CN111777760A - Polylysine separation and purification process and application - Google Patents

Polylysine separation and purification process and application Download PDF

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Publication number
CN111777760A
CN111777760A CN202010637151.3A CN202010637151A CN111777760A CN 111777760 A CN111777760 A CN 111777760A CN 202010637151 A CN202010637151 A CN 202010637151A CN 111777760 A CN111777760 A CN 111777760A
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polylysine
solution
concentration
eluent
exchange resin
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赵兵
常森林
赵庆生
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Institute of Process Engineering of CAS
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    • C08G69/00—Macromolecular compounds obtained by reactions forming a carboxylic amide link in the main chain of the macromolecule
    • C08G69/46—Post-polymerisation treatment
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
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    • A23B4/14—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
    • A61K8/84—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds obtained by reactions otherwise than those involving only carbon-carbon unsaturated bonds
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    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
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    • A61Q19/00—Preparations for care of the skin
    • 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
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Abstract

The invention provides a polylysine separation and purification process and application, wherein the process reduces the difficulty of the subsequent polylysine separation and purification through reasonable microfiltration treatment; polylysine purity is increased by an improvement of the elution process after adsorption on the ion exchange resin. The process is simple in overall operation and environment-friendly, the recovery rate of the para-polylysine is high, and the purity of the isolated and purified poly-lysine is excellent. In addition, the invention organically combines the separation and purification process and the application of polylysine to form a complete ecological industrial chain and provides a good idea for the overall layout of polylysine.

Description

Polylysine separation and purification process and application
Technical Field
The invention belongs to the technical field of fermentation engineering, and relates to a polylysine separation and purification process and application thereof.
Background
Polylysine has broad-spectrum antibacterial property, has good inhibition effect on gram-positive bacteria, gram-negative bacteria, saccharomycetes, mold, bacteriophage and the like, has the advantages of wide antibacterial spectrum, strong antibacterial ability, high temperature resistance (120 ℃), good water solubility, no influence on food flavor, low use amount, high safety and the like compared with the traditional chemical preservatives such as benzoic acid, sorbic acid and the like, and is a natural fermented food preservative with excellent preservative property and huge commercial potential. In addition, polylysine is also widely used in the fields of medicine, chemical engineering, electronic materials, bioengineering, and the like.
At present, the production of polylysine in Japan is at the leading level in the world, and the consumption of polylysine in China has strong dependence on the import of Japan. In the eighties of the last century, japan and korea have begun to use as food additives, and the us FDA approved for use in 2003. However, polylysine is approved only by the national committee in 2014 since the research of polylysine in China is late. In recent years, several enterprises in our country have tried production, but behind Japan, the problems of low fermentation level, low total product yield, high production cost, poor quality and the like exist, the mechanism is lack of deep research, and the development and application of polylysine in our country are greatly limited. Therefore, the method has great significance in carrying out process research aiming at the problems of serious pollution, low product purity, low yield, poor color and the like in the subsequent separation and purification process of the fermentation liquor.
In addition, polylysine can be mixed with other preservatives for use, and the antibacterial effect is greatly improved through the synergistic interaction, so that the antiseptic effect can be improved, and the dosage of the preservatives can be reduced, thereby achieving the purposes of reducing the production cost and expanding the application range.
Disclosure of Invention
Aiming at the technical problems in the prior art, the invention provides a polylysine separation and purification process and application thereof, wherein the process has the advantages of high product recovery rate, less environmental pollution and high product quality; and the application provides a thought for the application of the polylysine and has a vivid guiding function for the wide application of the polylysine.
In order to achieve the purpose, the invention adopts the following technical scheme:
one of the purposes of the invention is to provide a polylysine separation and purification process, which comprises the following steps:
(1) carrying out solid-liquid separation and microfiltration on the fermentation liquor to obtain polylysine permeate;
(2) adsorbing the polylysine permeate obtained in the step (1) by using ion exchange resin, and eluting by using a solvent after adsorption to obtain polylysine eluent;
(3) desalting the polylysine eluent obtained in the step (2), and concentrating and drying after desalting to obtain the polylysine.
As a preferred technical scheme of the invention, the solid-liquid separation in the step (1) comprises a continuous mode or a batch mode.
Preferably, the continuous solid-liquid separation is a butterfly centrifuge.
Preferably, the batch solid-liquid separation is plate and frame filtration.
Preferably, the pH is adjusted to 6-9 or not adjusted before microfiltration, and the adjusted pH can be 6.5, 7, 7.5, 8 or 8.5, etc., but is not limited to the recited values, and other values not recited in the range of the values are also applicable.
In the invention, before microfiltration in the step (1), the pH of the clear liquid of the fermentation liquor after solid-liquid separation is adjusted, so that polylysine is changed from a positively charged state to an uncharged or negatively charged state, the electrostatic adsorption effect of polylysine and other impurities is weakened, and the difficulty of subsequent separation and purification of polylysine is reduced. In the step (3), the quality of the polylysine solution obtained in the step (2) of the process meets the requirement without an additional activated carbon decoloring unit, so that the loss of the polylysine is reduced, and the recovery rate of the polylysine product is improved.
As a preferred technical scheme of the invention, the ion exchange resin in the step (2) comprises a strong acid cation exchange resin or a weak acid cation exchange resin, and is preferably a weak acid cation exchange resin.
Preferably, the weakly acidic cation exchange resin comprises any one or a combination of at least two of D111, D113, D151, D152, 110, 111, 112, 122, 116 or IRC-50.
Preferably, the ion exchange resin is washed with water to a pH of 10 or less, such as 9, 8, 7, 6, 5, 4, or 3, before use, but is not limited to the recited values, and other values not recited within the range of values are equally applicable.
Preferably, the pH of the polylysine permeate is adjusted to 6 to 9, such as 6.5, 7, 7.5, 8, or 8.5, prior to the adsorption treatment, but is not limited to the recited values, and other values not recited within this range are also applicable.
In a preferred embodiment of the present invention, the elution treatment in step (2) includes washing and eluting the ion exchange resin after the adsorption treatment with a solvent.
