CN110204752A - A kind of preparation method being packed for soybean protein film - Google Patents
A kind of preparation method being packed for soybean protein film Download PDFInfo
- Publication number
- CN110204752A CN110204752A CN201910463027.7A CN201910463027A CN110204752A CN 110204752 A CN110204752 A CN 110204752A CN 201910463027 A CN201910463027 A CN 201910463027A CN 110204752 A CN110204752 A CN 110204752A
- Authority
- CN
- China
- Prior art keywords
- soybean protein
- packed
- parts
- graphene oxide
- protein film
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J5/00—Manufacture of articles or shaped materials containing macromolecular substances
- C08J5/18—Manufacture of films or sheets
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2389/00—Characterised by the use of proteins; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2405/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2401/00 or C08J2403/00
- C08J2405/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K3/00—Use of inorganic substances as compounding ingredients
- C08K3/02—Elements
- C08K3/08—Metals
- C08K2003/0856—Iron
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K2201/00—Specific properties of additives
- C08K2201/011—Nanostructured additives
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K3/00—Use of inorganic substances as compounding ingredients
- C08K3/02—Elements
- C08K3/04—Carbon
- C08K3/042—Graphene or derivatives, e.g. graphene oxides
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K3/00—Use of inorganic substances as compounding ingredients
- C08K3/02—Elements
- C08K3/04—Carbon
- C08K3/046—Carbon nanorods, nanowires, nanoplatelets or nanofibres
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K5/00—Use of organic ingredients
- C08K5/16—Nitrogen-containing compounds
- C08K5/34—Heterocyclic compounds having nitrogen in the ring
- C08K5/3467—Heterocyclic compounds having nitrogen in the ring having more than two nitrogen atoms in the ring
- C08K5/3477—Six-membered rings
- C08K5/3492—Triazines
- C08K5/34922—Melamine; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K9/00—Use of pretreated ingredients
- C08K9/04—Ingredients treated with organic substances
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K9/00—Use of pretreated ingredients
- C08K9/08—Ingredients agglomerated by treatment with a binding agent
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Manufacturing & Machinery (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Biotechnology (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Materials Engineering (AREA)
- Polymers & Plastics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Carbon And Carbon Compounds (AREA)
Abstract
The invention discloses a kind of preparation methods for being packed for soybean protein film, belong to field of packaging material.After the present invention first mixes soybean protein and water, is digested with endopeptidase, and adjust pH to acidity, obtain acid enzymolysis liquid；After graphene oxide and water are mixed dispersion again, kayexalate is added, graphene oxide dispersion is made；Then acidity enzymatic hydrolysis drop is added in graphene oxide dispersion, and is concentrated under reduced pressure, concentrate is made；Using rice husk as raw material, is mixed after micronization processes with nanometer iron powder, and carbonize and further heat up reaction through heating, carbonized material is made；After finally concentrate, reduced sugar, Tang's chemotherapy, melamine and preservative are stirred, message film forming, then through hot pressing, cooling, winding is to get soybean protein film is packed for.Gained of the invention is packed for soybean protein film with excellent mechanical property.
Description
Technical field
The invention discloses a kind of preparation methods for being packed for soybean protein film, belong to field of packaging material.
Background technique
In recent years, tight caused by the increasingly exhausted and modern chemical industry with non-renewable resources such as petroleum, coal, natural gases
Environmental pollution constantly aggravates again, and more sight are focused on a kind of environment amenable natural polymer by researchers
On.Soybean protein (SPI) as one of them, by various amino acid through peptide bond be connected form, have it is renewable, can biology drop
Solution, cheap, the advantages that biocompatibility is excellent, thus be widely studied.In addition, soybean protein is because of excellent film forming,
By the concern in the fields such as macromolecule, packaging material.Soybean protein is a kind of vegetable protein of super quality and competitive price, abundance,
Application potential is big.Different from polysaccharide natural polymer, there are a large amount of hydrogen bonds, ionic bond and hydrophobic in soybean protein molecule
Key, good filming performance and mechanical property can be had by making it that small molecule plasticizer only be added under conditions of being not required to modified.
But the disadvantages of that there are intensity is lower for pure soybean protein film, and water resistance is poor, and microbial stability is bad, being unable to satisfy makes
With requiring, synthesis high molecular material need to be added it is modified and prepare laminated film for using.
Currently, soybean protein isolate is increasingly becoming a kind of novel plastic-substitute.Have researcher using soybean, starch,
Potato is raw material, and the edible package film formula container succeeded in developing has entered mature commercialization stage.
In edibility antistaling film, soybean protein film is always the fresh-keeping material of a new generation that expert has an optimistic view of
Material.Because soybean protein has good film forming characteristics, oxygen resistance is good, and air penetrability is lower than zein film, and has good
Good elasticity, toughness and preferable tensile strength.Meanwhile it also has certain moisture resistance and antibiotic and sterilizing ability, is suitable for
Food packaging is able to maintain its superior flavor especially to the higher food containing grease.
Current ferulic acid is a kind of phenolic acid for being widely present in plant kingdom in the prior art, it can make protein and more
Crosslinking is generated between sugar, polysaccharide and polysaccharide, it is not only safe and non-toxic, but also there are many unique functions.Ferulic acid is used for edibility
The preparation of soybean protein film not only can be used as crosslinking agent, but also the effects of can play anti-oxidant and anti-corrosion.Gelatin adds water as plasticising
Fracture tensile strength, extensibility and the elongation at break of soybean protein can be improved in agent, and improves flexibility, bending strength, toughness
And impact strength, glass transition temperature, the applicability of expansion polymer at a lower temperature are reduced, is improved to various base-materials
Bonding, increase or decrease the sealing of film, improve greasy property and reduce friction, reduce electrostatic charging ability, improve table
Face gloss and appearance.
