CN105713194A - Method for purifying epsilon-PL (epsilon-polylysine) - Google Patents
Method for purifying epsilon-PL (epsilon-polylysine) Download PDFInfo
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- CN105713194A CN105713194A CN201610190931.1A CN201610190931A CN105713194A CN 105713194 A CN105713194 A CN 105713194A CN 201610190931 A CN201610190931 A CN 201610190931A CN 105713194 A CN105713194 A CN 105713194A
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G69/00—Macromolecular compounds obtained by reactions forming a carboxylic amide link in the main chain of the macromolecule
- C08G69/02—Polyamides derived from amino-carboxylic acids or from polyamines and polycarboxylic acids
- C08G69/08—Polyamides derived from amino-carboxylic acids or from polyamines and polycarboxylic acids derived from amino-carboxylic acids
- C08G69/10—Alpha-amino-carboxylic acids
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Abstract
The invention discloses a method for purifying epsilon-PL (epsilon-polylysine). The method comprises steps as follows: a crude epsilon-PL product with the mass concentration being 75% or above is added to deionized water, an epsilon-PL aqueous solution with the mass concentration being 30%-50% is prepared, an organic solvent with the final volume concentration being 1%-10% is added to the aqueous solution, the mixture is subjected to a heating reflux reaction for 2 h, then slowly cooled to subzero 5 DEG C, stirred for crystallization and filtered, a filter cake a is obtained and dried, and epsilon-PL is obtained. With the adoption of the method, high-purity epsilon-PL (with the purity higher than 99.5%) can be obtained, PL products lost in the technology can be recycled, and high-purity epsilon-PL solids can be obtained after vacuum drying, are convenient to store and transport and can be better applied to fields of cosmetics, medicines and the like.
Description
(1) technical field
The present invention relates to a kind of method purifying epsilon-polylysine.
(2) background technology
Within 1977, Japanese scholars S.Shima and H.Sakai is screening from microorganism
During Dragendo～Positive (being abbreviated as DP) material, find a strain actinomycetes No.346 energy
Produce a large amount of and stable DP materials, by the analysis of acid hydrolysis products and structural analysis, it was demonstrated that
This DP material is a kind of homotype monomer-polymer containing 25 30 lysine residues, referred to as ε-
Poly-D-lysine (ε-PL).
Epsilon-polylysine is a kind of polypeptide with bacteriostasis efficacy, and this biological preservative is in the eighties
With regard to first Application in food antiseptic.Epsilon-polylysine can be decomposed into lysine in human body, and lysine
It is one of 8 kinds of aminoacid of needed by human, is also that countries in the world allow the amino of strengthening in food
Acid.Therefore epsilon-polylysine is a kind of auxotype antibacterial, and safety is higher than other chemical preservatives,
Its acute oral toxicity is 5g/kg.Epsilon-polylysine antimicrobial spectrum is wide, for the sharp-pointed false silk of Saccharomyces
Yeast, method Rhodotorula sp, product film pichia yeast, Flos Rosae Rugosae shadow yeast；In gram positive bacteria
Heat-resisting Bacillus stearothermophilus, Bacillus coagulans, bacillus subtilis；Product in gram negative bacteria
Moral bacillus, escherichia coli etc. have significantly suppression and killing action.Polylysine is to gram sun
The micrococcus luteus of property, Lactobacillus bulgaricus, hot streptococcus, gram-negative escherichia coli, sramana
Salmonella and saccharomycetic growth have obvious inhibition, and polylysine and acetic acid complex reagent are to hay
Bacillus has obvious inhibiting effect.
The mechanism of action of epsilon-polylysine is mainly manifested in following 3 aspects:
(1) cell wall and cell membrane system are acted on；
(2) hereditary material or hereditary particle structure are acted on；
(3) enzyme or functional protein are acted on.The bacteriostasis property of polylysine is studied, finds
ε-PL not only can suppress the G that thermostability is stronger+Micrococcus luteus, and to other natural antiseptic agent (as
Nisin) G of suppression it is difficult to-Escherichia coli, Salmonella fungistatic effect the best, also simultaneously
Lactobacillus bulgaricus, streptococcus thermophilus, saccharomycetic growth can be suppressed.But it is single use ε-PL
Time inconspicuous to bacillus subtilis, aspergillus niger suppression, use ε-PL and acetic acid Combined Processing, right
Bacillus subtilis inhibitory action strengthens, and the ε-PL after high-temperature process still has antibacterial work to micrococcus luteus
Property.
