CN114990091A - Preparation method and application of biological enzyme preparation for ecological leather - Google Patents
Preparation method and application of biological enzyme preparation for ecological leather Download PDFInfo
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- CN114990091A CN114990091A CN202210656206.4A CN202210656206A CN114990091A CN 114990091 A CN114990091 A CN 114990091A CN 202210656206 A CN202210656206 A CN 202210656206A CN 114990091 A CN114990091 A CN 114990091A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
- C12N9/18—Carboxylic ester hydrolases (3.1.1)
- C12N9/20—Triglyceride splitting, e.g. by means of lipase
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
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- C—CHEMISTRY; METALLURGY
- C14—SKINS; HIDES; PELTS; LEATHER
- C14C—CHEMICAL TREATMENT OF HIDES, SKINS OR LEATHER, e.g. TANNING, IMPREGNATING, FINISHING; APPARATUS THEREFOR; COMPOSITIONS FOR TANNING
- C14C11/00—Surface finishing of leather
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/38—Pseudomonas
Abstract
The invention provides a preparation method and application of a biological enzyme preparation for ecological leather, belonging to the technical field of leather processing, wherein the preparation method of the biological enzyme preparation comprises the following steps: s1, pouring the culture solution into a stirring container, adding the culture solution, placing the culture solution into pseudomonas capable of producing lipase, uniformly mixing, stirring and placing for 21-24 hours to obtain a mixed solution A, adding the mixed solution into the fermentation culture solution, and stirring and mixing again to obtain a mixed solution B for later use; s2, placing the mixed solution B into a centrifuge for centrifugation, and collecting supernatant to obtain a lipase solution; s3, dripping the lipase solution into the calcium chloride solution, standing for 2-3h to form gel beads; in the scheme, the lipase prepared by taking the culture solution as the carrier has the advantages of high purity, good quality, high activity, strong permeability and higher catalytic activity function, can catalyze various esterification and transesterification reactions, and can be better applied to leather prepared from animal fur.
Description
Technical Field
The invention belongs to the technical field of leather processing, and particularly relates to a preparation method and application of a biological enzyme preparation for ecological leather.
Background
The ecological leather is a leather product with various ecological indexes meeting the requirements of ecological standards. The ecological indexes comprise the hexavalent chromium content, the formaldehyde content, the carcinogenic aromatic amine content and the pentachlorophenol content in the leather; with the improvement of living standard, people have stronger health consciousness, and pay more and more attention to green, natural and pollution-free products, which is a main factor considered by consumers, so the ecological leather will become the mainstream of the leather industry development in the future.
In the prior art, when the leather is produced, the animal fur contains certain grease components, so that the quality of the prepared leather is poor, and even the physical function of a user can be damaged.
Disclosure of Invention
The invention aims to provide a preparation method and application of a bio-enzyme preparation for ecological leather, and aims to solve the problems that in the prior art, when leather is prepared and produced, animal fur contains certain grease components, so that the quality of the prepared leather is poor, and even the harm to the body function of a user is possibly caused.
In order to achieve the purpose, the invention provides the following technical scheme:
a preparation method of a biological enzyme preparation for ecological leather comprises the following steps:
s1, pouring the culture solution into a stirring container, adding the culture solution, placing the culture solution into pseudomonas capable of producing lipase, uniformly mixing, stirring and placing for 21-24 hours to obtain a mixed solution A, adding the mixed solution into the fermentation culture solution, and stirring and mixing again to obtain a mixed solution B for later use;
s2, placing the mixed solution B into a centrifuge for centrifugation, and collecting supernatant to obtain a lipase solution;
s3, dripping the lipase solution into the calcium chloride solution, standing for 2-3h to form gel beads;
s4, adding the lipase solution obtained in the step S3 into a polyethylene glycol solution to precipitate lipase; centrifuging and collecting precipitate; and (4) dialyzing the precipitate, and freeze-drying to obtain the lipase.
