the increase of enzyme amount.Through SPSS, analyze, definite addition of usining alkali protease is 0.6%(7500 U/g) as optimum addition.
The 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 enzymolysis time result.Experimental result is known, and in the time of 15～45 min, yield is in rising trend, and during 45～105 min, yield declines.Analyze former because: 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 due to selectivity and the high efficiency of enzyme, and the residual etc. of the mixed enzyme used of hydrolyzed starch considers, at enzymolysis time, continue overtime, dietary fiber yield declines again to some extent, and the present invention determines that protease hydrolyzed time 45min is for well.
embodiment 2pumpkin dietary fiber optimum preparation condition
On the basis of single factor experiment, for obtaining the optimised process of preparing 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 impacts that factor is prepared pumpkin dietary fiber of integrated survey, pumpkin total dietary fiber for evaluation result (TDF) yield represents.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, from the result of analysis of variance table shown in table 3, can find out, the addition of mixed enzyme has larger impact (P=0.000) with mixed enzyme enzymolysis time to pumpkin total dietary fiber (TDF) yield, next is protease hydrolyzed time (P=0.001), mixed enzyme ratio is on result impact little (P=0.072), and protease addition affects also little (P=0.128) to result.
In conjunction with mean estimated statement 4 and paired comparisons table table 5 in single factor statistics scale, can find A
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
1for the best.
In conjunction with mean estimated statement 6 and paired comparisons table table 7 in single factor statistics scale, can find B
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).
In conjunction with mean estimated statement 8 and paired comparisons table table 9 in single factor statistics scale, can find E
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: the actual conditions by above orthogonal experiments in conjunction with pumpkin dietary fiber experiment, the optimum extraction process that summary obtains pumpkin dietary fiber is A
1b
3c
4d
4e
4be that mixed enzyme addition is that the addition of 0.5%(AMS and carbohydrase is respectively 276.675U/g and 2062.5U/g), mixed enzyme enzymolysis time is 90min, ratio is 1:7, 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%.
In the present embodiment, pumpkin dietary fiber prepares by following preparation method:
(1) get fresh pumpkin, clean, go seed, adopt tissue mashing machine to smash to obtain pumpkin fruit to pieces;
(2) in the pumpkin fruit after smashing to pieces, add the phosphate buffer that the pH value of 0.2M is 6, the mass ratio of pumpkin fruit and phosphate buffer is 1:6;
Consisting of of phosphate buffer: contain 0.2M sodium hydrogen phosphate 123ml, 0.2M sodium dihydrogen phosphate 877ml in 1L phosphate buffer;
(3) in step (2), mixed in the pumpkin fruit of phosphate buffer and added mixed enzyme, the amount that mixed enzyme adds is determined according to 0.5% of phosphate buffer and pumpkin fruit gross weight; In mixed enzyme, AMS and carbohydrase weight ratio are that 1:7(is that AMS and carbohydrase are respectively 216 U/g and 2398 U/g), in 60 ℃ of enzymolysis 90min, after boiling the enzyme that goes out, be cooled to room temperature, with the NaOH of 2M, regulate the pH value to 10 of enzymolysis liquid to obtain enzymolysis liquid A;
(4) in step (3) gained enzymolysis liquid A, add alkali protease, 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 obtain enzymolysis liquid B;
(5) enzymolysis liquid B is filtered, filter the filter residue obtaining and be washed till neutrality with clear water, freeze drying (subzero 45 ℃ of freeze-drying, freeze drier), pulverizes, and obtains insoluble diedairy fiber (IDF);
Enzymolysis liquid B is filtered to the filtrate obtain by 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, after precipitation process, filter, filter residue freezing (with aforementioned) be dry, pulverize, obtain soluble dietary fiber (SDF).
The 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 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, with pipette, accurately pipetting 10 mL distilled water adds wherein, after shaking up, under room temperature, place 24 h, then the absolute ethyl alcohol that adds 5 times of volumes, reads the volume of the water-soluble dietary fiber (SDF) precipitating in liquid.
