CN105368905A - Microwave-assisted method for preparing pea protein polypeptides - Google Patents
Microwave-assisted method for preparing pea protein polypeptides Download PDFInfo
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- CN105368905A CN105368905A CN201510905986.1A CN201510905986A CN105368905A CN 105368905 A CN105368905 A CN 105368905A CN 201510905986 A CN201510905986 A CN 201510905986A CN 105368905 A CN105368905 A CN 105368905A
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Abstract
The invention belongs to the technical field of bioengineering, and particularly relates to a microwave-assisted method for preparing pea protein polypeptides. The microwave-assisted method for preparing the pea protein polypeptides includes steps of dilute alkali liquor adding, microwave treatment, coarse filtration, primary enzymatic hydrolysis, primary microwave treatment, secondary enzymatic hydrolysis, secondary microwave treatment, enzyme inactivation, chitosan adding, rough filtration, micro-filtration, ultra-filtration, nano-filtration, concentration and low-temperature vacuum drying. The microwave-assisted method for extracting the pea protein polypeptides has the advantages of high yield and purity, low solvent consumption and cost and easiness in industrial production. Besides, the microwave-assisted method is short in integral technological process and time, low in energy consumption, little in pollution and free of pollutant emission, toxic solvents can be omitted, and accordingly the purpose of clean production can be achieved.
Description
Technical field
The invention belongs to technical field of bioengineering, be specifically related to a kind of method that microwave-assisted prepares Semen Pisi sativi protein polypeptide.
Background technology
The annual vine crop of pea (Pisumsativum), the multiple pea of pinniform, small pea is avette, opens white or lilac flower, high 90-180cm, all without hair.Small pea Long Circle to oval, long 3-5cm, wide 1-2cm, Quan Yuan; Holder pea pea shape, avette, base portion ear surrounds pea handle.Pod oblong, long 5-10cm, inside has hard papery to serve as a contrast skin; Seed is circular, 2-10, dark green, becomes yellow after dry.The flowering fruit bearing stage 4-5 month.The multiple pea of even number pinniform, top tendril is Pea tendril, and holder pea is avette.Floral white or red-purple, single raw or 1-3 to be arranged in total shape armpit raw, have whisker inside style, cleisogamy, petal butterfly shape.Pod oblong or flat, be divided into soft pod and hard pod according to inside with or without internal layer keratin film and thickness thereof.Tender pod and seed edible.
Pea has been the world the 4th pea legume crop now.The dry pea cultivated area in the whole world 621.43 ten thousand hectares, total product 955.82 ten thousand tons; Whole world Semen Pisi sativi cultivated area 224.13 ten thousand hectares, total product 1697.50 ten thousand tons.The same year, the dry pea cultivated area of China 94.00 ten thousand hectares, total product 119.00 ten thousand tons; Semen Pisi sativi cultivated area 129.59 ten thousand hectares, total product 1027.43 ten thousand tons.China's pea cultivated area and total product account for global 15.13% and 12.45% respectively, and Semen Pisi sativi cultivated area and total product account for global 57.82% and 60.53% respectively.China is only second to Canadian second-biggest-in-the-world pea producing country, in world's pea is produced, occupy very important status.
Fresh pea nutritious, every 100 grams containing 7.2 grams, protein, heat 80 kilocalories, is equivalent to the nutritive value with amount bean curd.Particularly the content of vitamin B group is very high, as 18 times that VITMAIN B1 (0.54 milligram/100 grams) is bean curd, Lin Suanna Vitamin B2 Sodium Phosphate and vitamin PP are 2.5 times and 14 times of bean curd respectively, also have the nutritive ingredients such as more carotene, vitamins C and inorganic salt.Just owing to being rich in the various nutritive substances of needed by human body in pea, especially containing high-quality protein, can improve resistance against diseases and the rehabilitation ability of body, patient and old man and child will have some more.Can be used as green manure and feed in addition.
Pea peptide is obtained by enzymolysis Semen Pisi sativi protein, and taste is gentle, inexpensive, can be used for infant formula.
