CN106084023A - A kind of method extracting ice structural protein from cold ground winter wheat - Google Patents
A kind of method extracting ice structural protein from cold ground winter wheat Download PDFInfo
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- CN106084023A CN106084023A CN201610726901.8A CN201610726901A CN106084023A CN 106084023 A CN106084023 A CN 106084023A CN 201610726901 A CN201610726901 A CN 201610726901A CN 106084023 A CN106084023 A CN 106084023A
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- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
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Abstract
A kind of method extracting ice structural protein from cold ground winter wheat, relates to a kind of method extracting ice structural protein from cold ground winter wheat.The present invention is to solve that in prior art, ice structural protein extraction raw material is limited, extraction process is complicated, and the ice structural protein that relatively costly, extraction obtains can not be in frozen food and the most wide variety of problem.Extracting method: one, the preparation of ice hockey, a diameter of 2cm of ice hockey；Two, cold ground winter wheat flour is then centrifuged for the extraction of phosphate buffered solution mix and blend；Three, add ice hockey ice to carry, then leach ice hockey and naturally melt；Four, answering of winter wheat ISPs of cold ground carries.The present invention high and cold winter wheat that has drawn from is food source, and safety is guaranteed；Extraction ratio is 1.63%～4.56%, and ice structural protein prepared by the present invention has stronger Activity of Antifreeze.The present invention is be applicable to the ice structural protein extracted cold ground winter wheat.
Description
Technical field
The present invention relates to a kind of method extracting ice structural protein from cold ground winter wheat.
Background technology
Ice structural protein (ice structuring proteins, ISPs), is also called and does not freezes albumen, antifreeze protein
(antifreeze proteins, AFPs) is a class by some Fish, insecticide, plant, fungus and antibacterial is the extraneous ring of opposing
Polypeptide produced by the stress of border, has and stops ice crystal formation, modification ice crystal form and the ability of suppression recrystallization, and it has
Certain heat stagnation activity, i.e. can reduce the freezing point of solution, but the impact on fusing point is less with non-colligative property form, so that molten
Between freezing point and the fusing point of liquid, difference occurs.The distinctive character of ice structural protein ensures biology existence under cryogenic.
2006, China's Ministry of Public Health was announced ice structural protein and can be applied in chilled food as new food additive；
Limited owing to extracting ISPs raw material, mostly extract from cold water fish, the factors such as extraction process is complicated, cause its cost
Higher, expensive, not in frozen food and the most commonly used, the development to ISPs causes restriction.How to make
Produce the main task that simple, the cheap ISPs of extraction process is ISPs product development.The structure of plant ISPs and function
Complex, there is low grade fever stickiness (the most about 0.1～0.5 DEG C) and high recrystallization inhibition activity (exceeds Fish and insecticide ISPs
About 10～100 times) feature, a small amount of plant ISPs just has the brilliant inhibitory activity of higher weight.Document was only mentioned by auxiliary
The method that trash ice extracts ice structural protein from the Folium Ligustri Lucidi, but there is uncertain and shortcoming, complex process in the method, need into
Row is cold-induced, and extract impurity is many, and trash ice do not carries out strict control, so can not strictly control extraction ratio or yield with
And quality；The protein product of the Folium Ligustri Lucidi will could express through low temperature induction, synthesize and accumulate.Accumulate in vivo and disappear
Except there being seasonal variety.Typically disappearing in summer, autumn occurs, after Cold exposed, winter reaches summit.
Semen Tritici aestivi is the generalized grain crop that beats the world, the grain variety that Ye Shi China is important, occupies weight in national economy
Want status.Semen Tritici aestivi can be divided into two types according to habit: winter wheat and spring wheat.Winter wheat is relative to nutrition for spring wheat
Phase is long, from the trophophase to poor growth in reproduction period, and one very long winter low temperature stage of experience therebetween, until under the long-day in spring
Just bloom.The proportion of winter wheat plantation is more than spring wheat in the world.Winter wheat quality is good, and yield is high, but weather severe cold reduces
The geographical distribution region of winter wheat suitable planting, makes the agricultural productive force receive the biggest loss.The Northeast's winter temperature
Reaching subzero less than 30 DEG C, autumn, the winter wheat of plantation to experience a severe winter completed, and winter wheat is cold-resistant in growth course
Very competent, its seedling can pass the winter, and illustrates that the protein in its seedling has preferable cold tolerance.