In a preferred embodiment of the present invention, the culture solution in the stirring vessel in step S1 is 20L.
In a preferred embodiment of the present invention, the ratio of the mixed solution a to the fermentation culture solution in step S1 is 10: 1.
In a preferred embodiment of the present invention, the fermentation broth in step S1 is prepared by mixing yeast extract, peptone, NaCl, vegetable oil and water in any ratio.
In a preferred embodiment of the present invention, a high-speed refrigerated centrifuge is used in step S2.
In a preferred embodiment of the present invention, the high-speed refrigerated centrifuge is centrifuged at 10800g for 15min at 4 ℃.
In a preferred embodiment of the present invention, the centrifugation step S4 is performed at 10000g for 10min at 4 ℃.
The application of the biological enzyme preparation for ecological leather comprises the following steps:
firstly, putting the skin embryo into water, adding lipase, and soaking;
step two, performing drum throwing on the backwater leather blank obtained in the step one, and vibrating and softening the backwater leather blank to obtain vibrating and softening leather so as to enable the leather blank to return to a flat state;
and step three, polishing, spraying paint, rolling surface oil and drying the leather blank to obtain the ecological leather degreased by the lipase.
In a preferred embodiment of the invention, the ratio of the skin germ, the lipase and the water in the first step is 1 (0.5-1) to 5.
As a preferable scheme of the invention, in the first step, the skin embryo is soaked in water with lipase for 20-24 h.
Compared with the prior art, the invention has the beneficial effects that:
in the scheme, the lipase prepared by taking the culture solution as the carrier has the advantages of high purity, good quality, high yield, high activity, strong permeability and higher catalytic activity function, and can catalyze various esterification and transesterification reactions; in the leather applied to the preparation of the animal fur, as the animal fur contains certain grease components, the lipase disclosed by the invention can better hydrolyze grease molecules, so that a grease layer can be quickly removed, the harm to a human body can be reduced, the generation of polluted wastewater can be reduced, the effect is better, and the application prospect is wide.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention and not to limit the invention. In the drawings:
FIG. 1 is a flow chart of a method for preparing a bio-enzyme preparation for ecological leather in the invention;
FIG. 2 is a flow chart of the application of the bio-enzyme preparation for ecological leather in the invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
Referring to fig. 1, the present invention provides the following technical solutions:
a preparation method of a biological enzyme preparation for ecological leather comprises the following steps:
s1, pouring the culture solution into a stirring container, adding the culture solution, placing the culture solution into pseudomonas capable of producing lipase, uniformly mixing, stirring and placing for 21-24 hours to obtain a mixed solution A, adding the mixed solution into the fermentation culture solution, stirring and mixing again to obtain a mixed solution B for later use;
s2, placing the mixed solution B into a centrifuge for centrifugation, and collecting supernatant to obtain a lipase solution;
s3, dripping the lipase solution into the calcium chloride solution, standing for 2-3h to form gel beads;
s4, adding the lipase solution obtained in the step S3 into a polyethylene glycol solution to precipitate lipase; centrifuging and collecting precipitate; and (4) dialyzing the precipitate, and freeze-drying to obtain the lipase.
In the specific embodiment of the invention, a single colony of pseudomonas capable of producing lipase is placed in a culture solution for soaking, the culture solution is placed in a stirring container and is stirred for 23 hours all the time, so that lipase produced by pseudomonas can be better fused with the culture solution to obtain a mixed solution A, then the mixed solution A is placed in a fermentation culture solution for mixing, so that the lipase in the mixed solution A can be fermented to obtain a mixed solution B, the mixed solution B is placed in a high-speed freezing centrifuge for centrifugation to obtain a supernatant, namely a lipase solution, the lipase solution is then dripped into a calcium chloride solution, and the mixture is kept stand for 2-3 hours until gel globules are formed; adding the lipase solution into the polyethylene glycol solution to precipitate the lipase; centrifuging and collecting precipitate; the lipase prepared by using the culture solution as a carrier has the advantages of high purity, good quality, high yield, high activity, strong permeability and higher catalytic activity, can catalyze various esterification and transesterification reactions, and can be better applied to leather prepared from animal fur.