Mechanical property is bad in use for the traditional soybean protein film that is packed at present, and especially puncture can be compared with
Difference after leading to packaging, is easy the puncture by sharppointed article and influences the shelf life of interior packing article, thus also need to its into
Row research.
Summary of the invention
The present invention solves the technical problem of: for Traditional Packing soybean protein film mechanical property in use
Can be bad, especially puncture can be poor, after leading to packaging, is easy the puncture by sharppointed article and influences interior packing object
The drawbacks of shelf life of product, provides a kind of preparation method for being packed for soybean protein film.
In order to solve the above-mentioned technical problem, the technical scheme adopted by the invention is that:
(1) according to parts by weight, 40~60 portions of soybean proteins, 100~200 parts of water, 2~4 parts of endopeptidases, first by soybean are successively taken
Albumen is dispersed in water, and adds endopeptidase, cooling after constant temperature stirring digests, and adjusts pH to 4.0~4.2, obtains acid enzymatic hydrolysis
Liquid；
It (2) is in mass ratio after 1:10~1:20 is mixed by graphene oxide and water, ultrasonic disperse adds graphene oxide matter
The kayexalate of 0.1~0.2 times of amount, after being stirred to react, obtains graphene oxide dispersion；
(3) acid enzymolysis liquid is slowly added dropwise into graphene oxide dispersion, after being added dropwise, continues to be stirred to react, with
After be concentrated under reduced pressure, obtain concentrate；
(4) after rice husk being soaked in water, freezing squeezing is carried out, then through vacuum freeze drying, obtains a nanometer rice hull cellulose, then by gained
Nanometer rice hull cellulose and nanometer iron powder are after 100:1~150:1 is stirred in mass ratio, under inert gas shielding state,
Slowly heating charing, is further heated to 1500~1550 DEG C, after insulation reaction, and cooling obtains carbonized material；
(5) according to parts by weight, 80~100 portions of concentrates are successively taken, 10~15 parts of reduced sugars, 10~20 parts of carbonized materials, 4~8
Part melamine, 0.3~0.5 portion of preservative, after being uniformly mixed, casting film-forming, then through hot pressing, cooling, winding is to get packet
Dress uses soybean protein film.
Step (1) endopeptidase is trypsase, chymotrypsin, any one in elastoser.
Step (3) is described to be slowly added dropwise are as follows: is added dropwise with 3~5mL/min rate.
Step (3) described reduced pressure are as follows: in temperature be 75~85 DEG C, pressure be 400~420mmHg under the conditions of, decompression
45~60min is concentrated.
Step (4) inert gas is nitrogen, helium, any one in argon gas.
Step (4) the slow heating are as follows: be warming up to 480~500 DEG C with 0.6~0.8 DEG C/min rate program.
Step (5) reduced sugar is glucose, fructose, galactolipin, lactose, any one in maltose.
Step (5) preservative is potassium sorbate, sodium benzoate, in dehydroactic acid sodium, butyl p-hydroxybenzoate
Any one.
Step (2) described graphene oxide can also be pretreatment graphene oxide；The pretreatment graphene oxide
Preprocessing process are as follows: according to parts by weight, successively take 10~20 parts of graphene oxides, 60~80 parts of toluene di-isocyanate(TDI)s, 30
~50 parts of acetone, after being stirred, 3~5h of heating reflux reaction after washing and vacuum drying, obtains pretreatment oxygen using filter
Graphite alkene.
The chitosan of concentrate quality 3~5% can also be added in step (5) described concentrate；The chitosan is de- second
The chitosan that acyl degree is 90~95%.
The beneficial effects of the present invention are:
(1) technical solution of the present invention digests soybean protein first with endopeptidase, to make to possess complicated tertiary structure
Soybean protein be dissociated into linear polypeptide structure, and in acid condition, on the amino band in linear polypeptide structural molecule structure
Like charges and it is mutually exclusive, make linear polypeptide structure in aqueous solution can fine dispersion；
(2) technical solution of the present invention is handled the graphene oxide after ultrasound removing by using kayexalate,
Utilize the graphene area of the conjugation in the benzene ring structure and graphene oxide molecular structure in kayexalate molecular structure
It is adsorbed because of π-π interaction, while kayexalate can introduce sulfonic acid group in graphene-structured, to make
Graphene oxide structure has more negative electrical charges, because mutually exclusive with negative electrical charge of the same race, after effectively avoiding removing
Single-layer oxidized graphite alkene is reunited again；
(3) present invention is using occurring effectively absorption between linear polypeptide and single-layer oxidized graphite alkene with opposite charges, together
When, the polypeptide of linear structure is easy to wind on the corner angle or bulge-structure at single-layer oxidized graphite alkene edge and be affiliated to, to make
Mechanical snap is formed between the two, effectively improves binding force between the two, product mechanical property is enable effectively to be promoted, separately
Outside, due to the introducing of single-layer oxidized graphite alkene, puncture of the sharppointed article to packaging film is effectively prevented；And monolithic layer aoxidizes stone
It is again mutually discrete between black alkene, it has ensured the gas permeability of film, the shelf life of interior packing article is enable effectively to extend；In addition,
Under the action of melamine, as crosslinking agent, the protein after activating enzymatic hydrolysis crosslinks again, forms new cross-linked network
Network structure, and contained a small amount of nanometer iron powder can be changed into free metal ion under concentrate weak acid environment in carbonized material, and
It is chelated with polypeptide structure, to form ionomer, promotes packaging film internal crosslinking network density further, product
Mechanical property is further promoted；
(4) technical solution of the present invention makes rice hull cellulose by introducing carbonized material, and the heating rate in strict control carbonization process
In carbonization process, internal organic matter char shrinkage forms stress, and the keratinization silica of external package is as supporting layer,
It is shunk under stress and forms pleated structure, to promote carbonized material surface roughness, be conducive to linear cross-linked network structure
Winding be affiliated to, further improve the mechanical snap power between filler and cross-linked network structure, avoid two in loading process
Person occurs relative displacement and product mechanical property is caused to decline；
(5) technical solution of the present invention is by being aided with reduced sugar, and in hot pressing process, reduced sugar can be with active linear polypeptide
Between condensation reaction occur form schiff bases structure, and schiff bases structure and unstable, can also further occurrence complexity Mei La
Moral reaction, so that active linear polysaccharide structure be made finally to be changed into more stable compound, improves the stable storage of product
Property, performance permanently effective holding when enabling product as packaging film.
