CN114908133A - Method for improving yield of reducing sugar by hydrolyzing corn straws through biological combined ammoniation pretreatment - Google Patents
Method for improving yield of reducing sugar by hydrolyzing corn straws through biological combined ammoniation pretreatment Download PDFInfo
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- CN114908133A CN114908133A CN202210412143.8A CN202210412143A CN114908133A CN 114908133 A CN114908133 A CN 114908133A CN 202210412143 A CN202210412143 A CN 202210412143A CN 114908133 A CN114908133 A CN 114908133A
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- 240000008042 Zea mays Species 0.000 title claims abstract description 70
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 title claims abstract description 70
- 235000002017 Zea mays subsp mays Nutrition 0.000 title claims abstract description 70
- 235000005822 corn Nutrition 0.000 title claims abstract description 70
- 239000010902 straw Substances 0.000 title claims abstract description 66
- 238000000034 method Methods 0.000 title claims abstract description 29
- 235000000346 sugar Nutrition 0.000 title claims abstract description 27
- 230000003301 hydrolyzing effect Effects 0.000 title claims abstract description 16
- 240000001462 Pleurotus ostreatus Species 0.000 claims abstract description 30
- 235000001603 Pleurotus ostreatus Nutrition 0.000 claims abstract description 30
- 238000000855 fermentation Methods 0.000 claims abstract description 13
- 230000004151 fermentation Effects 0.000 claims abstract description 13
- ATRRKUHOCOJYRX-UHFFFAOYSA-N Ammonium bicarbonate Chemical compound [NH4+].OC([O-])=O ATRRKUHOCOJYRX-UHFFFAOYSA-N 0.000 claims abstract description 10
- 239000001099 ammonium carbonate Substances 0.000 claims abstract description 10
- 235000012501 ammonium carbonate Nutrition 0.000 claims abstract description 10
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 9
- 238000003860 storage Methods 0.000 claims abstract description 6
- 230000007613 environmental effect Effects 0.000 claims abstract description 5
- 238000010563 solid-state fermentation Methods 0.000 claims abstract description 4
- 239000000843 powder Substances 0.000 claims description 41
- 108010059892 Cellulase Proteins 0.000 claims description 37
- 229940106157 cellulase Drugs 0.000 claims description 37
- 239000000243 solution Substances 0.000 claims description 33
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 33
- 239000007788 liquid Substances 0.000 claims description 18
- 238000005303 weighing Methods 0.000 claims description 15
- 238000001035 drying Methods 0.000 claims description 13
- 230000001954 sterilising effect Effects 0.000 claims description 13
- 235000007685 Pleurotus columbinus Nutrition 0.000 claims description 11
- 230000007062 hydrolysis Effects 0.000 claims description 11
- 238000006460 hydrolysis reaction Methods 0.000 claims description 11
- 239000007787 solid Substances 0.000 claims description 11
- 230000000694 effects Effects 0.000 claims description 10
- 239000007974 sodium acetate buffer Substances 0.000 claims description 10
- BHZOKUMUHVTPBX-UHFFFAOYSA-M sodium acetic acid acetate Chemical compound [Na+].CC(O)=O.CC([O-])=O BHZOKUMUHVTPBX-UHFFFAOYSA-M 0.000 claims description 10
- 238000003756 stirring Methods 0.000 claims description 8
- 239000001963 growth medium Substances 0.000 claims description 6
- 238000012258 culturing Methods 0.000 claims description 5
- 238000004659 sterilization and disinfection Methods 0.000 claims description 4
- 230000003213 activating effect Effects 0.000 claims description 3
- 238000009630 liquid culture Methods 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 108090000790 Enzymes Proteins 0.000 claims description 2
- 102000004190 Enzymes Human genes 0.000 claims description 2
- 241000233866 Fungi Species 0.000 claims description 2
- 238000005119 centrifugation Methods 0.000 claims description 2
- 238000012824 chemical production Methods 0.000 claims description 2
- 229940088598 enzyme Drugs 0.000 claims description 2
- 238000004321 preservation Methods 0.000 claims 1
- 229920002678 cellulose Polymers 0.000 abstract description 9
- 239000001913 cellulose Substances 0.000 abstract description 9
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 abstract description 6
- 239000002028 Biomass Substances 0.000 abstract description 4
- 239000002351 wastewater Substances 0.000 abstract description 4
- 235000014633 carbohydrates Nutrition 0.000 abstract description 3
- 150000001720 carbohydrates Chemical class 0.000 abstract description 3
- 239000003112 inhibitor Substances 0.000 abstract description 3
- 238000005406 washing Methods 0.000 abstract description 3
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 abstract description 2
- 239000003513 alkali Substances 0.000 abstract description 2
- 235000011114 ammonium hydroxide Nutrition 0.000 abstract description 2