At present, on market, major part polylysine product is presented in hydrochlorate, and alkaline form
Polylysine product less, but epsilon-polylysine is compared with hydrochlorate, has higher biological live
Property, but due to the limitation of traditional handicraft, the production of high-purity epsilon-polylysine is the most difficult, but
The further raising required along with additive, it is necessary to improve the purity of epsilon-polylysine, one can be entered
Step is widely used in food, cosmetics and field of medicaments.Therefore, preparation high-purity epsilon-polylysine
Can the most efficiently utilize the produced polylysine of fermentation, improve production efficiency, produce higher warp
Ji benefit.
(3) summary of the invention
It is an object of the present invention to provide a kind of method purifying epsilon-polylysine, can obtain purity 99% with
On epsilon-polylysine product, for medical treatment and cosmetic industry.
The technical solution used in the present invention is:
The present invention provides a kind of method purifying epsilon-polylysine, and described method is: by mass concentration
The epsilon-polylysine crude product of more than 75% joins in deionized water, is configured to mass concentration 30-50%
Epsilon-polylysine aqueous solution, in aqueous solution add volume final concentration 1-10% organic solvent, add
Hot reflux reaction 2h, slow cooling is to-5 DEG C subsequently, stirred crystallization, filters, it is thus achieved that filter cake a, will
Filter cake a is dried, it is thus achieved that epsilon-polylysine；Described organic solvent is in methanol, dehydrated alcohol or acetone
One.
Further, described cooling rate is 3-10 DEG C/h.
Further, the described stirred crystallization time is 48h.
Further, in described epsilon-polylysine crude product, epsilon-polylysine mass content is 75%-95%.
Further, described filter cake a ice absolute ethanol washing, take the filter cake b after washing and add anhydrous
Ethanol continues to be heated to reflux 2h, and then slow cooling is to-15 DEG C, stirred crystallization, filters, it is thus achieved that filter
Cake c, by filter cake c absolute ethanol washing 2-3 time, is dried, it is thus achieved that epsilon-polylysine.
Further, described filter cake a ice absolute ethanol washing 2-3 time.
Further, the dehydrated alcohol volumetric usage in described addition filter cake b is calculated as with filter cake b mass
10-20ml/g。
Further, the speed being cooled to-15 DEG C described in is 5-15 DEG C/h.
Further, described being dried is vacuum drying 24-48h at 30-50 DEG C.
Filter cake a of the present invention, filter cake b and filter cake c are filtration gained filter cake, in order to distinguish not
Naming with the filter cake of step acquisition, letter itself does not has implication.
The beneficial effects are mainly as follows: employing the inventive method, available high-purity ε-
Polylysine (higher than 99.5%), the equal recoverable of polylysine product lost in the process,
After vacuum dried, available high-purity epsilon-polylysine solid, it is simple to store and transport, more may be used
It is applied to cosmetics and medicine and other fields.
(4) accompanying drawing explanation
Fig. 1 epsilon-polylysine standard substance chromatogram.
The chromatogram of the epsilon-polylysine product of Fig. 2 embodiment 27 preparation.
(5) detailed description of the invention
Below in conjunction with specific embodiment, the present invention is described further, but protection scope of the present invention
It is not limited to that:
Embodiment 1:
By 100g epsilon-polylysine crude product (80%) (purchased from Zhejiang Silver Elephant Bioengineering Co., Ltd.)
It is dissolved in deionized water the aqueous solution being configured to mass concentration 30%, is heated to reflux 2h, make ε-poly-
Lysine fully dissolves, and slow cooling is to-5 DEG C (5 DEG C/h) subsequently, and stirred crystallization 48h, without crystal
Separate out.
Embodiment 2:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 50%, is heated to reflux 2h, makes epsilon-polylysine fully dissolve, and slow cooling is extremely subsequently
-5 DEG C (5 DEG C/h), stirred crystallization 48h, separate out without crystal.
Embodiment 3:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the absolute methanol of volume final concentration 10%, continues heating
Backflow 2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirring knot subsequently
Brilliant 48h, solution turned cloudy, obtain a small amount of solid.