Specific embodiment
According to parts by weight, 10~20 parts of graphene oxides are successively taken, 60~80 parts of toluene di-isocyanate(TDI)s, 30~50 parts
Acetone, after being stirred, 3~5h of heating reflux reaction after washing and vacuum drying, obtains pretreatment graphite oxide using filter
Alkene；According to parts by weight, 40~60 portions of soybean proteins, 100~200 parts of water, 2~4 parts of endopeptidases, first by soybean protein are successively taken
It is poured into No. 1 beaker with water mixing, after being dispersed with stirring 20~30min with glass bar, adds endopeptidase, then No. 1 beaker is moved
Test the speed constant temperature blender with magnetic force to digital display, in temperature be 32~35 DEG C, under the conditions of revolving speed is 400~600r/min, constant temperature stirring
It is cooling after digesting 30~45min, and the hydrochloric acid for being 8~10% with mass fraction adjusts material pH to 4.0~4.2 in No. 1 beaker,
Obtain acid enzymolysis liquid；Graphene oxide will be pre-processed and water is that 1:10~1:20 mixing is poured into No. 2 beakers, and is incited somebody to action in mass ratio
No. 2 beakers move in ultrasonic disperse instrument, under the conditions of supersonic frequency is 45~50kHz after 20~40min of ultrasonic disperse, then to 2
Be added in number beaker and pre-process 0.1~0.2 times of graphene oxide quality of kayexalate, with blender with 200~
300r/min revolving speed is stirred to react 20~30min, obtains graphene oxide dispersion；Graphene oxide dispersion immigration band is stirred again
It mixes in the four-hole boiling flask of device, under the conditions of revolving speed is 300~500r/min, while stirring by dropping funel with 3~5mL/min
Acid enzymolysis liquid is added dropwise into four-hole boiling flask for rate, after adding to acidity enzymatic hydrolysis drop, continues to be stirred to react 20~30min,
Material in four-hole boiling flask is transferred in Rotary Evaporators again, in temperature be 75~85 DEG C, pressure be 400~420mmHg under the conditions of,
45~60min is concentrated under reduced pressure, obtains concentrate；It is in mass ratio after 1:5~1:10 mixes 2~3h of immersion, by rice by rice husk and water
Shell takes out, and uses 20~30s of liquid nitrogen frozen, then by the rice husk after freezing in pressure to squeeze under 2.7~3.2MPa, must press
Material is squeezed, then gained is squeezed into material vacuum freeze drying, obtains a nanometer rice hull cellulose；Then nanometer rice hull cellulose and nanometer iron powder are pressed
Mass ratio is after 100:1~150:1 is stirred, to be transferred in tube furnace, and be passed through with 200~300mL/min rate into furnace
Inert gas is warming up to 480~500 DEG C with 0.6~0.8 DEG C/min rate program, keeps the temperature charcoal under inert gas shielding state
After changing 2~3h, continue to further heat up with 8~10 DEG C/min rate to 1500~1550 DEG C, after 1~3h of insulation reaction, with furnace
It is cooled to room temperature, discharges, obtain carbonized material；According to parts by weight, 80~100 portions of concentrates are successively taken, 10~15 parts of reduced sugars, 10
~20 parts of carbonized materials, 4~8 parts of melamines, 0.3~0.5 portion of preservative, and the deacetylation of concentrate quality 3~5% is added
For 90~95% chitosan, after being stirred 45~60min with blender with 600~800r/min revolving speed, standing and defoaming is obtained
It is transferred in casting machine hopper at film liquid, then by gained at film liquid, control scraper knife saw is 2~3mm, after casting film-forming, in pressure
For 8~10MPa, under the conditions of temperature is 95~100 DEG C, 10~20min of hot pressing, then through cooling, wind to get soybean egg is packed for
Tunica albuginea.The endopeptidase is trypsase, chymotrypsin, any one in elastoser.The inert gas be nitrogen,
Any one in helium, argon gas.The reduced sugar is glucose, fructose, galactolipin, lactose, any one in maltose.
The preservative is potassium sorbate, sodium benzoate, any one in dehydroactic acid sodium, butyl p-hydroxybenzoate.