Pea taste is sweet, property is put down, and returns spleen, stomach warp; There is gas in benefit, antidiarrheal dysentery, adjust that battalion defends, effect of diuresis, subduing inflammation, solution mammary calculus poison; Cure mainly that beriberi, carbuncle are swollen, galactostasis, taste discomfort, hiccup vomiting, trusted subordinate's distending pain, thirstyly let out the illnesss such as dysentery.It also has that treatment diabetes, beautification function, sterilization are antitoxin, cancer preventing and treating, hypotensive, protect cardiovascular, eyeshield, replenish the calcium, regulate physique, control the purposes such as deficient qi and blood, antianaphylaxis.
The information being disclosed in this background technology part is only intended to increase the understanding to general background of the present invention, and should not be regarded as admitting or imply in any form that this information structure has been prior art that persons skilled in the art are known.
Summary of the invention
In order to overcome the deficiencies in the prior art, the invention provides a kind of method that microwave-assisted prepares Semen Pisi sativi protein polypeptide.
For solving the problems of the technologies described above, the present invention by the following technical solutions:
Microwave-assisted prepares a method for Semen Pisi sativi protein polypeptide, comprises the steps:
(1) be raw material with pea separation protein, add deionized water by solid-to-liquid ratio 1:15, fully after mixing, with sig water adjust pH to 9.0, microwave extraction 15min, temperature controls at 55-60 DEG C, be incubated 50-60min afterwards, obtain modified pea protein isolate solution;
(2) modified pea protein isolate solution is cooled to 33-38 DEG C, adjust pH to 9.0, adds the trypsinase of substrate quality 4%, microwave extraction 15min, temperature controls at 55-60 DEG C, and pH value remains on more than 8.0, obtains trypsin hydrolyzing solution;
(3) trypsin hydrolyzing solution is cooled to 33-38 DEG C, adjust pH to 7.6, adds the compound protease of substrate quality 2%, microwave extraction 10min, temperature controls at 55-60 DEG C, when pH value is down to 7.2, enzymolysis stops, and obtains composite protease hydrolysis solution;
(4) continued to be warming up to 85-90 DEG C by composite protease hydrolysis solution, when the time reaches 5min, the enzyme that goes out has been lived, and obtains the enzymic hydrolysis solution that goes out;
(5) the enzymic hydrolysis solution that will go out is cooled to 33-38 DEG C, adjust pH is to 4.8-5.0, add the Sales glycan of 0.01%, filter with 200 order tripod pendulum type batch centrifugals after suitably precipitating, cut 2000-10000 molecular weight ultrafiltration Fibrous membrane filtration with hurdle, then cut the nanofiltration of 200-1000 molecular weight and RO membrane filtration with hurdle and concentrate;
(6) by the Semen Pisi sativi protein enzymic hydrolysis solution adjust pH to 7.0 after filtering and concentrating, carry out concentrating low-temperature vacuum-drying, obtain Semen Pisi sativi protein polypeptide products.
As preferably, the sig water described in step (1) is for be adjusted to 8.5-9.0 weakly alkaline solution with NaOH solution by material liquid PH value.
As preferably, the alkaline enzymolysis pH value condition described in step (2) is 8.0-8.5.
As preferably, the complex enzyme hydrolysis pH value described in step (3) is 7.0-7.5.
As preferably, the compound protease described in step (4) is made up of alkaline enzyme, neutral enzymatic and food flavor enzyme, and the mass ratio of 3 kinds of enzymes is 3:1:1.
As preferably, the frequency of step (1), (2) and the microwave extraction device described in (3) is 2450MHz, and power is 20KW.
Compared with prior art, the present invention has following beneficial effect:
1. method of the present invention improves enzymolysis process and the technology of pea protein, solve pea protein and not easily digested and assimilated problem by animal, by the preparation of Bian microwave-assisted enzymolysis technology, there is bioactive oligopeptides, not only improve the digestibility of Semen Pisi sativi protein, and expand Semen Pisi sativi protein Application Areas.Although Semen Pisi sativi protein is by the byproduct in traditional food bean vermicelli production process, differentiation becomes pea deep processing industry, but still belong to simple foodstuff additive industry, industrial chain is short, competitive power far lags behind the industry giant of French Luo Gaite, Belgian section rope carat Semen Pisi sativi protein in the world, therefore, technological innovation, products innovation must be passed through, continuous prolongation pea industrial chain, comprehensive raising pea industry comprehensive benefit, growth of agricultural efficiency could be realized, increasing peasant income, rural area are flourishing, could-be with in a road construction and given play to powerful vitality.