Specifically, referring to fig. 1, the volume of the culture solution in the stirred vessel in the step S1 is 20L.
Specifically, referring to fig. 1, the ratio of the mixed solution a to the fermentation culture solution in the step S1 is 10: 1.
Specifically, referring to fig. 1, the fermentation broth in step S1 is prepared by mixing yeast extract, peptone, NaCl, vegetable oil and water in any ratio.
Specifically, referring to fig. 1, in step S2, a high-speed freezing centrifuge is used.
Specifically, referring to FIG. 1, the high speed refrigerated centrifuge is centrifuged at 10800g for 15min at 4 ℃.
Specifically, referring to FIG. 1, in step S4, centrifugation is performed at 10000g for 10min at 4 ℃.
Example 2
The embodiment 2 provides an application of the bio-enzyme preparation for ecological leather, and specifically further illustrates how the lipase prepared in the embodiment 1 is applied to the working principle or method of the preparation of ecological leather, as follows:
the application of the biological enzyme preparation for ecological leather comprises the following steps:
step one, placing the skin embryo in water, adding lipase, soaking, wherein the ratio of the skin embryo to the lipase to the water is 1:0.5:5, and placing the skin embryo in the water with the lipase for soaking for 24 hours;
step two, performing drum throwing on the backwater leather blank obtained in the step one, and vibrating and softening the backwater leather blank to obtain vibrating and softening leather so as to enable the leather blank to return to a flat state;
and step three, polishing, spraying paint, rolling surface oil and drying the leather blank to obtain the ecological leather degreased by the lipase.
The lipase disclosed by the invention can better hydrolyze grease molecules, so that a grease layer can be quickly removed, the harm to a human body and the generation of polluted wastewater can be reduced, the effect is better, and the application prospect is wide.
Example 3
The embodiment 3 provides an application of the bio-enzyme preparation for ecological leather, and specifically further illustrates how the lipase prepared in the embodiment 1 is applied to the working principle or method of ecological leather preparation, and the preparation method is the same as that of the embodiment 2 except that the weight parts of the skin germ, the lipase and the water are different from each other and the soaking time is different from each other, and specifically comprises the following steps:
the application of the biological enzyme preparation for the ecological leather comprises the following steps:
step one, placing the skin embryo in water, adding lipase, soaking, wherein the ratio of the skin embryo to the lipase to the water is 1:0.75:5, and placing the skin embryo in the water with the lipase for soaking for 22 hours;
step two, performing drum throwing on the backwater leather blank obtained in the step one, and vibrating and softening the backwater leather blank to obtain vibrating and softening leather so as to enable the leather blank to return to a flat state;
and step three, polishing, spraying paint, rolling surface oil and drying the leather blank to obtain the ecological leather degreased by the lipase.
Example 4
This embodiment 4 provides an application of the bio-enzyme preparation for ecological leather, and specifically further illustrates how the lipase prepared in example 1 is applied to the working principle or method of ecological leather preparation, and the preparation method is the same as that of example 2 except that the weight ratio of the skin germ, the lipase and the water and the soaking time are different, and specifically includes the following steps:
the application of the biological enzyme preparation for the ecological leather comprises the following steps:
step one, placing the skin embryo in water, adding lipase, soaking, wherein the ratio of the skin embryo to the lipase to the water is 1:1:5, and placing the skin embryo in the water with the lipase for soaking for 20 hours;
step two, performing drum throwing on the backwater leather blank obtained in the step one, and vibrating and softening the backwater leather blank to obtain vibrating and softening leather so as to enable the leather blank to return to a flat state;
and step three, polishing, spraying paint, rolling surface oil and drying the leather blank to obtain the ecological leather degreased by the lipase.
