CN112522342B - Method for efficiently performing enzymolysis on straws - Google Patents
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Abstract
The invention belongs to the technical field of biology, and particularly relates to a method for efficiently performing enzymolysis on straws, which comprises the following steps: (1) straw pretreatment: crushing and sieving straws, putting the straws into a steamer, steaming the straws by using water vapor, adding a mixed solution consisting of potassium hydrogen persulfate and boric acid, keeping the temperature and standing the straws, adding the straws into deionized water, keeping the temperature and standing the straws, drying the straws in an oven to constant weight, and performing high-pressure steam sterilization treatment to obtain pretreated straws; (2) compound enzymolysis culture: and (3) placing the pretreated straws in a culture medium containing trichoderma reesei and rhizopus oryzae for culture, thus obtaining the straw. The method for efficiently hydrolyzing the straws overcomes the problem of low enzymolysis effect of the traditional single cellulase, further improves the accessibility by a pretreatment method, fully utilizes the straws, enhances the enzyme activity by a culture medium containing hydrolyzed fish protein powder and cottonseed protein, and has higher economic value.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a method for efficiently performing enzymolysis on straws.
Background
The straw is the general term of the stem and leaf (ear) part of the mature crop, and comprises the rest part of the crop such as wheat, rice, corn, potatoes, rape and the like after the seeds are harvested. In times of lower agricultural production, straw can be used for bedding, feeding livestock, and for composting.
The straw is composed of three components of cellulose, hemicellulose and lignin, the cellulose is in a wrapping sheath of the lignin, the lignin is used as a high-molecular polymer highly resistant to biodegradation, the higher the degree of polymerization and the degree of crystallinity are, the more serious the wrapping phenomenon of the lignin is, so that the biodegradation of the cellulose is more difficult, and experiments prove that the enzymolysis rate of the straw which is not pretreated is only about 20%. The problem of lignin shielding can be solved through a pretreatment mode, the degree of polymerization and the degree of crystallinity of natural cellulose are reduced, and preconditions are created for subsequent enzymolysis of fibers, but different pretreatment methods inevitably produce different enzymolysis effects, and a method capable of greatly improving the enzymolysis effect needs to be found.
The use of modern chemical fertilizers greatly reduces the agricultural requirements on fertilizers made of straws, the treatment of the straws becomes a difficult problem, the straws are not treated in time and can influence the sowing of autumn-sown crops such as wheat, and in order to treat the straws, the phenomena of large-scale straw burning and repeated prohibition often occur, so that the air quality is polluted, and meanwhile, the great economic loss is caused.
The regeneration and use of the straws provides a new idea for solving the problem, intensive research is carried out at home and abroad at the present stage, certain results are obtained, but large-scale industrial production is still difficult to realize at present, saccharification and fermentation are one of the mature ways in the current straw treatment, however, better effects cannot be obtained in the aspects of pretreatment mode, enzymolysis conditions and the like, and the method can be particularly applied to industrial production stably and reliably.
The Chinese patent application CN104561129A discloses an enzymolysis saccharification treatment method of straws, the pretreatment process is simple, and the problem of lignin covering cannot be well solved, so that the final enzymolysis effect can be influenced.
Chinese patent CN109182418B discloses a method for microbial enzymatic hydrolysis and saccharification of straws, which can obtain a relatively effective enzymatic hydrolysis and saccharification effect, but a used fermentation medium is too complex and has high cost, and the pretreated straws are not subjected to sterilization operation, so that the risk of producing mixed bacteria is generated.
Disclosure of Invention
The invention aims to provide a method for obviously enhancing the accessibility of enzymolysis straws by effective pretreatment and obtaining higher enzymolysis conversion rate by a method combining enzymolysis and fermentation culture, particularly the method has simple steps, easily obtains used raw materials, is suitable for large-scale production in large industry and has higher economic value and environmental protection value.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a method for efficiently performing enzymolysis on straws comprises the following steps:
s1, pretreatment of straws;
s2, compound enzymolysis culture: and (3) placing the pretreated straws in a culture medium containing trichoderma reesei and rhizopus oryzae, uniformly stirring, and performing stacking culture to obtain the straw.
Further, the preprocessing of step S1 includes the following steps:
1) Crushing the straws, sieving the crushed straws with a 30-mesh sieve, and putting the crushed straws into a steamer to be steamed for 2 to 3 hours by using water vapor to obtain steamed straws A;
2) Adding the straw A obtained in the step 1) into a mixed solution prepared from potassium hydrogen persulfate and boric acid, wherein the added mass of the mixed solution is 2-3 times of that of the straw A, and standing for 1-2h at the temperature of 65-80 ℃ to obtain an acidified straw B;
3) Adding the straw B obtained in the step 2) into deionized water, preserving heat and standing for 10-30min at 40-60 ℃ to obtain straw C, drying the straw C to constant weight in a drying oven at 100-150 ℃, and then performing high-pressure steam sterilization treatment to obtain the finally pretreated straw D.