Accurately take 0.1g insoluble diedairy fiber powder (IDF) and be placed in tool plug scale test tube, add 10 mL distilled water, mix standing 18h under rear room temperature, read volume after sample sedimentation.
Expansive force=[volume after swelling (mL)-dry product volume (mL)]/sample dry weight (g)
(2) hold oily power: take 1.0 g dietary fibers in 50 mL centrifuge tubes, add peanut oil 10 mL, 37 ℃ of standing l h, the centrifugal 20min of 4000 r/min, discards oil and the residue on upper strata, with filter paper, blots free oil, weighs.
Hold oily power (g/g)=[heavy (the g)-sample dry weight (g) of sample oil]/sample dry weight (g)
(3) retention ability: take 1 g water-soluble dietary fiber powder (SDF) and put into beaker, then add water 100 mL, at room temperature, after electromagnetic agitation 24 h, add the absolute ethyl alcohol precipitation of 5 times of volumes, suction filtration, claims quality.Calculate retention ability.
Accurately take 0.8g sample (IDF), be placed in test tube, add 40mL distilled water, mix standing 20h under rear room temperature, 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) absorption of pumpkin dietary fiber to cholesterol:
Get the yolk of commercially available fresh hen egg, with 9 times of amount distilled water, fully beat into emulsion.Get respectively the 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 shaking table 37 ℃ of vibration 3 h, centrifugal 20min(precipitation dietary fiber under 4000r/min), draw 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 accompanying drawing 7.
Dietary fiber is to the adsorbance of cholesterol (A)=(before absorption in egg yolk liquid cholesterol amount-absorption afterwards cholesterol amount in supernatant)/dietary fiber weight.Pumpkin dietary fiber of the present invention the results are shown in shown in accompanying drawing 8 adsorption experiment of cholesterol.As seen from the figure, pumpkin dietary fiber is better than soybean dietary fiber far away to cholesterol adsorption capacity, and wherein the adsorption effect of insoluble diedairy fiber is best, reaches 18.365mg/g, and the adsorption effect in pH=7 environment is better than pH=2's.
(5) absorption of pumpkin dietary fiber to natrium nitrosum:
A. NO
2 -the formulation of calibration curve
Press 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), in 9 50 mL colorimetric cylinders, adds respectively 2 mL 0.4% sulfanilic acid solution, mixes, place after 3 ~ 5min, add respectively l mL 0.2% hydrochloride naphthodiamide solution, add water to scale, mix, standing 15 min, use 1cm cuvette, zero pipe of take is reference, measures 538nm wavelength place absorbance on spectrophotometer, drawing standard curve, is shown in shown in accompanying drawing 9.
B. adsorption effect is measured
It is that pH value is respectively 7.0 and 2.0 (regulating pH value with hydrochloric acid and NaOH respectively, simulation small intestine and gastric environment) that absorption environment is set, and reaction is carried out in 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 under 37 ℃ of constant temperature, electromagnetic agitation reaction, respectively gets the concentration that lmL sample liquid is measured NO2-at set intervals, respectively does blank assay contrast simultaneously, and experimental result is shown in shown in accompanying drawing 10 ~ accompanying drawing 13.As seen from the figure, pumpkin dietary fiber, in pH=2, has suction-operated to nitrite, there is no suction-operated in pH=7.The suction-operated of insoluble diedairy fiber (IDF) is greater than soluble dietary fiber (SDF) (adsorption time of solubility will be grown), and the adsorption capacity of two kinds of pumpkin dietary fibers is all greater than soybean dietary fiber.
(6) cation exchange capacity (CEC)
Take 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 and removes unnecessary acid with distilled water, shifts residue till (the AgNO3 solution with 10% is identified and no longer contained chlorion in triangular flask, vacuum drying), the NaCl solution that adds 100 mL 15%, by magnetic stirrer, makes indicator with 0.5% phenolphthalein, with the slow titration of 0.01 mol/L NaOH solution, when solution is micro-, stop titration when red, jolting, drips after fading again, until jolting 5min still colour-fast be terminal, record pH value in titration process.Make V
naOHwith pH value graph of a relation.With distilled water, replace HCl, measure the amount of the blank NaOH consuming.Pumpkin dietary fiber cation exchange capacity (CEC) experimental result is shown in shown in accompanying drawing 14 and accompanying drawing 15, as seen from the figure, in titration curve, dietary fiber curvilinear motion is relatively slow, cation exchange capacity (CEC) is poor, this may be because leaching process has destruction to a certain degree to the side-chain radical structure in 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 soybean insoluble diedairy fiber.