- 230000001988 toxicity Effects 0.000 abstract description 2
- 231100000419 toxicity Toxicity 0.000 abstract description 2
- 239000008367 deionised water Substances 0.000 description 6
- 229910021641 deionized water Inorganic materials 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 238000002156 mixing Methods 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 229920002488 Hemicellulose Polymers 0.000 description 3
- 239000002029 lignocellulosic biomass Substances 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 230000007797 corrosion Effects 0.000 description 2
- 238000005260 corrosion Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000003912 environmental pollution Methods 0.000 description 2
- 229920005610 lignin Polymers 0.000 description 2
- 238000009629 microbiological culture Methods 0.000 description 2
- 238000002203 pretreatment Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000002002 slurry Substances 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000002551 biofuel Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/02—Monosaccharides
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P2201/00—Pretreatment of cellulosic or lignocellulosic material for subsequent enzymatic treatment or hydrolysis
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/59—Biological synthesis; Biological purification
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Abstract
The invention discloses a method for improving the yield of reducing sugar by hydrolyzing corn straws through biological combined ammoniation pretreatment. Relates to the technical field of biomass energy. The method comprises the following steps: solid state fermentation of oyster mushroom; and (4) ammoniation pretreatment. Compared with the traditional biological pretreatment, the method shortens the fermentation time, improves the enzymolysis rate and reduces the loss of carbohydrate. The ammonium carbonate is used as an ammoniating agent, is different from ammonia water, has lower price, lower toxicity and corrosivity, does not need special equipment in storage and transportation, and has higher safety. Compared with the alkali pretreatment mainly comprising sodium hydroxide, the method has the advantages of milder condition, no need of washing, no generation of a large amount of wastewater, low requirement on equipment and environmental friendliness. The loss of cellulose components is small, the yield of reducing sugar is high, no inhibitor is produced, and the subsequent fermentation is facilitated.
Description
Technical Field
The invention relates to the technical field of biomass energy, in particular to a method for improving the yield of reducing sugar by hydrolyzing corn straws through biological combined ammoniation pretreatment.
Background
Currently, energy shortage has become a critical issue in the world, and biomass energy, as a renewable and recyclable energy source available worldwide, is one of the important solutions to the energy problem. Lignocellulosic biomass has become a major source of biofuels due to its advantages of low cost, wide sources, large reserves, and being renewable. Lignocellulosic biomass is composed primarily of cellulose, hemicellulose, and lignin, wherein cellulose and hemicellulose are potential sources of fermentable sugars, and lignin intertwines between cellulose and hemicellulose to form a natural physical barrier, thereby reducing the accessibility of cellulase to cellulose. In order to achieve high value utilization of lignocellulosic biomass, it is necessary to adopt effective pretreatment methods to overcome the recalcitrance of natural biomass and increase the accessibility of cellulose to cellulase hydrolysis.
The pretreatment methods generally employed mainly include three types, i.e., chemical, physical and biological pretreatment. Biological pretreatment is concerned about due to the characteristics of environmental protection, simple operation and the like, but the defects of long fermentation period, large carbohydrate loss, low sugar yield and the like of the treatment technology still exist. The alkaline pretreatment in the chemical pretreatment has the defects of environmental pollution, inhibitor generation, easy corrosion of equipment and the like.
Therefore, the problem to be solved by the technical personnel in the field is how to provide a method for improving the hydrolysis of corn straws to produce reducing sugar.
Disclosure of Invention
In view of this, the invention provides a method for improving the yield of reducing sugar by corn straw hydrolysis through biological combined ammoniation pretreatment. Solves the problems of long time consumption, low efficiency, high compound loss, corrosion-resistant equipment required by the traditional treatment, wastewater generation, complex post-treatment process, environmental pollution and the like.