Further, the preparation method of the mixed solution in the step 2) comprises the following steps: mixing potassium hydrogen persulfate and boric acid according to the mass ratio of 1-3:1, and dissolving the mixture in deionized water to obtain the potassium hydrogen persulfate/boric acid composite material.
Further, the dosage of the trichoderma reesei in the step S2 is 200-300 ten thousand u/kg, and the dosage of the rhizopus oryzae is 80-120 ten thousand u/kg.
Further, the culture medium of step S2 comprises: 0.3-0.5g/L CaCl 2 0.1-0.3g/L MgCl 2 0.5-2g/L peptone, 1-2g/L hydrolyzed fish protein powder, 1-2g/L cottonseed protein and the balance buffer solution.
Further, the buffer is disodium hydrogen phosphate-citric acid buffer, and the pH value of the buffer is 5-6.
Furthermore, the mass relationship between the addition amount of the culture medium and the added straw D is that 30-45mL of culture medium needs to be added into 1g of straw D.
Further, the culturing time of the step S2 is 36-48h, and the culturing temperature is 35-45 ℃.
The hydrolyzed fish protein powder is Purpu Le Zhuang hydrolyzed fish protein powder produced by the Hippon group, and the cottonseed protein is Pulepian cottonseed protein produced by the Hippon group.
Compared with the prior art, the invention has the following advantages:
(1) The invention carries out thorough and sufficient pretreatment on the straws, firstly crushes and screens the straws, carries out primary treatment by steam cooking, then carries out deep destruction on the straws by a mixed solution prepared by potassium hydrogen persulfate and boric acid, then puts the treated straws in deionized water for heat preservation treatment, and then carries out drying and sterilization, thereby obviously improving the accessibility of straw enzymolysis.
(2) Experiments fully prove that the method can effectively remove the lignin in the straws, and the removal rate can reach 82%.
(3) After sufficient pretreatment, the treated straws are subjected to combined enzymolysis through trichoderma reesei and rhizopus oryzae, and CaCl is added into a culture medium 2 、MgCl 2 The components such as peptone, hydrolyzed fish protein powder, cottonseed protein and the like generate synergistic effect to promote enzymolysis effect, generate extremely high enzyme activity, and simultaneously, a buffer system formed by disodium hydrogen phosphate-citric acid is added to ensure effective enzymolysis at a stable pH value.
(4) Experiments prove that the method for efficiently performing enzymolysis on the straws has a very high enzymolysis effect compared with the prior art, the content of reducing sugar in filtrate after enzymolysis can reach 320mg/g, and the method has a great economic value.
Detailed Description
The present invention will be further explained by way of specific embodiments in the form of examples. The scope of the above-described subject matter of the present invention is not limited to the following examples.
The following examples used the known and commercially available raw materials such as oxone, boric acid, disodium hydrogen phosphate-citric acid buffer, and the like, and all of them were commercially available, and Trichoderma reesei and Rhizopus oryzae were purchased from Shandongtai Biotech Co., ltd.
Example 1 method for efficiently performing enzymolysis on straws
A method for efficiently performing enzymolysis on straws comprises the following steps:
s1, pretreatment of straws:
1) Crushing straws, sieving with a 30-mesh sieve, and putting into a steamer to be steamed for 2 hours by using steam to obtain steamed straws A;
2) Adding the straw A obtained in the step 1) into a mixed solution prepared from potassium hydrogen persulfate and boric acid, wherein the added mass of the mixed solution is 2 times of that of the straw A, and standing for 1h at the temperature of 65 ℃ to obtain an acidified straw B; the preparation method of the mixed solution comprises the following steps: mixing potassium hydrogen persulfate and boric acid according to the mass ratio of 1:1, and dissolving the mixture in deionized water with the mass 5 times that of the mixture to obtain the potassium hydrogen persulfate/boric acid composite material;
3) Adding the straw B obtained in the step 2) into deionized water with the mass being 10 times that of the straw B, keeping the temperature and standing for 10min at 40 ℃ to obtain straw C, drying the straw C to constant weight in a 100 ℃ drying oven, and then performing high-pressure steam sterilization treatment, wherein the temperature during sterilization is 121 ℃, the pressure is 100kPa, and the sterilization time is 20min to obtain the finally pretreated straw D;
s2, compound enzymolysis culture: placing the pretreated straw D in a culture medium containing trichoderma reesei and rhizopus oryzae, wherein the dosage of the trichoderma reesei is 200 ten thousand u/kg, the dosage of the rhizopus oryzae is 80 ten thousand u/kg, the mass relation between the addition amount of the culture medium and the added straw D is that 1g of straw D needs to be added into 30mL of the culture medium, stacking and culturing for 36h after uniform stirring, and the temperature during culturing is 35 ℃ to obtain the straw D; the culture medium comprises: 0.3g/L of CaCl 2 0.1g/L of MgCl 2 0.5g/L peptone, 1g/L hydrolyzed fish protein powder, 1g/L cottonseed protein, and the balance of disodium hydrogen phosphate-citric acid buffer solution with pH of 5.