2. the method for Semen Pisi sativi protein polypeptide that the present invention extracts has yield high (than tradition raising 5-8%), purity high (oligopeptides ratio reaches more than 70%), solvent consumption is few, cost is low and be easy to the advantage of suitability for industrialized production.
3. the whole technical process of the present invention is short, the time is short (about quite tradition 1/3), and energy consumption is low, pollution less, does not use noxious solvent, non-pollutant discharge, reaches cleaner production target.
Accompanying drawing explanation
Fig. 1 is the process flow sheet that microwave-assisted of the present invention prepares Semen Pisi sativi protein polypeptide.
Embodiment
Below in conjunction with specific embodiment, elaboration detailed is further done to the present invention, but embodiments of the present invention are not limited to the scope that embodiment represents.These embodiments only for illustration of the present invention, but not for limiting the scope of the invention.In addition, after reading content of the present invention, those skilled in the art can do various amendment to the present invention, and these equivalent variations fall within appended claims limited range of the present invention equally.
The experimental technique used in following embodiment if no special instructions, is ordinary method.The material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
embodiment 1:
Microwave-assisted prepares a method for Semen Pisi sativi protein polypeptide, comprises the steps:
(1) be raw material with pea separation protein, add deionized water by solid-to-liquid ratio 1:15, fully after mixing, with sig water adjust pH to 9.0, microwave extraction 15min, temperature controls, at 55 DEG C, to be incubated 50min afterwards, obtains modified pea protein isolate solution; Described sig water is for be adjusted to 8.5 weakly alkaline solutions with NaOH solution by material liquid pH value; The frequency of described microwave extraction device is 2450MHz, and power is 20KW;
(2) modified pea protein isolate solution is cooled to 33-38 DEG C, adjust pH to 9.0, adds the trypsinase of substrate quality 4%, microwave extraction 15min, temperature controls at 55 DEG C, and pH value remains on more than 8.0, obtains trypsin hydrolyzing solution; Alkalescence enzymolysis pH value condition is 8.0; The frequency of described microwave extraction device is 2450MHz, and power is 20KW;
(3) trypsin hydrolyzing solution is cooled to 33-38 DEG C, adjust pH to 7.6, adds the compound protease of substrate quality 2%, microwave extraction 10min, temperature controls at 55 DEG C, and when pH value is down to 7.2, enzymolysis stops, and obtains composite protease hydrolysis solution; Complex enzyme hydrolysis pH value is 7.0; Described compound protease is made up of alkaline enzyme, neutral enzymatic and food flavor enzyme, and the mass ratio of 3 kinds of enzymes is 3:1:1; The frequency of described microwave extraction device is 2450MHz, and power is 20KW;
(4) continued to be warming up to 85 DEG C, when the time reaches 5min by composite protease hydrolysis solution, the enzyme that goes out has been lived, and obtains the enzymic hydrolysis solution that goes out;
(5) the enzymic hydrolysis solution that will go out is cooled to 33 DEG C, adjust pH to 4.8, add the Sales glycan of 0.01%, filter with 200 order tripod pendulum type batch centrifugals after suitably precipitating, cut 2000-10000 molecular weight ultrafiltration Fibrous membrane filtration with hurdle, then cut the nanofiltration of 200-1000 molecular weight and RO membrane filtration with hurdle and concentrate;
(6) by the Semen Pisi sativi protein enzymic hydrolysis solution adjust pH to 7.0 after filtering and concentrating, carry out concentrating low-temperature vacuum-drying, obtain Semen Pisi sativi protein polypeptide products.