Preferably, the solvent used for the washing comprises any one of water, an ethanol solution, an acetic acid solution, a urea solution, a sodium chloride solution or a sodium acetate solution, or a combination of at least two of them, as typical but non-limiting examples: a combination of water and an ethanol solution, a combination of an ethanol solution and an acetic acid solution, a combination of an acetic acid solution and a urea solution, a combination of a urea solution and a sodium chloride solution, a combination of a sodium chloride solution and a sodium acetate solution, or a combination of water, an ethanol solution and an acetic acid solution, and the like. The combination may be washed by mixing the solvents, or the combination may be washed by separately using the solvents.
Preferably, the mass concentration of the ethanol solution is 10-70%, more preferably 30-60%, and the dosage of the ethanol solution is 0-4 BV. The mass concentration of the ethanol solution may be 20%, 30%, 40%, 50% or 60%, and the amount of the ethanol solution may be 0.5BV, 1BV, 1.5BV, 2BV, 2.5BV, 3BV or 3.5BV, but is not limited to the above-mentioned values, and other values not listed in the above-mentioned ranges of values are also applicable.
Preferably, the concentration of the acetic acid solution is 0.05-2 mol/L, further preferably 0.1-1 mol/L, and the dosage of the acetic acid solution is 0-4 BV; the concentration of the acetic acid solution may be 0.08mol/L, 0.1mol/L, 0.2mol/L, 0.5mol/L, 0.8mol/L, 1mol/L, 1.2mol/L, 1.5mol/L or 1.8mol/L, etc., and the amount of the acetic acid solution may be 0.5BV, 1BV, 1.5BV, 2BV, 2.5BV, 3BV or 3.5BV, etc., but not limited to the values listed, and other values not listed in the above ranges of values are also applicable.
Preferably, the concentration of the urea solution is 0.1-1 mol/L, further preferably 0.1-0.5 mol/L, and the dosage of the urea solution is 0-4 BV; the concentration of the urea solution may be 0.2mol/L, 0.3mol/L, 0.4mol/L, 0.5mol/L, 0.6mol/L, 0.7mol/L, 0.8mol/L, or 0.9mol/L, etc., and the amount of the urea solution may be 0.5BV, 1BV, 1.5BV, 2BV, 2.5BV, 3BV, or 3.5BV, etc., but is not limited to the values listed, and other values not listed in the above ranges of values are also applicable.
Preferably, the concentration of the sodium chloride solution is 0.1-2 mol/L, further preferably 0.5-1.5 mol/L, and the using amount of the sodium chloride solution is 0-3 BV; the concentration of the sodium chloride solution may be 0.2mol/L, 0.5mol/L, 0.8mol/L, 1mol/L, 1.2mol/L, 1.5mol/L or 1.8mol/L, etc., and the amount of the sodium chloride solution may be 0.5BV, 1BV, 1.5BV, 2BV or 2.5BV, etc., but is not limited to the values listed, and other values not listed within the above ranges of values are also applicable.
Preferably, the concentration of the sodium acetate solution is 0.1-2 mol/L, further preferably 0.1-1 mol/L, and the using amount of the sodium chloride solution is 0-3 BV; the concentration of the acetic acid solution may be 0.2mol/L, 0.5mol/L, 0.8mol/L, 1mol/L, 1.2mol/L, 1.5mol/L or 1.8mol/L, etc., and the amount of the acetic acid solution may be 0.5BV, 1BV, 1.5BV, 2BV or 2.5BV, etc., but is not limited to the values listed, and other values not listed in the above ranges of values are also applicable.
Preferably, the solvent used for elution comprises a hydrochloric acid solution and/or a sodium hydroxide solution.
Preferably, the concentration of the hydrochloric acid is 0.05-0.5 mol/L, further preferably 0.1-0.3 mol/L, and the dosage of the hydrochloric acid solution is 0-5 BV; the concentration of hydrochloric acid may be 0.08mol/L, 0.1mol/L, 0.2mol/L, 0.3mol/L or 0.4mol/L, etc., and the amount of the hydrochloric acid solution may be 0.5BV, 1BV, 1.5BV, 2BV, 2.5BV, 3BV, 3.5BV, 4BV or 4.5BV, etc., but is not limited to the values listed, and other values not listed in the above ranges of values are also applicable.
Preferably, the concentration of the sodium hydroxide solution is 0.05-0.5 mol/L, further preferably 0.1-0.3 mol/L, and the using amount of the sodium hydroxide solution is 0-4 BV; the concentration of the sodium hydroxide solution may be 0.08mol/L, 0.1mol/L, 0.2mol/L, 0.3mol/L, or 0.4mol/L, etc., and the amount of the sodium hydroxide solution may be 0.5BV, 1BV, 1.5BV, 2BV, 2.5BV, 3BV, or 3.5BV, etc., but is not limited to the values listed, and other values not listed in the above numerical ranges may be used as well.
In the invention, after the polylysine solution is adsorbed by the ion exchange resin in the step (2), the elution procedure is improved, and the ion exchange resin is washed by selecting a proper solvent, so that the selective elution of impurities adsorbed on the ion exchange resin is preferentially realized; and then the adsorbed polylysine is eluted from the ion exchange resin by replacing the solvent, thereby achieving the purpose of further improving the purity of the polylysine.
As a preferred technical scheme of the invention, before the desalting treatment in the step (3), the pH of the polylysine eluent is adjusted to 5.5.
Preferably, the desalting treatment in step (3) is performed by using a membrane device.
Preferably, the membrane device adopts a roll membrane filter membrane core.
Preferably, the molecular weight of the core of the roll-up membrane filter membrane is not more than 5000, such as 1000, 1500, 2000, 2500, 3000, 3500, 4000 or 4500, and the like, but is not limited to the recited values, and other values not recited in the range of the values are also applicable.
Preferably, the end point of the desalting in step (3) is at a conductivity of the polylysine eluate of less than 200mS/cm, such as 50mS/cm, 80mS/cm, 100mS/cm, 120mS/cm, 150mS/cm or 180mS/cm, but not limited to the recited values, and other non-recited values within this range are equally applicable.
As a preferable technical scheme of the invention, the concentration in the step (3) comprises primary concentration and reduced pressure concentration which are sequentially carried out.
Preferably, the preliminary concentration is carried out in the membrane plant.
Preferably, the temperature of the concentration under reduced pressure is not more than 60 ℃, such as 10 ℃, 20 ℃, 30 ℃, 40 ℃ or 50 ℃, etc., but is not limited to the recited values, and other unrecited values within the range of values are also applicable.