Example 2 method for efficiently performing enzymolysis on straws
A method for efficiently carrying out enzymolysis on straws comprises the following steps:
s1, pretreatment of straws:
1) Crushing the straws, sieving the crushed straws with a 30-mesh sieve, and then putting the crushed straws into a steamer to be steamed for 3 hours by using water vapor to obtain steamed straws A;
2) Adding the straw A obtained in the step 1) into a mixed solution prepared from potassium hydrogen persulfate and boric acid, wherein the added mass of the mixed solution is 3 times of that of the straw A, and standing for 2 hours at the temperature of 80 ℃ to obtain an acidified straw B; the preparation method of the mixed solution comprises the following steps: mixing potassium hydrogen persulfate and boric acid according to the mass ratio of 3:1, and dissolving the mixture in deionized water with the mass 5 times that of the mixture to obtain the potassium hydrogen persulfate/boric acid composite material;
3) Adding the straw B obtained in the step 2) into deionized water with the mass being 10 times that of the straw B, keeping the temperature and standing for 30min at the temperature of 60 ℃ to obtain straw C, drying the straw C to constant weight in a drying oven at the temperature of 150 ℃, and then performing high-pressure steam sterilization treatment, wherein the temperature during sterilization is 121 ℃, the pressure is 100kPa, and the sterilization time is 20min to obtain the finally pretreated straw D;
s2, compound enzymolysis culture: placing the pretreated straw D in a culture medium containing trichoderma reesei and rhizopus oryzae, wherein the dosage of the trichoderma reesei is 300 ten thousand u/kg, the dosage of the rhizopus oryzae is 120 ten thousand u/kg, the mass relation between the addition amount of the culture medium and the added straw D is that 1g of straw D needs to be added into 45mL of the culture medium, stacking and culturing for 48h after uniform stirring, and the temperature during culturing is 45 ℃ to obtain the straw D; the culture medium comprises: 0.5g/L of CaCl 2 0.3g/L of MgCl 2 2g/L peptone, 2g/L hydrolyzed fish protein powder, 2g/L cottonseed protein and the balance of disodium hydrogen phosphate-citric acid buffer solution with the pH value of 6.
Example 3 method for efficiently carrying out enzymolysis on straws
A method for efficiently performing enzymolysis on straws comprises the following steps:
s1, pretreatment of straws:
1) Crushing straws, sieving with a 30-mesh sieve, and putting the straws into a steamer to be steamed for 2 hours to obtain steamed straws A;
2) Adding the straw A obtained in the step 1) into a mixed solution prepared from potassium hydrogen persulfate and boric acid, wherein the added mass of the mixed solution is 3 times of that of the straw A, and standing for 1h at the temperature of 70 ℃ to obtain an acidified straw B; the preparation method of the mixed solution comprises the following steps: mixing potassium hydrogen persulfate and boric acid according to the mass ratio of 2:1, and dissolving the mixture in deionized water with the mass 5 times that of the mixture to obtain the potassium hydrogen persulfate/boric acid composite material;
3) Adding the straw B obtained in the step 2) into deionized water with the mass being 10 times that of the straw B, keeping the temperature and standing for 20min at 50 ℃ to obtain straw C, drying the straw C to constant weight in a 120 ℃ drying oven, and then performing high-pressure steam sterilization treatment, wherein the temperature during sterilization is 121 ℃, the pressure is 100kPa, and the sterilization time is 20min to obtain the finally pretreated straw D;
s2, compound enzymolysis culture: placing the pretreated straw D in a culture medium containing trichoderma reesei and rhizopus oryzae, wherein the dosage of the trichoderma reesei is 300 ten thousand u/kg, the dosage of the rhizopus oryzae is 100 ten thousand u/kg, the mass relation between the addition amount of the culture medium and the added straw D is that 1g of straw D needs to be added into 40mL of the culture medium, stacking and culturing for 40h after uniform stirring, and the temperature during culturing is 45 ℃ to obtain the straw D; the culture medium comprises: 0.4g/L of CaCl 2 0.3g/L of MgCl 2 2g/L peptone, 2g/L hydrolyzed fish protein powder, 2g/L cottonseed protein and the balance of disodium hydrogen phosphate-citric acid buffer solution with the pH value of 6.
