CN211856480U - Reaction device and equipment for testing lignocellulose components of edible fungus culture substrate - Google Patents
Reaction device and equipment for testing lignocellulose components of edible fungus culture substrate Download PDFInfo
- Publication number
- CN211856480U CN211856480U CN201922339448.7U CN201922339448U CN211856480U CN 211856480 U CN211856480 U CN 211856480U CN 201922339448 U CN201922339448 U CN 201922339448U CN 211856480 U CN211856480 U CN 211856480U
- Authority
- CN
- China
- Prior art keywords
- edible mushroom
- residue
- mixed solution
- culture medium
- edible
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
Reaction unit and equipment are used in testing of edible mushroom culture substrate lignocellulose component, and the reaction unit includes an electric heating constant temperature water bath and a desk centrifuge. The electric heating constant-temperature water bath kettle is provided with a centrifuge tube for accommodating a first edible mushroom culture medium mixed liquid. And separating a first residue from the first edible mushroom culture medium mixed solution by the table centrifuge, wherein the first residue is mixed with a sulfuric acid solution to form a second edible mushroom culture medium mixed solution, and the second edible mushroom culture medium mixed solution is placed in the electric heating constant-temperature water bath kettle and is separated into a second supernatant and a second residue by the table centrifuge.
Description
Technical Field
The utility model relates to a culture medium's biotechnology field especially relates to a reaction unit and equipment are used in test of domestic fungus culture medium lignocellulose component.
Background
The earth has abundant lignocellulose renewable resources. The dry matter of the plants is generated in the amount of 1.55 x10 every year all over the world11Ton, wherein the total amount of cellulose and hemicellulose is about 8.5X 1010Ton. The degradation and conversion of lignocellulose by using fungi is an effective way for solving the energy shortage crisis. The edible fungi has strong degradation capability on lignocellulose, and simultaneously solves the problems of food shortage, energy crisis, environmental pollution and the like faced by the current. In recent years, in terms of lignocellulose activity and genomics, quantitative determination of each component of lignocellulose is very important in research processes for comparing the influence of different strains, different raw materials and different cultivation modes on lignocellulose degradation efficiency.
Lignocellulose is mainly composed of cellulose, hemicellulose and lignin. Cellulose is a macromolecular polysaccharide composed of D-glucose with beta-1, 4 glycosidic bonds. Further, cellulose (cellulose) is a class of organic compounds having the general chemical formula (C)6H10O5) n is a number of hundredsTo several thousand linear chains (glycosidic linkages) of β (1 → 4) -linked D-glucose units. Hemicellulose is a polymer composed of several different types of monosaccharides including xylose, arabinose, galactose, etc. Lignin is an aromatic polymer formed from phenylpropane held together by different types of chains. The composition and proportions of these polymers vary from plant species to plant species. The traditional method for measuring each component of lignocellulose mainly comprises a gravimetric method and an acid hydrolysis chemical sugar determination method, but the methods have the defects of long measuring period, complicated steps and low accuracy of measuring results. The National Renewable Energy Laboratory (NREL) of america hydrolyzes samples stepwise using 72% concentrated sulfuric acid and 4% dilute sulfuric acid, and then determines the monosaccharide content in the filtrate using a High Performance Liquid Chromatography (HPLC) method. The method is widely applied to the pretreatment of plant lignocellulose and the detection of the energy regeneration process due to small error and stable and reliable result.
However, in the process of measuring lignocellulose in a culture substrate of Agaricus bisporus (Agaricus bisporus) by using the NREL established method, the hydrolysis reaction of the method needs to be carried out in a pressure container, the potential overflow of sulfuric acid exists in the high-temperature and high-pressure hydrolysis process, and the hydrolysis reaction system is large, so that the method is not beneficial to measuring large-batch samples. Firstly, the dilute acid hydrolysis research of Zhangping lignocellulose biomass provides that within a certain range, the acid concentration, the temperature and the reaction time can be improved, so that the hydrolysis rate can be improved. Therefore, the utility model provides an improve equipment and method to edible mushroom culture medium lignocellulose component survey to above-mentioned each argument to improve lignocellulose component survey mode effectively.
Disclosure of Invention
An advantage of the utility model is that it provides a reaction unit and equipment are used in test of domestic fungus culture medium lignocellulose component, wherein reduces sulphuric acid effectively and spills over the risk and increase the speed of sulphuric acid hydrolysis reaction. In particular, the utility model utilizes lower temperature to avoid sulfuric acid to overflow, and utilizes the concentration of higher sulfuric acid to improve the rate of hydrolysis.
The utility model discloses an advantage lies in that it provides a reaction unit and equipment are used in test of domestic fungus culture medium lignocellulose component, wherein at the dilute sulphuric acid hydrolysis sample in-process, replaces high-pressure vessel with the water bath and reduces sulphuric acid and spill over the risk to improve the speed of sulphuric acid concentration acceleration sulphuric acid hydrolysis reaction.