The ammonia has low toxicity and corrosivity, strong volatility and easy recycling, and the enzymolysis rate of the cellulose can be improved by changing the crystallinity and the form of the cellulose. The ammoniating agent adopted by the invention has more advantages in the aspects of safety, transportation, storage and the like, and the yield of reducing sugar treated by a small amount of ammonium carbonate after the corn straws are subjected to solid state fermentation of the oyster mushroom can be obviously improved. In addition, the invention reduces the discharge cost of a large amount of washing wastewater, does not need large-scale equipment, has simple operation and is more beneficial to commercial production.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for improving the yield of reducing sugar by hydrolyzing corn straws through biological combined ammoniation pretreatment comprises the following steps:
(1) solid state fermentation of oyster mushroom: weighing a certain amount of corn straw powder, adjusting the water content, sterilizing, inoculating oyster mushroom seed liquid, fermenting, culturing and drying to obtain the corn straw powder subjected to biological pretreatment;
(2) ammoniation pretreatment: weighing a certain amount of biologically pretreated corn straw powder, an ammoniating agent and water, uniformly stirring, storing under a closed condition, drying after ammoniation is finished to obtain the ammoniated pretreated corn straw powder, and adding cellulase for hydrolysis to obtain a reducing sugar solution.
Further, deionized water is used for adjusting the moisture content in the step (1); the fermentation culture and storage equipment is a constant temperature incubator;
step (2) sealing conditions: placing in a container tightly sealed.
Preferably: the grain size of the corn straw powder in the step (1) is 20-40 meshes;
adjusting the water content to 80%;
and (3) sterilization: adopting high-pressure sterilization: sterilizing at 121 deg.C under 100kPa for 40 min;
the strain of the oyster mushroom: pleurotus ostreatus (Pleurotus ostreatus) with accession number of CGMCC 5.930;
further, Pleurotus ostreatus (Pleurotus ostreatus) collection number CGMCC 5.930 was purchased from China general microbiological culture Collection center.
The preparation method of the oyster mushroom seed liquid comprises the following steps: inoculating Pleurotus ostreatus on PDA culture medium, activating for 7d, inoculating 3 fungus cakes with diameter of 0.5cm on PDA liquid culture medium, culturing at 25 deg.C and rotation speed of 160r/min in dark for 7 d; when in use, homogenate for 5-10 s;
the mass ratio of the inoculated oyster mushroom seed liquid is 10 percent;
temperature of fermentation: keeping the temperature at 28 ℃ for 14-28 days.
Preferably: the mass ratio of the corn straw powder subjected to biological pretreatment in the step (2) to the ammoniating agent is 1: 0.05-0.2, wherein the solid content of the biologically pretreated corn straw powder is 30-70%;
and (3) storage: keeping the temperature at 30 ℃ for 7-11 days;
the cellulase comprises: 0.01g/mL of cellulase solution;
and (3) cellulase activity: 100 FPU/g;
adding 0.05moL/L acetic acid-sodium acetate buffer solution with pH value of 4.8 for hydrolysis;
the volume ratio of the cellulase solution to the acetic acid-sodium acetate buffer solution is 3: 7;
the mass ratio of the corn straw powder subjected to ammoniation pretreatment to the cellulase is as follows: 0.2 g: 30 mg;
conditions of hydrolysis: hydrolyzing in a water bath shaker at 50 deg.C and rotation speed of 150r/min for 48 h.
Further, the solid content of the biologically pretreated corn straw powder is equal to the mass of the biologically pretreated corn straw powder/(the mass of the biologically pretreated corn straw powder + the mass of water);
the enzyme activity of the used cellulase is 100FPU/g, each gram of corn straws needs 15FPU cellulase (namely, 15FPU/g) during enzymolysis, and the cellulase is prepared into 0.01g/mL solution and added into an enzymolysis system during use, namely, 3mL (0.01g/mL) of cellulase solution and 7mL of cellulase solution with the pH value of 4.8(0.05 mol.L) are added into 0.2g of corn straws -1 ) Acetic acid-sodium acetate buffer solution.
Preferably: the ammoniating agent in the step (2) is ammonium carbonate.
Preferably, the following components: the step (2) also comprises centrifugation after hydrolysis: 10000r/min, 10 min.
The invention also provides the application of the method in biological energy, environmental protection and chemical production.
According to the technical scheme, compared with the prior art, the invention discloses a method for improving the yield of reducing sugar by hydrolyzing corn straws through biological combined ammoniation pretreatment, and the technical effects are as follows:
1. compared with the traditional biological pretreatment, the method shortens the fermentation time, improves the enzymolysis rate and reduces the loss of carbohydrate.
2. The ammonium carbonate is used as an ammoniating agent, is different from ammonia water, has lower price, lower toxicity and corrosivity, does not need special equipment in storage and transportation, and has higher safety.
3. Compared with the alkali pretreatment mainly comprising sodium hydroxide, the method has the advantages of milder condition, no need of washing, no generation of a large amount of wastewater, low requirement on equipment and environmental friendliness.
