WO2016061827A1 - Biological degumming method for ramie - Google Patents

Abstract

The present invention relates to a biological degumming method for ramie. The method comprises the following steps: using bacillus (Bacillus sp. HG-28) as a strain; obtaining a degumming bacteria solution F used for biological degumming of ramie after a strain activation step, a step of expanded culture and a dilution step are performed; and then, conducting biological degumming by immersing raw ramie into the degumming bacteria solution F. By implementing the present invention, energy consumption is low, pollution is low, the biological degumming needs a short time and a simple operation process, the degumming efficiency is high, and the strain culture needs a short time and low cost; the obtained degummed ramie has high quality, a low residual gum content and high breaking strength, is superior to requirements on appearance and technical specifications for degummed ramie at a first level in national standards GB/T 20793-2006 of China; and the prospective of large-scale production is wide.

Description

 Ramie biological degumming method Technical field

The present invention belongs to the field of biotechnology, and more particularly to a method for performing ramie biological degumming using Bacillus sp. HG-28.

Background technique

Castor, the scientific name Boehmeria nivea (L.) Gaud., is an economic crop that provides important textile fibers. Ramie single fiber is long, strong in strength, fast in moisture absorption and moisture absorption, and good in thermal conductivity. It is an excellent textile raw material and medical material, and has a wide range of uses. Ramie fiber can be purely spun, or blended with cotton, wool, silk, etc., and its fabric has the advantages of anti-corrosion, antibacterial, breathable, and stiff. China is a big country producing ramie fiber, its output accounts for more than 95% of the world, and it has great demand in the international market. It is a characteristic industry with competitive advantages in the international market.

At present, the main bottleneck problem that restricts the development of ramie industry is how to improve the quality of ramie fiber to meet the high-end market demand at home and abroad under the premise of reducing the production cost of ramie fiber and the pollution problem in the production process. Because ramie contains gum as the total weight 19% to 30% Left and right, before the textile processing, degumming should be carried out. At present, almost all manufacturers use chemical degumming method in the process of ramie degumming. First, soak the ramie with acid, then simmer the ramie with lye. After scouring, the ramie is tortured by the torture machine. Then, it is rinsed with bleach, acid neutralized, then washed, dehydrated, oiled, and dried to obtain finished ramie fine dry hemp. The chemical degumming of ramie requires the use of a large amount of acid and alkali, and the amount of wastewater generated is large and difficult to handle, causing a large amount of pollution to the environment. Therefore, at present, many ramie primary processing enterprises are forced to shut down due to the production of wastewater polluting water bodies and failing to meet the standards. As a result, the demand for ramie raw materials is reduced, the income of farmers' ramie is not guaranteed, the enthusiasm for planting numbness is hit, and the entire industry is facing difficulties. Therefore, the problem of energy saving and emission reduction technology in the process of ramie degumming has become a bottleneck restricting the sustainable development of the ramie industry.

In recent years, in order to reduce or cure the pollution caused by degumming, it has been recognized that the use of microorganisms for ramie degumming is the main development direction in the future, and the research on ramie biodegumming technology and related process exploration have been carried out.

 Liu Ziwei et al. in CN1371990A The patent discloses a strain capable of producing bast fiber degumming enzyme and its application in ramie and marijuana degumming. The strain has extensive culture conditions, rapid growth and reproduction, and can produce more degumming under the same culture condition. Enzyme, good degumming effect, and It can be completed in 2 to 6 hours, the degumming rate is 95%, the fiber dispersion is 100%, the fiber is not damaged, and there is no environmental pollution. However, the two-stage seed culture time of the strain is relatively long (accumulated 16 to 28 cumulatively) Hours), after the end of seed culture, it needs to be fermented for another 12 to 18 hours before it can be used for ramie degumming. The production cycle is too long; and the residual rate of fine dry hemp products is high (2.18%), and the breaking strength is too small ( 4.30) cN/dtex) can only meet the requirements of GB/T 20793-2006 second-class products, and is not suitable for large-scale production of high-quality ramie fine dry hemp.