Preferably, the pressure for concentration under reduced pressure is not greater than-0.08 MPa, such as-0.08 MPa, -0.085MPa, -0.09MPa, -0.095MPa, or-0.1 MPa, but not limited to the recited values, and other values not recited within the range are equally applicable.
Preferably, the concentration of the polylysine in the solution after the concentration under reduced pressure is 5 to 30 wt%, such as 6 wt%, 10 wt%, 15 wt%, 20 wt% or 25 wt%, but not limited to the recited values, and other values not recited in the range of the recited values are also applicable.
In a preferred embodiment of the present invention, the drying method in step (3) includes any one of spray drying, freeze drying and vacuum pulse drying, and preferably spray drying.
Preferably, the air inlet temperature of the spray drying is 120-200 ℃, and the air outlet temperature is 70-100 ℃. The inlet temperature may be 130 ℃, 140 ℃, 150 ℃, 160 ℃, 170 ℃, 180 ℃ or 190 ℃, and the outlet temperature may be 75 ℃, 80 ℃, 85 ℃, 90 ℃ or 95 ℃, but is not limited to the values listed, and other values not listed in the above numerical ranges are also applicable. As a preferable technical scheme of the invention, the polylysine separation and purification process comprises the following steps:
(1) carrying out solid-liquid separation and microfiltration on the fermentation liquor to obtain polylysine permeate;
the solid-liquid separation comprises a continuous type or an intermittent type, and the pH is adjusted to 6-9 or not adjusted before microfiltration;
(2) adsorbing the polylysine permeate obtained in the step (1) by using ion exchange resin, and eluting by using a solvent after adsorption to obtain polylysine eluent;
the ion exchange resin comprises a strong-acid cation exchange resin and a weak-acid cation exchange resin, and the ion exchange resin is washed by water until the pH value is less than or equal to 10 before use;
before the adsorption treatment, adjusting the pH value of the polylysine permeation solution to 6-9;
the solvent comprises any one or the combination of at least two of water, hydrochloric acid, ethanol solution, acetic acid solution, sodium hydroxide solution, urea solution, sodium chloride solution or sodium acetate solution;
(3) adjusting the pH value of the polylysine eluent obtained in the step (2) to 5.5, desalting the polylysine eluent obtained in the step (2) in membrane equipment until the conductivity of the polylysine eluent is less than 200mS/cm, wherein the membrane equipment adopts a roll-type membrane filter membrane core, and concentrating and drying after desalting to obtain the polylysine.
The second purpose of the invention is to provide an application of polylysine prepared by the polylysine separation and purification process, wherein the polylysine is applied to the field of food additives or the field of daily chemical products.
As a preferable technical scheme of the invention, the field of the food additive comprises biological preservatives in the fields of meat products, wheaten foods or beverages.
Preferably, the-polylysine is used synergistically with one or a combination of at least two of natamycin, nisin, citric acid, acetic acid, chitosan.
Preferably, the polylysine is used as a food additive in an amount of 0.01 to 0.3g/kg, such as 0.02g/kg, 0.05g/kg, 0.1g/kg, 0.15g/kg, 0.2g/kg or 0.25g/kg, but not limited to the recited values, and other values within this range are equally applicable.
Preferably, the field of daily chemical products comprises additives or effective ingredients of the polylysine for daily chemical products.
Preferably, the polylysine is used as an additive for household chemicals to inhibit product spoilage.
Preferably, the-polylysine inhibits acne propionibacterium, staphylococcus aureus, streptococcus hemolyticus, and pseudomonas aeruginosa.
Preferably, the polylysine is used as an additive for daily chemical products in an amount of 0.01 to 0.3g/L, such as 0.02g/L, 0.05g/L, 0.1g/L, 0.15g/L, 0.2g/L, or 0.25g/L, but not limited to the recited values, and other values within the range are also applicable.
Compared with the prior art, the invention at least has the following beneficial effects:
(1) the invention provides a polylysine separation and purification process, the purity of the polylysine prepared by the process is more than 95%, and the recovery rate is more than 80%;
(2) the invention provides a polylysine separation and purification process, which is verified by a series of amplification experiments from laboratory level to industrial production level, and the result shows that the extraction process can achieve the purposes of improving the product recovery rate, reducing the environmental pollution and improving the product quality;
(3) the invention provides a process for preparing polylysine by a polylysine separation and purification process, which refines the application of polylysine in the field of food and daily chemical products, exploits the depth and the breadth of the application of polylysine, and improves the application prospect and the range of polylysine;
(4) the invention organically combines the extraction process and the application of polylysine to form a complete ecological industrial chain and provides a good idea for the overall layout of polylysine.
Drawings
FIG. 1 is a liquid chromatogram of an original fermentation broth;
FIG. 2a is a liquid chromatogram of a polylysine product obtained in example 1 of the present invention;
FIG. 2b is a liquid chromatogram of a polylysine product obtained in example 2 of the present invention;
FIG. 3 is a diagram showing the products of polylysine obtained in examples 1 and 2 of the present invention;
FIG. 4 is a plot of the time of flight of a polylysine product prepared in accordance with the present invention;
FIG. 5 shows H of a polylysine product prepared according to the present invention1-NMR spectrum;
FIG. 6 is a SEM image of spray-dried and freeze-dried polylysine products prepared in accordance with the present invention;
FIG. 7 is a diagram showing the inhibition effect of the-polylysine product on acne vulgaris.
The present invention is described in further detail below. The following examples are merely illustrative of the present invention and do not represent or limit the scope of the claims, which are defined by the claims.
Detailed Description
The technical scheme of the invention is further explained by the specific implementation mode in combination with the attached drawings.
