CN104250856A - Preparation method of hibiscus cannabinus fiber by using lysine bacillus DY2 strain for retting - Google Patents
Preparation method of hibiscus cannabinus fiber by using lysine bacillus DY2 strain for retting Download PDFInfo
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- CN104250856A CN104250856A CN201410334135.1A CN201410334135A CN104250856A CN 104250856 A CN104250856 A CN 104250856A CN 201410334135 A CN201410334135 A CN 201410334135A CN 104250856 A CN104250856 A CN 104250856A
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- retted fibre
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Abstract
The invention relates to a preparation method of hibiscus cannabinus fiber by using lysine bacillus strain DY2 strain for retting. The method includes steps of: (1) culturing lysine bacillus strain DY2 strains in a beef extract peptone medium, and adding a cryopreservation buffer; (2) inoculating the bacterium ring obtained in the step (1) to a hibiscus cannabinus bran medium, and shaking for culture; (3) inoculating the medium obtained in the step (2) to a hibiscus cannabinus bran fermentation culture medium, and conducting shake-flask fermentation to obtain a fermented bacteria liquid; (4) making a retting pool; and (5) sending the fermented bacteria liquid prepared in the step (3) to the retting pool for retting treatment, so as to prepare the hibiscus cannabinus fiber. The invention has the advantages of simple process and short production flow, and is applicable to large-scale industrialized production.
Description
Technical field
The invention belongs to retted fibre field, particularly a kind of method utilizing lysine bacillus DY2 bacterial strain retted fibre to prepare bastose.
Background technology
Fiber crops are one of textile raw materials, and its kind mainly contains flax, ramie, hemp, bluish dogbane, jute, bluish dogbane, sisal hemp, abaca, nettle and piemarker etc.Flax fibre is the general designation of the fiber obtained from various numb plant, has the advantage that moisture absorption, ventilative, heat radiation and other natural fabrics such as antibacterial are incomparable.China is as one of the abundantest country of hemp resource in the world, and the annual flax fibre produced and goods thereof, except the demand meeting domestic market, also as the export of commodities in large amounts U.S., West Europe and country in Southeast Asia, earn foreign exchange nearly 10,000,000,000 dollars every year.Reduce discharging policy and consumer's back to nature at national energy-saving, pursue under green consumption idea promotes, bast-fibre green processing technologies has become the study hotspot of field of textiles.
Retted fibre utilizes bacterium and moisture to the effect of plant, dissolves or corrodes the cellular connective tissue of major part and colloid that are enclosed in outside bast fibre bundle, thus impelling the process that fiber is separated with straw.In the bubble method of extensive use, straw is immersed in water, and water permeation, to the middle body of straw, makes inner cell expansion, bursts outermost layer, thus increases the absorption to moisture and bacterium.Process of retting flax with is the processing technology of producing good fiber quality, and the bacterial classification selection of retted fibre process is the key of retted fibre.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of method utilizing lysine bacillus DY2 bacterial strain retted fibre to prepare bastose, and the method is simple, production procedure is short, be applicable to large-scale industrial production.
A kind of method utilizing lysine bacillus DY2 bacterial strain retted fibre to prepare bastose of the present invention, comprising:
(1) the lysine bacillus DY2 bacterial strain of preservation is cultivated in beef-protein medium, add frozen buffer solution;
(2) in bluish dogbane bran mass, bacterium one ring of step (1) gained is accessed, Shaking culture; The addition of bacterium one ring is that every 1000ml bacterium liquid adds 200mg;
(3) culture medium inoculated of step (2) gained is entered bluish dogbane wheat bran fermentation medium, Shaking culture obtains zymocyte liquid;
(4) retted fibre pond is made;
(5) used by the zymocyte liquid of preparation in step (3) spray process to send into retted fibre pond and do retted fibre process, obtained bastose after retted fibre.
The 16Sr DNA sequence dna of the lysine bacillus DY2 bacterial strain in described step (1) is as shown in SEQ ID NO:1.
Consisting of of beef-protein medium in described step (1): every 1000ml culture medium comprises beef extract 3g, peptone 10g, sodium chloride 5g and water 1000ml.