In the scheme, the lipase prepared by taking the culture solution as a carrier has the advantages of high purity, good quality, high yield, high activity, strong permeability and higher catalytic activity function, and can catalyze various esterification and transesterification reactions; in the leather applied to the preparation of the animal fur, as the animal fur contains certain grease components, the lipase disclosed by the invention can better hydrolyze grease molecules, so that a grease layer can be quickly removed, the harm to a human body can be reduced, the generation of polluted wastewater can be reduced, the effect is better, and the application prospect is wide.
Example 5
Preferably, in the application of the biological enzyme preparation for the ecological leather,
1) the method is used for degreasing softened sheep wool clothing leather and comprises the following steps:
the liquor ratio is 150%, the temperature is 38-40 ℃, the ammonium-free deliming agent is 0.3-0.5%, the rotation is carried out for 40 minutes, lipase is added for 0.3-1.5%, the degreasing agent is added for 0.5-1.0%, the rotation is carried out for 30-60 minutes, after the degreasing effect is achieved, the water is drained, the water is washed by 200% warm water at 38-40 ℃ for 10 minutes, the water is drained, the water is washed by 200% water at 28-30 ℃ for 10 minutes, the water is drained, the degreasing is finished, and the next operation is carried out;
due to the large oil content of the sheep skin, the degreasing difficulty is high. The biological lipase prepared by the method has good degreasing effect, can reduce the dosage of a degreasing agent by 30-50%, completely saves kerosene, saves tanning cost and reduces environmental pollution at the same time.
2) The soaking water for the salted cattle wet skin comprises the following specific steps:
main soaking: the liquid ratio is 200%, the water temperature is 18-22 ℃, 0.3-0.6% of soda ash and 0.05-0.15% of bactericide are added, the rotation is carried out for 20 minutes, 0.03-0.5% of lipase and 0.1-0.3% of soaking auxiliary agent are added, the rotation is carried out for 30 minutes, the rotation is carried out for 5-10 minutes every hour for 12-16 hours, the rotation is carried out for 20 minutes the next day, the soaking and softening effect is checked, the hip cut is required to be white without yellow core, the skin is unfolded without greasy feeling, and after the soaking and degreasing effect is achieved, the water is drained and the skin is taken out, and the meat removing operation is carried out.
The lipase is used for main soaking of the salted and wet cowhide, the dosage of the degreasing agent can be saved by 50-80%, the soaking time is shortened by 4-6 hours, the COD content of the wastewater is reduced by 20-40%, and the soaked cowhide treated by the lipase is easy to remove meat and hair roots are also appropriately loosened, thereby being beneficial to the subsequent unhairing process.
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A preparation method of a biological enzyme preparation for ecological leather is characterized by comprising the following steps:
s1, pouring the culture solution into a stirring container, adding the culture solution, placing the culture solution into pseudomonas capable of producing lipase, uniformly mixing, stirring and placing for 21-24 hours to obtain a mixed solution A, adding the mixed solution into the fermentation culture solution, and stirring and mixing again to obtain a mixed solution B for later use;
s2, placing the mixed solution B into a centrifuge for centrifugation, and collecting supernatant to obtain a lipase solution;
s3, dripping the lipase solution into the calcium chloride solution, standing for 2-3h to form gel beads;
s4, adding the lipase solution obtained in the step S3 into a polyethylene glycol solution to precipitate lipase; centrifuging and collecting precipitate; and (4) dialyzing the precipitate, and freeze-drying to obtain the lipase.
2. The method for preparing bio-enzyme preparation for ecological leather, according to claim 1, wherein the culture solution in the stirring container in step S1 is 20L.
3. The method for preparing the bio-enzyme preparation for ecological leather, according to claim 1, wherein the ratio of the mixed solution A to the fermentation culture solution in the step S1 is 10: 1.
4. The preparation method and application of the bio-enzyme preparation for ecological leather according to claim 1, wherein the fermentation culture solution in the step S1 is prepared by mixing yeast extract, peptone, NaCl, vegetable oil and water in any proportion.