To better illustrate the invention and to facilitate the understanding of the technical solutions thereof, typical but non-limiting examples of the invention are as follows:
example 1
The embodiment provides a polylysine separation and purification process, which comprises the following steps:
(1) performing solid-liquid separation on the fermentation liquor with the polylysine concentration of 30g/L by adopting plate-and-frame filtration, and performing microfiltration on the fermentation liquor by adopting 300KD microfiltration membrane equipment to obtain polylysine permeate;
(2) adjusting the pH value of the polylysine permeate obtained in the step (1) to 7, adopting weak acid cation exchange resin IRC50 washed by water until the pH value is less than or equal to 10 to adsorb the polylysine permeate, washing the resin with purified water after the adsorption is saturated until an effluent liquid is clear, then eluting with 4BV0.2 mol/L sodium hydroxide solution, and collecting an eluent when polylysine can be detected in an eluent to obtain polylysine eluent;
(3) adjusting the pH value of the polylysine eluent obtained in the step (2) to 5.5 by using 1mol/L sodium hydroxide solution, desalting the polylysine eluent in a membrane filtration device with a membrane core of 800D until the conductivity of the polylysine eluent is less than 200mS/cm, carrying out reduced pressure concentration on the desalted polylysine solution at 60 ℃, stopping concentration when the mass concentration of the polylysine is 20%, and freeze-drying the concentrated solution to obtain the polylysine.
The liquid chromatogram of the prepared polylysine is shown in figure 2a, and the recovery rate and purity of the polylysine product are respectively 87.1% and 95.8% through liquid chromatogram detection.
Example 2
The embodiment provides a polylysine separation and purification process, which comprises the following steps:
(1) performing solid-liquid separation on the fermentation liquor with the polylysine concentration of 30g/L by adopting plate-and-frame filtration, and performing microfiltration on the fermentation liquor by adopting 300KD microfiltration membrane equipment to obtain polylysine permeate;
(2) adjusting the pH value of the polylysine permeate obtained in the step (1) to 9, performing adsorption treatment on the polylysine permeate by using weak-acid cation exchange resin IRC50 washed by water until the pH value is less than or equal to 10, after the adsorption is saturated, washing the resin by using purified water until an effluent liquid is clear, washing the resin by using a 3BV 60% ethanol solution, then washing by using water until the effluent liquid is clear, collecting the ethanol eluent to recover the ethanol, then eluting by using a 4BV0.2 mol/L sodium hydroxide solution, and when the polylysine can be detected in an eluent, collecting the eluent to obtain a polylysine eluent;
(3) adjusting the pH value of the polylysine eluent obtained in the step (2) to 5.5 by using 1mol/L sodium hydroxide solution, desalting the polylysine eluent in a membrane filtration device with a membrane core of 800D until the conductivity of the polylysine eluent is less than 200mS/cm, carrying out reduced pressure concentration on the desalted polylysine solution at 60 ℃, stopping concentration when the mass concentration of the polylysine is 20%, and freeze-drying the concentrated solution to obtain the polylysine.
The liquid chromatogram of the prepared polylysine is shown in fig. 2b, and the recovery rate and purity of the polylysine product are respectively 88.2% and 98.1% through liquid chromatogram detection.
Example 3
The embodiment provides a polylysine separation and purification process, which comprises the following steps:
(1) performing solid-liquid separation on the fermentation liquor with the polylysine concentration of 10g/L by adopting plate-and-frame filtration, and performing microfiltration on the fermentation liquor by adopting 300KD microfiltration membrane equipment to obtain polylysine permeate;
(2) adjusting the pH value of the polylysine permeate obtained in the step (1) to 8, adopting weak acid cation exchange resin IRC50 washed by water until the pH value is less than or equal to 10 to adsorb the polylysine permeate, after adsorption saturation, washing the resin by purified water until effluent is clear, washing the resin by 3BV0.4 mol/L urea solution, then washing by water until the effluent is clear, then eluting the resin by 4BV0.05 mol/L sodium hydroxide solution, and when polylysine can be detected in eluent, collecting eluent to obtain polylysine eluent;
(3) adjusting the pH value of the polylysine eluent obtained in the step (2) to 5.5 by using a sodium hydroxide solution, desalting the polylysine eluent in a membrane filtration device with a membrane core of 800D until the conductivity of the polylysine eluent is less than 200mS/cm, carrying out reduced pressure concentration on the desalted polylysine solution at 60 ℃, stopping concentration when the mass concentration of the polylysine is 5%, and carrying out spray drying on the concentrated solution to obtain the polylysine.
Liquid chromatography detection shows that the recovery rate of the polylysine product is 85.7 percent, and the purity is 96.2 percent.
Example 4
The embodiment provides a polylysine separation and purification process, which comprises the following steps:
(1) performing solid-liquid separation on fermentation liquor with the polylysine concentration of 20g/L by adopting plate-and-frame filtration, adjusting the pH of the fermentation liquor to 7, and performing microfiltration on the fermentation liquor by adopting 300KD microfiltration membrane equipment to obtain polylysine permeate;
(2) adjusting the pH value of the polylysine permeate obtained in the step (1) to 8, performing adsorption treatment on the polylysine permeate by using weak-acid cation exchange resin D113 which is washed by water until the pH value is less than or equal to 10, after the adsorption is saturated, washing the resin by using purified water until effluent is clear, washing the resin by using 4BV 30% ethanol solution, then washing by using water until the effluent is clear, collecting ethanol eluent to recover ethanol, eluting the resin by using 2BV 0.5mol/L sodium hydroxide solution, and when polylysine can be detected in the eluent, collecting the eluent to obtain polylysine eluent;
(3) adjusting the pH value of the polylysine eluent obtained in the step (2) to 5.5 by using a sodium hydroxide solution, desalting the polylysine eluent in a membrane filtration device with a membrane core of 2000D until the conductivity of the polylysine eluent is less than 200mS/cm, carrying out reduced pressure concentration on the desalted polylysine solution at 60 ℃, stopping concentration when the mass concentration of polylysine is 20%, and carrying out spray drying on the concentrated solution to obtain the polylysine.
Liquid chromatography detection shows that the recovery rate of the polylysine product is 90.1 percent, and the purity is 95.2 percent.
Example 5
The embodiment provides a polylysine separation and purification process, which comprises the following steps:
(1) performing solid-liquid separation on the fermentation liquor with the polylysine concentration of 30g/L by adopting plate-and-frame filtration, and performing microfiltration on the fermentation liquor by adopting 300KD microfiltration membrane equipment to obtain polylysine permeate;
(2) adjusting the pH value of the polylysine permeate obtained in the step (1) to 8, performing adsorption treatment on the polylysine permeate by using weak-acid cation exchange resin D113 which is washed by water until the pH value is less than or equal to 10, after the adsorption is saturated, washing the resin by using purified water until the effluent is clear, washing the resin by using 4BV0.2 mol/L acetic acid solution, then washing the resin by using water until the effluent is clear, washing the resin by using 2BV 50% ethanol solution, then washing the resin by using water until the effluent is clear, collecting the ethanol eluate to recover the ethanol, then eluting the resin by using 3BV 0.2mol/L sodium hydroxide solution, and collecting the eluate when polylysine can be detected in the eluate to obtain polylysine eluate;
(3) adjusting the pH value of the polylysine eluent obtained in the step (2) to 5.5 by using a sodium hydroxide solution, desalting the polylysine eluent in a membrane filtration device with a membrane core of 3000D until the conductivity of the polylysine eluent is less than 200mS/cm, carrying out reduced pressure concentration on the desalted polylysine solution at 60 ℃, stopping concentration when the mass concentration of polylysine is 30%, and carrying out spray drying on the concentrated solution to obtain the polylysine.