Consisting of of frozen buffer solution in described step (1): every frozen buffer solution of 100ml comprises glycerine 50ml and water 50ml; Addition is that every 1000ml bacterium liquid adds 1ml.
Bacterial strain preservation condition in described step (1) is-80 DEG C, pH nature; Condition of culture is 37 DEG C, pH nature, incubation time 12 hours.
Consisting of of bluish dogbane bran mass in described step (2): every 1000ml culture medium comprises bluish dogbane skin 200g, wheat bran 50g, brewer's wort 50ml and water 950ml; Consisting of of bluish dogbane wheat bran fermentation medium in step (3): every 1000ml culture medium comprises bluish dogbane skin 100g, wheat bran 100g, brewer's wort 200ml and water 800ml, and inoculum concentration is that every 1000ml bacterium liquid is inoculated into 30L bluish dogbane wheat bran fermentation medium.
Condition of culture in described step (2) and (3) is 37 DEG C, pH nature, incubation time 24 hours.
It is 15-20 centimetre that the fiber crops in the retted fibre pond in described step (4) plow spacing, and numb layer thickness is 2-2.5 centimetre.
Retted fibre condition in described step (5) is natural temperature (about 20-30 degree Celsius), pH nature, retted fibre time 10-20 days; The technique of spray process is specially every 48 hours to retted fibre region sprinkling bacterium liquid 6 ~ 10ml.
Obtained bastose in described step (5) is applied to and makes gunnysack, burlap, carpet, rope, paper, fiberboard or construction material.
The preserving number of lysine bacillus DY2 bacterial strain of the present invention is CGMCC No.9120, preservation day is on May 4th, 2014, suggestion Classification And Nomenclature is: lysine bacillus (Lysinibacillus sp.), depositary institution's title: CGMCC China Committee for Culture Collection of Microorganisms common micro-organisms center, preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica.
According to 16sR DNA sequencing Blast and cluster analysis result, this bacterium and lysine bacillus similitude are 99%, in conjunction with microscopy analysis, name this bacterial strain to be lysine bacillus (Lysinibacillus sp.) DY2 bacterial strain.During 16S rDNA checks order, the forward primer 27F adopted, reverse primer 1492R, be international primer, synthesized by Shanghai Sangon Biological Engineering Technology And Service Co., Ltd.
This strain culturing condition is extensive, good heat resistance, and growth and breeding is fast, effectively can remove the non-cellulose colloid in bastose, and enzyme is lived stable, bacterium, the equal avirulence of enzyme.This bacterial strain and culture systems can be directly used in bluish dogbane retted fibre.
beneficial effect
(1) bacterial strain of the present invention and culture systems, bluish dogbane retted fibre can be directly used in after testing, have the retted fibre cycle short, the features such as retted fibre is respond well, bacterial classification is not easily contaminated, processing cost is low, fiber quality is good, retted fibre mild condition, and contamination resistance is strong, heat resistance is good, the advantages such as non-environmental-pollution;
(2) preparation method of the present invention has the features such as simple, the applicable large-scale industrial production of technique; Utilize this system to carry out flax degumming and there is retted fibre time short-sighted weather condition, generally can shorten features such as 2-3 days (normal harvest season).
Accompanying drawing explanation
Fig. 1 is bacterial strain uses therefor microscope stained photographs (oily mirror) of the present invention;
Fig. 2 a-j is the DNA of bacteria collection of illustrative plates that the present invention adopts primer 2 7F to produce;
Fig. 3 a-j is the DNA of bacteria collection of illustrative plates that the present invention adopts primer 1492R to produce.
Detailed description of the invention
Below in conjunction with specific embodiment, set forth the present invention further.Should be understood that these embodiments are only not used in for illustration of the present invention to limit the scope of the invention.In addition should be understood that those skilled in the art can make various changes or modifications the present invention, and these equivalent form of values fall within the application's appended claims limited range equally after the content of having read the present invention's instruction.
Embodiment 1
The retted fibre time is fixed according to bluish dogbane harvest time, is generally mid or late September to mid or late October.