An advantage of the utility model is that it provides a reaction unit and equipment are used in test of domestic fungus culture medium lignocellulose component, wherein the utility model discloses replace 121 ℃ in the NREL with 100 ℃ dilute sulfuric acid to reduce sulphuric acid and spill over the risk. Also, this problem is ameliorated by increasing the concentration of sulfuric acid, since a decrease in temperature will result in a decrease in the rate of hydrolysis. In other words, the present invention improves the reaction system of hydrolysis at 121 ℃ of 0.3g sample in 87ml 4% (w/w) sulfuric acid in NREL method to hydrolysis at 100 ℃ of 0.1g sample in 10ml 15.4% (w/w) sulfuric acid, so as to effectively avoid sulfuric acid overflow and increase the reaction rate of sulfuric acid hydrolysis.
The utility model has the advantages of it provides a reaction unit and equipment are used in test of domestic fungus culture medium lignocellulose component, wherein through quick solid-liquid separation in a centrifuging tube, improves survey efficiency. Particularly, the whole system of the hydrolysis reaction is reduced due to the increase of the concentration of the sulfuric acid, so that the solid-liquid separation can be quickly carried out in a centrifugal tube, and the measurement efficiency is improved.
One advantage of the present invention is that it provides an edible mushroom cultivation substrate lignocellulose component testing apparatus and method, wherein the optimum hydrolysis time of different edible mushroom cultivation substrates is provided under the condition of fixed sulfuric acid concentration and reaction temperature.
The other advantages and features of the invention will be fully apparent from the following detailed description and realized by means of the instruments and combinations particularly pointed out in the appended claims.
According to the utility model discloses, can realize aforementioned purpose and other purposes and advantage the utility model discloses a reaction unit is used in test of domestic fungus culture medium lignocellulose component, include:
an electric heating constant temperature water bath pot, which is provided with a centrifuge tube for accommodating a first edible mushroom culture medium mixed solution;
and the table centrifuge is used for separating a first residue from the first edible mushroom culture medium mixed solution, wherein the first residue is mixed with a sulfuric acid solution to form a second edible mushroom culture medium mixed solution, the second edible mushroom culture medium mixed solution is placed in the electric heating constant-temperature water bath kettle, and a second supernatant and a second residue are separated from the second edible mushroom culture medium mixed solution by the table centrifuge.
In some embodiments, the centrifuge apparatus further comprises an actuating jaw device that grips and positions the centrifuge tube between the electrically heated thermostat water bath and the table centrifuge.
The utility model also provides an edible mushroom culture medium lignocellulose component test equipment, it includes:
a reaction unit for testing lignocellulose components in edible fungus culture substrate comprises:
an electric heating constant temperature water bath pot, which is provided with a centrifuge tube for accommodating a first edible mushroom culture medium mixed solution; and
a desk centrifuge, which separates a first residue from the cooled first edible mushroom culture medium mixed solution and carries out centrifugal washing;
an electric heating air blast drying box, which dries the washed first residue, wherein the dried first residue is mixed with sulfuric acid solution to form a second edible mushroom cultivation substrate mixed solution, the second edible mushroom cultivation substrate mixed solution is placed in the electric heating constant-temperature water bath kettle, and the second edible mushroom cultivation substrate mixed solution is separated into a second supernatant and a second residue by the table centrifuge;
a liquid phase anion chromatograph which tests the monosaccharide content of the second supernatant and calculates cellulose and hemicellulose; and
a lignin reaction unit which tests and calculates acid insoluble lignin from the second residue.
In some embodiments, the reaction apparatus for testing the lignocellulose component in the edible fungus cultivation substrate further comprises an actuating clamping jaw device which clamps and places the centrifuge tube between the devices.
In some embodiments, the second edible fungi culture medium mixed solution is placed in the electric heating constant-temperature water bath for heat preservation, so that cellulose and hemicellulose in the edible fungi culture medium sample are hydrolyzed into arabinose, galactose, glucose, xylose and mannose under the catalysis of sulfuric acid.
In some embodiments, the lignin reaction apparatus comprises a sand core funnel, a box-type electric resistance furnace, a circulating water type multipurpose vacuum pump, and an oven, wherein the second residue is filtered by the sand core funnel and washed with water by the circulating water type multipurpose vacuum pump, the sand core funnel with the residue left is placed in the oven and weighed after cooling, and the sand core funnel is placed in the box-type electric resistance furnace to be burned and weighed after cooling, wherein the centrifuge tube is moved between the apparatuses by the actuating jaws.
Further objects and advantages of the invention will be fully apparent from the ensuing description and drawings.
These and other objects, features and advantages of the present invention will become more fully apparent from the following detailed description, the accompanying drawings and the appended claims.
Drawings
FIG. 1 is a logic diagram of an apparatus for determining the lignocellulosic component of an edible fungus culture medium according to a preferred embodiment of the present invention.
Fig. 2 is a schematic perspective view of an electric heating thermostat water bath of an apparatus for measuring the lignocellulose component in the edible fungus cultivation substrate according to a preferred embodiment of the present invention.