4. The loss of cellulose components is small, the yield of reducing sugar is high, no inhibitor is produced, and the subsequent fermentation is facilitated.
Detailed Description
The following will clearly and completely describe the technical solutions in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The embodiment of the invention discloses a method for improving the yield of reducing sugar by hydrolyzing corn straws through biological combined ammoniation pretreatment.
The starting materials and apparatus not mentioned in the examples are all customary commercial starting materials and apparatus, for example: pleurotus ostreatus (Pleurotus ostreatus) with the collection number of CGMCC 5.930 is purchased from China general microbiological culture Collection center; and will not be described in detail herein.
The preparation method of the oyster mushroom seed liquid comprises the following steps: inoculating Pleurotus ostreatus on PDA culture medium, activating for 7d, inoculating 3 bacterial cakes with diameter of 0.5cm on PDA liquid culture medium with hole puncher, culturing at 25 deg.C and rotation speed of 160r/min in dark place for 7 d; when the slurry is used, the slurry is homogenized for 5-10 s.
Example 1
Weighing 10g of 20-40-mesh corn straw, adding deionized water into a 250mL conical flask to adjust the water content to 80%, sterilizing with high-pressure steam (the pressure is 100kPa, the temperature is 121 ℃) for 40min, inoculating 1mL of oyster mushroom liquid seed liquid, placing in a constant-temperature incubator at 28 ℃ for 21 days, and drying after fermentation is finished to obtain the corn straw powder subjected to biological pretreatment;
according to 50% solid content { solid content ═ mass of biologically pretreated corn straw powder/(mass of biologically pretreated corn straw powder + mass of water) }, dissolving ammonium carbonate accounting for 10% (w/w) of the mass of the biologically pretreated corn straw powder in required water, mixing with the biologically pretreated corn straw powder, uniformly stirring, placing in a closed container, storing in a constant-temperature incubator at 30 ℃ for 11 days, and drying after the stirring is finished;
weighing 0.2g of aminated corn stalks, 3mL of 0.01g/mL cellulase solution (cellulase activity: 100FPU/g) and 7mL of cellulase solution with the pH value of 4.8(0.05 mol.L) -1 ) The acetic acid-sodium acetate buffer solution is put into a 50mL centrifuge tube, hydrolyzed in a water bath shaker at 50 ℃ and 150r/min for 48h, the product is centrifuged (10000r/min, time 10min), and the yield of reducing sugar is measured by a DNS method to be 311 mg/g.
Example 2
Weighing 10g of 20-40-mesh corn straw, adding deionized water into a 250mL conical flask to adjust the water content to 80%, sterilizing with high-pressure steam (the pressure is 100kPa, the temperature is 121 ℃) for 40min, inoculating 1mL of oyster mushroom liquid seed liquid, placing in a constant-temperature incubator at 28 ℃ for 14 days, and drying after fermentation is finished to obtain the corn straw powder subjected to biological pretreatment;
dissolving 15% (w/w) ammonium carbonate, which is based on the mass of the biologically pretreated corn straw powder, in required water according to 50% of solid content { solid content ═ mass of the biologically pretreated corn straw powder/(mass of the biologically pretreated corn straw powder + mass of water) }, mixing the dissolved ammonium carbonate with the biologically pretreated corn straw powder, uniformly stirring the mixture, placing the mixture in a closed container, storing the mixture in a constant-temperature incubator at 30 ℃ for 7 days, and drying the mixture after the mixture is dried;
weighing 0.2g of aminated corn stalks, 3mL of 0.01g/mL cellulase solution (cellulase activity: 100FPU/g) and 7mL of cellulase solution with the pH value of 4.8(0.05 mol.L) -1 ) The acetic acid-sodium acetate buffer solution is put into a 50mL centrifuge tube, hydrolyzed in a water bath shaker at 50 ℃ and 150r/min for 48h, the product is centrifuged (10000r/min, time 10min), and the yield of the reducing sugar is measured by a DNS method to be 301 mg/g.
