CN101265470A - Inducement and preparation of S. keratinase and method for sorting wool by using the same - Google Patents
Inducement and preparation of S. keratinase and method for sorting wool by using the same Download PDFInfo
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- CN101265470A CN101265470A CNA2008100372598A CN200810037259A CN101265470A CN 101265470 A CN101265470 A CN 101265470A CN A2008100372598 A CNA2008100372598 A CN A2008100372598A CN 200810037259 A CN200810037259 A CN 200810037259A CN 101265470 A CN101265470 A CN 101265470A
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Abstract
The invention relates to a method for inducing and preparing an S. keratinase and a method for processing wool. The method for preparing the S. keratinase comprises the steps of using Stenotrophomonas maltophilia DHHJ as the original strain; culturing for 2-4 days as a generation at the constant temperature of 30-50DEG C when the pH value is equal to 7.5 by using 1-3g of wool, 0.2-1g of sodium chloride, 0.2-1g of disodium hydrogen phosphate, 0.01-0.1g of monopotassium phosphate and 50-200ml of water; continuously inducing and domesticating; culturing for 2-4days at the constant temperature of 30-50 DEG C when the pH value is equal to 7.5 by using 1-3g of chicken feather meal (or other feather meal), 0.2-1g of sodium chloride, 0.2-1g of disodium hydrogen phosphate, 0.01-0.1g of monopotassium phosphate and 50-200ml of water after efficient and stable strains are obtained so as to obtain enzyme culture solution; subjecting the culture solution to centrifugation, salting out, standing, centrifugation and purification to obtain the S. keratinase. The wool processing technique comprises the step of processing the wool and the textile thereof for 60-120min by using purified and concentrated S. keratinase with the pH value being equal to 7.8, the temperature being 20-50 DEG C, and enzyme concentration being 2-5% (owf). The S. keratinase can directly act on the wool so as to eliminate the chemical agent processing step, and be used for wool shrink-resistant processing with more environmental friendliness and high efficiency. The processed wool with enzyme has the same characteristics as cashmere in many aspects, and can be used for the processing and the production of cashmere imitation products.
Description
Technical field
The invention belongs to the field with the wool adjustment method of producing of microbial acclimation, enzyme, particularly relate to a kind of method of inducing, produce and using its sorting wool of S. M-Zyme
Background technology
Wool and fabric thereof can improve its felting resistance by arrangement, can also improve and improve woolen gloss and whiteness, feel, reduce the sensation of pricking of human body skin etc.
At present, the woolen felt proofing mostly adopts chlorination-resin technology, it can effectively improve woolen felt proofing performance and wearing comfort, be a kind of technology of comparative maturity, but it should be noted that carcinogenic character, non-biodegradation that adsorbability halogenide AOX that it produces had are very big to environmental hazard: will produce the AOX of 350g with chlorination-resin art breading 1t wool.In today of universe's pay attention to day by day environmental problem, the space of the industry of this high pollution and technology existence is more and more littler, so it is imperative to develop a kind of felt proofing technology of green.And enzyme is as protein, can be by Natural Degradation; And react single-minded, side reaction is few, does not produce pollution in theory, has obtained in recent years widely using in textile industry, is considered to substitute the method for the tool potentiality of chlorination of wool felt proofing technology.