Fig. 3 is a perspective view of a table centrifuge of an apparatus for measuring lignocellulose components in edible fungus cultivation substrate according to a preferred embodiment of the present invention.
Fig. 4 is a schematic perspective view of a reaction device for testing the lignocellulose component in the edible fungus cultivation substrate of the edible fungus cultivation substrate lignocellulose component testing device according to a preferred embodiment of the present invention. Illustrating the movement of the centrifuge tube by actuating the jaw mechanism.
Fig. 5 is a schematic perspective view of a box-type resistance furnace of the edible fungus cultivation substrate lignocellulose component measuring device according to a preferred embodiment of the present invention.
Fig. 6 is a perspective view schematically illustrating an oven of the edible fungus cultivation substrate lignocellulose component measuring apparatus according to a preferred embodiment of the present invention.
Fig. 7 is a perspective view of a high performance liquid anion chromatograph of the edible fungus culture medium lignocellulose component measuring device according to a preferred embodiment of the present invention.
Fig. 8 is a perspective view schematically illustrating an apparatus for determining the lignocellulose component in the edible fungus cultivation substrate according to a preferred embodiment of the present invention. Illustrating the movement of the centrifuge tube by actuating the jaw mechanism.
Detailed Description
The following description is presented to disclose the invention so as to enable any person skilled in the art to practice the invention. The preferred embodiments in the following description are given by way of example only, and other obvious variations will occur to those skilled in the art. The basic principles of the invention, as defined in the following description, may be applied to other embodiments, variations, modifications, equivalents and other technical solutions without departing from the spirit and scope of the invention.
It will be understood by those skilled in the art that in the present disclosure, the terms "longitudinal," "lateral," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," and the like are used in a generic and descriptive sense only and not for purposes of limitation, as the terms are used in the description to indicate that the referenced device or element must have the specified orientation, be constructed and operated in the specified orientation, and not for the purposes of limitation.
It is understood that the terms "a" and "an" should be interpreted as meaning that a number of one element or element is one in one embodiment, while a number of other elements is one in another embodiment, and the terms "a" and "an" should not be interpreted as limiting the number.
Referring to fig. 1 to 8, an apparatus and a method for testing the lignocellulose component of an edible fungus cultivation substrate are provided according to a preferred embodiment of the present invention, so as to be suitable for mass measurement of samples. The edible fungus culture substrate lignocellulose component measuring equipment 1 comprises a reaction device 100 for testing the edible fungus culture substrate lignocellulose component, an electronic balance 200, a vortex mixer 300, an electric heating blowing drying box 400, a lignin reaction device 500 and a high performance liquid anion chromatograph 600.
The reaction device 100 for testing the lignocellulose component of the edible fungus culture medium comprises an electric heating constant-temperature water bath 101 and a table centrifuge 102, wherein the table centrifuge 102 comprises at least one centrifuge tube 1021 for accommodating an edible fungus culture medium sample and distilled water and forming a first edible fungus culture medium mixed solution, the edible fungus culture medium sample is 0.1g, the distilled water is 10ml, then the centrifuge tube 1021 accommodating the first edible fungus culture medium mixed solution is placed in the electric heating constant-temperature water bath 101 at 100 ℃, then the table centrifuge 102 is utilized to form a first supernatant and a first residue, and then the first residue and a sulfuric acid solution are mixed to form a second edible fungus culture medium mixed solution, wherein the concentration of the sulfuric acid solution is 10ml 15.4% (w/w). Then, the second edible mushroom culture medium mixed solution is placed in the electric heating constant-temperature water bath 101 at the temperature of 100 ℃, and then a second supernatant and a second residue are formed by using the table centrifuge 102. The cellulose and hemicellulose are detected and calculated by the high performance liquid anion chromatograph 600. Acid-insoluble lignin was detected and calculated by the lignin reaction apparatus 500.
It should be noted that, as shown in fig. 4, the reaction apparatus 100 for testing the lignocellulose component in the edible fungus cultivation substrate further includes an actuating clamping jaw device 103 for clamping the centrifuge tube 1021 between the electric heating constant temperature water bath 101 and the table centrifuge 102. The actuating jaw device 103 has a six-axis moving module, and can move and rotate in XYZ axes as required to transfer the centrifugal tube 1021 to each apparatus.