Summary of the invention
The present invention is to solve that in prior art, ice structural protein extraction raw material is limited, extraction process is complicated, relatively costly,
Extracting the ice structural protein obtained can not be in frozen food and the most wide variety of problem, it is proposed that a kind of little from the cold ground winter
The method extracting ice structural protein in wheat.
The present invention extracts the method for ice structural protein from cold ground winter wheat and sequentially includes the following steps:
One, the preparation of ice hockey
Place glass bead in each ice hole site of silica gel ice tray, to the note distillation of ice hole, be subsequently placed in the refrigerator of-18 DEG C
Freezing, obtains ice hockey；
A diameter of 1cm of described glass bead；A diameter of 2cm of the described ice hockey obtained；Described silica gel ice tray be heatproof be-
The silica gel ice tray of 40 DEG C, silica gel ice tray is tabular, a length of 24cm, a width of 7.3cm, plate is provided with 10 × 2 spherical ice holes, often
The internal diameter in individual ice hole is 2cm；
Two, the extraction of cold ground winter wheat ISPs
It is 1g:(10～30mL by solid-liquid ratio) cold ground winter wheat flour is mixed with phosphate buffered solution, at room temperature bar
Under part, stirring extracts 0.5～3.0h, then with centrifuge 14～16min, obtains supernatant；
The pH of described phosphate buffered solution is 7.2～8.0, and phosphate concentration is 60～140mmol/L；Described stirring carries
Using magnetic stirring apparatus to carry out when taking, the rotating speed of magnetic stirring apparatus is 800～1000r/min；Described centrifugal time centrifuge turn
Speed is 8000～10000r/min；
Three, ice hockey extracts
Adding the ice hockey of step one preparation in step 2 supernatant, put in the refrigerator of-18 DEG C, ice carries 2～4min, this
Shi Bingqiu absorption ice structural protein reaches saturated, leaches ice hockey juxtaposition and the most naturally melts and obtain extracting solution, leaches ice hockey
After obtain remaining supernatant；The ice hockey amount of described addition supernatant is every 50mL supernatant 6～15；
Four, answering of winter wheat ISPs of cold ground carries
In remaining supernatant, add the ice hockey of step one preparation, repetition step 3 2～4 times, collect and extract every time
The extracting solution arrived, i.e. completes.Now in step 2, the ISPs in supernatant is all adsorbed by ice hockey substantially；
This law possesses following beneficial effect:
1, the present invention has drawn from high and cold winter wheat, and the ice structural protein of extraction is food source, for being applied to frozen food
On safety guaranteed；
2, the extraction ratio of the present invention cold ground winter wheat ISPs ice structural protein is 1.63%～4.56%；Wherein extraction ratio
For: the quality of the ice structural protein finally given accounts for the percentage ratio of the gross mass of the multiple proteins contained in raw material.
3, carry out cold ground winter wheat ice structural protein extracting solution concentrating, lyophilization obtains cold ground winter wheat ice structure egg
White lyophilized powder, redissolves lyophilized powder, solution carries out ice crystal observation, and contrasts with distilled water, sucrose solution, and result shows
Showing, cold ground winter wheat ice structural protein solution ice crystal amasss little, form of diverse, has preferable ice crystal and modifies characteristic；
4, the heat stagnation of the cold ground winter wheat ice structural protein extracting solution of differential scanning calorimeter detection present invention extraction is taked
Activity, extract concentration is 30mg/mL, and heat stagnation activity is 0.27～0.33 DEG C, shows that heat stagnation activity is notable, it follows that this
The cold ground winter wheat ice structural protein of bright preparation has stronger Activity of Antifreeze；
5, being observed by ice crystal, winter wheat ISPs redissolution liquid ice crystal in cold ground prepared by the inventive method is evenly distributed, form
Different, illustrate that it has Activity of Antifreeze.