Embodiment 4:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the acetone of volume final concentration 10%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, filters, obtains solid, and 35 DEG C of vacuum drying 24h obtain final products, calculate epsilon-polylysine
Absolute yield is 78%, and product purity is 93%.
Embodiment 5:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the ethanol of volume final concentration 10%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, filters, obtains solid, and 35 DEG C of vacuum drying 24h obtain final products, and epsilon-polylysine is absolute
Yield is 82%, and product purity is 92%.
Embodiment 6:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The solution of 25%, in this solution, adds the ethanol of volume final concentration 10%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, filters, obtains solid, and 35 DEG C of vacuum drying 24h obtain final products, calculate polylysine exhausted
Being 80% to yield, product purity is 95%.
Embodiment 7:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The solution of 15%, in this solution, adds the ethanol of volume final concentration 10%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, filters, obtains solid, and 35 DEG C of vacuum drying 24h obtain final products, and epsilon-polylysine is absolute
Yield is 62%, and product purity is 96%.
Embodiment 8:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The solution of 30%, in this solution, adds the ethanol of volume final concentration 1%, continues to be heated to reflux 2h,
Making epsilon-polylysine fully dissolve, slow cooling is extremely-5 DEG C (5 DEG C/h) subsequently, stirred crystallization 48h,
Filtering, obtain solid, 35 DEG C of vacuum drying 24h obtain final products, and epsilon-polylysine is definitely received
Rate is 34%, and product purity is 95%.
Embodiment 9:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the ethanol of volume final concentration 3%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, filters, obtains solid, and 35 DEG C of vacuum drying 24h obtain final products, and epsilon-polylysine is absolute
Yield is 74%, and product purity is 96%.
Embodiment 10:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the ethanol of volume final concentration 6%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, filters, obtains solid, and 35 DEG C of vacuum drying 24h obtain final products, and epsilon-polylysine is absolute
Yield is 78%, and product purity is 96%.
Embodiment 11:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the ethanol of volume final concentration 8%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, obtains solid, and 35 DEG C of vacuum drying 24h obtain final products, and epsilon-polylysine absolute yield is
78%, product purity is 94%.
Embodiment 12:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The solution of 30%, in this solution, adds the ethanol of volume final concentration 6%, continues to be heated to reflux 2h,
Making epsilon-polylysine fully dissolve, slow cooling is extremely-5 DEG C (10 DEG C/h) subsequently, stirred crystallization 48h,
Filtering, obtain solid, 35 DEG C of vacuum drying 24h obtain final products, calculate epsilon-polylysine exhausted
Being 68% to yield, product purity is 90%.
Embodiment 13:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the ethanol of volume final concentration 6%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (3 DEG C/h), stirred crystallization 48 subsequently
H, obtains solid, and 35 DEG C of vacuum drying 24h obtain final products, and epsilon-polylysine absolute yield is
80%, product purity is 95%.
Embodiment 14:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The solution of 40%, in this solution, adds the ethanol of volume final concentration 6%, continues to be heated to reflux 2h,
Making epsilon-polylysine fully dissolve, slow cooling is extremely-5 DEG C (5 DEG C/h) subsequently, stirred crystallization 48h,
Obtaining solid, 35 DEG C of vacuum drying 24h obtain final products, and epsilon-polylysine absolute yield is 86%,
Product purity is 94%.
Embodiment 15:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the ethanol of volume final concentration 6%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, is subsequently filtered out solid, add take 50mL ice dehydrated alcohol divide 2-3 time wash this solid, by gained
The vacuum drying of wet solid fully removes moisture.Subsequently, take above-mentioned gained solid 20g, add 400mL
Dehydrated alcohol, is heated to reflux 2h so that it is fully dissolve, and then slow cooling is to-15 DEG C (5 DEG C/h),
Stirred crystallization 36h, leaches gained crystal, and with and washing with alcohol 2-3 time, 35 DEG C of vacuum drying
24h obtains final products, epsilon-polylysine product purity 99.7%, yield 66%.