Example 1
According to parts by weight, 10~20 parts of graphene oxides are successively taken, 60~80 parts of toluene di-isocyanate(TDI)s, 30~50 part third
Ketone, after being stirred, 3~5h of heating reflux reaction after washing and vacuum drying, obtains pretreatment graphene oxide using filter；
According to parts by weight, 60 portions of soybean proteins, 200 parts of water are successively taken, soybean protein and water are first mixed and pour into No. 1 by 4 parts of endopeptidases
In beaker, after being dispersed with stirring 30min with glass bar, endopeptidase is added, then No. 1 beaker is moved into the digital display constant temperature magnetic force that tests the speed and is stirred
Mix device, in temperature be 35 DEG C, it is cooling after constant temperature stirring enzymatic hydrolysis 45min under the conditions of revolving speed is 600r/min, and use mass fraction
Material pH to 4.2 in No. 1 beaker is adjusted for 10% hydrochloric acid, obtains acid enzymolysis liquid；Graphene oxide will be pre-processed and water presses quality
Than being poured into No. 2 beakers for 1:20 mixing, and No. 2 beakers are moved in ultrasonic disperse instrument, under the conditions of supersonic frequency is 50kHz
It after ultrasonic disperse 40min, then is added into No. 2 beakers and pre-processes 0.2 times of graphene oxide quality of kayexalate, used
Blender is stirred to react 30min with 300r/min revolving speed, obtains graphene oxide dispersion；Graphene oxide dispersion is moved into again
In the four-hole boiling flask of belt stirrer, under the conditions of revolving speed is 500r/min, while stirring by dropping funel with 5mL/min rate
Acid enzymolysis liquid is added dropwise into four-hole boiling flask, after adding to acidity enzymatic hydrolysis drop, continues to be stirred to react 30min, then by four mouthfuls
Material is transferred in Rotary Evaporators in flask, is 85 DEG C in temperature, under the conditions of pressure is 420mmHg, 60min is concentrated under reduced pressure, obtains
Concentrate；It is in mass ratio after 1:10 is mixed and impregnated 3h, rice husk to be taken out, and with liquid nitrogen frozen 30s, then will be cold by rice husk and water
Rice husk after jelly, to be squeezed under 3.2MPa, must squeeze material, then gained is squeezed material vacuum freeze drying, obtain nanometer in pressure
Rice hull cellulose；It is in mass ratio then after 150:1 is stirred, to be transferred in tube furnace by nanometer rice hull cellulose and nanometer iron powder,
And inert gas is passed through into furnace with 300mL/min rate, under inert gas shielding state, with 0.8 DEG C/min rate program
It is warming up to 500 DEG C, after heat preservation charing 3h, continues to further heat up with 10 DEG C/min rate to 1550 DEG C, after insulation reaction 3h, with
Furnace is cooled to room temperature, and discharging obtains carbonized material；According to parts by weight, 100 portions of concentrates, 15 parts of reduced sugars, 20 parts of charings are successively taken
Material, 8 parts of melamines, 0.5 portion of preservative, and the chitosan that the deacetylation that concentrate quality 5% is added is 90%, use blender
After being stirred 60min with 800r/min revolving speed, standing and defoaming obtains into film liquid, then gained is transferred to casting machine hopper at film liquid
In, control scraper knife saw is 3mm, after casting film-forming, in pressure be 10MPa, under the conditions of temperature is 100 DEG C, hot pressing 20min, then
Through cooling, wind to get soybean protein film is packed for.The endopeptidase is trypsase.The inert gas is nitrogen.It is described
Reduced sugar is glucose.The preservative is potassium sorbate.
Example 2
According to parts by weight, 60 portions of soybean proteins, 200 parts of water are successively taken, soybean protein and water first are mixed by 4 parts of endopeptidases
Enter in No. 1 beaker, after being dispersed with stirring 30min with glass bar, add endopeptidase, then No. 1 beaker is moved into digital display and is tested the speed constant temperature
Magnetic stirring apparatus is 35 DEG C in temperature, and under the conditions of revolving speed is 600r/min, after constant temperature stirring enzymatic hydrolysis 45min, cooling must be digested
Liquid；It is in mass ratio that 1:20 mixing is poured into No. 2 beakers, and No. 2 beakers are moved to ultrasonic disperse instrument by graphene oxide and water
In, under the conditions of supersonic frequency is 50kHz after ultrasonic disperse 40min, then graphene oxide quality 0.2 is added into No. 2 beakers
Kayexalate again, is stirred to react 30min with blender with 300r/min revolving speed, obtains graphene oxide dispersion；Again
Graphene oxide dispersion is moved into the four-hole boiling flask of belt stirrer, under the conditions of revolving speed is 500r/min, is led to while stirring
It crosses dropping funel and enzymolysis liquid is added dropwise into four-hole boiling flask with 5mL/min rate, after enzymolysis liquid is added dropwise, continue to be stirred to react
30min, then material in four-hole boiling flask is transferred in Rotary Evaporators, it is 85 DEG C in temperature, under the conditions of pressure is 420mmHg, subtracts
Pressure concentration 60min, obtains concentrate；It is in mass ratio after 1:10 mixes immersion 3h, rice husk to be taken out, and use liquid nitrogen by rice husk and water
30s is freezed, then by the rice husk after freezing in pressure is to be squeezed under 3.2MPa, must squeeze material, then vacuum is expected into gained squeezing
Freeze-drying, obtains a nanometer rice hull cellulose；Then nanometer rice hull cellulose and nanometer iron powder are stirred in mass ratio for 150:1
Afterwards, it is transferred in tube furnace, and inert gas is passed through into furnace with 300mL/min rate, under inert gas shielding state, with
0.8 DEG C/min rate program is warming up to 500 DEG C, after heat preservation carbonizes 3h, and continuation is further heated up with 10 DEG C/min rate to 1550
DEG C, it after insulation reaction 3h, cools to room temperature with the furnace, discharges, obtain carbonized material；According to parts by weight, 100 portions of concentrates are successively taken,
15 parts of reduced sugars, 20 parts of carbonized materials, 8 parts of melamines, 0.5 portion of preservative, and the deacetylation that concentrate quality 5% is added is
90% chitosan, after being stirred 60min with blender with 800r/min revolving speed, standing and defoaming obtains into film liquid, then by gained
It is transferred in casting machine hopper at film liquid, control scraper knife saw is 3mm, is 10MPa in pressure, temperature is 100 DEG C after casting film-forming
Under the conditions of, hot pressing 20min, then through cooling, wind to get soybean protein film is packed for.The endopeptidase is trypsase.It is described
Inert gas is nitrogen.The reduced sugar is glucose.The preservative is potassium sorbate.