The liquid chromatography detection shows that the recovery rate of the polylysine product is 81.3 percent, and the purity is 97.6 percent.
Example 6
The embodiment provides a polylysine separation and purification process, which comprises the following steps:
(1) performing solid-liquid separation on the fermentation liquor with the polylysine concentration of 20g/L by adopting butterfly centrifugation, adjusting the pH of the fermentation liquor to 8 by using sodium hydroxide, and performing microfiltration on the fermentation liquor by adopting 300KD microfiltration membrane equipment to obtain polylysine permeate;
(2) adjusting the pH value of the polylysine permeate obtained in the step (1) to 8, performing adsorption treatment on the polylysine permeate by using weak-acid cation exchange resin D151 which is washed by water until the pH value is less than or equal to 10, after the adsorption is saturated, washing the resin by using purified water until effluent is clear, washing the resin by using 3BV 70 wt% ethanol solution, then washing the resin by using water until the effluent is clear, collecting ethanol eluent to recover ethanol, washing the resin by using 1BV 2mol/L sodium chloride solution, then washing the resin by using water until the effluent is clear, then eluting the resin by using 3BV 0.3mol/L sodium hydroxide solution, and when polylysine can be detected in the eluent, collecting the eluent to obtain polylysine eluent;
(3) adjusting the pH value of the polylysine eluent obtained in the step (2) to 5.5 by using a sodium hydroxide solution, desalting the polylysine eluent in a membrane filtration device with a membrane core of 3000D until the conductivity of the polylysine eluent is less than 200mS/cm, carrying out reduced pressure concentration on the desalted polylysine solution at 60 ℃, stopping concentration when the mass concentration of polylysine is 30%, and carrying out spray drying on the concentrated solution to obtain the polylysine.
Liquid chromatography detection shows that the recovery rate of the polylysine product is 85.7 percent, and the purity is 98.8 percent.
Example 7
The embodiment provides a polylysine separation and purification process, which comprises the following steps:
(1) performing solid-liquid separation on the fermentation liquor with the polylysine concentration of 10g/L by adopting plate-and-frame filtration, and performing microfiltration on the fermentation liquor by adopting 300KD microfiltration membrane equipment to obtain polylysine permeate;
(2) adjusting the pH value of the polylysine permeate obtained in the step (1) to 7, performing adsorption treatment on the polylysine permeate by using weak-acid cation exchange resin D151 which is washed by water until the pH value is less than or equal to 10, after the adsorption is saturated, washing the resin by using purified water until effluent is clear, washing the resin by using 2BV 1mol/L sodium acetate solution, then washing by using water until the effluent is clear, then eluting the resin by using 4BV 0.15mol/L sodium hydroxide solution, and when polylysine can be detected in the eluent, collecting eluent to obtain polylysine eluent;
(3) adjusting the pH value of the polylysine eluent obtained in the step (2) to 5.5 by using a sodium hydroxide solution, desalting the polylysine eluent in a membrane filtration device with a membrane core of 5000D until the conductivity of the polylysine eluent is less than 200mS/cm, carrying out reduced pressure concentration on the desalted polylysine solution at 60 ℃, stopping concentration when the mass concentration of polylysine is 30%, and freeze-drying the concentrated solution to obtain the polylysine.
The liquid chromatography detection shows that the recovery rate of the polylysine product is 81.6 percent, and the purity is 97.0 percent.
Example 8
The embodiment provides a polylysine separation and purification process, which comprises the following steps:
(1) performing solid-liquid separation on the fermentation liquor with the polylysine concentration of 40g/L by adopting plate-and-frame filtration, adjusting the pH of the fermentation liquor to 9 by using sodium hydroxide, and performing microfiltration on the fermentation liquor by adopting 300KD microfiltration membrane equipment to obtain polylysine permeate;
(2) adsorbing the polylysine permeate obtained in the step (1) by using weak-acid cation exchange resin IRC50 washed by water until the pH value is less than or equal to 10, after the adsorption is saturated, washing the resin by using purified water until an effluent liquid is clear, washing the resin by using 2BV 0.5mol/L urea solution, then washing by using water until the effluent liquid is clear, eluting the resin by using 4BV0.2 mol/L sodium hydroxide solution, and when polylysine can be detected in an elution solution, collecting the elution solution to obtain polylysine eluent;
(3) adjusting the pH value of the polylysine eluent obtained in the step (2) to 5.5 by using a sodium hydroxide solution, desalting the polylysine eluent in a membrane filtration device with a membrane core of 800D until the conductivity of the polylysine eluent is less than 200mS/cm, carrying out reduced pressure concentration on the desalted polylysine solution at 60 ℃, stopping concentration when the mass concentration of polylysine is 30%, and freeze-drying the concentrated solution to obtain the polylysine.
The liquid chromatography detection shows that the recovery rate of the polylysine product is 81.8 percent, and the purity is 95.4 percent.