embodiment 2:
Microwave-assisted prepares a method for Semen Pisi sativi protein polypeptide, comprises the steps:
(1) be raw material with pea separation protein, add deionized water by solid-to-liquid ratio 1:15, fully after mixing, with sig water adjust pH to 9.0, microwave extraction 15min, temperature controls, at 60 DEG C, to be incubated 60min afterwards, obtains modified pea protein isolate solution; Described sig water is for be adjusted to 9.0 weakly alkaline solutions with NaOH solution by material liquid PH value; The frequency of described microwave extraction device is 2450MHz, and power is 20KW;
(2) modified pea protein isolate solution is cooled to 38 DEG C, adjust pH value to 9.0, add the trypsinase of substrate quality 4%, microwave extraction 15min, temperature controls at 60 DEG C, and pH value remains on more than 8.0, obtains trypsin hydrolyzing solution; Alkalescence enzymolysis pH value condition is 8.5; The frequency of described microwave extraction device is 2450MHz, and power is 20KW;
(3) trypsin hydrolyzing solution is cooled to 38 DEG C, adjust pH to 7.6, adds the compound protease of substrate quality 2%, microwave extraction 10min, temperature controls at 60 DEG C, and when pH value is down to 7.2, enzymolysis stops, and obtains composite protease hydrolysis solution; Complex enzyme hydrolysis pH value is 7.5; Described compound protease is made up of alkaline enzyme, neutral enzymatic and food flavor enzyme, and the mass ratio of 3 kinds of enzymes is 3:1:1; The frequency of described microwave extraction device is 2450MHz, and power is 20KW;
(4) continued to be warming up to 90 DEG C, when the time reaches 5min by composite protease hydrolysis solution, the enzyme that goes out has been lived, and obtains the enzymic hydrolysis solution that goes out;
(5) the enzymic hydrolysis solution that will go out is cooled to 38 DEG C, adjust pH to 5.0, add the Sales glycan of 0.01%, filter with 200 order tripod pendulum type batch centrifugals after suitably precipitating, cut 2000-10000 molecular weight ultrafiltration Fibrous membrane filtration with hurdle, then cut the nanofiltration of 200-1000 molecular weight and RO membrane filtration with hurdle and concentrate;
(6) by the Semen Pisi sativi protein enzymic hydrolysis solution adjust pH to 7.0 after filtering and concentrating, carry out concentrating low-temperature vacuum-drying, obtain Semen Pisi sativi protein polypeptide products.
embodiment 3:
Microwave-assisted prepares a method for Semen Pisi sativi protein polypeptide, comprises the steps:
(1) be raw material with pea separation protein, add deionized water by solid-to-liquid ratio 1:15, fully after mixing, with sig water adjust pH to 9.0, microwave extraction 15min, temperature controls, at 57 DEG C, to be incubated 55min afterwards, obtains modified pea protein isolate solution; Described sig water is for be adjusted to 8.8 weakly alkaline solutions with NaOH solution by material liquid pH value; The frequency of described microwave extraction device is 2450MHz, and power is 20KW;
(2) modified pea protein isolate solution is cooled to 35 DEG C, adjust pH to 9.0, adds the trypsinase of substrate quality 4%, microwave extraction 15min, temperature controls at 57 DEG C, and pH value remains on more than 8.0, obtains trypsin hydrolyzing solution; Alkalescence enzymolysis pH value condition is 8.3; The frequency of described microwave extraction device is 2450MHz, and power is 20KW;
(3) trypsin hydrolyzing solution is cooled to 35 DEG C, adjust pH to 7.6, adds the compound protease of substrate quality 2%, microwave extraction 10min, temperature controls at 58 DEG C, and when pH value is down to 7.2, enzymolysis stops, and obtains composite protease hydrolysis solution; Complex enzyme hydrolysis pH value is 7.3; Described compound protease is made up of alkaline enzyme, neutral enzymatic and food flavor enzyme, and the mass ratio of 3 kinds of enzymes is 3:1:1; The frequency of described microwave extraction device is 2450MHz, and power is 20KW;
(4) continued to be warming up to 88 DEG C, when the time reaches 5min by composite protease hydrolysis solution, the enzyme that goes out has been lived, and obtains the enzymic hydrolysis solution that goes out;
(5) the enzymic hydrolysis solution that will go out is cooled to 35 DEG C, adjust pH to 4.9, add the Sales glycan of 0.01%, filter with 200 order tripod pendulum type batch centrifugals after suitably precipitating, cut 2000-10000 molecular weight ultrafiltration Fibrous membrane filtration with hurdle, then cut the nanofiltration of 200-1000 molecular weight and RO membrane filtration with hurdle and concentrate;
(6) by the Semen Pisi sativi protein enzymic hydrolysis solution adjust pH to 7.0 after filtering and concentrating, carry out concentrating low-temperature vacuum-drying, obtain Semen Pisi sativi protein polypeptide products.