Example 3
Weighing 10g of 20-40-mesh corn straw, adding deionized water into a 250mL conical flask to adjust the water content to 80%, sterilizing with high-pressure steam (the pressure is 100kPa, the temperature is 121 ℃) for 40min, inoculating 1mL of oyster mushroom liquid seed liquid, placing in a constant-temperature incubator at 28 ℃ for 28 days, and drying after fermentation is finished to obtain the corn straw powder subjected to biological pretreatment;
dissolving 15% (w/w) ammonium carbonate, which is based on the mass of the biologically pretreated corn straw powder, in required water according to 50% of solid content { solid content ═ mass of the biologically pretreated corn straw powder/(mass of the biologically pretreated corn straw powder + mass of water) }, mixing with the biologically pretreated corn straw powder, uniformly stirring, placing in a closed container, storing in a 30 ℃ constant-temperature incubator for 11 days, and drying after the stirring is finished;
weighing 0.2g of aminated corn stalks, 3mL of 0.01g/mL cellulase solution (cellulase activity: 100FPU/g) and 7mL of cellulase solution with the pH value of 4.8(0.05 mol.L) -1 ) The acetic acid-sodium acetate buffer solution is put into a 50mL centrifuge tube, hydrolyzed in a water bath shaker at 50 ℃ and 150r/min for 48h, the product is centrifuged (10000r/min, time 10min), and the yield of reducing sugar is 374mg/g measured by a DNS method.
Example 4
Weighing 10g of 20-40-mesh corn straw, adding deionized water into a 250mL conical flask to adjust the water content to 80%, sterilizing with high-pressure steam (the pressure is 100kPa, the temperature is 121 ℃) for 40min, inoculating 1mL of oyster mushroom liquid seed liquid, placing in a constant-temperature incubator at 28 ℃ for 28 days, and drying after fermentation is finished to obtain the corn straw powder subjected to biological pretreatment;
according to 50% solid content { solid content ═ mass of biologically pretreated corn straw powder/(mass of biologically pretreated corn straw powder + mass of water) }, dissolving ammonium carbonate accounting for 10% (w/w) of the mass of the biologically pretreated corn straw powder in required water, mixing with the biologically pretreated corn straw powder, uniformly stirring, placing in a closed container, storing for 9 days in a 30 ℃ constant-temperature incubator, and drying after the mixing is finished;
weighing 0.2g of aminated corn stalks, 3mL of 0.01g/mL cellulase solution (cellulase activity: 100FPU/g) and 7mL of cellulase solution with the pH value of 4.8(0.05 mol.L) -1 ) The acetic acid-sodium acetate buffer solution is put into a 50mL centrifuge tube, hydrolyzed in a water bath shaker at 50 ℃ and 150r/min for 48h, the product is centrifuged (10000r/min, time 10min), and the yield of reducing sugar is measured by a DNS method to be 328 mg/g.
Comparative experiment
1. Compared with the raw material which is not subjected to ammoniation pretreatment:
weighing 0.2g of corn straw and 3mL of 0.01g/mL cellulase solution (cellulase activity: 100FPU/g), putting the corn straw and the cellulase solution into a 50mL centrifuge tube, hydrolyzing the corn straw and the cellulase solution in a water bath shaking table with the temperature of 50 ℃ and the speed of 150r/min for 48h, centrifuging the product, sucking the supernatant, and measuring the yield of reducing sugar by a DNS method to be 205 mg/g.
2. Adjusting the process steps and the raw material ratio:
weighing 10g of 20-40-mesh corn straw, adding deionized water into a 250mL conical flask to adjust the water content to 80%, sterilizing with high-pressure steam (the pressure is 100kPa, the temperature is 121 ℃) for 40min, inoculating 1mL of oyster mushroom liquid seed liquid, placing in a constant-temperature incubator at 28 ℃ for 28 days, and drying after fermentation is finished to obtain the biological pretreatment corn straw powder.
Weighing 0.2g of biologically pretreated corn straw powder, 3mL of 0.01g/mL cellulase solution (cellulase activity: 100FPU/g) and 7mL of acetic acid-sodium acetate buffer solution with the pH value of 4.8(0.05 mol.L < -1 >) into a 50mL centrifuge tube, hydrolyzing in a water bath shaking table with the temperature of 50 ℃ and the pH value of 150r/min for 48h, centrifuging the product, and absorbing the supernatant to measure the yield of the reducing sugar to be 210mg/g by a DNS method.
The embodiments in the present description are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments are referred to each other.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (6)
1. A method for improving the yield of reducing sugar by hydrolyzing corn straws through biological combined ammoniation pretreatment is characterized by comprising the following steps:
(1) solid state fermentation of oyster mushroom: weighing a certain amount of corn straw powder, adjusting the water content, sterilizing, inoculating oyster mushroom seed liquid, fermenting, culturing and drying to obtain the corn straw powder subjected to biological pretreatment;
(2) ammoniation pretreatment: weighing a certain amount of biologically pretreated corn straw powder, an ammoniating agent and water, uniformly stirring, storing under a closed condition, drying after ammoniation is finished to obtain the ammoniated pretreated corn straw powder, and adding cellulase for hydrolysis to obtain a reducing sugar solution.