Example 3
According to parts by weight, 60 portions of soybean proteins, 200 parts of water are successively taken, soybean protein and water first are mixed by 4 parts of endopeptidases
Enter in No. 1 beaker, after being dispersed with stirring 30min with glass bar, add endopeptidase, then No. 1 beaker is moved into digital display and is tested the speed constant temperature
Magnetic stirring apparatus, in temperature be 35 DEG C, it is cooling after constant temperature stirring enzymatic hydrolysis 45min under the conditions of revolving speed is 600r/min, and use matter
It measures the hydrochloric acid that score is 10% and adjusts material pH to 4.2 in No. 1 beaker, obtain acid enzymolysis liquid；Graphene oxide and water are pressed into quality
Than being poured into No. 2 beakers for 1:20 mixing, and No. 2 beakers are moved in ultrasonic disperse instrument, under the conditions of supersonic frequency is 50kHz
After ultrasonic disperse 40min, graphene oxide dispersion is obtained；Graphene oxide dispersion is moved into the four-hole boiling flask of belt stirrer again
In, under the conditions of revolving speed is 500r/min, acid is added dropwise with 5mL/min rate into four-hole boiling flask by dropping funel while stirring
Property enzymolysis liquid, to acidity enzymatic hydrolysis drop add after, continue to be stirred to react 30min, then material in four-hole boiling flask is transferred to rotation
In evaporimeter, it is 85 DEG C in temperature, under the conditions of pressure is 420mmHg, 60min is concentrated under reduced pressure, obtains concentrate；Rice husk and water are pressed
Mass ratio is after 3h is impregnated in 1:10 mixing, rice husk to be taken out, and with liquid nitrogen frozen 30s, then be in pressure by the rice husk after freezing
It is squeezed under 3.2MPa, material must be squeezed, then gained is squeezed into material vacuum freeze drying, obtain a nanometer rice hull cellulose；It will then receive
Rice rice hull cellulose and nanometer iron powder are after 150:1 is stirred, to be transferred in tube furnace in mass ratio, and with 300mL/min rate
It is passed through inert gas into furnace, under inert gas shielding state, is warming up to 500 DEG C with 0.8 DEG C/min rate program, keeps the temperature charcoal
After changing 3h, continues to further heat up with 10 DEG C/min rate to 1550 DEG C, after insulation reaction 3h, cools to room temperature with the furnace, discharge,
Obtain carbonized material；According to parts by weight, 100 portions of concentrates are successively taken, 15 parts of reduced sugars, 20 parts of carbonized materials, 8 parts of melamines, 0.5
Part preservative, after being stirred 60min with blender with 800r/min revolving speed, standing and defoaming obtains into film liquid, then gained is formed a film
Liquid is transferred in casting machine hopper, and control scraper knife saw is 3mm, is 10MPa in pressure, temperature is 100 DEG C of conditions after casting film-forming
Under, hot pressing 20min, then through cooling, wind to get soybean protein film is packed for.The endopeptidase is trypsase.The inertia
Gas is nitrogen.The reduced sugar is glucose.The preservative is potassium sorbate.
Example 4
According to parts by weight, 60 portions of soybean proteins, 200 parts of water are successively taken, soybean protein and water first are mixed by 4 parts of endopeptidases
Enter in No. 1 beaker, after being dispersed with stirring 30min with glass bar, add endopeptidase, then No. 1 beaker is moved into digital display and is tested the speed constant temperature
Magnetic stirring apparatus, in temperature be 35 DEG C, it is cooling after constant temperature stirring enzymatic hydrolysis 45min under the conditions of revolving speed is 600r/min, and use matter
It measures the hydrochloric acid that score is 10% and adjusts material pH to 4.2 in No. 1 beaker, obtain acid enzymolysis liquid；Graphene oxide and water are pressed into quality
Than being poured into No. 2 beakers for 1:20 mixing, and No. 2 beakers are moved in ultrasonic disperse instrument, under the conditions of supersonic frequency is 50kHz
After ultrasonic disperse 40min, then 0.2 times of graphene oxide quality of kayexalate is added into No. 2 beakers, uses blender
It is stirred to react 30min with 300r/min revolving speed, obtains graphene oxide dispersion；Graphene oxide dispersion is moved into band stirring again
In the four-hole boiling flask of device, under the conditions of revolving speed is 500r/min, pass through dropping funel while stirring with 5mL/min rate to four mouthfuls
Acid enzymolysis liquid is added dropwise in flask, after adding to acidity enzymatic hydrolysis drop, continues to be stirred to react 30min, then will be in four-hole boiling flask
Material is transferred in Rotary Evaporators, is 85 DEG C in temperature, under the conditions of pressure is 420mmHg, 60min is concentrated under reduced pressure, obtains concentrate；
According to parts by weight, successively take 100 portions of concentrates, 15 parts of reduced sugars, 8 parts of melamines, 0.5 portion of preservative, with blender with
After 800r/min revolving speed is stirred 60min, standing and defoaming obtains into film liquid, then gained is transferred in casting machine hopper at film liquid,
Control scraper knife saw is 3mm, after casting film-forming, in pressure be 10MPa, under the conditions of temperature is 100 DEG C, hot pressing 20min, then through cold
But, winding is to get soybean protein film is packed for.The endopeptidase is trypsase.The inert gas is nitrogen.The reduction
Sugar is glucose.The preservative is potassium sorbate.