But present this type of studied some difficulties that run into more or less: the woolen scale layer is the major cause that wool fabric produces felting, and it accounts for wool total amount 10%.Woolen scale top layer is made up of lipoid layer and the egg white layer under it.The lipoid layer has hydrophobic interaction, and cystine is more in the protein layer under it, makes it have a large amount of two sulfur-crosslinked and acid amides crosslinked [1～2] (referring to N L R King and J H Bradbury.Aust.J.Biol.Sci..1968 (21): 375 and C M Carr.I H Leaverand A E Hughes.Test.Res.J..1986 (56): 457).This makes the scale top layer have very strong chemical resistance, cause wool to be difficult to be subjected to the direct effect [3～4] of enzyme (referring to " textile journal " 2002.2: " proteolytic enzyme is to the woolen Study on mechanism " and J D Leeder.Wool Sci.Rev..1986 (63): 13), so the anti-felting effect very limited [5] of directly using the acquisition of protease treatment wool is (referring to Middlbrook W R, Phillips H. " The application of enzymes to the productionof shrinkage-resistant wool and mixture fabrics " [J] .J.S.D.C, 1941, (57): 137-144.).Someone thinks, carry out modification with the protease treatment wool, pre-treatment (oxidation or reduction) is must step, the proteolytic enzyme of also not finding at present can be hydrolyzed to wool without pre-treatment [6] (referring to " Jiangsu silk " 2006.5: " proteolytic enzyme is in the application aspect the Wool fiber modification ").Arrangement is a prerequisite with chemical pre-treatment often so present enzyme is to wool, and it mainly breaks disulfide bond crosslinking by the method (as H2O2) of oxidation, makes it to form the electrophilic sulfonic acid group.And then use biological enzyme to handle it, to obtain anti-preferably felting performance.Like this, used the initial purpose of biological enzyme not only not reach, compared a step with chlorination-resin method many on the contrary, its industrial prospect is had a greatly reduced quality.
So this achievement in research has certain breakthrough meaning.
Summary of the invention
Technical problem to be solved by this invention provides a kind of method of inducing, produce and using its sorting wool of S. M-Zyme, and present method can directly directly be handled wool, and cost is low, and is free from environmental pollution.
The method of inducing, produce and use its sorting wool of a kind of S. M-Zyme of the present invention comprises:
(1) producing of S. M-Zyme: to have a liking for maltose oligotrophy Zymomonas mobilis (Stenotrophomonas maltophilia) DHHJ is original strain (inoculum size 3～5%), use wool 1～3g, 0.2～1g sodium-chlor, 0.2～1g Sodium phosphate dibasic, 0.01～0.1g potassium primary phosphate, 50～200ml water, at pH=7.5,30～50 ℃, constant temperature is cultivated 2～4 days down as a generation, constantly induces, tames.Wait to obtain efficient, stablize bacterial classification after, use 1～3g chicken feather powder (or other feather meals), 0.2～1g sodium-chlor, 0.2～1g Sodium phosphate dibasic, 0.01～0.1g potassium primary phosphate, 50～200ml water, at pH=7.5,30～50 ℃, constant temperature was cultivated 2～4 days down, obtained to contain the enzyme nutrient solution.This nutrient solution through centrifugal, saltout, leave standstill, more centrifugal, purify after, make the S. M-Zyme;
(2) wool treatment process: the S. M-Zyme that obtains is purified, concentrated, at pH=7.8,20～50 ℃, when enzyme concn is 2～5% (owf) to wool and fabric treating 60～120min thereof, use 280nm uv-spectrophotometric instrument and microscope to detect, and the wool fiber of handling through enzyme carried out the parametric measurement of reduction rate, felting rate, alkali solubility, intensity and whiteness, prove macroscopical treatment effect.
The described original strain of step (1) screens from septic chicken feather, has degradation of feather by using Keratin sulfate ability.
The preferred culture condition of step (1): at pH=7.5,40 ℃, constant temperature culture 3 days.
It is that inductor is tamed that step (1) adopts wool, and the employing chicken feather is that raw material is cultivated and produced.
The preferred treatment condition of step (2) are enzyme concn 3% (owf), and pH=7.8 handles 75min for 40 ℃.
The every index of woolen all is better than untreated wool after the experimental data display process that step (2) is measured, and is used for the processing and the production of more environmental protection and anti-felting arrangement of wool efficiently and imitative cashmere product.