Detailed description of the invention:
Technical solution of the present invention is not limited to act detailed description of the invention set forth below, also includes between each detailed description of the invention
Arbitrarily reasonable combination.
Detailed description of the invention one: present embodiment extracts the method for ice structural protein according to the following steps from cold ground winter wheat
Carry out:
One, the preparation of ice hockey
Place glass bead in each ice hole site of silica gel ice tray, note distilled water to ice hole, be subsequently placed in the refrigerator of-18 DEG C
Middle freezing, obtains ice hockey；
Two, the extraction of cold ground winter wheat ISPs
It is 1g:(10～30mL by solid-liquid ratio) cold ground winter wheat flour is mixed with phosphate buffered solution, at room temperature bar
Under part, stirring extracts 0.5～3.0h, then with centrifuge 14～16min, obtains supernatant；
Three, ice hockey extracts
Adding the ice hockey of step one preparation in step 2 supernatant, put in the refrigerator of-18 DEG C, ice carries 2～4min, this
Shi Bingqiu absorption ice structural protein reaches saturated, leaches ice hockey juxtaposition and the most naturally melts and obtain extracting solution, leaches ice hockey
After obtain remaining supernatant；
Four, answering of winter wheat ISPs of cold ground carries
In remaining supernatant, add the ice hockey of step one preparation, repetition step 3 2～4 times, collect and extract every time
The extracting solution arrived, i.e. completes.
Present embodiment possesses following beneficial effect:
1, present embodiment has drawn from high and cold winter wheat, and the ice structural protein of extraction is food source, for being applied to freezing
Safety on food is guaranteed；
2, the extraction ratio of present embodiment cold ground winter wheat ISPs ice structural protein is 1.63%～4.56%；
3, carry out cold ground winter wheat ice structural protein extracting solution concentrating, lyophilization obtains cold ground winter wheat ice structure egg
White lyophilized powder, redissolves lyophilized powder, solution carries out ice crystal observation, and contrasts with distilled water, sucrose solution, and result shows
Showing, cold ground winter wheat ice structural protein solution ice crystal amasss little, form of diverse, has preferable ice crystal and modifies characteristic；
4, the cold ground winter wheat ice structural protein extracting solution of differential scanning calorimeter detection present embodiment extraction is taked
Heat stagnation activity, extract concentration is 30mg/mL, and heat stagnation activity is 0.27～0.33 DEG C, shows that heat stagnation activity is notable, it follows that
Cold ground winter wheat ice structural protein prepared by present embodiment has stronger Activity of Antifreeze；
5, being observed by ice crystal, winter wheat ISPs redissolution liquid ice crystal in cold ground prepared by present embodiment method is evenly distributed,
Come in every shape, illustrate that it has Activity of Antifreeze.
Detailed description of the invention two: present embodiment is unlike detailed description of the invention one: glass bead described in step one
A diameter of 1cm；A diameter of 2cm of the described ice hockey obtained；Described silica gel ice tray be heatproof be the silica gel ice tray of-40 DEG C, silica gel
Ice tray is tabular, a length of 24cm, a width of 7.3cm, and plate is provided with 10 × 2 spherical ice holes, and the internal diameter in each ice hole is 2cm.
Other steps are identical with detailed description of the invention one with parameter.
Detailed description of the invention three: present embodiment is unlike detailed description of the invention one or two: phosphate described in rapid two
The pH of buffer solution is 7.2～8.0, and phosphate concentration is 60～140mmol/L.Other steps and parameter and detailed description of the invention
One or two is identical.
Detailed description of the invention four: present embodiment is unlike one of detailed description of the invention one to three: described in step 2
Using magnetic stirring apparatus to carry out when stirring is extracted, the rotating speed of magnetic stirring apparatus is 800～1000r/min.Other steps and parameter
Identical with one of detailed description of the invention one to three.
Detailed description of the invention five: present embodiment is unlike one of detailed description of the invention one to four: described in step 2
Time centrifugal, the rotating speed of centrifuge is 8000～10000r/min.Other steps and parameter and one of detailed description of the invention one to four phase
With.