embodiment4: tradition prepares the method for pea polypeptide
A method for pea separation protein enzyme-squash techniqued anti-oxidation peptide, step is:
(1) raw materials pretreatment: take pea separation protein as raw material, configuration quality concentration is the pea separation protein aqueous solution of 7.5%, and 90 DEG C of water-bath 15min carry out pre-treatment, cool for subsequent use;
(2) trypsin digestion: be placed on subsequently in constant temperature blender with magnetic force, solution is adjusted to temperature 40 DEG C, pH10 while stirring, according to being 6% add trypsinase than E/S at the bottom of enzyme, the work of trypsinase enzyme is 4 × 104U/g, start hydrolysis reaction, dripping 2MNaOH solution at any time keeps enzymolysis solution pH10 constant, after abundant enzyme digestion reaction 4h, boils 15min and to go out enzyme;
(3) neutral protease enzymolysis: after the cooling of step (2) product by 50 DEG C, pH7.5, E/S 6% add neutral protease, works of neutral protease enzyme is 1 × 105U/g, and hydrolysis 4h, boils the enzyme that goes out, with 2MHCl solution tune pH7 after cooling;
(4) centrifugal: step (3) product goes precipitation according to the centrifugal 15min of 8000r/min, and gained supernatant liquor is Semen Pisi sativi protein hydrolysate PPH, and the feed liquid spraying dry after filtering and concentrating, obtains anti-oxidation peptide crude product.
The beneficial effect of table 1 extracting method of the present invention
As known from Table 1, traditional method unavoidably uses a large amount of soda acid and solvent, and complex procedures, length consuming time, low conversion rate, weakness and the deficiencies such as impurity is many; And Bian microwave radiation exaraction of the present invention, do not need solvent, soda acid consumption is few, and enzymolysis time is short, and transformation efficiency is high, and prepares polypeptide for next step and create better condition.
Bian microwave-assisted of the present invention urgees zymolysis technique, with regard to enzymolysis time, namely microwave-assisted enzymolysis only completes with 30min, and traditional method 8-16h just can complete enzymic hydrolysis operation, greatly can shorten the production cycle and reduce production cost, and enzymolysis transformation efficiency is than passing blunderbuss height 5-8%.
The Bian of the present invention comparatively advanced membrane filtration in the current world and membrane filtration concentration technique ,-be to shorten the time, obtain product purity high; Two is that energy consumption is low, and blowdown is few.The present invention's microwave is separated with membrane filtration and is coupled, substitute single-use enzyme solution to prepare pea anti-oxidation peptide, technique is simple, and the time is short, and efficiency is high, produce and labor safety reliable.
Because preparation pea polypeptide enzymolysis time is short, membrane filtration separating energy consumption is low, and blowdown is few, reaches national cleaner production target call.Because technique is comparatively simple, easy to operate, environmental protection, suitable plant sizeization is produced and is adopted.
The aforementioned description to concrete exemplary of the present invention is to illustrate and the object of illustration.These descriptions not want the present invention to be defined as disclosed precise forms, and obviously, according to above-mentioned instruction, can much change and change.The object selected exemplary embodiment and describe is to explain certain principles of the present invention and practical application thereof, thus those skilled in the art can be realized and utilize various different exemplary of the present invention and various different selection and change.Scope of the present invention is intended to limited by claims and equivalents thereof.