Further, the edible mushroom culture substrate sample is weighed by the electronic balance 200, the weight of the sample is 0.1g, and the mass m is accurately recorded (accurate to 0.0001 g). Mixing the first edible mushroom cultivation substrate mixed solution and the second edible mushroom cultivation substrate mixed solution by the vortex mixer 300. The first residue is dried by the electrothermal blowing dry oven 400. Specifically, cellulose and hemicellulose are measured by the high performance liquid anion chromatograph 600; and the second residue will be further processed by the lignin reaction unit 500 to obtain acid insoluble lignin. The lignin reaction apparatus 500 comprises a sand core funnel 501, a box-type resistance furnace 502, a circulating water type multipurpose vacuum pump 503, and an oven 504. The second residue is filtered by the sand core funnel 501, washed by the circulating water type multipurpose vacuum pump 503 and dried by the oven 504, and then burned by the box-type resistance furnace 502, and the content of the acid-insoluble lignin is calculated. It should be noted that, as shown in fig. 8, the centrifuge tube 1021 is moved between apparatuses by the actuating jaw device 103. In addition, it is worth mentioning that a vacuum drying device may be used instead of the oven 504, and an oven may also be used instead of the oven 504, which is not a limitation of the present invention.
More specifically, 0.1g of the edible mushroom cultivation substrate sample is weighed by the electronic balance 200, the mass m of the edible mushroom cultivation substrate sample is accurately recorded to 0.0001g, and 0.1g of the edible mushroom cultivation substrate sample is placed in the centrifugal tube 1021, wherein the size of the centrifugal tube 1021 is 15 ml. Then, 10ml of distilled water is added to the centrifuge tube 1021, and the edible mushroom culture medium sample is uniformly mixed with the distilled water by shaking the vortex mixer 300 to form the first edible mushroom culture medium mixture. And placing the centrifuge tube 1021 containing the first edible mushroom culture medium mixed solution in the electric heating constant-temperature water bath kettle 101 at 100 ℃ for heat preservation for 2 hours. The first supernatant was then discarded, leaving the first residue, after allowing it to cool to 25 ℃ and placing the centrifuge tube 1021 in the tabletop centrifuge 102 for 10 minutes at 3000 rpm. The first residue was further washed with distilled water by centrifugation for 3 or 4 times, and then dried in the electric hot air drying oven 400 at 100 ℃ for 5 hours. Taking out and drying the first residue, adding 1ml of 72% sulfuric acid solution, shaking up by horizontal shaking of the vortex mixer 300, placing in the electric heating constant temperature water bath 101 at 30 ℃ for 1 hour, then 9mL of distilled water is added to dilute the sulfuric acid solution to 1.2 mol/L15.4% (w/w), then uniformly mixing the mixture by the vortex mixer 300 to form a second edible fungus culture medium mixed solution, and the mixed solution of the second edible fungus culture medium is placed in the electric heating constant-temperature water bath pot 101 for 2 hours at the temperature of 100 ℃, so that the cellulose and hemicellulose in the edible fungus culture substrate sample are hydrolyzed into arabinose, galactose, glucose, xylose and mannose under the catalysis of sulfuric acid, it is then re-cooled to 25 ℃ and centrifuged by the tabletop centrifuge 102 at 3000rpm for 10 minutes to form the second supernatant and the second residue. At this time, the monosaccharide content of the second supernatant is measured by the hplc 600 to measure cellulose and hemicellulose. And the second residue is subjected to an acid-insoluble lignin reaction. In particular, the mixed liquid of the second edible fungus culture medium is placed in 100 ℃ the electric heating constant temperature water bath 101 is 2 hours, wherein the utility model discloses will set up 6 gradients with 1.2mol/L sulfuric acid solution hydrolysis time, it is 0.5, 1.0, 1.5, 2.0, 2.5, 3.0 hours respectively to survey the change of different hydrolysis time cellulose, hemicellulose, the acid insoluble lignin content of each edible fungus culture medium, in order to confirm the most suitable hydrolysis time.
In the reaction of the insoluble lignin, the second residue was suction-filtered through the sand core funnel 501, and after the second residue on the sand core funnel 501 was washed to neutrality with the circulating water type multipurpose vacuum pump, the sand core funnel 501 with the residue left thereon was dried in the oven 504 at 100 ℃ for 3 hours, and after cooling, the weight M1 of the sand core funnel 501 was accurately weighed by the electronic balance 200. Then, the sand core funnel 501 is placed in the box type resistance furnace 502 to be burnt at the high temperature of 550 ℃ for 3 hours, after the sand core funnel 501 is completely cooled, the weight M2 of the sand core funnel 501 is accurately weighed by the electronic balance 200 again, and the difference value is calculated, namely M1-M2, so that the acid-insoluble lignin is obtained.
