CN108130354A - A kind of method that ultrasound, heat treatment auxiliary two enzymes method prepare corn peptide - Google Patents
A kind of method that ultrasound, heat treatment auxiliary two enzymes method prepare corn peptide Download PDFInfo
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Abstract
The present invention provides a kind of methods that ultrasound, heat treatment auxiliary two enzymes method prepare corn peptide, belong to biotechnology.The present invention is with corn protein powder（Containing 60 or so protein）For raw material, the physicochemical property of albumen can be changed based on ultrasonic wave and heating, improve protein digestion rate；First corn protein powder is pre-processed by the way of ultrasound and heat treatment, gained corn protein solution is digested through alpha-amylase and is purified, then in vacuum low-pressure state through hydrolysis by novo, go the processes such as bitter, filtering, micro- worry and concentration, vacuum drying through enzyme deactivation, centrifugation, decoloration again, finally more than 93% protein content corn Gly-His-Lys.The method provided by the invention for preparing corn peptide has operability strong, improves enzymolysis efficiency and rate, reduces enzymolysis time, reduces enzymolysis process non-enzymatic browning and bitter taste is formed；The corn peptide of preparation has higher protein content, and molecular weight is small, can be widely used in the developmental research of food, health products and medicine etc..
Description
Technical field：
The present invention relates to a kind of preparation methods of corn peptide, and in particular to a kind of ultrasound, heat treatment auxiliary two enzymes method prepare corn
The method of peptide, belongs to biotechnology.
Background technology：
Corn peptide be corn protein powder after various modes are degraded, the zein that is obtained by the processes such as isolating and purifying, refining
Catabolite, molecular weight be generally concentrated at 1000 hereinafter, activity it is higher, rich in glutamic acid, proline, leucine and alanine
And rich in hydrophobic amino acid.Since amino acid composition is special, corn peptide is made to have special physicochemical property and various biological work(
Energy.Corn peptide degrades to obtain through zein, and property is different from zein, and corn peptide dissociation group numbers are significantly larger than jade
Rice gluten, hydrophily increase thus have some good physicochemical properties：Good fluidity, viscosity stabilization, be affected by temperature it is small；
Dissolubility is good, and applicable pH range is wide；Retentiveness is good；It is not easy to agglomerate under the conditions of acid, isoelectric point is not easy to precipitate；Stronger emulsibility and
Foaming characteristic；Stability is good, also with the physiological activity such as no antigen, anti-oxidant, lowering blood pressure and blood fat.
Biologically active peptide preparation method common at present has gene engineering research, acid-hydrolysis method and enzymatic isolation method etc..Genetic engineering
The advantages of method is that purity is higher, and shortcoming is that its R＆D cycle is long, isolates and purifies difficulty；Though acid-hydrolysis method prepares active peptide letter
It is single cheap, but reaction condition is violent, amino acid be damaged it is serious, be also possible to generate noxious material.Therefore enzymatic isolation method production activity
Peptide becomes research emphasis, and this method working condition is mild, easily controllable and safe.But enzyme hydrolysis method be one kind be not thorough,
Incomplete method for hydrolysis, and traditional mode of action enzymolysis efficiency is not high, enzyme availability is not high.Using pretreatment albumen
Mode can improve enzymolysis efficiency and rate, and common processing mode has ultrasonic wave, heating, microwave, high pressure etc..Therefore, it is efficient
Rate prepares the corn peptide of high-purity, corn protein powder can be pre-processed when preparing corn peptide, and processing technology is done excellent
Change.
Invention content：
The rate and efficiency of corn peptide are prepared to improve enzyme process, the present invention provides a kind of ultrasound, heat treatment auxiliary two enzymes method systems
The method of standby corn peptide makes obtained more than 93% corn peptide protein content, available in food, health products and medicine etc..
The method that a kind of ultrasound of the present invention, heat treatment auxiliary two enzymes method prepare corn peptide includes the following steps：
（1）Corn protein powder crushed 20-60 mesh sieve and water is made into the protein solution that solid content is 10-12%, and stirring is equal
It is even.
（2）Corn protein solution carries out ultrasound pretreatment 20-40min, carries out heat treatment 20-40min.
（3）Protein solution after ultrasound, heat treatment, adjusts pH value 5.5-6.0, and 80-85 DEG C of temperature adds in high temperature and forms sediment
Powder enzyme Termamyl SC are digested, enzyme digestion reaction time 2h.
（4）Clear liquid and solid slag are detached with centrifugal process, solid slag is the zein that purity improves, and protein content is carried from 60%
Height is to more than 80%；Solid slag is made into the protein solution that solid content is 10-12% again.