Detailed description of the invention six: present embodiment is unlike one of detailed description of the invention one to five: described in step 3
The ice hockey amount adding supernatant is every 50mL supernatant 6～15.Other steps and parameter and detailed description of the invention one to five it
One is identical.
Detailed description of the invention seven: present embodiment is unlike one of detailed description of the invention one to six: described in step 3
The ice hockey amount adding supernatant is every 50mL supernatant 10.Other steps and parameter and one of detailed description of the invention one to six phase
With.
For checking beneficial effects of the present invention, carry out tests below:
Embodiment 1
The present embodiment extracts the method for ice structural protein from cold ground winter wheat and sequentially includes the following steps:
One, the preparation of ice hockey
Place glass bead in each ice hole site of silica gel ice tray, to the water filling of ice hole, be subsequently placed in the refrigerator of-18 DEG C cold
Freeze, obtain ice hockey；
A diameter of 1cm of described glass bead；A diameter of 2cm of the described ice hockey obtained；Described silica gel ice tray be heatproof be-
The silica gel ice tray of 40 DEG C, silica gel ice tray is tabular, a length of 24cm, a width of 7.3cm, plate is provided with 10 × 2 spherical ice holes, often
The internal diameter in individual ice hole is 2cm；
Two, the extraction of cold ground winter wheat ISPs
It is that cold ground winter wheat flour is mixed with phosphate buffered solution by 1g:15mL by solid-liquid ratio, stirs at ambient temperature
Mix extraction 2.0h, be then centrifuged 15min with centrifuge with the centrifugal speed of 10000r/min, obtain supernatant；The present embodiment is adopted
Be cold ground winter wheat powder " east agriculture winter wheat No. 1 ", be uniquely can be at the winter wheat variety of Heilongjiang Province's safe overwintering, albumen
Content is 12.25%；
The pH of described phosphate buffered solution is 7.8, and phosphate concentration is 100mmol/L；Described stirring uses when extracting
Magnetic stirring apparatus is carried out, and the rotating speed of magnetic stirring apparatus is 900r/min；
Three, ice hockey extracts
Adding the ice hockey of step one preparation in step 2 supernatant, put in the refrigerator of-18 DEG C, ice carries 2min, now
Ice hockey absorption ice structural protein reaches saturated, leaches ice hockey juxtaposition and the most naturally melts and obtain extracting solution, after leaching ice hockey
Obtain remaining supernatant；The ice hockey amount of described addition supernatant is every 50mL supernatant 10；
Four, answering of winter wheat ISPs of cold ground carries
In remaining supernatant, the ice hockey of addition step one preparation, repeats step 34 times, collects what extraction every time obtained
Extracting solution, i.e. completes.
The present embodiment possesses following beneficial effect:
1, the present embodiment has drawn from high and cold winter wheat, and the ice structural protein of extraction is food source, for being applied to freezing food
Safety on product is guaranteed；
2, the extraction ratio of the present embodiment cold ground winter wheat ISPs ice structural protein is 4.56%；
3, carry out cold ground winter wheat ice structural protein extracting solution concentrating, lyophilization obtains cold ground winter wheat ice structure egg
White lyophilized powder, redissolves lyophilized powder, solution carries out ice crystal observation, and contrasts with distilled water, sucrose solution, and result shows
Showing, cold ground winter wheat ice structural protein solution ice crystal amasss little, form of diverse, has preferable ice crystal and modifies characteristic；
4, differential scanning calorimeter is taked to detect the heat of the cold ground winter wheat ice structural protein extracting solution that the present embodiment extracts
Stagnant activity, extract concentration is 30mg/mL, and heat stagnation activity is 0.33 DEG C, shows that heat stagnation activity is notable, it follows that the present embodiment
The cold ground winter wheat ice structural protein of preparation has stronger Activity of Antifreeze；
5, being observed by ice crystal, winter wheat ISPs redissolution liquid ice crystal in cold ground prepared by the present embodiment method is evenly distributed, shape
State is different, illustrates that it has Activity of Antifreeze.