It is worth mentioning that the edible fungi cultivation substrate sample is implemented as cultivation substrate of the straw mushroom Agaricus bisporus (Agaricus bisporus), the straw mushroom volvacea (volvarialla volvacea), the wood mushroom Flammulina velutipes (Flammulina velutipes), the Hypsizygus marmoreus (Hypsizygus marmoreus), the mushroom (Lentinus edodes), the Pleurotus eryngii (Pleurotus eryngii), the Ganoderma lucidum (Ganoderma lucidum), wherein the cultivation substrate of the straw mushroom Agaricus bisporus (Agaricus bisporus), the straw mushroom volvacea (volvaria volvacea) is a cultivation material which is not sown after the secondary fermentation. The culture medium of the wood rotting fungus, namely the Flammulina velutipes, the Hypsizygus marmoreus, the Lentinus edodes, the Pleurotus eryngii and the lucid Ganoderma (Ganoderma lucidum) is a sterilized and unsown culture medium. The formula of the agaricus bisporus comprises 70% of wheat straw, 18% of chicken manure, 4% of rapeseed cake and 8% of gypsum. The straw mushroom is prepared from 80% of straw, 18% of needle mushroom dregs and 2% of lime. The formula of the flammulina velutipes is 35% of corncobs, 35% of rice bran, 10% of soybean hulls, 5% of cottonseed hulls and 5% of corn flour. The formula of the Hypsizygus marmoreus comprises 35% of sawdust, 20% of rice bran, 15% of corncob, 13% of cottonseed hull, 12% of bran and 5% of corn flour. The formula of the mushroom comprises 79% of wood chips, 20% of bran and 1% of gypsum. The formula of the pleurotus eryngii comprises 30% of wood chips, 25% of cottonseed hulls, 18% of corncobs, 15% of bran, 5% of corn flour, 5% of soybean flour and 1% of gypsum. The formula of the ganoderma lucidum is 39% of wood chips, 39% of cotton seed hulls, 20% of corn flour, 1% of gypsum and 1% of cane sugar. All the culture mediums are dried in the air and crushed and then screened by a 40-mesh sieve for later use.
The utility model is hydrolyzed by 1.2mol/L sulfuric acid solution and placed at 100 ℃ in the electric heating constant temperature water bath 101, the second edible fungus culture medium is mixed with the liquid. With the increase of the hydrolysis time, the cellulose content in the hydrolysate of the agaricus bisporus, the Hypsizygus marmoreus, the Pleurotus eryngii and the lucid ganoderma substrate shows a remarkable trend of increasing first and then decreasing. And after the cellulose content in the hydrolysate of the straw mushroom, the mushroom and the needle mushroom matrixes is increased to the maximum value along with the time extension, the subsequent cellulose content is firstly reduced and then increased. The edible fungi with the tendency of increasing and then decreasing the hemicellulose content in the substrate hydrolysate along with the change of time are mushroom, agaricus bisporus, hypsizygus marmoreus, flammulina velutipes and pleurotus eryngii, and the tendency of the change of the hemicellulose content of volvariella volvacea and ganoderma lucidum is not obvious. For acid-insoluble lignin, the content of the volvariella volvacea and agaricus bisporus matrix is basically maintained after being reduced to the minimum value along with the prolonging of the hydrolysis time, the pleurotus eryngii matrix has the trend of increasing, then reducing and then increasing, and except for the 3 matrixes, the content of the acid-insoluble lignin of other edible fungus matrixes is obviously reduced and then increased. Comprehensively considering the content changes of cellulose, hemicellulose and acid-insoluble lignin at different hydrolysis times, the optimum hydrolysis time of straw mushroom, shiitake mushroom, agaricus bisporus, flammulina velutipes and Hypsizygus marmoreus is determined to be 1.5 hours, and the optimum hydrolysis time of Pleurotus eryngii and Ganoderma lucidum is determined to be 2 hours.
It is worth mentioning that the utility model discloses a monosaccharide test method, it utilizes high performance liquid anion chromatograph 800, a chromatographic separation post, a guard column go on, wherein the column temperature is 30 ℃, the sample volume 25 uL, mobile phase and gradient elution condition are seen in table 1.
TABLE 1 mobile phase and gradient elution conditions
Table 1 Mobile phase and gradient elution condition
Mobile phase A: water; mobile phase B: 4nmol/L sodium hydroxide solution; mobile phase C: 1mol/L sodium acetate solution
Mobile phase A:H2O；mobile phase B:4nmol/L NaOH solution；mobile phaseC:1mol/L NaAC solution
In addition, the utility model also provides a monosaccharide recovery rate test method, wherein accurately weigh 177.9mg arabinose, 166.8mg galactose, 150.3mg glucose, 152.2mg xylose, 157.3mg mannose respectively and dissolve in 10mL ultrapure water, prepare monosaccharide standard solution of arabinose of concentration 118.2 μmol/mL, galactose of concentration 92.56 μmol/mL, glucose of concentration 83.41 μmol/mL, xylose of concentration 101.4 μmol/mL, mannose of concentration 87.29 μmol/mL respectively. Then, 0.2mL of each prepared monosaccharide standard solution was taken, and 1mL of the total monosaccharide standard solution was added with 1mL of 72% sulfuric acid solution and 8mL of ultrapure water, and hydrolyzed at 100 ℃ for 2 hours to measure the concentration of the hydrolyzed monosaccharide solution and calculate the monosaccharide recovery rate.
And calculating the recovery rates of arabinose, galactose, glucose, xylose and mannose and the content of each monosaccharide in the sample by referring to a method established by NREL, and calculating the content of cellulose, hemicellulose and lignin in the sample according to the following formula.
