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Fibrin enzymolysis auxiliary agent from stalk stuff of plant

A technology of plant straw and cellulase, which is applied to the preparation method of peptides, organic chemistry, peptides, etc., can solve problems such as hindering the large-scale utilization of natural cellulose resources, reducing the degradation efficiency of natural cellulose, and immature production technology. Achieve low price, reduce production cost, reduce usage and cost

Inactive Publication Date: 2007-12-26
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This dense aggregate state also makes it difficult for cellulase to access it, reducing the degradation efficiency of natural cellulose
Moreover, the specific activity of cellulase is generally low at present, and the production technology is not mature enough, so that the production cost of the enzyme is too high, and it also hinders the large-scale utilization of natural cellulose resources.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Ultrafinely pulverize the corn stalks to 50 meshes, add the corn stalks to water at a ratio of 1:10 by mass of the corn stalks and clear water, and wash them twice; filter them with nylon filter cloth after each cleaning, and finally The filter residue on the filter cloth is the obtained cell wall.

[0026] Add 0.1N NaOH solution (30mM L-ascorbic acid VC, mass percentage is 0.3% mercaptoethanol, 3mM PMSF and 8mM EDTA in this NaOH solution) by the mass ratio of cell wall and NaOH solution in the cell wall, Incubate at 50°C for 20 hours, and add PMSF every 4 hours to a final concentration of 3mM; after the extraction is complete, centrifuge at 2000 rpm for 10 minutes to take the supernatant;

[0027] Adjust the pH of the supernatant to 4 with hydrochloric acid, and add 1.5 times the volume of -20°C 95% V / V alcohol to shake well, and place it at 4°C for 2 hours; centrifuge at 8000 rpm for 15 minutes to obtain the precipitate. After the sediment is dried under low-temperat...

Embodiment 2

[0031] Superfinely pulverize the wheat straw to 40 meshes, add the wheat straw to the water at a ratio of 1:6 by weight of the wheat straw and clean water, and wash once; after each cleaning, filter with a nylon filter cloth, and finally The residue on the filter is the obtained cell wall.

[0032] Add 0.1N NaOH solution (containing 20mM L-ascorbic acid VC, mass percentage in this NaOH solution is 0.1% mercaptoethanol, 1mMPMSF and 5mM EDTA) in the ratio of 1: 6 by the mass ratio of cell wall and NaOH solution. Incubate at 30°C for 10 hours, and add PMSF every 3 hours to a final concentration of 1 mM; after extraction, centrifuge at 1000 rpm for 5 minutes to take the supernatant;

[0033] Adjust the pH of the supernatant to 4.5 with hydrochloric acid, add 1 volume of -20°C 95% (V / V) alcohol, shake well, and place it at 4°C for 1 hour; centrifuge at 3000 rpm for 10 minutes to obtain a precipitate After the precipitate is dried under low-temperature vacuum, the wheat straw cell ...

Embodiment 3

[0037] The sorghum straw is ultrafinely pulverized to 60 mesh, and the sorghum straw material is added to the water at a ratio of 1:12 by mass of the sorghum straw material and clear water, and washed three times; after each washing, it is filtered with a nylon filter cloth, Finally, the filter residue on the filter cloth is the obtained cell wall.

[0038]Add 0.5N NaOH solution (50mM L-ascorbic acid VC, mass percentage is 0.5% mercaptoethanol, 5mM PMSF and 10mM EDTA in this NaOH solution) by the mass ratio of cell wall and NaOH solution in the cell wall, Incubate at 70°C for 30 hours, and add PMSF every 5 hours to a final concentration of 5mM; after the extraction is completed, centrifuge at 3000 rpm for 15 minutes to take the supernatant;

[0039] Adjust the pH of the supernatant to 5.5 with hydrochloric acid, add 2 times the volume of -20°C 95% (V / V) alcohol, shake well, and place it at 4°C for 3 hours; centrifuge at 10,000 rpm for 20 minutes to obtain a precipitate After ...

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Abstract

This invention relates to a method for preparing enzymolysis aid of cellulose from crop straws. The method comprises: (1) washing crop straws with water, and filtering to obtain cell wall filter residue; (2) adding 0.1-0.5 mol / L NaOH solution containing 20-50 mM ascorbic acid Vc, 0.1-0.5 wt.% mercaptoethanol, 1-5 mM PMSF and 5-10 mM EDTA; (3) keeping at 30-70 deg.C for 10-30 h, adding PMSF every 3-5 h until the final concentration is 1-5 mM, and extracting; (4) centrifuging after extraction, and collecting the supernatant; (5) adjusting the pH value of the supernatant with HCl, adding ethanol, shaking uniformly, standing, centrifuging, collecting the precipitate, and vacuum-drying at low temperature to obtain enzymolysis aid of cellulose 10 wt.% of cell walls of crop straws. The method has such advantages as abundant raw materials, and low cost.

Description

technical field [0001] The invention relates to a cellulose enzymatic hydrolysis aid, in particular to a cellulose enzymatic hydrolysis aid derived from plant straw materials. Background technique [0002] Natural cellulose is the most abundant renewable resource on earth. In my country, the reserves are also very rich, with an annual output of 1.145 billion tons; among them, the output of straw and husks alone reaches 700 million tons; in addition, the amount of natural cellulose derived from forestry by-products, urban garbage and industrial waste is also very large. considerable. However, these resources have not been rationally developed for a long time, and most of them are incinerated except a small amount for papermaking and feed. This is not only a huge waste of natural resources, but also causes serious pollution to the environment. Cellulose hydrolysis represents one of the largest material flows in the global carbon cycle, and its potential importance in the con...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/36C07K1/14C07K2/00
Inventor 陈洪章卢迪韩业君
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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