 Liu Zhengchu et al. in CN101235357B The patent discloses a process for the rapid extraction of ramie fiber by the industrial fermentation of Erwinia. The strain is hobby mannose, and it is secreted for 7 to 9 hours to achieve key enzymes for secreting non-cellulosic degradation (pectinase, β- The peak of mannanase and xylanase). After the bacteria are activated for 5 to 6 hours, they are inoculated on the herbal fiber materials such as ramie and fermented 5 to 6 The hourly fermented forage turns blue, stripping and partially degrading the pectin, hemicellulose and part of the lignin associated with the fiber cells, and then removing the non-cellulosic residue attached to the fiber by washing with a high-pressure water column to achieve extraction. The purpose of pure fiber. However, the preparation of the activated bacterial liquid is long, the process is cumbersome, and the use of ultrafiltration membrane equipment is required, which is not conducive to the complete activation production process of the seed in the factory.

 Cao Junwei et al. in CN1177003C A new strain of Bacillus alkalophilus and its use in ramie degumming are disclosed in the patent. The technology provided by the invention has the advantages of short degumming period, high degumming efficiency, less fouling of bacteria, stable enzyme activity, low processing cost and good fiber quality. Wu Yu is waiting at CN101270343A A ramie degumming bacterium is disclosed in the patent. The degumming ability of the ramie degumming bacteria of the invention is qualitatively improved, and the hemp fiber is not damaged. After the degumming, the impurities are almost completely removed, and the degumming speed is at least doubled. Zhang Xingqun and so on CN101654660B The patent discloses a Bacillus subtilis strain having ricin degumming activity, and the preparation and application thereof, and the system using the strain as the core is used for ramie degumming, and the guar degumming using the system has a short degumming time and a ramie fiber dispersion rate. achieve 100%, degumming rate reaches 90 Above %, the quality of refined hemp reaches the level of chemical degumming. The above several methods have problems such as high price and long incubation time of the culture system, and all of them are only laboratory tests, and there is still a big gap from the actual application in industrial production.

The above problems in the prior art mainly lie in the relatively harsh culture conditions of the activation and amplification of the strain, the long incubation time, the long time of degumming, the difficulty in controlling, the excessive consumption of chemicals and water, and the ramie Product quality is not high. At present, the degumming microorganisms and biological degumming technology that are really used in industrial production are still not mature enough to completely solve the bottleneck problem. .

Summary of the invention

 The technical problem to be solved by the present invention is to provide a ramie biological degumming method, Solve the problems of serious pollution, strict process conditions, long production cycle and low product quality in the existing ramie degumming industry.

 The technical solution of the present invention to solve the above problems lies in:

 A ramie biological degumming method is provided, the method comprising the steps of:

S1, degumming bacterial culture: using Bacillus sp. HG-28 as a strain, through the strain activation step, expanding the cultivation step, and diluting the step, obtaining the degumming bacteria F for the ramie biological degumming;

S2, biological degumming: the ramie raw hemp into the hemp cage is immersed in the degumming bacteria F, the mass ratio of the ramie ramie to the degumming bacterium F is 1: 11 ~ 1 : 12 to 0.5 ~ 3 m ³ / h The ventilation is introduced into the air. After 4 to 12 hours of biological degumming, the fermenter is sealed and heated to raise the pressure to 0.1 to 0.15 MPa for 20 to 40 minutes to complete the degumming process.

 In the ramie biological degumming method of the present invention, the step S1 is further included before a strain preservation step, wherein the strain storage step is specifically:

Bacillus sp. HG-28 was inoculated into a 250 mL Erlenmeyer flask containing 50-100 mL of medium No. 1 and maintained at a temperature of 33-38 ° C for 90-180 r/min. , cultured on a shaker for 3 to 6 h to obtain bacterial liquid A. The bacterial solution A and the glycerin aqueous solution having a volume fraction of 50% were separately transferred into a 1 to 5 mL centrifuge tube in a ratio of 3:2, and uniformly mixed to obtain a bacterial solution B, and stored at 20 ° C for use;

 Wherein, the medium No. 1 comprises: 10 g/L peptone, 5 g/L yeast dipping powder, 10 g/L NaCl. After mixing with water and making up to volume, adjust the initial pH to 7 to 7.5.

 In the ramie biological degumming method of the present invention, the strain activating step further comprises:

 Move the centrifuge tube containing the bacterial solution B to room temperature for 5 to 15 minutes, then press the bacterial solution B to 0.1% to 5%. The inoculation amount is inoculated into a 1 L conical flask containing 200-400 mL of medium No. 1 or medium No. 2 and maintained at a temperature of 33 to 38 ° C for 90 to 180 r/min. The rotation speed was cultured on a shaker for 3 to 6 hours to obtain the bacterial liquid C;

 Wherein the second medium comprises: 10 g/L peptone, 5 g/L yeast dipping powder, 5 g/L NaCl. After mixing with water and constant volume, adjust the initial pH value from 6.8 to 7.7.