Example 5
According to parts by weight, 60 portions of soybean proteins, 200 parts of water are successively taken, soybean protein and water first are mixed by 4 parts of endopeptidases
Enter in No. 1 beaker, after being dispersed with stirring 30min with glass bar, add endopeptidase, then No. 1 beaker is moved into digital display and is tested the speed constant temperature
Magnetic stirring apparatus, in temperature be 35 DEG C, it is cooling after constant temperature stirring enzymatic hydrolysis 45min under the conditions of revolving speed is 600r/min, and use matter
It measures the hydrochloric acid that score is 10% and adjusts material pH to 4.2 in No. 1 beaker, obtain acid enzymolysis liquid；Graphene oxide and water are pressed into quality
Than being poured into No. 2 beakers for 1:20 mixing, and No. 2 beakers are moved in ultrasonic disperse instrument, under the conditions of supersonic frequency is 50kHz
After ultrasonic disperse 40min, then 0.2 times of graphene oxide quality of kayexalate is added into No. 2 beakers, uses blender
It is stirred to react 30min with 300r/min revolving speed, obtains graphene oxide dispersion；Graphene oxide dispersion is moved into band stirring again
In the four-hole boiling flask of device, under the conditions of revolving speed is 500r/min, pass through dropping funel while stirring with 5mL/min rate to four mouthfuls
Acid enzymolysis liquid is added dropwise in flask, after adding to acidity enzymatic hydrolysis drop, continues to be stirred to react 30min, then will be in four-hole boiling flask
Material is transferred in Rotary Evaporators, is 85 DEG C in temperature, under the conditions of pressure is 420mmHg, 60min is concentrated under reduced pressure, obtains concentrate；
It is in mass ratio after 1:10 is mixed and impregnated 3h, rice husk to be taken out, and with liquid nitrogen frozen 30s, then will be after freezing by rice husk and water
Rice husk is to be squeezed under 3.2MPa in pressure, must squeeze material, then gained is squeezed material vacuum freeze drying, obtain a nanometer rice husk fibre
Dimension；Then be in mass ratio by nanometer rice hull cellulose and nanometer iron powder after 150:1 is stirred, be transferred in tube furnace, and with
300mL/min rate is passed through inert gas into furnace, under inert gas shielding state, with the heating of 0.8 DEG C/min rate program
To 500 DEG C, after heat preservation charing 3h, continue to further heat up with 10 DEG C/min rate to 1550 DEG C, it is cold with furnace after insulation reaction 3h
But to room temperature, discharging obtains carbonized material；According to parts by weight, 100 portions of concentrates are successively taken, 20 parts of carbonized materials, 8 parts of melamines,
0.5 portion of preservative, after being stirred 60min with blender with 800r/min revolving speed, standing and defoaming obtains into film liquid, then by gained
It is transferred in casting machine hopper at film liquid, control scraper knife saw is 3mm, is 10MPa in pressure, temperature is 100 DEG C after casting film-forming
Under the conditions of, hot pressing 20min, then through cooling, wind to get soybean protein film is packed for.The endopeptidase is trypsase.It is described
Inert gas is nitrogen.The preservative is potassium sorbate.
Example 6
According to parts by weight, 60 portions of soybean proteins, 200 parts of water are successively taken, soybean protein and water first are mixed by 4 parts of endopeptidases
Enter in No. 1 beaker, after being dispersed with stirring 30min with glass bar, add endopeptidase, then No. 1 beaker is moved into digital display and is tested the speed constant temperature
Magnetic stirring apparatus, in temperature be 35 DEG C, it is cooling after constant temperature stirring enzymatic hydrolysis 45min under the conditions of revolving speed is 600r/min, and use matter
It measures the hydrochloric acid that score is 10% and adjusts material pH to 4.2 in No. 1 beaker, obtain acid enzymolysis liquid；Graphene oxide and water are pressed into quality
Than being poured into No. 2 beakers for 1:20 mixing, and No. 2 beakers are moved in ultrasonic disperse instrument, under the conditions of supersonic frequency is 50kHz
After ultrasonic disperse 40min, then 0.2 times of graphene oxide quality of kayexalate is added into No. 2 beakers, uses blender
It is stirred to react 30min with 300r/min revolving speed, obtains graphene oxide dispersion；Graphene oxide dispersion is moved into band stirring again
In the four-hole boiling flask of device, under the conditions of revolving speed is 500r/min, pass through dropping funel while stirring with 5mL/min rate to four mouthfuls
Acid enzymolysis liquid is added dropwise in flask, after adding to acidity enzymatic hydrolysis drop, continues to be stirred to react 30min, then will be in four-hole boiling flask
Material is transferred in Rotary Evaporators, is 85 DEG C in temperature, under the conditions of pressure is 420mmHg, 60min is concentrated under reduced pressure, obtains concentrate；
It is in mass ratio after 1:10 is mixed and impregnated 3h, rice husk to be taken out, and with liquid nitrogen frozen 30s, then will be after freezing by rice husk and water
Rice husk is to be squeezed under 3.2MPa in pressure, must squeeze material, then gained is squeezed material vacuum freeze drying, obtain a nanometer rice husk fibre
Dimension；Then be in mass ratio by nanometer rice hull cellulose and nanometer iron powder after 150:1 is stirred, be transferred in tube furnace, and with
300mL/min rate is passed through inert gas into furnace, under inert gas shielding state, with the heating of 0.8 DEG C/min rate program
To 500 DEG C, after heat preservation charing 3h, continue to further heat up with 10 DEG C/min rate to 1550 DEG C, it is cold with furnace after insulation reaction 3h
But to room temperature, discharging obtains carbonized material；According to parts by weight, 100 portions of concentrates are successively taken, 15 parts of reduced sugars, 20 parts of carbonized materials,
0.5 portion of preservative, after being stirred 60min with blender with 800r/min revolving speed, standing and defoaming obtains into film liquid, then by gained
It is transferred in casting machine hopper at film liquid, control scraper knife saw is 3mm, is 10MPa in pressure, temperature is 100 DEG C after casting film-forming
Under the conditions of, hot pressing 20min, then through cooling, wind to get soybean protein film is packed for.The endopeptidase is trypsase.It is described
Inert gas is nitrogen.The reduced sugar is glucose.The preservative is potassium sorbate.