Embodiment 16:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the ethanol of volume final concentration 6%, continues to be heated to reflux,
Making polylysine fully dissolve, slow cooling is extremely-5 DEG C (5 DEG C/h) subsequently, stirred crystallization 48h, with
After leach solid, add take 50mL ice dehydrated alcohol divide 2-3 time wash this solid, by wet for gained solid
Vacuum drying fully removes moisture.Subsequently, take above-mentioned gained solid 20g, add 400mL volume
Concentration 95% ethanol water, is heated to reflux 2h so that it is fully dissolve, and then slow cooling is extremely
-15 DEG C (5 DEG C/h), stirred crystallization 36h, gained crystal is leached, and with and washing with alcohol 2-3
Secondary, 35 DEG C of vacuum drying 24h obtain final products, product purity 98.2%, yield 52%.
Embodiment 17:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the ethanol of volume final concentration 6%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, is subsequently filtered out solid, add take 50mL ice dehydrated alcohol divide 2-3 time wash this solid, by gained
The vacuum drying of wet solid fully removes moisture.Subsequently, take above-mentioned gained solid 20g, add 400mL
Volumetric concentration 75% ethanol water, is heated to reflux 2h so that it is fully dissolve, then slow cooling
To-15 DEG C (5 DEG C/h), stirred crystallization 36h, gained crystal is leached, and by also washing with alcohol 2-3
Secondary, 35 DEG C of vacuum drying 24h obtain final products, product purity 96.1%, yield 50%.
Embodiment 18:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the ethanol of volume final concentration 6%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, is subsequently filtered out solid, add take 50mL ice dehydrated alcohol divide 2-3 time wash this solid, by gained
The vacuum drying of wet solid fully removes moisture.Subsequently, take above-mentioned gained solid 20g, add 400mL
Dehydrated alcohol, is heated to reflux 2h so that it is fully dissolve, and then slow cooling is to-15 DEG C (10 DEG C/h),
Stirred crystallization 36h, leaches gained crystal, and with and washing with alcohol 2-3 time, 35 DEG C of vacuum drying
24h obtains final products, product purity 99.6%, yield 65%.
Embodiment 19:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the ethanol of volume final concentration 6%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, is subsequently filtered out solid, add take 50mL ice dehydrated alcohol divide 2-3 time wash this solid, by gained
The vacuum drying of wet solid fully removes moisture.Subsequently, take above-mentioned gained solid 20g, add 400mL
Dehydrated alcohol, is heated to reflux 2h so that it is fully dissolve, and then slow cooling is to-15 DEG C (15 DEG C/h),
Stirred crystallization 36h, leaches gained crystal, and with and washing with alcohol 2-3 time, 35 DEG C of vacuum drying
24h obtains final products, product purity 99.1%, yield 64%.
Embodiment 20:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the ethanol of volume final concentration 6%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, is subsequently filtered out solid, add take 50mL ice dehydrated alcohol divide 2-3 time wash this solid, by gained
The vacuum drying of wet solid fully removes moisture.Subsequently, take above-mentioned gained solid 20g, add 400mL
Dehydrated alcohol, is heated to reflux 2h so that it is fully dissolve, and then slow cooling is to-15 DEG C (10 DEG C/h),
Stirred crystallization 36h, leaches gained crystal, and with and washing with alcohol 2-3 time, 40 DEG C of vacuum drying
24h obtains final products, product purity 99.6%, yield 66%, and product appearance is light yellow.
Embodiment 21:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the ethanol of volume final concentration 6%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, is subsequently filtered out solid, add take 50mL ice dehydrated alcohol divide 2-3 time wash this solid, by gained
The vacuum drying of wet solid fully removes moisture.Subsequently, take above-mentioned gained solid 20g, add 400mL
Dehydrated alcohol, is heated to reflux 2h so that it is fully dissolve, and then slow cooling is to-15 DEG C (10 DEG C/h),
Stirred crystallization 36h, leaches gained crystal, and with and washing with alcohol 2-3 time, 45 DEG C of vacuum drying
24h obtains final products, product purity 99.7%, yield 65%, and product appearance changes, table
Face presents buff.
Embodiment 22:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the ethanol of volume final concentration 6%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, is subsequently filtered out solid, add take 50mL ice dehydrated alcohol divide 2-3 time wash this solid, by gained
The vacuum drying of wet solid fully removes moisture.Subsequently, take above-mentioned gained solid 20g, add 400mL
Dehydrated alcohol, is heated to reflux 2h so that it is fully dissolve, and then slow cooling is to-15 DEG C (10 DEG C/h),
Stirred crystallization 36h, leaches gained crystal, and with and washing with alcohol 2-3 time, 50 DEG C of vacuum drying
24h obtains final products, product purity 99.6%, yield 57%, and product appearance changes, table
Face presents buff, and crystal generation dissolution phenomena, and bottom crystalline adheres to vessel surface, it is difficult to point
From.