 In the ramie biological degumming method of the present invention, the expanding the culturing step further comprises:

 Inoculate the bacterial solution C at a dose of 1% to 5% to 50 L containing 25 to 35 L of No. 2 medium. In the seed tank, at a temperature of 33 to 38 ° C, maintain a rotation speed of 90 to 180 r / min and a ventilation of 5 to 30 L / min, culture for 2 to 5 h to obtain a bacterial liquid. D ;

Then inoculate the bacterial solution D into a 1 t seed tank containing 300-700 kg of No. 2 medium or No. 3 medium, maintaining a rotation speed of 60 to 150 r/min at a temperature of 33 to 38 ° C and 0.5. The aeration volume of ~ 3 m ³ /h was cultured for 2 to 5 hours to obtain the bacterial solution E;

Wherein, the medium No. 3 comprises: 5 to 20 g/L of soybean meal, 5 to 15 g/L of bran, 0 to 5 g/L of NaCl, and 0 to 0.1 g/L of K ₂ HPO ₄ , 0 to 0.1 g/L of (NH ₄ ) ₂ SO ₄ . After mixing with water and making up to volume, adjust the initial pH to 6.8 to 7.7.

The invention has the following beneficial effects: the Bacillus strain Bacillus sp. HG-28 is used for ramie degumming, has low energy consumption and light pollution; the biological degumming time is short, the process is easy to operate, the degumming efficiency is high, and the time of cell culture is high. Short and low cost; the obtained ramie fine dry hemp has high quality, low residual glue rate and high breaking strength, which is superior to the national standard GB/T 20793-2006 "rice fine dry hemp" first-class fine dry hemp appearance and various technical indicators Large-scale production has broad prospects.

detailed description

The technical solutions in the embodiments of the present invention are clearly and completely described below. 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 obtained by those skilled in the art based on the embodiments of the present invention without creative efforts are within the scope of the present invention.

The Bacillus sp. HG-28 used in the present invention is isolated from the soil of the castor plantation in Xian'an District of Xianning County by using a selective medium, and is identified as Bacillus by 16 S rDNA sequence analysis and construction of a phylogenetic tree, and It is named Bacillus sp. HG-28. Bacillus sp. HG-28 was submitted to the China Center for Type Culture Collection (CCTCC, address Wuhan University, Wuhan, Hubei Province, China) on July 2, 2013. The deposit number is CCTCC No: M 2013308.

 Example 1

 Strain activation procedure: Move the centrifuge tube containing the bacterial solution B to room temperature for 5 minutes, then press the bacterial solution B to 0.1%. The inoculation amount was inoculated into a 1 L Erlenmeyer flask containing 200 No. 1 medium, and maintained at 180 r/min at a temperature of 38 ° C for 6 h on a shaker to obtain a bacterial solution C. The formulation of No. 1 medium is: 10 g/L peptone per unit volume of water, 5 g/L yeast dipping powder and 10 g/L NaCl, and the initial pH value is adjusted to 7 .

Expanding the culture step: Inoculum C was inoculated into a 50 L seed tank containing 35 L of No. 2 medium at a rate of 5%, maintaining a rotational speed of 90 r/min and 5 L/ at a temperature of 33 °C. The aeration of min was cultured for 5 h to obtain the bacterial solution D. Then, the bacterial solution D was inoculated into a 1 t seed tank containing 700 kg of the second medium, and kept at a temperature of 38 ° C for 60 r / min. The rotation speed and the aeration of 0.5 m ³ /h were cultured for 2 hours to obtain the bacterial solution E. The formulation of the second medium was: 10 g/L peptone per unit volume of water, 5 g/L yeast dipping powder, 5 g/L of NaCl and adjust the initial pH to 6.8.

 The dilution step includes: injecting 5.5t of water into the fermenter, maintaining the temperature at 34 ° C, adjusting the pH value to 6.8, and the bacterial solution E Inject the mixture into the fermenter at a volume fraction of 2%, and mix evenly to obtain the degumming bacteria F.

Biological degumming: The ramie raw hemp loaded into the cage is immersed in the degumming bacteria F. The mass ratio of the ramie ramie to the degumming bacterium F is 1:1:1, and the air is introduced into the air at a flow rate of 3 m ³ /h. After degumming for 12 h, the fermenter was sealed and heated to raise the pressure to 0.15 MPa for 40 min to end the degumming process.