Claims (7)
1. the method extracting ice structural protein from cold ground winter wheat, it is characterised in that the method sequentially includes the following steps:
One, the preparation of ice hockey
Place glass bead in each ice hole site of silica gel ice tray, note distilled water to ice hole, be subsequently placed in the refrigerator of-18 DEG C cold
Freeze, obtain ice hockey；
Two, the extraction of cold ground winter wheat ISPs
It is 1g:(10～30mL by solid-liquid ratio) by cold ground winter wheat flour and phosphate buffered solution mix and blend, at room temperature bar
Under part, stirring extracts 0.5～3.0h, then with centrifuge 14～16min, obtains supernatant；
Three, ice hockey extracts
Adding the ice hockey of step one preparation in step 2 supernatant, put in the refrigerator of-18 DEG C, ice carries 2～4min, now ice
Ball absorption ice structural protein reaches saturated, leaches ice hockey juxtaposition and the most naturally melts and obtain extracting solution, must after leaching ice hockey
To remaining supernatant；
Four, answering of winter wheat ISPs of cold ground carries
The ice hockey of addition step one preparation, repetition step 3 2～4 times in remaining supernatant, collect what extraction every time obtained
Extracting solution, i.e. completes.
A kind of method extracting ice structural protein from cold ground winter wheat the most according to claim 1, it is characterised in that: step
A diameter of 1cm of glass bead described in rapid；A diameter of 2cm of the described ice hockey obtained；Described silica gel ice tray is that heatproof is-40 DEG C
Silica gel ice tray, silica gel ice tray is tabular, a length of 24cm, a width of 7.3cm, and plate is provided with 10 × 2 spherical ice holes, Mei Gebing
The internal diameter in hole is 2cm.
A kind of method extracting ice structural protein from cold ground winter wheat the most according to claim 1, it is characterised in that: step
The pH of phosphate buffered solution described in rapid two is 7.2～8.0, and phosphate concentration is 60～140mmol/L.
A kind of method extracting ice structural protein from cold ground winter wheat the most according to claim 1, it is characterised in that: step
Stirring described in rapid two uses magnetic stirring apparatus to carry out when extracting, and the rotating speed of magnetic stirring apparatus is 800～1000r/min.
A kind of method extracting ice structural protein from cold ground winter wheat the most according to claim 1, it is characterised in that: step
When being centrifuged described in rapid two, the rotating speed of centrifuge is 8000～10000r/min.
A kind of method extracting ice structural protein from cold ground winter wheat the most according to claim 1, it is characterised in that: step
Adding the ice hockey amount of supernatant described in rapid three is every 50mL supernatant 6～15.
A kind of method extracting ice structural protein from cold ground winter wheat the most according to claim 1, it is characterised in that: step
The ice hockey amount adding supernatant described in rapid three is every 50mL supernatant 10.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610726901.8A CN106084023A (en) | 2016-08-25 | 2016-08-25 | A kind of method extracting ice structural protein from cold ground winter wheat |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610726901.8A CN106084023A (en) | 2016-08-25 | 2016-08-25 | A kind of method extracting ice structural protein from cold ground winter wheat |
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| CN106084023A true CN106084023A (en) | 2016-11-09 |
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| CN201610726901.8A Pending CN106084023A (en) | 2016-08-25 | 2016-08-25 | A kind of method extracting ice structural protein from cold ground winter wheat |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101684144A (en) * | 2008-09-26 | 2010-03-31 | 江南大学 | Method for preparing ice structure protein |
| CN104650199A (en) * | 2015-01-12 | 2015-05-27 | 江南大学 | Method for quickly and specifically purifying ice structure protein in barley grains |
-
2016
- 2016-08-25 CN CN201610726901.8A patent/CN106084023A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101684144A (en) * | 2008-09-26 | 2010-03-31 | 江南大学 | Method for preparing ice structure protein |
| CN104650199A (en) * | 2015-01-12 | 2015-05-27 | 江南大学 | Method for quickly and specifically purifying ice structure protein in barley grains |
Non-Patent Citations (2)
| Title |
|---|
| 刘尚 等: ""一种女贞叶抗冻蛋白的分离纯化"", 《植物学通报》 * |
| 张超 等: ""特异性亲和吸附法纯化冬小麦麸皮抗冻蛋白"", 《中国粮油学报》 * |
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Application publication date: 20161109 |
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