Comparative example: the production of Shanghai packaging material Co., Ltd is packed for soybean protein film.
1 to 6 gained of example is packed for soybean protein film and comparative example product carries out performance detection, specific detection method is such as
Under:
Sample is made to the sample of 80mm × 80mm, selects the calm and peaceful testing machine Co., Ltd BCJ-3A plastic film punching in Jilin Province
It hits testing machine and carries out confrontation pendulum impact energy test experiments.Specific testing result is as shown in table 1:
Table 1: performance detection table
| Detection content | Example 1 | Example 2 | Example 3 | Example 4 | Example 5 | Example 6 | Comparative example |
| Impact strength/J | 0.70 | 0.52 | 0.45 | 0.48 | 0.49 | 0.51 | 0.21 |
| Tensile strength/MPa | 6.91 | 4.86 | 4.16 | 4.24 | 4.50 | 4.73 | 2.42 |
| Elongation at break/% | 72.25 | 64.62 | 60.48 | 63.84 | 61.04 | 62.47 | 53.74 |
By 1 testing result of table it is found that present invention gained is packed for soybean protein film with excellent mechanical property.
Claims (10)
1. a kind of preparation method for being packed for soybean protein film, it is characterised in that specific preparation step are as follows:
(1) according to parts by weight, 40~60 portions of soybean proteins, 100~200 parts of water, 2~4 parts of endopeptidases, first by soybean are successively taken
Albumen is dispersed in water, and adds endopeptidase, cooling after constant temperature stirring digests, and adjusts pH to 4.0~4.2, obtains acid enzymatic hydrolysis
Liquid；
It (2) is in mass ratio after 1:10~1:20 is mixed by graphene oxide and water, ultrasonic disperse adds graphene oxide matter
The kayexalate of 0.1~0.2 times of amount, after being stirred to react, obtains graphene oxide dispersion；
(3) acid enzymolysis liquid is slowly added dropwise into graphene oxide dispersion, after being added dropwise, continues to be stirred to react, with
After be concentrated under reduced pressure, obtain concentrate；
(4) after rice husk being soaked in water, freezing squeezing is carried out, then through vacuum freeze drying, obtains a nanometer rice hull cellulose, then by gained
Nanometer rice hull cellulose and nanometer iron powder are after 100:1~150:1 is stirred in mass ratio, under inert gas shielding state,
Slowly heating charing, is further heated to 1500~1550 DEG C, after insulation reaction, and cooling obtains carbonized material；
(5) according to parts by weight, 80~100 portions of concentrates are successively taken, 10~15 parts of reduced sugars, 10~20 parts of carbonized materials, 4~8
Part melamine, 0.3~0.5 portion of preservative, after being uniformly mixed, casting film-forming, then through hot pressing, cooling, winding is to get packet
Dress uses soybean protein film.
2. a kind of preparation method for being packed for soybean protein film according to claim 1, it is characterised in that step (1) is described
Endopeptidase is trypsase, chymotrypsin, any one in elastoser.
3. a kind of preparation method for being packed for soybean protein film according to claim 1, it is characterised in that step (3) is described
It is slowly added dropwise are as follows: be added dropwise with 3~5mL/min rate.
4. a kind of preparation method for being packed for soybean protein film according to claim 1, it is characterised in that step (3) is described
Be concentrated under reduced pressure are as follows: in temperature be 75~85 DEG C, pressure be 400~420mmHg under the conditions of, be concentrated under reduced pressure 45~60min.
5. a kind of preparation method for being packed for soybean protein film according to claim 1, it is characterised in that step (4) is described
Inert gas is nitrogen, helium, any one in argon gas.
6. a kind of preparation method for being packed for soybean protein film according to claim 1, it is characterised in that step (4) is described
Slowly heating are as follows: be warming up to 480~500 DEG C with 0.6~0.8 DEG C/min rate program.
7. a kind of preparation method for being packed for soybean protein film according to claim 1, it is characterised in that step (5) is described
Reduced sugar is glucose, fructose, galactolipin, lactose, any one in maltose.
8. a kind of preparation method for being packed for soybean protein film according to claim 1, it is characterised in that step (5) is described
Preservative is potassium sorbate, sodium benzoate, any one in dehydroactic acid sodium, butyl p-hydroxybenzoate.