 After the biological degumming is completed, after the process of copying, bleaching, oiling, drying, etc., the fineness of the ramie single fiber after degumming is 5.26 dtex. The breaking strength of the bundle fiber is 5.02 cN/dtex, the residual rubber ratio is 1.75%, the whiteness is 51 degree, the pH value is 6.89, and the moisture regain rate is 9.13%.

 Example 2

 Strain activation procedure: Move the centrifuge tube containing the bacterial solution B to room temperature for 15 min, then press the bacterial solution B at 5%. The inoculation amount was inoculated into a 1 L conical flask containing 400 mL of No. 1 medium, and maintained at 90 r/min at 33 ° C for 3 h to obtain a bacterial solution C. The formulation of No. 1 medium is: 10 g/L peptone per unit volume of water, 5 g/L yeast dipping powder and 10 g/L NaCl, and the initial pH value is adjusted to 7.2. .

Expanding the culture step: Inoculum C was inoculated into a 50 L seed tank containing 25 L of No. 2 medium at a temperature of 38 ° C, maintained at 180 r/min and 30 L/ at a temperature of 38 ° C. The ventilation rate of min was cultured for 2 h to obtain the bacterial liquid D; the medium of the second medium was prepared by mixing 10 g/L peptone per unit volume of water, 5 g/L yeast dipping powder, and 5 g/L NaCl. And adjust the initial pH to 7.2. Then, inoculum D was inoculated into a 1 t seed tank containing 300 kg of medium No. 3, maintained at a temperature of 33 ° C, maintained at a speed of 150 r / min and aeration of 3 m ³ / h, and cultured for 5 h. To obtain the bacterial liquid E; the medium of the third medium is prepared by mixing 5 g/L of soybean meal, 15 g/L of bran, 5 g/L of NaCl per unit volume of water, and adjusting the initial pH value to 6.8. .

 The dilution step includes: injecting 5.5t of water into the fermenter, keeping the temperature at 37 ° C, adjusting the pH value to 7.5, and the bacterial solution E Inject the fermenter in a volume fraction of 6% and mix well to obtain the degumming bacteria F.

Biological degumming: The ramie raw hemp loaded into the cage is immersed in the degumming bacteria F. The mass ratio of the ramie ramie to the degumming bacterium F is 1: 12, and the air is introduced into the air at a ventilation of 0.5 m ³ /h. After degumming for 4 h, the fermenter was sealed and heated to raise the pressure to 0.1 MPa for 20 min to complete the degumming process.

 After the biological degumming is completed, after the process of copying, bleaching, oiling, drying, etc., the fineness of the ramie single fiber after degumming is 5.19 dtex. The breaking strength of the bundle fiber is 5.05 cN/dtex, the residual gel ratio is 1.63%, the whiteness is 49 degrees, the pH is 7.01, and the moisture regain is 9.06%.

 Example 3

 Strain activation procedure: Move the centrifuge tube containing the bacterial solution B to room temperature for 10 min, then press the bacterial solution B at 3%. The inoculation amount was inoculated into a 1 L conical flask containing 300 mL of No. 2 medium, and maintained at a temperature of 37 ° C for 120 h/min, and cultured on a shaker for 4 h to obtain a bacterial solution. C; No. 2 medium is formulated by mixing 10 g/L peptone per unit volume of water, 5 g/L yeast dipping powder, 5 g/L NaCl, and adjusting the initial pH value to 7.7. .

Expanding the culture step: Inoculating the bacterial solution C into a 50 L seed tank containing 30 L of No. 2 medium at a temperature of 37 ° C, maintaining a rotation speed of 120 r/min and 20 L/min aeration, cultured for 4 h, to obtain bacterial solution D; then inoculum D was inoculated into a 1 t seed tank containing 500 kg of medium No. 3, maintained at a temperature of 37 ° C, maintained at 120 r / The rotation speed of min and the ventilation of 1.5 m ³ /h were cultured for 4 h to obtain the bacterial liquid E. The formulation of the third medium was: 20 g/L of soy meal, 5 bran, 0.1 per unit volume of water. g/L of K ₂ HPO ₄ , 0.1 g/L of (NH ₄ ) ₂ SO ₄ , and adjusting the initial pH to 7.7.

 The dilution step includes: injecting water into the fermenter, keeping the temperature at 36 ° C, adjusting the pH value to 7.0, and pressing the bacterial solution E to 4%. The volume fraction is injected into the fermenter and mixed evenly to obtain the degummed bacterial liquid F.