（5）The protein solution prepared is adjusted into pH7.0-8.0, alkali protease is added in, it is advance to be then pumped into temperature
It is adjusted to be hydrolyzed in 50-60 DEG C of vacum reaction tank, progressive rolling movement low pressure hydrolysis 2.0-3.5h.
（6）Retort temperature is increased to 95-97 DEG C, enzyme deactivation 30min；After temperature is down to 15-20 DEG C in tank, blowing.
（7）After feed liquid centrifugation, activated carbon, 50-65 DEG C of set temperature, stirring decoloration 1h, then through taking out are added in supernatant
Filter is concentrated after micro-filtration, then vacuum dried obtains corn Gly-His-Lys.
Further, the ultrasonic pretreatment condition is that initial temperature is 20 DEG C, supersonic frequency 40kHz, impulse ultrasound
Working time is 10s and intermittent time 3s, ultrasonic time 20-40min；Heat treatment condition is that heat treatment temperature is 121 DEG C,
Processing time is 20-40min.
Further, the optimal pH of the alpha-amylase Termamyl SC is 5.5-6.0, and temperature is 70-85 DEG C, is added
Dosage is the 0.1-0.15% of corn protein powder dry weight.
Further, the additive amount of the alkali protease is the 1-1.2% of zein dry weight, and is reacted in vacuum reaction
Tank mesolow carries out, and feed liquid wants progressive rolling movement to increase reaction rate.
Further, the additive amount of the activated carbon is the 0.5-1% of solution weight.
Beneficial effects of the present invention：
The present invention improves enzymolysis process corn protein powder using ultrasound and the pretreatment mode of heat treatment processing corn protein powder
Enzymolysis efficiency and rate, reduce enzymolysis time.
The present invention digests corn protein powder first with alpha-amylase, to improve zein powder purity, makes corn egg
Protein content is increased to more than 80% from 60% in white powder；And more than 95% enzymolysis process albumen yield, make loss is mainly starch
And glucide.
The present invention, alkali protease carry out the hydrolysis of zein in vacuum tank, can avoid in hydrolytic process
The formation of non-enzymatic browning and some bad flavor substances.
Corn peptide prepared by the present invention, more than 93% protein content, bitter taste is extremely low, available for food, health products and medicine
Product etc..
Specific embodiment：
Below by specific embodiment, the present invention will be further described, but the present invention is not limited only to this.
Embodiment 1：
（1）Corn protein powder crushed 20-60 mesh sieve and water is made into the protein solution that solid content is 10-12%, and stirring is equal
It is even.
（2）The corn protein solution stirred evenly first carries out ultrasound pretreatment 20min, then carries out heat treatment 20min.
（3）Pretreated protein solution, adjusts pH value 5.5, and 85 DEG C of temperature adds in corn protein powder dry weight 0.1%
Alpha-amylase Termamyl SC carry out enzyme digestion reaction 2h.
（4）Centrifuge 4000r/min detaches clear liquid and solid slag 15min, collects solid slag and is made into solid content again as 10-
12% protein solution.
（5）Regulatory protein pH value of solution 8.0 adds in the alkali protease of corn protein powder dry weight 1-1.2%, is then pumped into
Temperature has been adjusted to be hydrolyzed in 55 DEG C of vacum reaction tank in advance, progressive rolling movement low pressure hydrolysis 2.0-3.5h.
（6）Retort temperature is increased to 95 DEG C, enzyme deactivation 30min；Recirculated water quickly reduces temperature in tank and, to 15-20 DEG C, protects
Hold blowing after 10-15min.
（7）Feed liquid collects supernatant after 4000r/min centrifuges 15min, adjusts temperature 60 C, adds in solution weight
The activated carbon of 0.5-1%, is stirred to react 1h；Then through filtering, concentrate after micro-filtration, then vacuum dried corn Gly-His-Lys are obtained.
Embodiment 2：
（1）Corn protein powder crushed 20-60 mesh sieve and water is made into the protein solution that solid content is 10-12%, and stirring is equal
It is even.
（2）The corn protein solution stirred evenly first carries out heat treatment 20min, and ultrasound pretreatment 20min is carried out after cooling.
（3）Pretreated protein solution, adjusts pH value 5.5, and 85 DEG C of temperature adds in corn protein powder dry weight 0.1%
Alpha-amylase Termamyl SC carry out enzyme digestion reaction 2h.
（4）Centrifuge 4000r/min detaches clear liquid and solid slag 15min, collects solid slag and is made into solid content again as 10-
12% protein solution.