Embodiment 23:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the ethanol of volume final concentration 6%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, is subsequently filtered out solid, add take 50mL ice dehydrated alcohol divide 2-3 time wash this solid, by gained
The vacuum drying of wet solid fully removes moisture.Subsequently, take above-mentioned gained solid 20g, add 400mL
Dehydrated alcohol, is heated to reflux 2h so that it is fully dissolve, and then slow cooling is to-15 DEG C (10 DEG C/h),
Stirred crystallization 36h, leaches gained crystal, and with and washing with alcohol 2-3 time, 35 DEG C of vacuum drying
36h obtains final products, product purity 99.7%, yield 64%, and product appearance changes, table
Face presents yellow.
Embodiment 24:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the ethanol of volume final concentration 6%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, is subsequently filtered out solid, add take 50mL ice dehydrated alcohol divide 2-3 time wash this solid, by gained
The vacuum drying of wet solid fully removes moisture.Subsequently, take above-mentioned gained solid 20g, add 400mL
Dehydrated alcohol, is heated to reflux 2h so that it is fully dissolve, and then slow cooling is to-15 DEG C (10 DEG C/h),
Stirred crystallization 36h, leaches gained crystal, and with and washing with alcohol 2-3 time, 35 DEG C of vacuum drying
48h obtains final products, product purity 99.6%, yield 54%, and product appearance surface presents yellow,
A small amount of crystal melting also sticks together at vessel surface.
Embodiment 25:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the ethanol of volume final concentration 6%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, is subsequently filtered out solid, add take 50mL ice dehydrated alcohol divide 2-3 time wash this solid, by gained
The vacuum drying of wet solid fully removes moisture.Subsequently, take above-mentioned gained solid 20g, add 300mL
Dehydrated alcohol, is heated to reflux 2h so that it is fully dissolve, and then slow cooling is to-15 DEG C (10 DEG C/h),
Stirred crystallization 36h, leaches gained crystal, and with and washing with alcohol 2-3 time, 35 DEG C of vacuum drying
24h obtains final products, product purity 99.4%, yield 68%, and product appearance is light yellow.
Embodiment 26:
100g epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the ethanol of volume final concentration 6%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, is subsequently filtered out solid, add take 50mL ice dehydrated alcohol divide 2-3 time wash this solid, by gained
The vacuum drying of wet solid fully removes moisture.Subsequently, take above-mentioned gained solid 20g, add 500mL
Dehydrated alcohol, is heated to reflux 2h so that it is fully dissolve, and then slow cooling is to-15 DEG C (10 DEG C/h),
Stirred crystallization 36h, leaches gained crystal, and with and washing with alcohol 2-3 time, 35 DEG C of vacuum drying
24h obtains final products, product purity 99.6%, yield 58%, and product appearance is light yellow.
Embodiment 27:
10kg epsilon-polylysine crude product (80%) is dissolved in deionized water and is configured to mass concentration
The aqueous solution of 30%, in this solution, adds the ethanol of volume final concentration 6%, continues to be heated to reflux
2h, makes epsilon-polylysine fully dissolve, and slow cooling is to-5 DEG C (5 DEG C/h), stirred crystallization 48 subsequently
H, is subsequently filtered out solid, add take 5L ice dehydrated alcohol divide 2-3 time wash this solid, gained is wet
Solid vacuum drying fully removes moisture.Subsequently, in above-mentioned solid 6.0kg, 100L is added anhydrous
Ethanol, is heated to reflux 2h so that it is fully dissolve, and then slow cooling is to-15 DEG C (5 DEG C/h), stirs
Mix crystallization 36h, gained crystal is leached, and with also washing with alcohol 2-3 time, 35 DEG C of vacuum drying
24h obtains final products, product purity 99.7%, yield 66%, and product chromatogram is shown in Fig. 2, marks
Quasi-product spectrogram is shown in Fig. 1.