Adopting chicken feather is the bacterial isolates (denomination of invention: a kind of bacterial strain and application thereof of producing M-Zyme that raw material screening is had a liking for maltose oligotrophy Zymomonas mobilis (Stenotrophomonas maltophilia) DHHJ, the patent No.: 2007101719394): a large amount of feathers that will collect from market are embedded in outdoor than enrichment culture the damp soil, until feather tangible degraded is arranged, during keep competent moisture.Get above-mentioned rotten chicken feather 1g in enrichment medium 1 (extractum carnis 0.5g, peptone 1g, sodium-chlor 0.5g, pH7.4-7.6,100mL) in, in 28 ℃, 120 commentaries on classics/min constant temperature culture 48h.Get again its nutrient solution 2mL change over to enrichment medium 2 (Zulkovsky starch 2g, feather meal a 2g, sodium-chlor 0.5g, pH7.4-7.6,100mL) in, 28 ℃, the further enrichment culture 48h of 120r/min.Getting whole pregnant solution 100 μ L coats phosphoric acid and clears up protein culture medium (Zulkovsky starch 2g, phosphoric acid digestible protein 2.5g, agar powder 2g, sodium-chlor 0.5g, pH7.4-7.6,100mL) flat board, filter out the single strain of growth, in contrast with beef-protein medium (perfect medium).The bacterial classification that primary dcreening operation is obtained is selected respectively and is connected to phosphoric acid and clears up dull and stereotyped and feather meal plate culture medium (the Zulkovsky starch 2g of protein culture medium, feather meal b2g, agar powder 2g, sodium-chlor 0.5g, pH 7.4-7.6,100mL) flat board is in 33 ℃ of following constant temperature culture 48h, growth on controlled observation two flat boards, selected effective strain.Again these effective strains are coated phosphoric acid and clear up the protein culture medium flat board, repeat to do several times further separation and purification.
Wherein, the treatment process of feather meal: feather meal a cleans chicken feather with washing composition, after clear water is cleaned, boiled 30 minutes with 0.05% sodium hydroxide solution and 0.05% hydrochloric acid soln respectively, fully clean with clear water again, in 80 ℃ of oven dry 48h, with pulverizer with its separated pulverizing after, cross 60 mesh sieves; Feather meal b cleans chicken feather with washing composition, and clear water is cleaned the back in Autoclave inner high voltage boiling 1h, and is fully clean with clear water again, in 80 ℃ of oven dry 48h, with pulverizer with its separated pulverizing after, cross 100 mesh sieves.
Beneficial effect
1, the S. M-Zyme produced of the present invention can directly act on wool, has saved the chemical reagent treatment step, has avoided contaminate environment;
2, enzyme extraction process using chicken feather is a main raw material, saves cost, and is efficiently feasible, is easy to industrialization promotion;
3, the anti-felting property of woolen, whiteness, the physical strength of enzyme processing are improved; Feel is improved, and the pungency decline to human body skin has the many-sided advantage of cashmere.
Description of drawings
Fig. 1 has a liking for the S. M-Zyme of maltose oligotrophy Zymomonas mobilis generation to wool treatment effect microscope comparison diagram (left side figure is the wool that is untreated, and right figure is the wool after handling);
Fig. 2 has a liking for the S. M-Zyme of maltose oligotrophy Zymomonas mobilis generation to wool treatment effect microscope comparison diagram (left side figure is the wool that is untreated, and right figure is the wool after handling).
Embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in and limit the scope of the invention.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after the content of having read the present invention's instruction, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
Embodiment 1
To have a liking for maltose oligotrophy Zymomonas mobilis (Stenotrophomonas maltophilia) DHHJ is original strain, use wool to be inductor, determine optimal culture condition by orthogonal experiment, and it is induced, tames cultivation: wool 2g, sodium-chlor 0.4g, Sodium phosphate dibasic 0.4g, potassium primary phosphate 0.03g, water 100ml, pH=7.5,40 ℃, constant temperature culture 2～3 days.
After domestication for several times, obtain efficient, stable bacterial classification, use following cultivation ratio to carry out amplification culture: chicken feather 2g, sodium-chlor 0.4g, Sodium phosphate dibasic 0.4g, potassium primary phosphate 0.03g, water 100ml, pH=7.5,40 ℃, constant temperature culture 72 hours obtains containing the nutrient solution of enzyme.
Embodiment 2
The nutrient solution that obtaining of obtaining is contained enzyme through centrifugal, saltout, leave standstill, more centrifugal, purify, concentrate after, wool is handled, used 280nm uv-spectrophotometric instrument and microscope to detect, obtain optimal treatment condition: enzyme concn 3% (owf), pH=7.8 handles 75min for 40 ℃.