Biological degumming: The ramie ramie loaded into the cage is immersed in the degumming bacterium F, and the mass ratio of the ramie ramie to the degumming bacterium F is 1: 12, and the air is introduced into the air at 1.5 m ³ /h. After degumming for 6 h, the fermenter was sealed and heated to raise the pressure to 0.12 MPa for 30 min to complete the degumming process.

 After the biological degumming is completed, after the process of copying, bleaching, oiling, drying, etc., the fineness of the ramie single fiber after degumming is 5.12 dtex. The breaking strength of the bundle fiber is 5.15 cN/dtex, the residual gel ratio is 1.53%, the whiteness is 51 degrees, the pH is 6.98, and the moisture regain is 9.21%.

Claims (5)

1. A ramie biological degumming method, characterized in that the method comprises the following steps:

   S1, degumming bacterial culture: using Bacillus sp. HG-28 as a strain, through the strain activation step, expanding the culture step, and diluting the step, obtaining the degumming bacteria F for the ramie biological degumming;

   S2, biological degumming: the ramie raw hemp into the hemp cage is immersed in the degumming bacteria F, the mass ratio of the ramie ramie to the degumming bacterium F is 1: 11 ~ 1 : 12 to 0.5 ~ 3 m ³ / h The ventilation is introduced into the air. After 4 to 12 hours of biological degumming, the fermenter is sealed and heated to raise the pressure to 0.1 to 0.15 MPa for 20 to 40 minutes to complete the degumming process.
2. The ramie biological degumming method according to claim 1, wherein said step S1 further comprises a strain preservation step, wherein the strain storage step is specifically:

   Bacillus sp. HG-28 was inoculated into a 250 mL Erlenmeyer flask containing 50-100 mL of medium No. 1 and maintained at a temperature of 33-38 ° C for 90-180 r/min. Incubate for 3 to 6 hours on a shaker to obtain the bacterial solution A. Transfer the bacterial solution A to the glycerol aqueous solution with a volume fraction of 50% in a ratio of 3:2 and mix it into a 1 to 5 mL centrifuge tube to obtain a bacterial solution B. , − kept at 20 °C for standby;

   Wherein the first medium comprises: 10 g/L peptone, 5 g/L yeast dipping powder, 10 g/L NaCl .
3. The ramie biological degumming method according to claim 2, wherein the step of activating the strain further comprises:

   Move the centrifuge tube containing the bacterial solution B to room temperature for 5 to 15 minutes, then press the bacterial solution B to 0.1% to 5%. The inoculation amount is inoculated into a 1 L conical flask containing 200-400 mL of medium No. 1 or medium No. 2 and maintained at a temperature of 33 to 38 ° C for 90 to 180 r/min. The rotation speed was cultured on a shaker for 3 to 6 hours to obtain the bacterial liquid C;

   Wherein the second medium comprises: 10 g/L peptone, 5 g/L yeast dipping powder, 5 g/L NaCl .
4. The ramie biological degumming method according to claim 3, wherein the step of expanding the culture further comprises:

   Inoculate the bacterial solution C in a 50 L seed tank containing 25 to 35 L of No. 2 medium at an inoculation amount of 1% to 5%, at 33 ～ At a temperature of 38 ° C, maintain a rotational speed of 90 ~ 180 r / min and a ventilation of 5 ~ 30 L / min, culture for 2 ~ 5 h, to obtain bacterial liquid D ;

   Then inoculate the bacterial solution D into a 1 t seed tank containing 300-700 kg of No. 2 medium or No. 3 medium, maintaining a rotation speed of 60 to 150 r/min at a temperature of 33 to 38 ° C and 0.5. The aeration volume of ~ 3 m ³ /h was cultured for 2 to 5 hours to obtain the bacterial solution E;

   Wherein, the medium No. 3 comprises: 5-20 g/L of soybean meal, 5-15 g/L of bran, 0-5 g/L of NaCl, 0 to 0.1 g/L of K ₂ HPO ₄ , 0 to 0.1 g/L of (NH ₄ ) ₂ SO ₄ .
5. The ramie biological degumming method according to claim 4, wherein the diluting step further comprises:

   Inject water into the fermenter, keep the temperature at 34 ~ 37 °C, adjust the pH value to 6.8 ~ 7.5, add the bacteria solution E And the mixture is uniformly mixed to obtain the degumming liquid F, and the bacterial liquid E accounts for 2% to 6% of the degumming liquid F.