Example 9
The embodiment provides a polylysine separation and purification process, which comprises the following steps:
(1) performing solid-liquid separation on the fermentation liquor with the polylysine concentration of 30g/L by adopting plate-and-frame filtration, adjusting the pH of the fermentation liquor to 9 by using sodium hydroxide, and performing microfiltration on the fermentation liquor by adopting 300KD microfiltration membrane equipment to obtain polylysine permeate;
(2) adsorbing the polylysine permeate obtained in the step (1) by using weak-acid cation exchange resin IRC50 washed by water until the pH value is less than or equal to 10, washing the resin by using purified water until an effluent is clear after the adsorption is saturated, washing the resin by using 1BV 1mol/L urea solution, then washing the resin by using water until the effluent is clear, washing the resin by using 3BV 0.5mol/L sodium chloride solution, then washing the resin by using water until the effluent is clear, then eluting the resin by using 5BV 0.15mol/L sodium hydroxide solution, and collecting an eluent to obtain polylysine eluent when polylysine can be detected in an eluent;
(3) adjusting the pH value of the polylysine eluent obtained in the step (2) to 5.5 by using a sodium hydroxide solution, desalting the polylysine eluent in a membrane filtration device with a membrane core of 2000D until the conductivity of the polylysine eluent is less than 200mS/cm, carrying out reduced pressure concentration on the desalted polylysine solution at 60 ℃, stopping concentration when the mass concentration of polylysine is 15%, and carrying out spray drying on the concentrated solution to obtain the polylysine.
Liquid chromatography detection shows that the recovery rate of the polylysine product is 82.8 percent, and the purity is 96.6 percent.
Example 10
The embodiment provides a polylysine separation and purification process, which comprises the following steps:
(1) performing solid-liquid separation on the fermentation liquor with the polylysine concentration of 30g/L by adopting plate-and-frame filtration, adjusting the pH of the fermentation liquor to 8 by using sodium hydroxide, and performing microfiltration on the fermentation liquor by adopting 300KD microfiltration membrane equipment to obtain polylysine permeate;
(2) adsorbing the polylysine permeate obtained in the step (1) by using a weakly acidic cation exchange resin D113 which is washed by water until the pH value is less than or equal to 10, washing the resin by using purified water after the adsorption is saturated until an effluent liquid is clear, washing the resin by using 1BV of 1.5mol/L sodium acetate solution, then washing the resin by using water until the effluent liquid is clear, washing the resin by using 3BV of 0.3mol/L sodium chloride solution, then washing the resin by using water until the effluent liquid is clear, then eluting the resin by using 5BV of 0.15mol/L sodium hydroxide solution, and collecting an eluent when polylysine can be detected in an eluent to obtain polylysine eluent;
(3) adjusting the pH value of the polylysine eluent obtained in the step (2) to 5.5 by using a sodium hydroxide solution, desalting the polylysine eluent in a membrane filtration device with a membrane core of 5000D until the conductivity of the polylysine eluent is less than 200mS/cm, carrying out reduced pressure concentration on the desalted polylysine solution at 60 ℃, stopping concentration when the mass concentration of polylysine is 10%, and carrying out spray drying on the concentrated solution to obtain the polylysine.
Liquid chromatography detection shows that the recovery rate of the polylysine product is 83.1 percent, and the purity is 98.1 percent.
Example 11
The embodiment provides a polylysine separation and purification process, which comprises the following steps:
(1) performing solid-liquid separation on the fermentation liquor with the polylysine concentration of 20g/L by adopting plate-and-frame filtration, adjusting the pH of the fermentation liquor to 9 by using sodium hydroxide, and performing microfiltration on the fermentation liquor by adopting 300KD microfiltration membrane equipment to obtain polylysine permeate;
(2) adsorbing the polylysine permeate obtained in the step (1) by using a weak-acid cation exchange resin D113 which is washed by water until the pH value is less than or equal to 10, after the adsorption is saturated, washing the resin by using purified water until an effluent liquid is clear, washing the resin by using a 1BV 1mol/L acetic acid solution, then washing by using water until the effluent liquid is clear, eluting the resin by using a 3BV 0.2mol/L hydrochloric acid solution, and when polylysine can be detected in an eluent, collecting the eluent to obtain polylysine eluent;
(3) adjusting the pH value of the polylysine eluent obtained in the step (2) to 5.5 by using a sodium hydroxide solution, desalting the polylysine eluent in a membrane filtration device with a membrane core of 3000D until the conductivity of the polylysine eluent is less than 200mS/cm, carrying out reduced pressure concentration on the desalted polylysine solution at 60 ℃, stopping concentration when the mass concentration of polylysine is 30%, and carrying out spray drying on the concentrated solution to obtain the polylysine.
The liquid chromatography detection shows that the recovery rate of the polylysine product is 87.0 percent, and the purity is 95.3 percent.
Example 12
This example was carried out under the same conditions as example 11 except that the pH was not adjusted before the microfiltration in step (1), but the pH of the filtrate was adjusted to 9 after the microfiltration in step (2).
The liquid chromatography detection shows that the recovery rate of the polylysine product is 81.1 percent, and the purity is 95.0 percent.
Example 13
The embodiment provides a polylysine separation and purification process, which comprises the following steps:
(1) performing solid-liquid separation on the fermentation liquor with the polylysine concentration of 40g/L by adopting plate-and-frame filtration, adjusting the pH of the fermentation liquor to 8 by using sodium hydroxide, and performing microfiltration on the fermentation liquor by adopting 300KD microfiltration membrane equipment to obtain polylysine permeate;
(2) adsorbing the polylysine permeate obtained in the step (1) by using weak-acid cation exchange resin IRC50 washed by water until the pH value is less than or equal to 10, after the adsorption is saturated, washing the resin by using purified water until an effluent is clear, washing the resin by using 3BV0.1 mol/L acetic acid solution, then washing the resin by using water until the effluent is clear, washing the resin by using 3BV 20 wt% ethanol solution, then washing the resin by using water until the effluent is clear, collecting the ethanol eluent to recover ethanol, then eluting the resin by using 5BV 0.1mol/L hydrochloric acid solution, and when polylysine can be detected in an elution solution, collecting the eluent to obtain polylysine eluent;
(3) adjusting the pH value of the polylysine eluent obtained in the step (2) to 5.5 by using a sodium hydroxide solution, desalting the polylysine eluent in a membrane filtration device with a membrane core of 5000D until the conductivity of the polylysine eluent is less than 200mS/cm, carrying out reduced pressure concentration on the desalted polylysine solution at 60 ℃, stopping concentration when the mass concentration of polylysine is 10%, and carrying out spray drying on the concentrated solution to obtain the polylysine.
Liquid chromatography detection shows that the recovery rate of the polylysine product is 82.4 percent, and the purity is 98.1 percent.
Example 14
This example was carried out under the same conditions as example 13 except that the pH was not adjusted before the microfiltration in step (1), but the pH of the filtrate was adjusted to 8 after the microfiltration in step (2).