Measure parameter values such as reduction rate, felting rate, alkali solubility, intensity, whiteness, prove that it is to woolen macroscopic view treatment effect.
Embodiment 3
Behind S. M-Zyme processing wool fiber, analyze the variation of reduction rate, felting rate, alkali solubility, intensity and whiteness.The S. M-Zyme is discussed to the woolen mode of action, the influence coefficient of fibre-tendering and fabric intensity and the feasibility of large-scale application are probed into.
One, reduction rate is measured
The mensuration of fabric reduction rate: the sample of enzyme being handled front and back dries to constant weight in 105 ℃ of baking ovens.
Reduction rate (%)=(1-handles the preceding dry fabric weight of back dry fabric weight/processing) * 100
Two, the felting rate is measured
The mensuration of felting rate: soap flakes 1g/L, bath raio 1: 30,30 ℃ of washing lotion temperature, washing time 1h, oven dry, treat to be calculated as follows after the fabric moisture balance:
Felting rate (%)=(1-washes the preceding fabric area of back fabric area/wash) * 100
Three, alkali solubility is measured
The damage of wool fiber adopts alkali solubility method to measure:
Alkali solubility (%)=(2.000 * (1-G) W)/(2.000 * (1-G)) * 100
In the formula, G-water ratio (%), residual samples weighed (g) after the W-alkaline purification
Four, strength detection
Fabric intensity is measured: measure by standard method on strength tester.
Five, measuring brightness
The mensuration of whiteness: on intellectual colourity whiteness instrument, measure by standard method.
From the said determination result as can be seen:
1, reduction rate is compared obviously with control group, but shows that to the stable tendency that to a certain degree presents enzyme has its treating part and treatment effect better to wool, and the decomposition by enzyme has improved water-soluble protein content in the solution, just shows as the increase of reduction rate.Increase weight-loss ratio in time and be half n type curve, comparing with control group is more than 50 times of no enzymatic reaction, and obvious wool processing power is arranged.
2, compare with control group, through experiment test repeatedly, the wool felt shrinkage after enzyme is handled prolonged and significantly reduces with the reaction times, and 6 percentage points of decreased average have the using value of pair actual treatment.
3, owing to having the outer scale layer, wool itself is insoluble in alkaline solution, therefore peelling off the top layer scale layer can improve the woolen caustic solubility, experimental data shows that the wool fiber after enzyme is handled more is soluble in alkaline solution, and solubleness is proportionate in time, these data illustrate that equally wool through handling, has reached felt proofing, improved flexible purpose.
4, the wool intensity contrast situation that records by tension test shows that wool has a little damage, and intensity reduces, but degree of injury according to domestic and international technical literature record, this experiment woolen intensity has reached good level, and intensity is reduced in the excellent scope, and does not have the apparent damage phenomenon of rupture.Enzyme processes and displays wool intensity is half u type with the processing time, and its fall is minimum.
5, whiteness does not have obvious decline, at tolerance interval, and can carry out the resin bleaching after enzyme is handled usually and handle, and experiment conclusion is had no effect.
In sum, S. M-Zyme has tangible processing power to wool, the every index of woolen all is better than untreated wool after the multinomial experimental data display process, can also improve and improve woolen gloss, dough kneading sensation and feel, minimizing is to the sensation of pricking of human body skin, have great application prospect, this technology should be committed to as early as possible the actual production life.