Cellulose content ═ glucose x0.9x 100%
Hemicellulose (arabinose + xylose) x0.88x100% + (galactose + mannose) x0.9x100%
In addition, it is worth mentioning that the average recovery rates of arabinose, galactose, glucose, xylose and mannose after the monosaccharide standard solution was treated at 100 ℃ for 2 hours were 100.2%, 100.9%, 100.6%, 101.0% and 101.2%, respectively, indicating that monosaccharide was not lost under the reaction conditions.
In addition, the utility model provides a test method of edible mushroom culture medium lignocellulose component, it includes following step:
(A) mixing an edible fungus culture medium sample and distilled water to form a first edible fungus culture medium mixed solution, and placing the first edible fungus culture medium mixed solution in an electric heating constant-temperature water bath kettle 101 at 100 ℃;
(B) forming a first supernatant and a first residue from the first edible mushroom cultivation substrate mixture by a desktop centrifuge 102;
(C) mixing the first residue with a sulfuric acid solution to form a second edible mushroom culture medium mixed solution, and placing the mixed solution in an electric heating constant-temperature water bath kettle 101 at 100 ℃;
(D) forming a second supernatant and a second residue from the second edible mushroom cultivation substrate mixture by the tabletop centrifuge 102; and
(E) determining the cellulose, hemicellulose and acid insoluble lignin reaction test of the second supernatant and the second residue.
According to the step (A), the edible mushroom culture medium sample is 0.1g, the distilled water is 10ml, and the edible mushroom culture medium sample is placed in a 15ml centrifuge tube 1021.
According to the step (A), the first edible mushroom culture medium mixed solution is placed in the electric heating constant-temperature water bath 101 at the temperature of 100 ℃ for 2 hours.
According to step (B), the desktop centrifuge 102 is centrifuged at 3000rpm for 10 minutes while the first edible mushroom culture medium mixture is cooled to 25 ℃.
According to step (C) further comprising:
(C1) washing the first residue with water for 3 or 4 times, and drying at 100 deg.C for 5 hr;
(C2) adding 1ml of 72% sulfuric acid solution to the dried first residue, and standing in the electric-heating constant-temperature water bath 101 at 30 ℃ for 1 hour; and
(C3) adding 9mL of distilled water to dilute the sulfuric acid solution to 1.2 mol/L15.4% (w/w) to form a second edible mushroom culture medium mixed solution; and
(C4) and placing the mixed solution of the second edible fungus culture medium in the electric heating constant-temperature water bath 101 at 100 ℃ for 2 hours.
According to the step (C), hydrolyzing the cellulose and hemicellulose in the edible fungus culture medium sample into arabinose, galactose, glucose, xylose and mannose under the catalysis of sulfuric acid.
According to step (D), the second edible mushroom cultivation substrate mixture is cooled to 25 ℃, and the tabletop centrifuge 102 is centrifuged at 3000rpm for 10 minutes.
According to the step (E), the high performance liquid anion chromatograph 600 is used to measure the monosaccharide content of the second supernatant, and the cellulose and hemicellulose content is calculated, wherein the calculation formula is:
cellulose content ═ glucose x0.9x 100%.
Hemicellulose is (arabinose + xylose) x0.88x100% + (galactose + mannose) x0.9x100%.
The acid insoluble lignin reaction test further comprising:
(E1) suction-filtering the second residue;
(E2) washing the second residue to neutrality;
(E3) drying the second residue and weighing it by weight M1;
(E4) burning at high temperature, and weighing the weight M2; and
(E5) the difference, M1-M2, was calculated.
According to step (E1), the second residue is suction filtered with a sand core funnel 501.
The second residue, which was suction filtered and washed with water, was dried at 100 ℃ for 3 hours according to the step (E3), and the weight M1 of the sand core funnel 501 was accurately weighed with an electronic balance 200 after cooling.
According to the step (E4), the sand core funnel 501 is placed in a box-type resistance furnace 502 and burned at a high temperature of 550 ℃ for 3 hours, and after complete cooling, the weight M2 of the sand core funnel 501 is accurately weighed again by the electronic balance 200.
Calculating the acid insoluble lignin content according to step (E) with the formula:
in addition, the utility model provides a test method of edible mushroom culture medium lignocellulose component, it includes following step:
(a) weighing an edible mushroom culture medium sample in a centrifugal tube 1021;
(b) adding distilled water to form a first edible mushroom culture medium mixed solution in the centrifuge tube 1021, and placing in an electric heating constant temperature water bath 101;
(c) centrifuging the first edible mushroom culture medium mixed solution by using a desktop centrifuge 102 to obtain a first residue;
(d) washing and drying the first residue with water;
(e) mixing the dried first residue with a sulfuric acid solution and standing;
(f) diluting the solution of step (e) to form a second edible mushroom cultivation substrate mixed solution, and placing the second edible mushroom cultivation substrate mixed solution in an electric heating constant temperature water bath 101;
(g) centrifuging the second edible mushroom culture medium mixed solution by using the desktop centrifuge 102 to obtain a second supernatant and a second residue; and
(h) determining the cellulose, hemicellulose and acid insoluble lignin reaction test on the second supernatant.