Retting flax method:
(1) the lysine bacillus DY2 bacterial strain preserved is cultivated in beef-protein medium (every 1000ml culture medium comprises beef extract 3g, peptone 10g, sodium chloride 5g and water 1000ml), add frozen buffer solution (the frozen buffer solution of every 100ml comprises glycerine 50ml and water 50ml), condition of culture is 37 DEG C, pH nature, incubation time 12 hours;
(2) in bluish dogbane bran mass (every 1000ml culture medium comprises bluish dogbane skin 200g, wheat bran 50g, brewer's wort 50ml and water 950ml) 1000ml, access bacterium one ring, Shaking culture, condition of culture is 37 DEG C, pH nature, incubation time 24 hours;
(3) culture medium inoculated is entered bluish dogbane wheat bran fermentation medium (every 1000ml culture medium comprises bluish dogbane skin 100g, wheat bran 100g, brewer's wort 200ml and water 800ml), Shaking culture obtains zymocyte liquid, condition of culture is 37 DEG C, pH nature, incubation time 24 hours;
(4) preparation in retted fibre pond: fresh hide is separated from kenaf whole stalk (bast has caused many wounds because of mechanical presses, the impact that re-packs, and is beneficial to degumming bacterium invasion, but does not destroy fiber) with 4HB-800 type red ramie peeler; Afterbody end is neat (can be got rid of as degree during to wash fiber crops, about 20 is one), ending is held together together, straight, fold becomes three to four layers, and (retted fibre tank depth is similar, numb is just in time flooded during immersion) or doubling, often fiber crops middle part by tail place fresh hide binding, with between be connected with the fresh hide of binding, arrange in order, arrange tightly; General each retted fibre pond is thrown in all in batches, is convenient to wash fiber crops in batches; Throw in good fiber crops after then by fiber crops being all pressed in water, substantially do not surface and be advisable.Finally use thicker plastic sheeting (transparent) by all numb covering, then compress plastic sheeting, in case numb skin is dried up.Fiber crops time spacing 15-20 centimetre.Fiber crops layer thickness is generally advisable at 2-2.5 centimetre.
(5) use spray process to enter retted fibre pond the zymocyte liquid of preparation and do retted fibre process, obtaining after retted fibre can for textile fabrics bastose, and retted fibre condition is natural temperature, pH nature; The technique of spray process is specially every 48 hours to retted fibre region sprinkling bacterium liquid 8ml.
Complete the retting time: look temperature fixed, general mid or late September, the retting time extended gradually to mid or late October, if temperature Change is little, then the retting time is substantially identical.10-20 days is about at bluish dogbane normal harvest season.
Claims (10)
1. utilize lysine bacillus DY2 bacterial strain retted fibre to prepare a method for bastose, comprising:
(1) the lysine bacillus DY2 bacterial strain of preservation is cultivated in beef-protein medium, add frozen buffer solution;
(2) in bluish dogbane bran mass, bacterium one ring of step (1) gained is accessed, Shaking culture;
(3) culture medium inoculated of step (2) gained is entered bluish dogbane wheat bran fermentation medium, Shaking culture obtains zymocyte liquid;
(4) retted fibre pond is made;
(5) used by the zymocyte liquid of preparation in step (3) spray process to send into retted fibre pond and do retted fibre process, obtained bastose after retted fibre.
2. a kind of method utilizing lysine bacillus DY2 bacterial strain retted fibre to prepare bastose according to claim 1, is characterized in that: the 16Sr DNA sequence dna of the lysine bacillus DY2 bacterial strain in described step (1) is as shown in SEQ ID NO:1.
3. a kind of method utilizing lysine bacillus DY2 bacterial strain retted fibre to prepare bastose according to claim 1, is characterized in that: consisting of of the beef-protein medium in described step (1): every 1000ml culture medium comprises beef extract 3g, peptone 10g, sodium chloride 5g and water 1000ml.
4. a kind of method utilizing lysine bacillus DY2 bacterial strain retted fibre to prepare bastose according to claim 1, is characterized in that: consisting of of the frozen buffer solution in described step (1): every frozen buffer solution of 100ml comprises glycerine 50ml and water 50ml; Addition is that every 1000ml bacterium liquid adds 1ml.
5. a kind of method utilizing lysine bacillus DY2 bacterial strain retted fibre to prepare bastose according to claim 1, is characterized in that: the bacterial strain preservation condition in described step (1) is-80 DEG C, pH nature; Condition of culture is 37 DEG C, pH nature, incubation time 12 hours.
6. a kind of method utilizing lysine bacillus DY2 bacterial strain retted fibre to prepare bastose according to claim 1, it is characterized in that: consisting of of the bluish dogbane bran mass in described step (2): every 1000ml culture medium comprises bluish dogbane skin 200g, wheat bran 50g, brewer's wort 50ml and water 950ml; Consisting of of bluish dogbane wheat bran fermentation medium in step (3): every 1000ml culture medium comprises bluish dogbane skin 100g, wheat bran 100g, brewer's wort 200ml and water 800ml, and inoculum concentration is that every 1000ml bacterium liquid is inoculated into 30L bluish dogbane wheat bran fermentation medium.
7. a kind of method utilizing lysine bacillus DY2 bacterial strain retted fibre to prepare bastose according to claim 1, it is characterized in that: the condition of culture in described step (2) and (3) is 37 DEG C, pH nature, incubation time 24 hours.
8. a kind of method utilizing lysine bacillus DY2 bacterial strain retted fibre to prepare bastose according to claim 1, is characterized in that: it is 15-20 centimetre that the fiber crops in the retted fibre pond in described step (4) plow spacing, and numb layer thickness is 2-2.5 centimetre.
9. a kind of method utilizing lysine bacillus DY2 bacterial strain retted fibre to prepare bastose according to claim 1, is characterized in that: the retted fibre condition in described step (5) is natural temperature, pH nature, retted fibre time 10-20 days; The technique of spray process is specially every 48 hours to retted fibre region sprinkling bacterium liquid 6 ~ 10ml.
10. a kind of method utilizing lysine bacillus DY2 bacterial strain retted fibre to prepare bastose according to claim 1, is characterized in that: the obtained bastose in described step (5) is applied to and makes gunnysack, burlap, carpet, rope, paper, fiberboard or construction material.
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Cited By (1)
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| CN105567605A (en) * | 2016-02-19 | 2016-05-11 | 黑龙江省科学院大庆分院 | Lysinibacillus sp. and application thereof, degumming auxiliary containing lysinibacillus sp. and preparation method of degumming auxiliary |
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| CN1374398A (en) * | 2002-04-12 | 2002-10-16 | 山东省纤维检验局 | Prepn and use of enzyme prepn for use in hemp degluing industrial process |
| CN101654660A (en) * | 2009-07-14 | 2010-02-24 | 东华大学 | Bacillus subtillis strain containing ramie degumming activity, preparation and application thereof |
| CN102002467A (en) * | 2010-08-13 | 2011-04-06 | 东华大学 | Bacillus cereus DA3 strain and preparation method and application thereof |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1374398A (en) * | 2002-04-12 | 2002-10-16 | 山东省纤维检验局 | Prepn and use of enzyme prepn for use in hemp degluing industrial process |
| CN101654660A (en) * | 2009-07-14 | 2010-02-24 | 东华大学 | Bacillus subtillis strain containing ramie degumming activity, preparation and application thereof |
| CN102002467A (en) * | 2010-08-13 | 2011-04-06 | 东华大学 | Bacillus cereus DA3 strain and preparation method and application thereof |
Non-Patent Citations (1)
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| 胡开辉等: ""红麻脱胶菌的筛选及其产酶条件"", 《河南科技大学学报(自然科学版)》, vol. 27, no. 5, 30 October 2006 (2006-10-30), pages 79 - 82 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105567605A (en) * | 2016-02-19 | 2016-05-11 | 黑龙江省科学院大庆分院 | Lysinibacillus sp. and application thereof, degumming auxiliary containing lysinibacillus sp. and preparation method of degumming auxiliary |
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