（5）Regulatory protein pH value of solution 8.0 adds in the alkali protease of corn protein powder dry weight 1-1.2%, is then pumped into
Temperature has been adjusted to be hydrolyzed in 55 DEG C of vacum reaction tank in advance, progressive rolling movement low pressure hydrolysis 2.0-3.5h.
（6）Retort temperature is increased to 95 DEG C, enzyme deactivation 30min；Recirculated water quickly reduces temperature in tank and, to 15-20 DEG C, protects
Hold blowing after 10-15min.
（7）Feed liquid collects supernatant after 4000r/min centrifuges 15min, adjusts temperature 60 C, adds in solution weight
The activated carbon of 0.5-1%, is stirred to react 1h；Then through filtering, concentrate after micro-filtration, then vacuum dried corn Gly-His-Lys are obtained.
Embodiment 3：
Using without ultrasound and heat treatment procedure prepared by corn peptide for compare, to corn peptide obtained in Examples 1 and 2 into
Row degree of hydrolysis（DH）It is analyzed with protein content.
Degree of hydrolysis test is that the reaction solution of 125uL and 2.0mL 0.2mol/L pH8.2 are added in blank glass pipe
Phosphate buffer, oscillation are uniformly mixed, and 2,4, the 6- trinitrobenzene sulfonic acid for then adding in 1.0mL 0.01% are sufficiently mixed, will be mixed
It closes liquid and is placed in 50 DEG C of water-bath to be protected from light to heat and react 30min, after adding in the 0.1M sodium sulfites termination reaction of 2.0mL, cooling
15min measures light absorption value in 420nm, and blank is that distilled water surrogate response liquid is used under same operation.Free amino group content according to
The standard curve of L-Leu content determines（Y=6.8119x+0.0083, coefficient R2=0.992）
DH（%）=（Lt-Lo）/（Lmax-Lo）*100
In formula：Free amino group content when Lt represents to digest the t times in sample；Lo represents zero moment free amino group content；Lmax tables
Show free amino group content in complete hydrolysis liquid.
Protein content is detected using Kjeldahl's method, conversion coefficient 6.24.
Degree of hydrolysis and protein content analysis result table
| Sample ID | Degree of hydrolysis % | Protein content % | Average molecular weight |
| Control sample | 11.3 | 89.2 | 891 |
| 1 sample of embodiment | 13.4 | 93.8 | 877 |
| 2 sample of embodiment | 12.8 | 93.5 | 873 |
By table as it can be seen that ultrasound and heat treatment corn protein powder help to improve protein content and hydrolytic process hydrolysis in corn peptide
Efficiency；In addition, ultrasound and heat treatment sequence have an impact hydrolysis effect, Grape berry is than first Grape berry after first ultrasound pretreatment
Ultrasound pretreatment improves percent hydrolysis and becomes apparent from afterwards.
Claims (7)
1. a kind of method that ultrasound, heat treatment auxiliary two enzymes method prepare corn peptide, which is characterized in that using corn protein powder as original
Material, processing step include：
（1）Corn protein powder crushed 20-60 mesh sieve and water is made into the protein solution that solid content is 10-12%, and stirring is equal
It is even；
（2）Corn protein solution carries out ultrasound pretreatment 20-40min, carries out heat treatment 20-40min；
（3）Protein solution after ultrasound, heat treatment, adjusts pH value 5.5-6.0, and 80-85 DEG C of temperature adds in alpha-amylase
Termamyl SC are digested, enzyme digestion reaction time 2h；
（4）Clear liquid and solid slag are detached with centrifugal process, solid slag is the zein that purity improves；Solid slag is made into solid content again to contain
Measure the protein solution for 10-12%；
（5）The protein solution prepared is adjusted into pH7.0-8.0, alkali protease is added in, is then pumped into temperature and is adjusted in advance
It is hydrolyzed in 50-60 DEG C of vacum reaction tank, progressive rolling movement low pressure hydrolysis 2.0-3.5h；
（6）Retort temperature is increased to 95-97 DEG C, enzyme deactivation 30min；Treat that temperature is down to 15-20 DEG C in tank, blowing；
（7）After feed liquid centrifugation, add in activated carbon in supernatant, 50-65 DEG C of set temperature, stirring decoloration 1h, then through filtering, it is micro-
It is concentrated after filter, then vacuum dried obtains corn Gly-His-Lys.