According to the step (a), the edible fungus culture medium sample is 0.1g, and the mass m of the edible fungus culture medium sample is accurate to 0.0001 g.
The centrifuge tube 1021 has a size of 15ml according to step (a).
According to step (b), the distilled water was 10 ml.
According to the step (b), 0.1g of the edible mushroom cultivation substrate sample and 10ml of the distilled water are shaken by a vortex mixer 300 to be uniformly mixed to form the first edible mushroom cultivation substrate mixture.
According to the step (b), the centrifuge tube 1021 containing the first edible mushroom cultivation matrix mixed solution is placed in the electric heating constant temperature water bath 101 with the temperature of 100 ℃ for heat preservation for 2 hours.
According to step (c), the desktop centrifuge 102 is centrifuged at 3000rpm for 10 minutes while the first edible mushroom culture substrate mixture is cooled to 25 ℃.
According to step (d), it is washed 3 or 4 times with distilled water by centrifugation and dried at 100 ℃ for 5 hours using an electrothermal blowing dry box 400.
According to step (e), the dried first residue and 1ml of 72% sulfuric acid solution were shaken up by a vortex mixer 300 and left in the electric thermostat water bath 101 at 30 ℃ for 1 hour.
According to step (f), 9mL of distilled water was added to dilute the sulfuric acid solution to 1.2 mol/L15.4% (w/w).
According to the step (f), the second edible mushroom culture medium mixed solution is placed in the electric heating constant-temperature water bath 101 at the temperature of 100 ℃ for 2 hours.
According to the step (f), hydrolyzing the cellulose and hemicellulose in the edible fungus culture medium sample into arabinose, galactose, glucose, xylose and mannose under the catalysis of a sulfuric acid solution.
According to step (g), the desktop centrifuge 102 is centrifuged at 3000rpm for 10 minutes while the second edible mushroom culture substrate mixture is cooled to 25 ℃.
According to step (h), the monosaccharide content of the second supernatant is determined and the cellulose and hemicellulose content therein is calculated using the hplc 600.
According to step (h), the cellulose content calculation formula: cellulose content ═ glucose x0.9x 100%.
According to step (h), the hemicellulose content calculation formula: hemicellulose is (arabinose + xylose) x0.88x100% + (galactose + mannose) x0.9x100%.
The acid insoluble lignin reaction test further comprising:
(h1) a sand core funnel 501 filters the second residue and washes the second residue to neutrality;
(h2) the sand core funnel with the residue left is placed in an oven 504 at 100 ℃ to dry for 3 hours.
(h3) Accurately weighing the weight M1 of the sand core funnel 501 after cooling;
(h4) placing the sand core funnel in a box type resistance furnace 502 at 550 ℃ and firing at high temperature for 3 hours; and
(h5) after complete cooling, the weight of the sand core funnel M2 was accurately weighed again.
According to step (h), calculating the content formula of the acid-insoluble lignin:
it will be understood by those skilled in the art that the embodiments of the present invention as described above and shown in the drawings are given by way of example only and are not limiting of the present invention.
The objects of the present invention have been fully and effectively accomplished. The functional and structural principles of the present invention have been shown and described in the embodiments without departing from the principles, embodiments of the present invention may have any deformation or modification.
Claims (6)
1. The reaction device for testing the lignocellulose component of the edible fungus culture substrate is characterized by comprising:
an electric heating constant temperature water bath pot, which is provided with a centrifuge tube for accommodating a first edible mushroom culture medium mixed solution;
and the table centrifuge is used for separating a first residue from the first edible mushroom culture medium mixed solution, wherein the first residue is mixed with a sulfuric acid solution to form a second edible mushroom culture medium mixed solution, the second edible mushroom culture medium mixed solution is placed in the electric heating constant-temperature water bath kettle, and a second supernatant and a second residue are separated from the second edible mushroom culture medium mixed solution by the table centrifuge.
2. The reaction device for testing the lignocellulose component in the edible fungus cultivation substrate as recited in claim 1, further comprising an actuating jaw device for grasping and placing the centrifuge tube between the electric heating constant temperature water bath and the table centrifuge.
3. The utility model provides an edible mushroom culture substrate lignocellulose component test equipment which characterized in that includes:
a reaction unit for testing lignocellulose components in edible fungus culture substrate comprises:
an electric heating constant temperature water bath pot, which is provided with a centrifuge tube for accommodating a first edible mushroom culture medium mixed solution; and
a desk centrifuge, which separates a first residue from the cooled first edible mushroom culture medium mixed solution and carries out centrifugal washing;
an electric heating air blast drying box, which dries the washed first residue, wherein the dried first residue is mixed with sulfuric acid solution to form a second edible mushroom cultivation substrate mixed solution, the second edible mushroom cultivation substrate mixed solution is placed in the electric heating constant-temperature water bath kettle, and the second edible mushroom cultivation substrate mixed solution is separated into a second supernatant and a second residue by the table centrifuge;
a liquid phase anion chromatograph which tests the monosaccharide content of the second supernatant and calculates cellulose and hemicellulose; and
a lignin reaction unit which tests and calculates acid insoluble lignin from the second residue.