Liquid chromatography detection shows that the recovery rate of the polylysine product is 80.3 percent, and the purity is 95.3 percent.
Example 15
The embodiment provides a polylysine separation and purification process, which comprises the following steps:
(1) performing solid-liquid separation on the fermentation liquor with the polylysine concentration of 30g/L by adopting plate-and-frame filtration, adjusting the pH of the fermentation liquor to 8 by using sodium hydroxide, and performing microfiltration on the fermentation liquor by adopting 300KD microfiltration membrane equipment to obtain polylysine permeate;
(2) adsorbing the polylysine permeate obtained in the step (1) by using weak-acid cation exchange resin IRC50 washed by water until the pH value is less than or equal to 10, washing the resin by using purified water until an effluent is clear after the adsorption is saturated, washing the resin by using 1BV 1mol/L urea solution, then washing the resin by using water until the effluent is clear, washing the resin by using 3BV 0.5mol/L sodium chloride solution, then washing the resin by using water until the effluent is clear, then eluting the resin by using 5BV 0.15mol/L sodium hydroxide solution, and collecting an eluent to obtain polylysine eluent when polylysine can be detected in an eluent;
(3) adjusting the pH value of the polylysine eluent obtained in the step (2) to 5.5 by using a sodium hydroxide solution, desalting the polylysine eluent in a membrane filtration device with a membrane core of 3000D until the conductivity of the polylysine eluent is less than 200mS/cm, carrying out reduced pressure concentration on the desalted polylysine solution at 60 ℃, stopping concentration when the mass concentration of the polylysine is 20%, and carrying out spray drying on the concentrated solution to obtain the polylysine.
Liquid chromatography detection shows that the recovery rate of the polylysine product is 82.1 percent, and the purity is 95.8 percent.
Example 16
Polylysine is used as a noodle preservative: the method comprises the steps of taking 100mL of purified water, adding 5g of polylysine, uniformly spraying the liquid onto the surface of 10kg of fresh wet noodles, taking fresh wet noodles sprayed with the same amount of clear water as a control, respectively and normally placing for a week, comparing the appearances and the smells of two types of noodles treated differently, wherein the appearances and the smells of the fresh noodles are basically not changed after an experimental group is placed for a week, and for the control group, the noodles are sour and mildewed after being placed for 4 days, which shows that the polylysine serving as a noodle preservative can effectively prolong the preservation period of the fresh noodles.
Example 17
Polylysine is used as a fruit and vegetable preservative: brushing fresh cherries with purified water, draining, soaking in an aqueous solution containing 5% of polylysine, 1% of citric acid and 0.5% of glycine, taking out after 30min, draining, putting into a sterile bag, storing in a refrigerator at 4 ℃, evaluating the quality (color, smell and state) of the cherries according to the interval days by taking untreated fresh cherries as a reference, and indicating that the shelf life of the cherries is prolonged from 10 days to 15 days. Therefore, the polylysine serving as the fruit and vegetable preservative can greatly prolong the storage period of the fruits and vegetables.
Example 18
Polylysine was evaluated as an effect of inhibiting acnes: under aseptic operation, taking the acne bacillus suspension with proper concentration, and uniformly coating the acne bacillus suspension on a culture medium by using a coating rod; the oxford cup is lightly placed on the surface of the solidified culture medium, and the oxford cup is lightly pressed to ensure good contact. 200 μ L of 0.2 mg/mL-polylysine solution was added to the Oxford cup, incubated at 37 ℃ for 48h and removed, and the difference between the white control and the sample treatment in each dish was measured, and the results are shown in FIG. 7. The experimental result shows that-polylysine has better inhibition effect on acne bacillus and can be used as a product aiming at the acne bacillus.
Comparative example 1
This comparative example was carried out under the same conditions as in example 2 except that the pH of the polylysine permeate obtained in step (1) was not adjusted to 9 in step (2).
Liquid chromatography detection shows that the recovery rate of the polylysine product is 90.5 percent, and the purity is 90.2 percent.
Comparative example 2
This comparative example was carried out under the same conditions as in example 2 except that the elution was carried out in step (2) directly using a sodium hydroxide solution.
Liquid chromatography detection shows that the recovery rate of the polylysine product is 89.1 percent, and the purity is 95.8 percent.
The spray drying conditions used in the above examples and comparative examples were inlet air temperature of 120 deg.C and outlet air temperature of 80 deg.C, and the freeze drying conditions were vacuum degree of-0.099 MPa and cold trap temperature of-45 deg.C.
According to the examples and the test results of the comparative example, the purity of the prepared polylysine is more than 95%, and the recovery rate is more than 80% by adopting the separation and purification process of the polylysine provided by the invention. As can be seen from examples 16 to 18, polylysine has a good effect in the fields of food preservation, fruit and vegetable preservation, and antimicrobial of daily necessities. As can be seen from the comparison between examples 11 and 12 and the comparison between examples 13 and 14, the purity and recovery rate of polylysine obtained by performing pH adjustment on the fermentation broth before microfiltration and then performing the subsequent steps are superior to those of polylysine obtained by performing pH adjustment on the polylysine permeate after microfiltration and then performing the subsequent steps. In contrast, when comparative example 1 is compared with example 2, it can be seen that if the pH of the polylysine permeate is not adjusted, the purity of the finally prepared polylysine is lowered; comparison of comparative example 2 with example 2 shows that elution of the ion exchange resin directly with the eluent also reduces the purity of the polylysine produced.
The applicant declares that the present invention illustrates the detailed structural features of the present invention through the above embodiments, but the present invention is not limited to the above detailed structural features, that is, it does not mean that the present invention must be implemented depending on the above detailed structural features. It should be understood by those skilled in the art that any modifications of the present invention, equivalent substitutions of selected components of the present invention, additions of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.
The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.
It should be noted that the various technical features described in the above embodiments can be combined in any suitable manner without contradiction, and the invention is not described in any way for the possible combinations in order to avoid unnecessary repetition.
In addition, any combination of the various embodiments of the present invention is also possible, and the same should be considered as the disclosure of the present invention as long as it does not depart from the spirit of the present invention.

Claims (10)

1. A polylysine separation and purification process is characterized by comprising the following steps:
(1) carrying out solid-liquid separation and microfiltration on the fermentation liquor to obtain polylysine permeate;
(2) adsorbing the polylysine permeate obtained in the step (1) by using ion exchange resin, and eluting by using a solvent after adsorption to obtain polylysine eluent;
(3) desalting the polylysine eluent obtained in the step (2), and concentrating and drying after desalting to obtain the polylysine.