In the present embodiment, pumpkin dietary fiber prepares by following preparation method:
(1) get fresh pumpkin, clean, go seed, adopt tissue mashing machine to smash to obtain pumpkin fruit to pieces;
(2) in the pumpkin fruit after smashing to pieces, add the phosphate buffer that the pH value of 0.2M is 6, the mass ratio of pumpkin fruit and phosphate buffer is 1:6;
(3) in step (2), mixed in the pumpkin fruit of phosphate buffer and added mixed enzyme, the amount that mixed enzyme adds is determined according to 1.1% of phosphate buffer and pumpkin fruit gross weight; In mixed enzyme, AMS and carbohydrase weight ratio are that 1:1(is that AMS and carbohydrase are respectively 1221.813U/g and 3021.334U/g), in 80 ℃ of enzymolysis 15min, after boiling the enzyme that goes out, be cooled to room temperature, with the NaOH of 2M, regulate the pH value to 10 of enzymolysis liquid to obtain enzymolysis liquid A;
(4) in step (3) gained enzymolysis liquid A, add alkali protease, the addition of alkali protease is 0.2% of enzymolysis liquid A weight, and 2500U/g, in 50 ℃ of enzymolysis 105min, boils the enzyme that goes out, and is cooled to room temperature and obtains enzymolysis liquid B;
(5) enzymolysis liquid B is filtered, filter the filter residue obtaining and be washed till neutrality with clear water, freeze drying (with reference to conventional or employing freeze drier freeze-drying at-45 ℃), pulverizes, and obtains insoluble diedairy fiber (IDF);
Enzymolysis liquid B is filtered to the filtrate obtain by 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, after precipitation process, filter, by 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 the 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.
In the present embodiment, pumpkin dietary fiber prepares by following preparation method:
(1) get fresh pumpkin, clean, go seed, adopt tissue mashing machine to smash to obtain pumpkin fruit to pieces;
(2) in the pumpkin fruit after smashing to pieces, add the phosphate buffer that the pH value of 0.2M is 5, the mass ratio of pumpkin fruit and phosphate buffer is 1:6;
(3) in step (2), mixed in the pumpkin fruit of phosphate buffer and added mixed enzyme, the amount that mixed enzyme adds is determined according to 1.0% of phosphate buffer and pumpkin fruit gross weight; In mixed enzyme, AMS and carbohydrase weight ratio are that 1:9(is that AMS and carbohydrase are respectively 222.148U/g and 4944.007U/g), in 40 ℃ of enzymolysis 105min, after boiling the enzyme that goes out, be cooled to room temperature, with the NaOH of 2M, regulate the pH value to 9 of enzymolysis liquid, obtain enzymolysis liquid A;
(4) in step (3) gained enzymolysis liquid A, add alkali protease, the addition of alkali protease is 1.0% of enzymolysis liquid A weight, and 12500U/g, in 40 ℃ of enzymolysis 15min, boils the enzyme that goes out, and is cooled to room temperature and obtains enzymolysis liquid B;
(5) enzymolysis liquid B is filtered, filter the filter residue obtaining and be washed till neutrality with clear water, freeze-drying, pulverizes, and obtains insoluble diedairy fiber (IDF);
Enzymolysis liquid B is filtered to the filtrate obtain by 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, after precipitation process, filter, by 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 the 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.