Embodiment 28:
In Example 27, gained epsilon-polylysine product 3.0kg, is dissolved in deionized water and is configured to
The aqueous solution of mass concentration 30%, in this solution, adds the ethanol of volume final concentration 6%, continues
Being heated to reflux 2h, make epsilon-polylysine fully dissolve, slow cooling is to-5 DEG C (5 DEG C/h) subsequently, stirs
Mix crystallization 48h, be subsequently filtered out solid, add take 2L ice dehydrated alcohol divide 2-3 time wash this solid, general
The vacuum drying of gained wet solid fully removes moisture.Subsequently, in above-mentioned solid 2kg add 30L without
Water-ethanol, is heated to reflux 2h so that it is fully dissolve, and then slow cooling is to-15 DEG C (5 DEG C/h),
Stirred crystallization 36h, leaches gained crystal, and with and washing with alcohol 2-3 time, 35 DEG C of vacuum drying
24h obtains final products, product purity 99.9%, yield 62%.
Claims (9)
1. the method purifying epsilon-polylysine, it is characterised in that described method is: by mass concentration
The epsilon-polylysine crude product of more than 75% joins in deionized water, be configured to the ε of mass concentration 30-50%-
Polylysine aqueous solution, adds the organic solvent of volume final concentration 1-10% in aqueous solution, is heated to reflux
Reaction 2h, slow cooling is to-5 DEG C subsequently, stirred crystallization, filters, it is thus achieved that filter cake a, is done by filter cake a
Dry, it is thus achieved that epsilon-polylysine；Described organic solvent is the one in methanol, ethanol or acetone.
2. the method purifying epsilon-polylysine as claimed in claim 1, it is characterised in that described cooling rate
For 3-10 DEG C/h.
3. the method purifying epsilon-polylysine as claimed in claim 1, it is characterised in that described stirred crystallization
Time is 48h.
4. the method purifying epsilon-polylysine as claimed in claim 1, it is characterised in that described ε-poly-bad ammonia
In acid crude, epsilon-polylysine mass content is 75%-95%.
5. the method purifying epsilon-polylysine as claimed in claim 1, it is characterised in that described filter cake a uses
Ice absolute ethanol washing, takes the addition dehydrated alcohol of the filter cake b after washing and continues to be heated to reflux 2h, then delay
Slowly-15 DEG C it are cooled to, stirred crystallization, filter, it is thus achieved that filter cake c, by filter cake c washing with alcohol 2-3 time,
It is dried, it is thus achieved that epsilon-polylysine.
6. the method purifying epsilon-polylysine as claimed in claim 5, it is characterised in that described filter cake a uses
Ice absolute ethanol washing 2-3 time.
7. the method purifying epsilon-polylysine as claimed in claim 5, it is characterised in that described dehydrated alcohol
Volumetric usage is calculated as 10-20ml/g with filter cake b mass.
8. the method purifying as claimed in claim 5 epsilon-polylysine, it is characterised in that described in be cooled to-15
DEG C speed be 5-15 DEG C/h.
9. the method purifying as claimed in claim 5 epsilon-polylysine, it is characterised in that described be dried be
24-48h it is vacuum dried at 30-50 DEG C.
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| CN110170305A (en) * | 2019-04-30 | 2019-08-27 | 浙江工业大学 | A kind of separating technology of polylysine |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1526698A (en) * | 2003-03-06 | 2004-09-08 | 四川三高生化股份有限公司 | Production process of high-purity DL-lysine |
| CN101665442A (en) * | 2009-10-09 | 2010-03-10 | 蚌埠丰原涂山制药有限公司 | Crystallization method of high purity DL-lysine |
| CN101701069A (en) * | 2009-09-22 | 2010-05-05 | 浙江银象生物工程有限公司 | Method for extracting epsilon-polylysine and salt thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1526698A (en) * | 2003-03-06 | 2004-09-08 | 四川三高生化股份有限公司 | Production process of high-purity DL-lysine |
| CN101701069A (en) * | 2009-09-22 | 2010-05-05 | 浙江银象生物工程有限公司 | Method for extracting epsilon-polylysine and salt thereof |
| CN101665442A (en) * | 2009-10-09 | 2010-03-10 | 蚌埠丰原涂山制药有限公司 | Crystallization method of high purity DL-lysine |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110170305A (en) * | 2019-04-30 | 2019-08-27 | 浙江工业大学 | A kind of separating technology of polylysine |
| CN110170305B (en) * | 2019-04-30 | 2022-05-24 | 浙江工业大学 | Polylysine separation process |
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