Claims (5)
1. the method for inducing, produce and use its sorting wool of a S. M-Zyme comprises:
(1) producing of S. M-Zyme: to have a liking for maltose oligotrophy Zymomonas mobilis (Stenotrophomonas maltophilia) DHHJ is original strain, inoculum size 3～5%, use wool 1～3g, 0.2～1g sodium-chlor, 0.2～1g Sodium phosphate dibasic, 0.01～0.1g potassium primary phosphate, 50～200ml water, at pH=7.5,30～50 ℃, constant temperature is cultivated 2～4 days down as a generation, constantly induces, tames; Wait to obtain efficient, stablize bacterial classification after, use 1～3g chicken feather powder or other feather meals, 0.2～1g sodium-chlor, 0.2～1g Sodium phosphate dibasic, 0.01～0.1g potassium primary phosphate, 50～200ml water, at pH=7.5,30～50 ℃, constant temperature was cultivated 2～4 days down, obtained to contain the enzyme nutrient solution; This nutrient solution through centrifugal, saltout, leave standstill, more centrifugal, purify after, make the S. M-Zyme;
(2) wool treatment process: the S. M-Zyme that obtains is purified, concentrated, at pH=7.8,20～50 ℃, when enzyme concn is 2～5% (owf) to wool and fabric treating 60～120min thereof, use 280nm uv-spectrophotometric instrument and microscope to detect, and the wool fiber of handling through enzyme carried out the parametric measurement of reduction rate, felting rate, alkali solubility, intensity and whiteness, prove macroscopical treatment effect.
2. the method for inducing, produce and use its sorting wool of S. M-Zyme according to claim 1 is characterized in that: the described original strain of step (1) screens from septic chicken feather, has degradation of feather by using Keratin sulfate ability.
3. the method for inducing, produce and use its sorting wool of S. M-Zyme according to claim 1 is characterized in that: the described optimal culture conditions of step (1): at pH=7.5, and 40 ℃, constant temperature culture 3 days.
4. according to the method for inducing, produce and use its sorting wool of claim 1 or 3 described S. M-Zymes, it is characterized in that: it is that inductor is tamed that step (1) adopts wool, and the employing chicken feather is that raw material is cultivated and produced.
5. the method for inducing, produce and use its sorting wool of S. M-Zyme according to claim 1 is characterized in that: the described treatment condition of step (2) are enzyme concn 3% (owf), and pH=7.8 handles 75min for 40 ℃.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102329751A (en) * | 2011-09-21 | 2012-01-25 | 江南大学 | Stenotrophomonas maltophilia for generating keratinase and application of stenotrophomonas maltophilia |
| CN104099371A (en) * | 2014-07-18 | 2014-10-15 | 辽宁师范大学 | Application of KAP26.1 gene as exogenous gene introduced into cashmere goat cells and used for improving wool fineness |
| CN105040454A (en) * | 2015-07-23 | 2015-11-11 | 东华大学 | Biological treatment method for wool blend fabric |
| CN107177986A (en) * | 2017-05-24 | 2017-09-19 | 无锡协新毛纺织股份有限公司 | A kind of method for carrying out feld proofing to wool fabric using modified protein enzyme |
| CN107723263A (en) * | 2017-10-24 | 2018-02-23 | 东华大学 | A kind of screening system for different keratin degrading ability microbial strains |
-
2008
- 2008-05-09 CN CNA2008100372598A patent/CN101265470A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102329751A (en) * | 2011-09-21 | 2012-01-25 | 江南大学 | Stenotrophomonas maltophilia for generating keratinase and application of stenotrophomonas maltophilia |
| CN102329751B (en) * | 2011-09-21 | 2013-07-24 | 江南大学 | Stenotrophomonas maltophilia for generating keratinase and application of stenotrophomonas maltophilia |
| CN104099371A (en) * | 2014-07-18 | 2014-10-15 | 辽宁师范大学 | Application of KAP26.1 gene as exogenous gene introduced into cashmere goat cells and used for improving wool fineness |
| CN105040454A (en) * | 2015-07-23 | 2015-11-11 | 东华大学 | Biological treatment method for wool blend fabric |
| CN107177986A (en) * | 2017-05-24 | 2017-09-19 | 无锡协新毛纺织股份有限公司 | A kind of method for carrying out feld proofing to wool fabric using modified protein enzyme |
| CN107177986B (en) * | 2017-05-24 | 2019-11-08 | 无锡协新毛纺织股份有限公司 | A method of feld proofing is carried out using modification proteases on wool fabric |
| CN107723263A (en) * | 2017-10-24 | 2018-02-23 | 东华大学 | A kind of screening system for different keratin degrading ability microbial strains |
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Open date: 20080917 |