2. the method that a kind of ultrasound according to claim 1, heat treatment auxiliary two enzymes method prepare corn peptide, feature exist
In：Step（2）In, ultrasonic pretreatment condition is, initial temperature is 20 DEG C, supersonic frequency 40kHz, impulse ultrasound work when
Between be 10s and intermittent time 3s, ultrasonic time 20-40min；Heat treatment condition is, heat treatment temperature is 121 DEG C, during processing
Between be 20-40min.
3. the method that a kind of ultrasound according to claim 1, heat treatment auxiliary two enzymes method prepare corn peptide, feature exist
In：Step（3）In, the optimal pH of alpha-amylase Termamyl SC used is 5.5-6.0, and temperature is 70-85 DEG C, addition
Measure the 0.1-0.15% for corn protein powder dry weight.
4. the method that a kind of ultrasound according to claim 1, heat treatment auxiliary two enzymes method prepare corn peptide, feature exist
In：Step（4）In, protein content is increased to more than 80% from 60% in the protein liquid after alpha-amylase enzymolysis purification.
5. the method that a kind of ultrasound according to claim 1, heat treatment auxiliary two enzymes method prepare corn peptide, feature exist
In：Step（5）With（6）In, the amount for adding in alkali protease is the 1-1.2% of zein dry weight；Contain on a small quantity in protein liquid
Reduced sugar, to avoid the formation of hydrolytic process non-enzymatic browning and some bad flavor substances, hydrolysis is in vacum reaction tank
Low pressure carries out, and feed liquid wants progressive rolling movement to increase reaction rate；After hydrolysis carries out 2.0-3.5h, retort passes through recirculated water
Fast cooling when temperature is down to 20 DEG C, keeps blowing after 10-15min.
6. the method that a kind of ultrasound according to claim 1, heat treatment auxiliary two enzymes method prepare corn peptide, feature exist
In：Step（7）In, the additive amount of activated carbon is the 0.5-1% of solution weight.
7. the corn peptide prepared according to claim 1-6, more than 93% protein content, bitter taste is extremely low, available for eating
Product, health products and drug etc..
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Cited By (7)
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|---|---|---|---|---|
| CN110747247A (en) * | 2018-07-23 | 2020-02-04 | 江苏挺卫实业有限公司 | Separation and extraction method of corn peptide for anti-alcohol beverage |
| CN110819675A (en) * | 2019-11-20 | 2020-02-21 | 吉林云步科技有限公司 | Preparation method for extracting high-activity small-molecule peptide from corn |
| CN110981979A (en) * | 2019-12-18 | 2020-04-10 | 内蒙古阜丰生物科技有限公司 | Method for improving corn starch yield |
| CN111505197A (en) * | 2020-05-28 | 2020-08-07 | 河南三方元泰检测技术有限公司 | Method for detecting protein content in food |
| CN111518858A (en) * | 2020-05-14 | 2020-08-11 | 武汉天天好生物制品有限公司 | Vegetable protein peptide and preparation method thereof |
| CN111631329A (en) * | 2020-05-09 | 2020-09-08 | 南宁荣港生物科技有限公司 | Egg polypeptide preparation method and egg polypeptide beverage |
| CN114073291A (en) * | 2020-08-13 | 2022-02-22 | 黑龙江省北大荒绿色健康食品有限责任公司 | Method for preparing protein peptide reinforced corn flour |
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| CN110747247A (en) * | 2018-07-23 | 2020-02-04 | 江苏挺卫实业有限公司 | Separation and extraction method of corn peptide for anti-alcohol beverage |
| CN110819675A (en) * | 2019-11-20 | 2020-02-21 | 吉林云步科技有限公司 | Preparation method for extracting high-activity small-molecule peptide from corn |
| CN110981979A (en) * | 2019-12-18 | 2020-04-10 | 内蒙古阜丰生物科技有限公司 | Method for improving corn starch yield |
| CN111631329A (en) * | 2020-05-09 | 2020-09-08 | 南宁荣港生物科技有限公司 | Egg polypeptide preparation method and egg polypeptide beverage |
| CN111518858A (en) * | 2020-05-14 | 2020-08-11 | 武汉天天好生物制品有限公司 | Vegetable protein peptide and preparation method thereof |
| CN111518858B (en) * | 2020-05-14 | 2022-10-11 | 武汉天天好生物制品有限公司 | Vegetable protein peptide and preparation method thereof |
| CN111505197A (en) * | 2020-05-28 | 2020-08-07 | 河南三方元泰检测技术有限公司 | Method for detecting protein content in food |
| CN114073291A (en) * | 2020-08-13 | 2022-02-22 | 黑龙江省北大荒绿色健康食品有限责任公司 | Method for preparing protein peptide reinforced corn flour |
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Application publication date: 20180608 |