2. The process of claim 1, wherein the solid-liquid separation of step (1) comprises a continuous or batch type;
preferably, the continuous solid-liquid separation is butterfly centrifugation;
preferably, the intermittent solid-liquid separation is plate-frame filtration;
preferably, the pH is adjusted to 6-9 or not adjusted before the microfiltration.
3. The process according to claim 1 or 2, wherein the ion exchange resin of step (2) comprises a strongly acidic cation exchange resin and a weakly acidic cation exchange resin, preferably a weakly acidic cation exchange resin;
preferably, the weakly acidic cation exchange resin comprises any one or a combination of at least two of D111, D113, D151, D152, 110, 111, 112, 122, 116 or IRC-50;
preferably, the ion exchange resin is washed with water to pH less than or equal to 10 before use;
preferably, the pH of the polylysine permeate is adjusted to 6-9 before the adsorption treatment.
4. The process according to any one of claims 1 to 3, wherein the elution treatment of step (2) comprises washing and eluting the ion exchange resin after the adsorption treatment with a solvent;
preferably, the solvent used for washing comprises any one of water, ethanol solution, acetic acid solution, urea solution, sodium chloride solution or sodium acetate solution or the combination of at least two of the above solutions;
preferably, the mass concentration of the ethanol solution is 10-70%, more preferably 30-60%, and the dosage of the ethanol solution is 0-4 BV;
preferably, the concentration of the acetic acid solution is 0.05-2 mol/L, further preferably 0.1-1 mol/L, and the dosage of the acetic acid solution is 0-4 BV;
preferably, the concentration of the urea solution is 0.1-1 mol/L, further preferably 0.1-0.5 mol/L, and the dosage of the urea solution is 0-4 BV;
preferably, the concentration of the sodium chloride solution is 0.1-2 mol/L, further preferably 0.5-1.5 mol/L, and the using amount of the sodium chloride solution is 0-3 BV;
preferably, the concentration of the sodium acetate solution is 0.1-2 mol/L, further preferably 0.1-1 mol/L, and the dosage of the sodium chloride solution is 0-3 BV.
Preferably, the solvent used for elution comprises a hydrochloric acid solution and/or a sodium hydroxide solution;
preferably, the concentration of the sodium hydroxide solution is 0.05-0.5 mol/L, further preferably 0.1-0.3 mol/L, and the using amount of the sodium hydroxide solution is 0-4 BV;
preferably, the concentration of the hydrochloric acid is 0.05-0.5 mol/L, further preferably 0.1-0.3 mol/L, and the dosage of the hydrochloric acid solution is 0-5 BV.
5. The process according to any one of claims 1 to 4, wherein, prior to the desalting treatment of step (3), the pH of the polylysine eluate is adjusted to 5.5;
preferably, the desalting treatment in the step (3) is carried out by using a membrane device;
preferably, the membrane equipment adopts a roll-up membrane filter membrane core;
preferably, the molecular weight of the roll-type membrane filter membrane core is not more than 5000;
preferably, the end point of the desalting treatment of step (3) is that the conductivity of the polylysine eluate is less than 200 mS/cm.
6. The process according to any one of claims 1 to 5, wherein the concentration in the step (3) comprises preliminary concentration and concentration under reduced pressure which are carried out in this order;
preferably, the preliminary concentration is carried out in the membrane plant;
preferably, the temperature of the reduced pressure concentration is not more than 60 ℃;
preferably, the concentration of the polylysine in the solution after the concentration under reduced pressure is 5 to 30 wt%.
7. The process according to any one of claims 1 to 6, wherein the drying method of step (3) comprises any one of spray drying, freeze drying or vacuum pulse drying, preferably spray drying;
preferably, the air inlet temperature of the spray drying is 120-200 ℃, and the air outlet temperature is 70-100 ℃.
8. The process according to any one of claims 1 to 7, characterized in that it comprises the following steps:
(1) carrying out solid-liquid separation and microfiltration on the fermentation liquor to obtain polylysine permeate;
the solid-liquid separation comprises a continuous type or an intermittent type, and the pH is adjusted to 6-9 or not adjusted before microfiltration;
(2) adsorbing the polylysine permeate obtained in the step (1) by using ion exchange resin, and eluting by using a solvent after adsorption to obtain polylysine eluent;
the ion exchange resin comprises strong-acid cation exchange resin or weak-acid cation exchange resin, and the ion exchange resin is washed by water until the pH value is less than or equal to 10 before use;
before the adsorption treatment, adjusting the pH value of the polylysine permeation solution to 6-9;
the solvent comprises any one or the combination of at least two of water, hydrochloric acid, ethanol solution, acetic acid solution, sodium hydroxide solution, urea solution, sodium chloride solution or sodium acetate solution;
(3) adjusting the pH value of the polylysine eluent obtained in the step (2) to 5.5, desalting the polylysine eluent obtained in the step (2) in membrane equipment until the conductivity of the polylysine eluent is less than 200mS/cm, wherein the membrane equipment adopts a roll-type membrane filter membrane core, and concentrating and drying after desalting to obtain the polylysine.
9. Use of polylysine prepared by the process of any one of claims 1 to 8 in the field of food additives or in the field of household chemicals.
10. Use according to claim 9, wherein the field of food additives comprises biological preservatives in the field of fruits and vegetables, meat products, pasta or beverages;
preferably, the polylysine is used synergistically with one or a combination of at least two of natamycin, nisin, citric acid, acetic acid, chitosan;
preferably, the dosage of the polylysine used as a food additive is 0.01-0.3 g/kg;
preferably, the field of daily chemical products comprises an additive or an active ingredient of the polylysine for daily chemical products;
preferably, the polylysine is used as an additive for household chemicals to inhibit product spoilage;
preferably, said-polylysine inhibits acne propionibacterium, staphylococcus aureus, streptococcus hemolyticus, and pseudomonas aeruginosa;
preferably, the dosage of the polylysine used as an additive of daily chemical products is 0.01-0.3 g/L.
CN202010637151.3A 2020-07-03 2020-07-03 Polylysine separation and purification process and application Pending CN111777760A (en)

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