In the present embodiment, pumpkin dietary fiber prepares by following preparation method:
(1) get fresh pumpkin, clean, go seed, adopt tissue mashing machine to smash to obtain pumpkin fruit to pieces;
(2) in the pumpkin fruit after smashing to pieces, add the phosphate buffer that the pH value of 0.2M is 6, the mass ratio of pumpkin fruit and phosphate buffer is 1:6;
(3) in step (2), mixed in the pumpkin fruit of phosphate buffer and added mixed enzyme, the amount that mixed enzyme adds is determined according to 0.5% of phosphate buffer and pumpkin fruit gross weight; In mixed enzyme, AMS and carbohydrase weight ratio are that 1:7(is that AMS and carbohydrase are respectively 216 U/g and 2398 U/g), in 60 ℃ of enzymolysis 90min, boil and be cooled to room temperature after the enzyme that goes out and obtain enzymolysis liquid A;
(4) in step (3) gained enzymolysis liquid A, add flavor protease, 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 obtain enzymolysis liquid B;
(5) enzymolysis liquid B is filtered, filter the filter residue obtaining and be washed till neutrality with clear water, freeze-drying, pulverizes, and obtains insoluble diedairy fiber (IDF);
Enzymolysis liquid B is filtered to the filtrate obtain by 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, after precipitation process, filter, by filter residue freeze-drying, pulverize, obtain soluble dietary fiber (SDF).
The 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 (4)
1. a preparation method for pumpkin dietary fiber, is characterized in that comprising the following steps:
(1) get fresh pumpkin, clean, remove the peel, go to smash to obtain pumpkin fruit to pieces after seed;
(2) in the pumpkin fruit after step (1) is smashed to pieces, adding pH value is 5～7 phosphate buffer, and the weight ratio of pumpkin fruit and phosphate buffer is 1:6, obtains 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 to the enzyme that goes out, suction filtration, obtain filtrate and filter residue;
With clear water, filter residue is washed till to neutrality, dries, obtain insoluble diedairy fiber;
Gained filtrate, with ethanolic solution suction filtration after precipitation process at room temperature, is dried filter residue to obtain water-soluble dietary fiber;
Wherein, the addition of the described AMS of step (3) and carbohydrase is respectively 608.685U/g and 4537.5U/g; Or the total addition level of described AMS and carbohydrase determines according to 0.5～1.1% of pumpkin fruit weight, the part by weight of described AMS and carbohydrase is 1:1～1:9; The time of the described enzymolysis of step (3) is 90min;
The addition of the described protease of step (4) is 7500U/g, and the time of the described enzymolysis of step (4) is 75min.
2. the preparation method of pumpkin dietary fiber according to claim 1, the part by weight that it is characterized in that described AMS and carbohydrase is 1:7.
3. the preparation method of pumpkin dietary fiber according to claim 1, is characterized in that the temperature of the described enzymolysis of step (3) is 40～80 ℃.
4. the pumpkin dietary fiber that described in claim 1～3 any one, preparation method prepares.
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| CN103719880B (en) * | 2014-01-17 | 2015-07-01 | 昆明医科大学 | Preparation method of high-activity purple sweet potato dietary fiber |
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| CN107874265A (en) * | 2017-10-11 | 2018-04-06 | 中国农业科学院农产品加工研究所 | A kind of method for extracting pomace dietary fibers |
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| CN108095130A (en) * | 2018-01-08 | 2018-06-01 | 中国农业科学院特产研究所 | Food-grade ginseng dietary fiber and preparation method thereof |
| CN109717484A (en) * | 2019-03-15 | 2019-05-07 | 南京中医药大学 | A kind of preparation method with the anti-dietary fiber to treat constipation |
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| CN110839874A (en) * | 2019-11-19 | 2020-02-28 | 广东工业大学 | Water-soluble dietary fiber and application |
| CN111264876A (en) * | 2020-02-20 | 2020-06-12 | 临沂大学 | Method for extracting garlic and radix astragali dietary fiber |
| CN112914112A (en) * | 2021-03-05 | 2021-06-08 | 华南农业大学 | Fingered citron dietary fiber with weight-losing and lipid-lowering functions and preparation method and application thereof |
| CN115137079A (en) * | 2022-07-08 | 2022-10-04 | 南开大学 | Buckwheat extract with function of regulating intestinal flora |
| CN115336616B (en) * | 2022-07-12 | 2023-05-30 | 广东省农业科学院蚕业与农产品加工研究所 | Application of insoluble dietary fiber in preparation of baked food |
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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 |