4. The edible mushroom culture substrate lignocellulose component testing apparatus as recited in claim 3, wherein the reaction device for edible mushroom culture substrate lignocellulose component testing further comprises an actuating jaw device which grips and places the centrifuge tube between the devices.
5. The edible fungus culture substrate lignocellulose component testing device as defined in claim 3, wherein the second edible fungus culture substrate mixed solution is placed in the electric heating constant-temperature water bath for heat preservation so as to hydrolyze cellulose and hemicellulose in the edible fungus culture substrate sample into arabinose, galactose, glucose, xylose and mannose under the catalysis of sulfuric acid.
6. The edible mushroom cultivation substrate lignocellulose component testing apparatus as recited in claim 4, wherein the lignin reaction device comprises a sand core funnel, a box-type resistance furnace, a circulating water type multipurpose vacuum pump, and an oven, wherein the second residue is filtered by the sand core funnel and washed with water by the circulating water type multipurpose vacuum pump, the sand core funnel with the residue left is placed in the oven and weighed after cooling, and the sand core funnel is placed in the box-type resistance furnace to be burned and weighed after cooling, wherein the centrifuge tube is moved between the devices by the actuating jaws.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201922339448.7U CN211856480U (en) | 2019-12-20 | 2019-12-20 | Reaction device and equipment for testing lignocellulose components of edible fungus culture substrate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201922339448.7U CN211856480U (en) | 2019-12-20 | 2019-12-20 | Reaction device and equipment for testing lignocellulose components of edible fungus culture substrate |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN211856480U true CN211856480U (en) | 2020-11-03 |
Family
ID=73216595
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201922339448.7U Active CN211856480U (en) | 2019-12-20 | 2019-12-20 | Reaction device and equipment for testing lignocellulose components of edible fungus culture substrate |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN211856480U (en) |
-
2019
- 2019-12-20 CN CN201922339448.7U patent/CN211856480U/en active Active
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Jung et al. | Comparison of lignin, cellulose, and hemicellulose contents for biofuels utilization among 4 types of lignocellulosic crops | |
| Saeman | Kinetics of wood saccharification-hydrolysis of cellulose and decomposition of sugars in dilute acid at high temperature | |
| CN105361187B (en) | A kind of modified grain dietary fiber and preparation method thereof | |
| Ibáñez et al. | Downscaled method using glass microfiber filters for the determination of Klason lignin and structural carbohydrates | |
| Lee et al. | Biological conversion assay using Clostridium phytofermentans to estimate plant feedstock quality | |
| CN113151006B (en) | Trichoderma reesei strain capable of producing cellulase with improved activity and application thereof | |
| CN105900984A (en) | Method for preparing pesticide by using biomass hydrothermal carbonization liquid phase and preparing organic fertilizers by using solid phase | |
| CN104770574A (en) | Method of preparing feather protein powder from keratinase | |
| CN111122726B (en) | Edible fungus culture medium lignocellulose component testing equipment and method thereof | |
| CN107090478B (en) | Method for extracting water-soluble dietary fiber from lentinus edodes stems | |
| CN211856480U (en) | Reaction device and equipment for testing lignocellulose components of edible fungus culture substrate | |
| CN104798981B (en) | A method of preparing feather albumen powder using alkali protease and keratinase | |
| CN109355329A (en) | A method of reduced sugar is prepared using tobacco rod as biorefinery raw material | |
| CN106699908A (en) | Method for extracting amylose from high amylose corn starch | |
| Xu et al. | Gamagrass varieties as potential feedstock for fermentable sugar production | |
| Haw et al. | Nuclear magnetic resonance study of autohydrolyzed and organosolv-treated lodgepole pinewood using carbon-13 with cross polarization and magic-angle spinning | |
| Tian et al. | Production of bioethanol from sweet potato tubers with different storage times | |
| CN111118087B (en) | Lignocellulose pretreatment system based on FeOCl and application thereof | |
| CN107356702B (en) | Method for detecting crude protein in castor seeds or castor cake meal | |
| KR101395053B1 (en) | Preparation Method of Glucose from Plant Biomass | |
| CN109358154A (en) | The measuring method of monosaccharide composition in a kind of acidic polysaccharose | |
| CN106226524A (en) | A kind of detection method of edible fungi dsRNA virus | |
| Li et al. | Ethanosolv pretreatment of bamboo with dilute acid for efficient enzymatic saccharification | |
| CN116144717A (en) | Method for preprocessing straw waste by using choline hydroxide eutectic solvent | |
| Templeton et al. | Biomass Compositional Analysis Method Variability (Poster) |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| GR01 | Patent grant | ||
| GR01 | Patent grant |