2. The method for improving the yield of reducing sugar by hydrolyzing corn straws through biological combined ammoniation pretreatment according to claim 1, which is characterized in that: the grain size of the corn straw powder in the step (1) is 20-40 meshes;
the moisture content is adjusted to 80%;
the sterilization comprises the following steps: adopting high-pressure sterilization: sterilizing at 121 deg.C under 100kPa for 40 min;
the strain of the oyster mushroom comprises the following components: pleurotus ostreatus (Pleurotus ostreatus) with preservation number of CGMCC 5.930;
the preparation method of the oyster mushroom seed liquid comprises the following steps: inoculating Pleurotus ostreatus on PDA culture medium, activating for 7d, inoculating 3 fungus cakes with diameter of 0.5cm on PDA liquid culture medium, culturing at 25 deg.C and rotation speed of 160r/min in dark for 7 d; when in use, homogenizing for 5-10 s;
the mass ratio of the oyster mushroom seed liquid to the oyster mushroom seed liquid is 10%;
temperature of fermentation: keeping the temperature at 28 ℃ for 14-28 days.
3. The method for improving the yield of reducing sugar by hydrolyzing corn straws through biological combined ammoniation pretreatment according to claim 1, which is characterized in that: the mass ratio of the corn straw powder subjected to biological pretreatment in the step (2) to the ammoniating agent is 1: 0.05 to 0.2 percent, the solid content of the biologically pretreated corn straw powder is 30 to 70 percent;
the storage comprises the following steps: keeping the temperature at 30 ℃ for 7-11 days;
the cellulase comprises: 0.01g/mL of cellulase solution;
the cellulase has the following enzyme activity: 100 FPU/g;
the hydrolysis also needs to add acetic acid-sodium acetate buffer solution with the pH value of 4.8 and 0.05 moL/L;
the volume ratio of the cellulase solution to the acetic acid-sodium acetate buffer solution is 3: 7;
the mass ratio of the corn straw powder subjected to ammoniation pretreatment to the cellulase is as follows: 0.2 g: 30 mg;
the conditions of the hydrolysis are as follows: hydrolyzing in a water bath shaker at 50 deg.C and rotation speed of 150r/min for 48 h.
4. The method for improving the yield of reducing sugar by hydrolyzing corn straws through biological combined ammoniation pretreatment according to claim 1, which is characterized in that: and (3) the ammoniating agent in the step (2) is ammonium carbonate.
5. The method for improving the yield of reducing sugar by hydrolyzing corn straws through biological combined ammoniation pretreatment according to claim 1, which is characterized in that: the step (2) also comprises centrifugation after hydrolysis: 10000r/min, 10 min.
6. Use of the method according to any one of claims 1 to 5 in bioenergy, environmental protection, and chemical production.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210412143.8A CN114908133B (en) | 2022-04-19 | 2022-04-19 | Method for improving reducing sugar produced by corn straw hydrolysis through biological combined ammoniation pretreatment |
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| CN202210412143.8A CN114908133B (en) | 2022-04-19 | 2022-04-19 | Method for improving reducing sugar produced by corn straw hydrolysis through biological combined ammoniation pretreatment |
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| CN114908133B CN114908133B (en) | 2024-02-13 |
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| CN107177637A (en) * | 2017-06-15 | 2017-09-19 | 沈阳农业大学 | The methods and applications pre-processed using microorganism to maize straw hydrolysis and saccharification |
| CN108148879A (en) * | 2018-03-04 | 2018-06-12 | 华中农业大学 | A kind of preprocess method of maize straw |
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| CA2565433A1 (en) * | 2006-10-25 | 2008-04-25 | Iogen Energy Corporation | Inorganic salt recovery during processing of lignocellulosic feedstocks |
| KR20150093483A (en) * | 2014-02-07 | 2015-08-18 | 한국과학기술원 | Methods for Pretreating Lignocellulosic Biomass |
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| CN107177637A (en) * | 2017-06-15 | 2017-09-19 | 沈阳农业大学 | The methods and applications pre-processed using microorganism to maize straw hydrolysis and saccharification |
| CN108148879A (en) * | 2018-03-04 | 2018-06-12 | 华中农业大学 | A kind of preprocess method of maize straw |
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