Claims (6)
1. microwave-assisted prepares a method for Semen Pisi sativi protein polypeptide, it is characterized in that, comprises the steps:
(1) be raw material with pea separation protein, add deionized water by solid-to-liquid ratio 1:15, fully after mixing, with sig water adjust pH to 9.0, microwave extraction 15min, temperature controls at 55-60 DEG C, be incubated 50-60min afterwards, obtain modified pea protein isolate solution;
(2) modified pea protein isolate solution is cooled to 33-38 DEG C, adjust pH to 9.0, adds the trypsinase of substrate quality 4%, microwave extraction 15min, temperature controls at 55-60 DEG C, and pH value remains on more than 8.0, obtains trypsin hydrolyzing solution;
(3) trypsin hydrolyzing solution is cooled to 33-38 DEG C, adjust pH to 7.6, adds the compound protease of substrate quality 2%, microwave extraction 10min, temperature controls at 55-60 DEG C, when pH value is down to 7.2, enzymolysis stops, and obtains composite protease hydrolysis solution;
(4) continued to be warming up to 85-90 DEG C by composite protease hydrolysis solution, when the time reaches 5min, the enzyme that goes out has been lived, and obtains the enzymic hydrolysis solution that goes out;
(5) the enzymic hydrolysis solution that will go out is cooled to 33-38 DEG C, adjust pH is to 4.8-5.0, add the Sales glycan of 0.01%, filter with 200 order tripod pendulum type batch centrifugals after suitably precipitating, cut 2000-10000 molecular weight ultrafiltration Fibrous membrane filtration with hurdle, then cut the nanofiltration of 200-1000 molecular weight and RO membrane filtration with hurdle and concentrate;
(6) by the Semen Pisi sativi protein enzymic hydrolysis solution adjust pH to 7.0 after filtering and concentrating, carry out concentrating low-temperature vacuum-drying, obtain Semen Pisi sativi protein polypeptide products.
2. microwave-assisted according to claim 1 prepares the method for Semen Pisi sativi protein polypeptide, it is characterized in that, the sig water described in step (1) is for be adjusted to 8.5-9.0 weakly alkaline solution with NaOH solution by material liquid PH value.
3. microwave-assisted according to claim 1 prepares the method for Semen Pisi sativi protein polypeptide, it is characterized in that, the alkaline enzymolysis pH value condition described in step (2) is 8.0-8.5.
4. microwave-assisted according to claim 1 prepares the method for Semen Pisi sativi protein polypeptide, it is characterized in that, the complex enzyme hydrolysis pH value described in step (3) is 7.0-7.5.
5. microwave-assisted according to claim 1 prepares the method for Semen Pisi sativi protein polypeptide, it is characterized in that, the compound protease described in step (4) is made up of alkaline enzyme, neutral enzymatic and food flavor enzyme, and the mass ratio of 3 kinds of enzymes is 3:1:1.
6. microwave-assisted according to claim 1 prepares the method for Semen Pisi sativi protein polypeptide, it is characterized in that, the frequency of step (1), (2) and the microwave extraction device described in (3) is 2450MHz, and power is 20KW.
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| CN105928750A (en) * | 2016-04-11 | 2016-09-07 | 沈阳师范大学 | Processing method of small red bean isolated protein with high solubility |
| CN106086138A (en) * | 2016-08-10 | 2016-11-09 | 柳江县渡庄生物科技有限公司 | A kind of method that microwave-assisted and membrane filtration prepare fresh water fin albumen oligopeptide |
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| CN109965221A (en) * | 2019-05-13 | 2019-07-05 | 河南省食品工业科学研究所有限公司 | A kind of processing method of peameal |
| CN110438188A (en) * | 2019-09-06 | 2019-11-12 | 广西民族大学 | A kind of preparation method of pea protein Gly-His-Lys |
| CN111018164A (en) * | 2020-03-09 | 2020-04-17 | 烟台双塔食品股份有限公司 | Method for extracting antibacterial peptide and albumin from pea bean serum wastewater |
| CN111018164B (en) * | 2020-03-09 | 2020-07-31 | 烟台双塔食品股份有限公司 | Method for extracting antibacterial peptide and albumin from pea bean serum wastewater |
| CN114316028A (en) * | 2021-12-30 | 2022-04-12 | 南宁市武鸣金三川农业科技有限公司 | Process for producing collagen polypeptide by physical and biological double-cutting one-step method |
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