9. a kind of preparation method for being packed for soybean protein film according to claim 1, it is characterised in that step (2) is described
Graphene oxide can also be pretreatment graphene oxide；The preprocessing process of the pretreatment graphene oxide are as follows: by weight
Number meter, successively takes 10~20 parts of graphene oxides, 60~80 parts of toluene di-isocyanate(TDI)s, and 30~50 parts of acetone are stirred
Afterwards, 3~5h of heating reflux reaction after washing and vacuum drying, obtains pretreatment graphene oxide using filter.
10. a kind of preparation method for being packed for soybean protein film according to claim 1, it is characterised in that step (5) institute
State the chitosan that concentrate quality 3~5% can also be added in concentrate；The chitosan is the shell that deacetylation is 90~95%
Glycan.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910463027.7A CN110204752A (en) | 2019-05-30 | 2019-05-30 | A kind of preparation method being packed for soybean protein film |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910463027.7A CN110204752A (en) | 2019-05-30 | 2019-05-30 | A kind of preparation method being packed for soybean protein film |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110204752A true CN110204752A (en) | 2019-09-06 |
Family
ID=67789618
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910463027.7A Pending CN110204752A (en) | 2019-05-30 | 2019-05-30 | A kind of preparation method being packed for soybean protein film |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110204752A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103183837A (en) * | 2012-12-14 | 2013-07-03 | 江南大学 | Preparation method of heat resistant soy protein/graphene oxide composite membrane |
| US20180105659A1 (en) * | 2015-04-23 | 2018-04-19 | Rowan University | Processes of Increasing Crystallinity Alignment of Protein Films and Products Thereof |
| CN108624285A (en) * | 2018-05-30 | 2018-10-09 | 王成 | A kind of preparation method of water-resistant type soy protein adhesives |
| CN108948389A (en) * | 2018-06-26 | 2018-12-07 | 常州五荣化工有限公司 | A kind of preparation method of gelatin-graphene oxide composite membrane |
| CN109817992A (en) * | 2019-01-12 | 2019-05-28 | 袁玲燕 | A kind of preparation method of the dedicated redox elctro-catalyst of magnesium air fuel cell |
-
2019
- 2019-05-30 CN CN201910463027.7A patent/CN110204752A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103183837A (en) * | 2012-12-14 | 2013-07-03 | 江南大学 | Preparation method of heat resistant soy protein/graphene oxide composite membrane |
| US20180105659A1 (en) * | 2015-04-23 | 2018-04-19 | Rowan University | Processes of Increasing Crystallinity Alignment of Protein Films and Products Thereof |
| CN108624285A (en) * | 2018-05-30 | 2018-10-09 | 王成 | A kind of preparation method of water-resistant type soy protein adhesives |
| CN108948389A (en) * | 2018-06-26 | 2018-12-07 | 常州五荣化工有限公司 | A kind of preparation method of gelatin-graphene oxide composite membrane |
| CN109817992A (en) * | 2019-01-12 | 2019-05-28 | 袁玲燕 | A kind of preparation method of the dedicated redox elctro-catalyst of magnesium air fuel cell |
Non-Patent Citations (1)
| Title |
|---|
| YUFEI HAN等: "Properties of Soy Protein Isolate Biopolymer Film Modified by Graphene", 《PLOYMERS》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chang et al. | Lime pretreatment of switchgrass | |
| Moniruzzaman et al. | Enzymatic hydrolysis of high-moisture corn fiber pretreated by AFEX and recovery and recycling of the enzyme complex | |
| CN111875908B (en) | Self-healing aerogel | |
| CN105192723B (en) | A kind of maca dietary fiber and its preparation method and application | |
| CN105670000B (en) | A kind of method for preparing hydrophobic protein | |
| CN110512109A (en) | A kind of preparation method of graphene enhancing titanium composite material | |
| CN108559564A (en) | A kind of preparation method of water repellent bio matter briquette binder | |
| CN110204752A (en) | A kind of preparation method being packed for soybean protein film | |
| CN113140717B (en) | Preparation method of sodium alginate-based double-network carbon aerogel negative electrode material for lithium ion battery | |
| CN109971123A (en) | A kind of preparation method of epoxy-boron nitride composite | |
| CN111303492B (en) | Waterproof ultra-light plant fiber composite material applied to degradable dinner plate and preparation method thereof | |
| CN115322417B (en) | Preparation method of high-modulus high-heat-resistance polyarylether nitrile self-reinforced film | |
| CN115819830A (en) | Preparation method of biomass-based nanofiber and MXene composite aerogel | |
| CN116426032A (en) | Bagasse cellulose aerogel and preparation method thereof | |
| CN110105523A (en) | Fireproof heated board polyurethane foam and its processing technology based on biomass material | |
| CN113501996B (en) | Flexible cellulose-based conductive composite film and preparation method and application thereof | |
| CN109880369A (en) | A kind of nano-TiO2Composite material and preparation method | |
| CN114874504A (en) | Light and heat-insulating polysaccharide-based composite aerogel taking wood waste as raw material and preparation method thereof | |
| CN108517233A (en) | A kind of composite coal binder | |
| CN109329755A (en) | A kind of production method of new modified bean dregs dietary fiber-albumen hypoglycemic formula powder | |
| CN1120074A (en) | Method of production for direct reduction furnace charge by cold-consolidation briquetting method | |
| CN108948389A (en) | A kind of preparation method of gelatin-graphene oxide composite membrane | |
| CN101357971B (en) | Organic solvent water absorbent and preparation method thereof | |
| CN101343418B (en) | Acylated soy protein biodegradable plastic and preparation thereof | |
| CN110584137B (en) | Method for preparing insoluble dietary fiber by using apple pomace |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |