CN113789576A - Flax biological degumming method based on solid fermentation method and alkaline composite strain system - Google Patents
Flax biological degumming method based on solid fermentation method and alkaline composite strain system Download PDFInfo
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- LUEWUZLMQUOBSB-FSKGGBMCSA-N (2s,3s,4s,5s,6r)-2-[(2r,3s,4r,5r,6s)-6-[(2r,3s,4r,5s,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5s,6r)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-4,5-dihydroxy-2-(hydroxymethyl)oxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@@H](O[C@@H]2[C@H](O[C@@H](OC3[C@H](O[C@@H](O)[C@@H](O)[C@H]3O)CO)[C@@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O LUEWUZLMQUOBSB-FSKGGBMCSA-N 0.000 claims description 14
- 229920001817 Agar Polymers 0.000 claims description 14
- 229920002581 Glucomannan Polymers 0.000 claims description 14
- 239000001888 Peptone Substances 0.000 claims description 14
- 108010080698 Peptones Proteins 0.000 claims description 14
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 14
- 230000004913 activation Effects 0.000 claims description 14
- 239000008272 agar Substances 0.000 claims description 14
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 14
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 14
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 14
- 239000007853 buffer solution Substances 0.000 claims description 14
- 239000004202 carbamide Substances 0.000 claims description 14
- 238000009960 carding Methods 0.000 claims description 14
- 238000009472 formulation Methods 0.000 claims description 14
- 229940046240 glucomannan Drugs 0.000 claims description 14
- 238000007654 immersion Methods 0.000 claims description 14
- 238000011081 inoculation Methods 0.000 claims description 14
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- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 14
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 14
- 239000000843 powder Substances 0.000 claims description 14
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 14
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 14
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 14
- 229920001221 xylan Polymers 0.000 claims description 14
- 150000004823 xylans Chemical class 0.000 claims description 14
- 239000002609 medium Substances 0.000 claims description 13
- 238000007865 diluting Methods 0.000 claims description 12
- 239000011248 coating agent Substances 0.000 claims description 9
- 238000000576 coating method Methods 0.000 claims description 9
- 238000002791 soaking Methods 0.000 claims description 8
- 230000003213 activating effect Effects 0.000 claims description 7
- 238000004061 bleaching Methods 0.000 claims description 7
- 238000010276 construction Methods 0.000 claims description 7
- 239000012153 distilled water Substances 0.000 claims description 7
- 230000002779 inactivation Effects 0.000 claims description 7
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 7
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 7
- 230000010355 oscillation Effects 0.000 claims description 7
- 239000008055 phosphate buffer solution Substances 0.000 claims description 7
- 238000003756 stirring Methods 0.000 claims description 7
- 239000006228 supernatant Substances 0.000 claims description 7
- 238000010257 thawing Methods 0.000 claims description 7
- 229940041514 candida albicans extract Drugs 0.000 claims description 6
- 230000001954 sterilising effect Effects 0.000 claims description 6
- 238000004659 sterilization and disinfection Methods 0.000 claims description 6
- 239000012138 yeast extract Substances 0.000 claims description 6
- 238000009395 breeding Methods 0.000 claims description 3
- 230000001488 breeding effect Effects 0.000 claims description 3
- 238000007710 freezing Methods 0.000 claims description 3
- 230000008014 freezing Effects 0.000 claims description 3
- 239000002504 physiological saline solution Substances 0.000 claims description 2
- 241000196324 Embryophyta Species 0.000 abstract 1
- 238000001816 cooling Methods 0.000 abstract 1
- 239000000758 substrate Substances 0.000 abstract 1
- 238000009875 water degumming Methods 0.000 abstract 1
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 238000005265 energy consumption Methods 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 239000004753 textile Substances 0.000 description 3
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 230000000593 degrading effect Effects 0.000 description 2
- QOSATHPSBFQAML-UHFFFAOYSA-N hydrogen peroxide;hydrate Chemical compound O.OO QOSATHPSBFQAML-UHFFFAOYSA-N 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
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- 239000000126 substance Substances 0.000 description 2
- 244000198134 Agave sisalana Species 0.000 description 1
- 241000185686 Apocynum venetum Species 0.000 description 1
- 240000008564 Boehmeria nivea Species 0.000 description 1
- 240000000491 Corchorus aestuans Species 0.000 description 1
- 235000011777 Corchorus aestuans Nutrition 0.000 description 1
- 235000010862 Corchorus capsularis Nutrition 0.000 description 1
- 229920002488 Hemicellulose Polymers 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
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Classifications
-
- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01C—CHEMICAL OR BIOLOGICAL TREATMENT OF NATURAL FILAMENTARY OR FIBROUS MATERIAL TO OBTAIN FILAMENTS OR FIBRES FOR SPINNING; CARBONISING RAGS TO RECOVER ANIMAL FIBRES
- D01C1/00—Treatment of vegetable material
- D01C1/02—Treatment of vegetable material by chemical methods to obtain bast fibres
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- Chemical & Material Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Textile Engineering (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a novel flax biological degumming method. The method is formed by combining three steps, and comprises the steps of evolutionary engineering cultivation of own bacteria of the bast fiber plants, breaking of a biological resistance barrier and biological degumming by a solid fermentation method. The method is inspirational from traditional rain and dew retting, and the invention moves the traditional few-water degumming mode into a manually controllable environment and realizes high-efficiency, water-saving and energy-saving degumming by modifying strains and substrates. In the invention, the deep cooling pretreatment of the flax, domestication of bacteria on the surface of the flax and the degumming process based on a solid fermentation method are all the bright points of the invention.
Description
Technical Field
The invention relates to a degumming processing method for fibrilia, in particular to a degumming processing method for flax fiber, belonging to the field of fibrilia textile processing.
Technical Field
Fibrilia is a natural fiber material with high strength and good moisture absorption and heat insulation performance, and is widely concerned in the fields of traditional textile processing and novel composite materials. The hemp fiber includes flax, ramie, sisal, jute, apocynum venetum and other varieties, and the natural fiber is acquired by necessarily going through a degumming process.
The degumming process is a process of removing impurities such as pectin, lignin, hemicellulose, lipid wax, protein, ash and the like among bast fibers, thereby opening fiber bundles and obtaining independent fibers. At present, the degumming mode used in China mainly comprises the steps of firstly using a biological method (water retting and rain and dew retting) to carry out primary colloid removal, and then using sodium hydroxide as a main material to boil and degum at high temperature and high pressure to obtain the required fiber. The method has the problems of high water consumption, difficult treatment of sewage caused by excessive use of chemicals and great energy consumption in the long-time high-temperature high-pressure alkali boiling process, and the problems become main factors for restricting the development of the linen textile industry.
The invention obtains inspiration from the degumming by the traditional rain and dew method, combines the solid fermentation method used for fermenting tea leaves and preparing special enzyme with biological degumming, and innovates a clean biological degumming mode with low water consumption, low energy consumption and no pollution. In order to solve the problems of low efficiency, poor quality and the like commonly existing in biological degumming, a clean pretreatment mode is used for breaking a flax biological resistance barrier, a freezing method is innovatively used for pretreating flax, three kinds of bacteria with specific degrading enzymes are cultivated by utilizing the characteristic that bacteria carried by the flax skin can survive in a dry environment on the surface of the flax skin and a microbial evolution engineering breeding mode, and an efficient composite bacteria degumming system suitable for biological degumming by a solid fermentation method is formed. The degumming mode formed comprehensively in the three aspects is a novel degumming method which is simple, convenient, efficient, clean and environment-friendly.
Disclosure of Invention
The invention provides a novel flax degumming method based on a composite strain system by a solid fermentation method. .
The technical scheme adopted by the invention is as follows:
a novel flax degumming method based on a composite strain system of a solid fermentation method comprises the steps of evolutionary engineering cultivation of self-carrying bacteria of hemp, breaking of a biological resistance barrier and a biological degumming process of the solid fermentation method.
The process for constructing the composite strain degumming system by the bast self-carrying strain evolution engineering breeding comprises the following steps:
1) carding and fluffing the flax bast fibers of the required degumming variety, placing the flax bast fibers in a bath ratio of 1: 30-60 into a conical flask with potassium dihydrogen phosphate buffer solution with the concentration of 0.5-1 g/L, culturing for 1-5 d in a constant temperature incubator at the temperature of 32-37 ℃, taking supernatant, adding physiological saline to dilute to 10 DEG2～105And then, coating the medium on a selective solid plate culture medium, and culturing for 1-3 d in a constant temperature incubator at the temperature of 32-37 ℃ to obtain the required bacterial colony. The formulation of the selective solid plate medium described in the above text is as follows: 2-5 g/L of needed degummed flax powder, 5g/L of NaCl and 20g/L of agar, wherein the culture medium is divided into two parts by using an ultrasonic oscillation instrument before sterilizationDispersing uniformly;
2) selecting strains with different forms from the bacterial colonies obtained in the step 1) to be inoculated into conical flasks with other three selective liquid culture media respectively, culturing for 1-3 d in a constant-temperature shaking table at the rotating speed of 100-250 rpm, and then diluting bacterial liquid in each conical flask by 102～105Coating the mixture on a nutrient solid plate culture medium, culturing for 1-3 d in a constant temperature incubator at the temperature of 32-37 ℃, and observing the growth condition of strains on each plate under a colony counter. The formula of the selective culture medium is as follows: 10g/L of nutrients (pectin, xylan, glucomannan), 1g/L of monopotassium phosphate, 5g/L of NaCl, 1g/L of ammonium sulfate were selected and dissolved using a sodium carbonate/sodium bicarbonate buffer solution with pH 10.0. The formulation of the nutrient solid culture medium in the steps is as follows: 5g/L of yeast extract powder, 10g/L of peptone, 5g/L of NaCl, 1g/L of monopotassium phosphate, 0.5g/L of magnesium sulfate heptahydrate and 20g/L of agar;
3) after the comparison in the step 2), acquiring (three) strains with the best growth conditions, domesticating by using a selective domestication culture medium, subculturing for 10-15 generations, storing the (three) strains for later use by using a glycerol tube method, and finishing the construction of an alkaline composite degumming strain system. The formula of the selected domestication culture medium in the steps is as follows: 10g/L of nutrient (pectin, xylan, glucomannan), 10g/L of peptone, 1g/L of potassium dihydrogen phosphate, 5g/L of NaCl, 1g/L of ammonium sulfate were selected and dissolved using a sodium carbonate/sodium bicarbonate buffer solution at pH 10.0.
The process of the biological resistance barrier disruption is as follows:
1) soaking needed degummed flax in water according to a bath ratio of 1: 30, adding 5g/L urea, stirring until the flax fiber is in a dispersed state, putting the flax fiber in a refrigerator at ultralow temperature of-50 ℃ for 0.5 to 2 hours, taking out the flax fiber, and naturally thawing for later use;
2) placing the hemp skin immersion liquid in the step 1) without replacement in an ultrasonic crusher, and repeating twice at the temperature of 30-55 ℃, the ultrasonic frequency of 40KHz and the ultrasonic frequency of 0.5 h;
3) pouring the hemp skin immersion liquid in the step 2), washing with cold water, washing with hot water, removing urea on the surface of the hemp skin, and then placing in a 55 ℃ drying oven for drying, and reserving a sample for later use.
The solid fermentation degumming engineering comprises the following steps:
1) paving flax: taking a proper square plastic container, placing the flax to be degummed after carding in the container, wherein the flax needs to be fluffy as much as possible, and placing the flax in a constant-temperature constant-humidity incubator with the humidity of 50-80% and the temperature of 30-37 ℃;
2) activating strains: inoculating the bred three specific strains into a liquid culture medium for activation, and carrying out 3-10 generations for later use;
3) turning hemp and inoculating bacteria: uniformly mixing the three bacteria solutions after activation in the step 2), and then uniformly mixing the three bacteria solutions with distilled water, wherein the ratio of the mixed bacteria is 15%. And (3) placing the prepared inoculation bacteria liquid into a sprinkling can, turning the flax in the container while sprinkling, ensuring that the inoculation of the flax bacteria liquid is uniform, and obtaining the flax bacteria liquid after the surface of the flax is uniformly hydrated. The degumming process is carried out for 3-5 d, and the step 3) is repeated every 12h until the degumming process is finished;
4) post-treatment (inactivation, bleaching): taking out the flax fibers treated in the step 3), washing with clean water, placing in a water bath kettle at 50-75 ℃ for treatment for 20-90 min, taking out, washing with cold water, washing with hot water, and drying in an oven to obtain the degummed flax fibers.
According to the invention, from the perspectives of low energy consumption, no alkali and low water consumption, enrichment, primary screening, secondary screening and domestication are carried out by using the characteristics that the strain on the surface of the hemp skin is suitable for a low-water environment and the components carried by the hemp skin can be used as a nutrient source in a microbial evolution engineering mode to obtain the strain containing the specific degrading enzyme suitable for the solid fermentation method under the low-water condition, so that a composite degumming strain system suitable for the method is constructed; the biological resistance barrier is broken by using a clean and pollution-free pretreatment mode, and a freezing method is innovatively used for pretreatment, so that the problems of low efficiency, poor quality and the like caused by biological degumming are solved; the traditional rain and dew method is used for retting flax to obtain inspiration, the solid fermentation method and the biological degumming are combined to construct a controllable and efficient biological degumming environment, and the flax fiber with good quality is obtained. The method is only tested at present, and the potential of the method can be specifically modified and used for each bast fiber needing degumming. The invention has the beneficial effects that: aiming at the current situation that the traditional chemical degumming method has high energy consumption, high pollution and high water consumption, the invention has obvious improvement on energy saving, water saving and cleaning, and no strong base such as NaOH and the like is used in the whole process; compared with the currently developed biological degumming method, the biological resistance barrier of flax is broken by utilizing a cleaning pretreatment mode, the problems of low quality and low efficiency frequently existing in biological degumming are solved, and the water and energy conservation are obviously improved on the basis.
Detailed Description
The invention is further described below by means of specific embodiments. Unless otherwise specified, technical means not described in the embodiments may be implemented in a manner well known to those skilled in the art. In addition, the embodiments should be considered illustrative, and not restrictive, of the scope of the invention, which is defined solely by the claims. It will be apparent to those skilled in the art that various modifications, substitutions, and improvements in the materials, amounts, dimensions, and shapes of the embodiments disclosed herein may be made without departing from the spirit and scope of the invention, and the invention is to be limited only by the specific parameters set forth herein as the scope of the invention is to be determined with the permissible error.
Example 1
1) Carding and fluffing required degummed flax bast fiber, placing the flax bast fiber in a bath ratio of 1: 60 into a conical flask with 1g/L potassium dihydrogen phosphate buffer solution, culturing for 3d in a constant temperature incubator at 37 ℃, and diluting the supernatant with normal saline to 10%3Then, the cells were spread on a selective solid plate medium and cultured in an incubator at 37 ℃ for 2d to obtain desired colonies. The formulation of the selective solid plate medium described in the above text is as follows: 5g/L of needed degummed flax powder, 5g/L of NaCl and 20g/L of agar, wherein an ultrasonic oscillation instrument is needed to uniformly disperse the culture medium before sterilization;
2) inoculating the bacterial colony obtained in the step 1) with strains of different forms into conical flasks with other three selective liquid culture media, culturing for 2d in a constant temperature shaking table at the rotating speed of 150rpm, and diluting the bacterial liquid in each conical flask by 103Plating on nutrient solid plateOn the medium, the culture was carried out at 37 ℃ for 2d in a constant temperature incubator, and the growth of the strain on each plate was observed in a colony counter. The formula of the selective culture medium is as follows: 10g/L of nutrients (pectin, xylan, glucomannan), 1g/L of monopotassium phosphate, 5g/L of NaCl, 1g/L of ammonium sulfate were selected and dissolved using a sodium carbonate/sodium bicarbonate buffer solution with pH 10.0. The formulation of the nutrient solid culture medium in the steps is as follows: 5g/L of yeast extract powder, 10g/L of peptone, 5g/L of NaCl, 1g/L of monopotassium phosphate, 0.5g/L of magnesium sulfate heptahydrate and 20g/L of agar;
3) after the comparison in the step 2), acquiring (three) strains with the best growth conditions, domesticating by using a selective domestication culture medium, subculturing for 10 generations, storing the (three) strains for later use by using a glycerol tube method, and finishing the construction of the alkaline composite degumming strain system. The formula of the selected domestication culture medium in the steps is as follows: 10g/L of nutrient (pectin, xylan, glucomannan), 10g/L of peptone, 1g/L of potassium dihydrogen phosphate, 5g/L of NaCl, 1g/L of ammonium sulfate were selected and dissolved using a sodium carbonate/sodium bicarbonate buffer solution at pH 10.0.
The process of the biological resistance barrier disruption is as follows:
1) soaking needed degummed flax in water at a bath ratio of 1: 30, adding 5g/L urea, stirring until the flax fiber is in a dispersed state, placing in a refrigerator at ultralow temperature of-50 ℃ for 0.5h, taking out, and naturally thawing for later use;
2) placing the hemp skin immersion liquid in the step 1) without replacement in an ultrasonic crusher, and repeating twice at 55 ℃, 40KHz ultrasonic frequency and 0.5h ultrasonic frequency;
3) pouring the hemp skin immersion liquid in the step 2), washing with cold water, washing with hot water, removing urea on the surface of the hemp skin, and then placing in a 55 ℃ drying oven for drying, and reserving a sample for later use.
The solid fermentation degumming engineering comprises the following steps:
1) paving flax: taking a proper square plastic container, placing the flax to be degummed after carding in the container, wherein the flax needs to be fluffy as much as possible, and placing the flax in a constant-temperature constant-humidity incubator with the humidity of 80% and the temperature of 32 ℃;
2) activating strains: inoculating the bred three specific strains into a liquid culture medium for activation, and standing for later use after 5 generations;
3) turning hemp and inoculating bacteria: uniformly mixing the three bacteria solutions after activation in the step 2), and then uniformly mixing the three bacteria solutions with distilled water, wherein the ratio of the mixed bacteria is 15%. And (3) placing the prepared inoculation bacteria liquid into a sprinkling can, turning the flax in the container while sprinkling, ensuring that the inoculation of the flax bacteria liquid is uniform, and obtaining the flax bacteria liquid after the surface of the flax is uniformly hydrated. The degumming process is carried out for 3.5d, and the step 3) is repeated every 12h until the degumming process is finished;
4) post-treatment (inactivation, bleaching): taking out the flax fibers treated in the step 3), washing with clear water, placing in 5% hydrogen peroxide aqueous solution at 75 ℃ for treatment for 45min, taking out, washing with cold water, washing with hot water, and drying in an oven to obtain the degummed flax fibers.
Example 2
1) Carding and fluffing required degummed flax bast fiber, placing into a conical flask with 1g/L potassium dihydrogen phosphate buffer solution at a bath ratio of 1: 40, culturing at 35 deg.C in a constant temperature incubator for 5d, collecting supernatant, diluting with normal saline to 10%3、104、105Then, the cells were spread on a selective solid plate medium and cultured in a thermostat incubator at 35 ℃ for 2d to obtain desired colonies. The formulation of the selective solid plate medium described in the above text is as follows: 2g/L of needed degummed flax powder, 5g/L of NaCl and 20g/L of agar, wherein an ultrasonic oscillation instrument is needed to uniformly disperse the culture medium before sterilization;
2) inoculating the bacterial colony obtained in the step 1) with strains of different forms into conical flasks with other three selective liquid culture media, culturing for 2d in a constant temperature shaking table at the rotating speed of 120rpm, and diluting the bacterial liquid in each conical flask by 104Coating on nutrient solid plate culture medium, culturing at 35 deg.C in constant temperature incubator for 2d, and observing the growth of strain on each plate under colony counter. The formula of the selective culture medium is as follows: 10g/L of nutrients (pectin, xylan, glucomannan), 1g/L of monopotassium phosphate, 5g/L of NaCl, 1g/L of ammonium sulfate were selected and dissolved using a sodium carbonate/sodium bicarbonate buffer solution with pH 10.0. The formulation of the nutrient solid culture medium in the steps is as follows: yeast5g/L of soaking powder, 10g/L of peptone, 5g/L of NaCl, 1g/L of monopotassium phosphate, 0.5g/L of magnesium sulfate heptahydrate and 20g/L of agar;
3) after the comparison in the step 2), acquiring (three) strains with the best growth conditions, domesticating by using a selective domestication culture medium, subculturing for 10 generations, storing the (three) strains for later use by using a glycerol tube method, and finishing the construction of the alkaline composite degumming strain system. The formula of the selected domestication culture medium in the steps is as follows: 10g/L of nutrient (pectin, xylan, glucomannan), 10g/L of peptone, 1g/L of potassium dihydrogen phosphate, 5g/L of NaCl, 1g/L of ammonium sulfate were selected and dissolved using a sodium carbonate/sodium bicarbonate buffer solution at pH 10.0.
The process of the biological resistance barrier disruption is as follows:
1) soaking needed degummed flax in water at a bath ratio of 1: 30, adding 5g/L urea, stirring until the flax fiber is in a dispersed state, placing in a refrigerator at ultralow temperature of-50 ℃ for 1h, taking out, and naturally thawing for later use;
2) placing the hemp skin immersion liquid in the step 1) in an ultrasonic crusher at 30 ℃, 40KHz ultrasonic frequency and 0.5h ultrasonic frequency, and repeating twice;
3) pouring the hemp skin immersion liquid in the step 2), washing with cold water, washing with hot water, removing urea on the surface of the hemp skin, and then placing in a 55 ℃ drying oven for drying, and reserving a sample for later use.
The solid fermentation degumming engineering comprises the following steps:
1) paving flax: taking a proper square plastic container, placing the flax to be degummed after carding in the container, and placing the flax in a constant-temperature constant-humidity incubator with the humidity of 60% and the temperature of 30 ℃ to ensure that the flax is fluffy as much as possible;
2) activating strains: inoculating the bred three specific strains into a liquid culture medium for activation, and standing by after 3 generations;
3) turning hemp and inoculating bacteria: uniformly mixing the three bacteria solutions after activation in the step 2), and then uniformly mixing the three bacteria solutions with distilled water, wherein the ratio of the mixed bacteria is 15%. And (3) placing the prepared inoculation bacteria liquid into a sprinkling can, turning the flax in the container while sprinkling, ensuring that the inoculation of the flax bacteria liquid is uniform, and obtaining the flax bacteria liquid after the surface of the flax is uniformly hydrated. The degumming process is carried out for 4d, and the step 3) is repeated every 12h until the degumming process is finished;
4) post-treatment (inactivation, bleaching): taking out the flax fibers treated in the step 3), washing with clean water, placing in a water bath kettle at 50 ℃, treating for 30min in 5% hydrogen peroxide water solution, taking out, washing with cold water, washing with hot water, and drying in an oven to obtain the degummed flax fibers.
Example 3
1) Carding and fluffing required degummed flax bast fiber, placing into a conical flask with 1g/L potassium dihydrogen phosphate buffer solution at a bath ratio of 1: 30, culturing at 35 deg.C in a constant temperature incubator for 3d, collecting supernatant, diluting with normal saline to 10%2And (5) coating the strain on a selective solid plate culture medium, and culturing for 3d in a constant temperature incubator at 35 ℃ to obtain the required bacterial colony. The formulation of the selective solid plate medium described in the above text is as follows: 3g/L of needed degummed flax powder, 5g/L of NaCl and 20g/L of agar, wherein an ultrasonic oscillation instrument is needed to uniformly disperse the culture medium before sterilization;
2) inoculating the bacterial colony obtained in the step 1) with strains of different forms into conical flasks with other three selective liquid culture media, culturing for 3d in a constant temperature shaking table at a rotating speed of 200rpm, and diluting the bacterial liquid in each conical flask by 102Coating the mixture on a nutrient solid plate culture medium, culturing for 3d in a constant temperature incubator at 32-37 ℃, and observing the growth condition of strains on each plate under a colony counter. The formula of the selective culture medium is as follows: 10g/L of nutrients (pectin, xylan, glucomannan), 1g/L of monopotassium phosphate, 5g/L of NaCl, 1g/L of ammonium sulfate were selected and dissolved using a sodium carbonate/sodium bicarbonate buffer solution with pH 10.0. The formulation of the nutrient solid culture medium in the steps is as follows: 5g/L of yeast extract powder, 10g/L of peptone, 5g/L of NaCl, 1g/L of monopotassium phosphate, 0.5g/L of magnesium sulfate heptahydrate and 20g/L of agar;
3) after the comparison in the step 2), acquiring (three) strains with the best growth conditions, domesticating by using a selective domestication culture medium, subculturing for 10 generations, storing the (three) strains for later use by using a glycerol tube method, and finishing the construction of the alkaline composite degumming strain system. The formula of the selected domestication culture medium in the steps is as follows: 10g/L of nutrient (pectin, xylan, glucomannan), 10g/L of peptone, 1g/L of potassium dihydrogen phosphate, 5g/L of NaCl, 1g/L of ammonium sulfate were selected and dissolved using a sodium carbonate/sodium bicarbonate buffer solution at pH 10.0.
The process of the biological resistance barrier disruption is as follows:
1) soaking needed degummed flax in water at a bath ratio of 1: 30, adding 5g/L urea, stirring until the flax fiber is in a dispersed state, placing in a refrigerator at ultralow temperature of-50 ℃ for 2h, taking out, and naturally thawing for later use;
2) placing the hemp skin immersion liquid in the step 1) without replacement in an ultrasonic crusher, and repeating twice at 55 ℃, 40KHz ultrasonic frequency and 0.5h ultrasonic frequency;
3) pouring the hemp skin immersion liquid in the step 2), washing with cold water, washing with hot water, removing urea on the surface of the hemp skin, and then placing in a 55 ℃ drying oven for drying, and reserving a sample for later use.
The solid fermentation degumming engineering comprises the following steps:
1) paving flax: taking a proper square plastic container, placing the flax to be degummed after carding in the container, and placing the flax in a constant-temperature constant-humidity incubator with the humidity of 75% and the temperature of 32 ℃ to be fluffy as much as possible;
2) activating strains: inoculating the bred three specific strains into a liquid culture medium for activation, and standing for later use after 10 generations;
3) turning hemp and inoculating bacteria: uniformly mixing the three bacteria solutions after activation in the step 2), and then uniformly mixing the three bacteria solutions with distilled water, wherein the ratio of the mixed bacteria is 15%. And (3) placing the prepared inoculation bacteria liquid into a sprinkling can, turning the flax in the container while sprinkling, ensuring that the inoculation of the flax bacteria liquid is uniform, and obtaining the flax bacteria liquid after the surface of the flax is uniformly hydrated. The degumming process is carried out for 3-5 d, and the step 3) is repeated every 12h until the degumming process is finished;
4) post-treatment (inactivation, bleaching): taking out the flax fibers treated in the step 3), washing with clean water, placing in a water bath kettle at 75 ℃ for treatment for 20min, taking out, washing with cold water, washing with hot water, and drying in an oven to obtain the degummed flax fibers.
Example 4
1) Carding and fluffing required degummed flax bast fiber, placing into a conical flask with potassium dihydrogen phosphate buffer solution with concentration of 0.5g/L at a bath ratio of 1: 60, culturing at 35 deg.C in a constant temperature incubator for 2d, collecting supernatant, diluting with normal saline to 10%2Then, the cells were spread on a selective solid plate medium and cultured in an incubator at 37 ℃ for 1 day to obtain desired colonies. The formulation of the selective solid plate medium described in the above text is as follows: 5g/L of needed degummed flax powder, 5g/L of NaCl and 20g/L of agar, wherein an ultrasonic oscillation instrument is needed to uniformly disperse the culture medium before sterilization;
2) inoculating the bacterial colony obtained in the step 1) with strains of different forms into conical flasks with other three selective liquid culture media, culturing for 1d in a constant temperature shaking table at a rotating speed of 200rpm, and diluting the bacterial liquid in each conical flask by 102Coating on nutrient solid plate culture medium, culturing at 37 deg.C for 1d in constant temperature incubator, and observing the growth of strain on each plate under colony counter. The formula of the selective culture medium is as follows: 10g/L of nutrients (pectin, xylan, glucomannan), 1g/L of monopotassium phosphate, 5g/L of NaCl, 1g/L of ammonium sulfate were selected and dissolved using a sodium carbonate/sodium bicarbonate buffer solution with pH 10.0. The formulation of the nutrient solid culture medium in the steps is as follows: 5g/L of yeast extract powder, 10g/L of peptone, 5g/L of NaCl, 1g/L of monopotassium phosphate, 0.5g/L of magnesium sulfate heptahydrate and 20g/L of agar;
3) after the comparison in the step 2), acquiring (three) strains with the best growth conditions, domesticating by using a selective domestication culture medium, subculturing for 15 generations, storing the (three) strains for later use by using a glycerol tube method, and finishing the construction of the alkaline composite degumming strain system. The formula of the selected domestication culture medium in the steps is as follows: 10g/L of nutrient (pectin, xylan, glucomannan), 10g/L of peptone, 1g/L of potassium dihydrogen phosphate, 5g/L of NaCl, 1g/L of ammonium sulfate were selected and dissolved using a sodium carbonate/sodium bicarbonate buffer solution at pH 10.0.
The process of the biological resistance barrier disruption is as follows:
1) soaking needed degummed flax in water at a bath ratio of 1: 30, adding 5g/L urea, stirring until the flax fiber is in a dispersed state, placing in a refrigerator at ultralow temperature of-50 ℃ for 1h, taking out, and naturally thawing for later use;
2) placing the hemp skin immersion liquid in the step 1) without replacement in an ultrasonic crusher, and repeating twice at 55 ℃, 40KHz ultrasonic frequency and 0.5h ultrasonic frequency;
3) pouring the hemp skin immersion liquid in the step 2), washing with cold water, washing with hot water, removing urea on the surface of the hemp skin, and then placing in a 55 ℃ drying oven for drying, and reserving a sample for later use.
The solid fermentation degumming engineering comprises the following steps:
1) paving flax: taking a proper square plastic container, placing the flax to be degummed after carding in the container, wherein the flax needs to be fluffy as much as possible, and placing the flax in a constant-temperature constant-humidity incubator with the humidity of 50% and the temperature of 30 ℃;
2) activating strains: inoculating the bred three specific strains into a liquid culture medium for activation, and standing by after 3 generations;
3) turning hemp and inoculating bacteria: uniformly mixing the three bacteria solutions after activation in the step 2), and then uniformly mixing the three bacteria solutions with distilled water, wherein the ratio of the mixed bacteria is 15%. And (3) placing the prepared inoculation bacteria liquid into a sprinkling can, turning the flax in the container while sprinkling, ensuring that the inoculation of the flax bacteria liquid is uniform, and obtaining the flax bacteria liquid after the surface of the flax is uniformly hydrated. The degumming process is carried out for 5d, and the step 3) is repeated every 12h until the degumming process is finished;
4) post-treatment (inactivation, bleaching): taking out the flax fibers treated in the step 3), washing with clean water, placing in a water bath kettle at 65 ℃ for treatment for 60min, taking out, washing with cold water, washing with hot water, and drying in an oven to obtain the degummed flax fibers.
Example 5
1) Carding and fluffing required degummed flax bast fiber, placing the flax bast fiber in a bath ratio of 1: 50 into a conical flask with 1g/L potassium dihydrogen phosphate buffer solution, culturing for 4d in a constant temperature incubator at 36 ℃, and diluting the supernatant with normal saline to 10%2Then spreading on selective solid plate culture medium, and culturing at 336 deg.C in constant temperature incubator for 3d to obtain desired bacteriaAnd (6) dropping. The formulation of the selective solid plate medium described in the above text is as follows: 5g/L of needed degummed flax powder, 5g/L of NaCl and 20g/L of agar, wherein an ultrasonic oscillation instrument is needed to uniformly disperse the culture medium before sterilization;
2) inoculating the bacterial colony obtained in the step 1) with strains of different forms into conical flasks with other three selective liquid culture media, culturing for 1d in a constant temperature shaking table at the rotating speed of 250rpm, and diluting the bacterial liquid in each conical flask by 102Coating on nutrient solid plate culture medium, culturing at 36 deg.C for 3d in constant temperature incubator, and observing growth of strain on each plate under colony counter. The formula of the selective culture medium is as follows: 10g/L of nutrients (pectin, xylan, glucomannan), 1g/L of monopotassium phosphate, 5g/L of NaCl, 1g/L of ammonium sulfate were selected and dissolved using a sodium carbonate/sodium bicarbonate buffer solution with pH 10.0. The formulation of the nutrient solid culture medium in the steps is as follows: 5g/L of yeast extract powder, 10g/L of peptone, 5g/L of NaCl, 1g/L of monopotassium phosphate, 0.5g/L of magnesium sulfate heptahydrate and 20g/L of agar;
3) after the comparison in the step 2), acquiring (three) strains with the best growth conditions, domesticating by using a selective domestication culture medium, subculturing for 15 generations, storing the (three) strains for later use by using a glycerol tube method, and finishing the construction of the alkaline composite degumming strain system. The formula of the selected domestication culture medium in the steps is as follows: 10g/L of nutrient (pectin, xylan, glucomannan), 10g/L of peptone, 1g/L of potassium dihydrogen phosphate, 5g/L of NaCl, 1g/L of ammonium sulfate were selected and dissolved using a sodium carbonate/sodium bicarbonate buffer solution at pH 10.0.
The process of the biological resistance barrier disruption is as follows:
1) soaking needed degummed flax in water at a bath ratio of 1: 30, adding 5g/L urea, stirring until the flax fiber is in a dispersed state, placing in a refrigerator at ultralow temperature of-50 ℃ for 0.5h, taking out, and naturally thawing for later use;
2) placing the hemp skin immersion liquid in the step 1) without replacement in an ultrasonic crusher, and repeating twice at 55 ℃, 40KHz ultrasonic frequency and 0.5h ultrasonic frequency;
3) pouring the hemp skin immersion liquid in the step 2), washing with cold water, washing with hot water, removing urea on the surface of the hemp skin, and then placing in a 55 ℃ drying oven for drying, and reserving a sample for later use.
The solid fermentation degumming engineering comprises the following steps:
1) paving flax: taking a proper square plastic container, placing the flax to be degummed after carding in the container, wherein the flax needs to be fluffy as much as possible, and placing the flax in a constant-temperature constant-humidity incubator with the humidity of 80% and the temperature of 36 ℃;
2) activating strains: inoculating the bred three specific strains into a liquid culture medium for activation, and standing for later use after 5 generations;
3) turning hemp and inoculating bacteria: uniformly mixing the three bacteria solutions after activation in the step 2), and then uniformly mixing the three bacteria solutions with distilled water, wherein the ratio of the mixed bacteria is 15%. And (3) placing the prepared inoculation bacteria liquid into a sprinkling can, turning the flax in the container while sprinkling, ensuring that the inoculation of the flax bacteria liquid is uniform, and obtaining the flax bacteria liquid after the surface of the flax is uniformly hydrated. The degumming process is carried out for 4d, and the step 3) is repeated every 12h until the degumming process is finished;
4) post-treatment (inactivation, bleaching): taking out the flax fibers treated in the step 3), washing with clean water, placing in a water bath kettle at 55 ℃, treating for 90min in 5% hydrogen peroxide water solution, taking out, washing with cold water, washing with hot water, and drying in an oven to obtain the degummed flax fibers.
Claims (1)
1. A novel flax degumming method based on solid fermentation method features that the composite degumming strain system is constructed by the evolution engineering of self-carrying bacteria, the biologic resistance barrier is broken by freezing and ultrasonic method, and the degumming process based on solid fermentation method is used.
The process for constructing the composite strain degumming system by the bast self-carrying strain evolution engineering breeding comprises the following steps:
1) carding and fluffing the flax bast fibers of the required degumming variety, placing the flax bast fibers in a bath ratio of 1: 30-60 into a conical flask with potassium dihydrogen phosphate buffer solution with the concentration of 0.5-1 g/L, culturing for 1-5 d in a constant temperature incubator at the temperature of 32-37 ℃, taking supernatant, adding physiological saline to dilute to 10 DEG2～105And then, coating the medium on a selective solid plate culture medium, and culturing for 1-3 d in a constant temperature incubator at the temperature of 32-37 ℃ to obtain the required bacterial colony. The formulation of the selective solid plate medium described in the above text is as follows: 2-5 g/L of needed degummed flax powder, 5g/L of NaCl and 20g/L of agar, wherein an ultrasonic oscillation instrument is used for uniformly dispersing the culture medium before sterilization:
2) selecting strains with different forms from the bacterial colonies obtained in the step 1) to be inoculated into conical flasks with other three selective liquid culture media respectively, culturing for 1-3 d in a constant-temperature shaking table at the rotating speed of 100-250 rpm, and then diluting bacterial liquid in each conical flask by 102～105Coating the mixture on a nutrient solid plate culture medium, culturing for 1-3 d in a constant temperature incubator at the temperature of 32-37 ℃, and observing the growth condition of strains on each plate under a colony counter. The formula of the selective culture medium is as follows: 10g/L of nutrients (pectin, xylan, glucomannan), 1g/L of monopotassium phosphate, 5g/L of NaCl, and 1g/L of ammonium sulfate were selected and dissolved using a sodium carbonate/sodium bicarbonate buffer solution with a pH of 10.0. The formulation of the nutrient solid culture medium in the steps is as follows: 5g/L of yeast extract powder, 10g/L of peptone, 5g/L of NaCl, 1g/L of monopotassium phosphate, 0.5g/L of magnesium sulfate heptahydrate and 20g/L of agar;
3) after the comparison in the step 2), acquiring (three) strains with the best growth conditions, domesticating by using a selective domestication culture medium, subculturing for 10-15 generations, storing the (three) strains for later use by using a glycerol tube method, and finishing the construction of an alkaline composite degumming strain system. The formula of the selected domestication culture medium in the steps is as follows: 10g/L of nutrient (pectin, xylan, glucomannan), 10g/L of peptone, 1g/L of potassium dihydrogen phosphate, 5g/L of NaCl, 1g/L of ammonium sulfate were selected and dissolved using a sodium carbonate/sodium bicarbonate buffer solution at pH 10.0.
The process of the biological resistance barrier disruption is as follows:
1) soaking needed degummed flax in water according to a bath ratio of 1: 30, adding 5g/L urea, stirring until the flax fiber is in a dispersed state, putting the flax fiber in a refrigerator at ultralow temperature of-50 ℃ for 0.5 to 2 hours, taking out the flax fiber, and naturally thawing for later use;
2) placing the hemp skin immersion liquid in the step 1) without replacement in an ultrasonic crusher, and repeating twice at the temperature of 30-55 ℃, the ultrasonic frequency of 40KHz and the ultrasonic frequency of 0.5 h;
3) pouring the hemp skin immersion liquid in the step 2), washing with cold water and hot water, removing urea on the surface of the hemp skin, and then placing in a 55 ℃ drying oven for drying, and reserving a sample for later use;
the solid fermentation degumming engineering comprises the following steps:
1) paving flax: taking a proper square plastic container, placing the flax to be degummed after carding in the container, wherein the flax needs to be fluffy as much as possible, and placing the flax in a constant-temperature constant-humidity incubator with the humidity of 50-80% and the temperature of 30-37 ℃;
2) activating strains: inoculating the bred three specific strains into a liquid culture medium for activation, and carrying out 3-10 generations for later use;
3) turning hemp and inoculating bacteria: uniformly mixing the three bacteria solutions after activation in the step 2), and then uniformly mixing the three bacteria solutions with distilled water, wherein the ratio of the mixed bacteria is 15%. And (3) placing the prepared inoculation bacteria liquid into a sprinkling can, turning the flax in the container while sprinkling, ensuring that the inoculation of the flax bacteria liquid is uniform, and obtaining the flax bacteria liquid after the surface of the flax is uniformly hydrated. The degumming process is carried out for 3-5 d, and the step 3) is repeated every 12h until the degumming process is finished;
4) post-treatment (inactivation, bleaching): taking out the flax fibers treated in the step 3), washing with clean water, placing in a water bath kettle at 50-75 ℃ for treatment for 20-90 min, taking out, washing with cold water, washing with hot water, and drying in an oven to obtain the degummed flax fibers.
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| CN115928288A (en) * | 2022-03-28 | 2023-04-07 | 宜兴市新东茂纺织科技有限公司 | High-density tencel flax fabric and preparation method thereof |
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