5. The method for preparing the bio-enzyme preparation for ecological leather, according to claim 1, wherein a high-speed refrigerated centrifuge is adopted in the step S2.
6. The method for preparing the bio-enzyme preparation for ecological leather, according to claim 5, wherein the high speed refrigerated centrifuge is used for centrifuging for 15min at 4 ℃ and 10800 g.
7. The method for preparing bio-enzyme preparation for ecological leather, according to claim 5, wherein in step S4, centrifugation is performed at 10000g for 10min at 4 ℃.
8. The application of the biological enzyme preparation for ecological leather is characterized by comprising the following steps:
firstly, putting a skin embryo into water, adding lipase, and soaking;
step two, performing drum throwing on the backwater leather blank obtained in the step one, and vibrating and softening the backwater leather blank to obtain vibrating and softening leather so as to enable the leather blank to return to a flat state;
and step three, polishing, spraying paint, rolling surface oil and drying the leather blank to obtain the ecological leather degreased by the lipase.
9. The application of the bio-enzyme preparation for ecological leather as claimed in claim 8, wherein the ratio of the leather germ, the lipase and the water in the first step is 1 (0.5-1) -1: 5.
10. The use of the bio-enzyme preparation for ecological leather, according to claim 8, wherein in the first step, the leather embryo is soaked in the water with lipase for 20-24 h.
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| CN202210656206.4A CN114990091A (en) | 2022-06-10 | 2022-06-10 | Preparation method and application of biological enzyme preparation for ecological leather |
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| CN202210656206.4A CN114990091A (en) | 2022-06-10 | 2022-06-10 | Preparation method and application of biological enzyme preparation for ecological leather |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1944629A (en) * | 2006-10-26 | 2007-04-11 | 中国农业科学院油料作物研究所 | Full cell catalyst of intracellular secretory lipase type and its preparing method |
| CN109735517A (en) * | 2019-03-12 | 2019-05-10 | 内蒙古博格农牧业开发有限责任公司 | Using the method for immobilization horse antibody extraction purification lipase |
| CN111575417A (en) * | 2020-05-21 | 2020-08-25 | 中牛集团有限公司 | Water-saving processing technology for cow leather and wet blue leather |
| CN112391376A (en) * | 2020-11-03 | 2021-02-23 | 天津科技大学 | Immobilized lipase hybrid nanoflower and preparation method and application thereof |
| CN113403298A (en) * | 2021-06-09 | 2021-09-17 | 万华化学集团股份有限公司 | Immobilization method of free enzyme |
| CN114196590A (en) * | 2021-12-23 | 2022-03-18 | 福瑞莱环保科技（深圳）股份有限公司 | Pseudomonas secreting lipase and application of pseudomonas in kitchen wastewater treatment |
-
2022
- 2022-06-10 CN CN202210656206.4A patent/CN114990091A/en active Pending
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| CN1944629A (en) * | 2006-10-26 | 2007-04-11 | 中国农业科学院油料作物研究所 | Full cell catalyst of intracellular secretory lipase type and its preparing method |
| CN109735517A (en) * | 2019-03-12 | 2019-05-10 | 内蒙古博格农牧业开发有限责任公司 | Using the method for immobilization horse antibody extraction purification lipase |
| CN111575417A (en) * | 2020-05-21 | 2020-08-25 | 中牛集团有限公司 | Water-saving processing technology for cow leather and wet blue leather |
| CN112391376A (en) * | 2020-11-03 | 2021-02-23 | 天津科技大学 | Immobilized lipase hybrid nanoflower and preparation method and application thereof |
| CN113403298A (en) * | 2021-06-09 | 2021-09-17 | 万华化学集团股份有限公司 | Immobilization method of free enzyme |
| CN114196590A (en) * | 2021-12-23 | 2022-03-18 | 福瑞莱环保科技（深圳）股份有限公司 | Pseudomonas secreting lipase and application of pseudomonas in kitchen wastewater treatment |
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