Comparative example 1, method for pretreating straw
A straw pretreatment method comprises the following steps:
s1, pretreatment of straws:
1) Crushing straws, sieving with a 30-mesh sieve, and putting the straws into a steamer to be steamed for 2 hours to obtain steamed straws A;
2) Adding the straw A obtained in the step 1) into a sulfuric acid solution with the mass fraction of 3%, wherein the mass of the added sulfuric acid solution is 3 times that of the straw A, and standing for 1h at the temperature of 70 ℃ to obtain acidified straw B;
3) Adding the straw B obtained in the step 2) into deionized water with the mass being 10 times that of the straw B, keeping the temperature and standing for 20min at 50 ℃ to obtain straw C, drying the straw C to constant weight in a 120 ℃ oven, and then performing high-pressure steam sterilization treatment at the temperature of 121 ℃, the pressure of 100kPa and the sterilization time of 20min to obtain the finally pretreated straw D.
Comparative example 1 is similar to example 3, except that, unlike the pretreatment method in example 3, comparative example 1 does not add a mixed solution consisting of oxone and boric acid, but is replaced with a sulfuric acid solution.
Comparative example 2, method for enzymolysis of straw
A method for efficiently performing enzymolysis on straws comprises the following steps:
s1, pretreatment of straws:
1) Crushing straws, sieving with a 30-mesh sieve, and putting into a steamer to be steamed for 2 hours by using steam to obtain steamed straws A;
2) Adding the straw A obtained in the step 1) into a mixed solution prepared from potassium hydrogen persulfate and boric acid, wherein the added mass of the mixed solution is 3 times of that of the straw A, and standing for 1h at the temperature of 70 ℃ to obtain an acidified straw B; the preparation method of the mixed solution comprises the following steps: mixing potassium hydrogen persulfate and boric acid according to the mass ratio of 2:1, and dissolving the mixture in deionized water with the mass 5 times that of the mixture to obtain the potassium hydrogen persulfate/boric acid composite material;
3) Adding the straw B obtained in the step 2) into deionized water with the mass being 10 times that of the straw B, keeping the temperature and standing for 20min at 50 ℃ to obtain straw C, drying the straw C to constant weight in a 120 ℃ drying oven, and then performing high-pressure steam sterilization treatment, wherein the temperature during sterilization is 121 ℃, the pressure is 100kPa, and the sterilization time is 20min to obtain the finally pretreated straw D;
s2, compound enzymolysis culture: placing the pretreated straw D in a culture medium containing trichoderma reesei and rhizopus oryzae, wherein the dosage of the trichoderma reesei is 300 ten thousand u/kg, the dosage of the rhizopus oryzae is 100 ten thousand u/kg, the mass relation between the addition amount of the culture medium and the added straw D is that 1g of straw D needs to be added into 40mL of culture medium, stacking and culturing for 40h after uniform stirring, and the temperature during culturing is 45 ℃ to obtain the straw D; the culture medium comprises: 0.4g/L of CaCl 2 0.3g/L of MgCl 2 5g/L of peptone and the balance of disodium hydrogen phosphate-citric acid buffer solution with pH 6.
Comparative example 2 similar to example 3, except that the hydrolyzed fish protein powder and cottonseed protein were not added to the medium of step S2 of comparative example 2, unlike example 3.
Comparative example 3, method for performing enzymolysis on straw
A method for enzymolysis of straws comprises the following steps:
s1, pretreatment of straws:
1) Crushing straws, sieving with a 30-mesh sieve, and putting the straws into a steamer to be steamed for 2 hours to obtain steamed straws A;
2) Adding the straw A obtained in the step 1) into a mixed solution prepared from potassium hydrogen persulfate and boric acid, wherein the added mass of the mixed solution is 3 times of that of the straw A, and standing for 1h at the temperature of 70 ℃ to obtain an acidified straw B; the preparation method of the mixed solution comprises the following steps: mixing potassium hydrogen persulfate and boric acid according to the mass ratio of 2:1, and dissolving the mixture in deionized water with the mass 5 times that of the mixture to obtain the potassium hydrogen persulfate/boric acid composite material;
3) Adding the straw B obtained in the step 2) into deionized water with the mass being 10 times that of the straw B, keeping the temperature and standing for 20min at 50 ℃ to obtain straw C, drying the straw C to constant weight in a 120 ℃ drying oven, and then performing high-pressure steam sterilization treatment, wherein the temperature during sterilization is 121 ℃, the pressure is 100kPa, and the sterilization time is 20min to obtain the finally pretreated straw D;
s2, compound enzymolysis culture: placing the pretreated straw D in a culture medium containing trichoderma reesei and rhizopus oryzae, wherein the dosage of the trichoderma reesei is 300 ten thousand u/kg, the dosage of the rhizopus oryzae is 100 ten thousand u/kg, the mass relation between the addition amount of the culture medium and the added straw D is that 1g of straw D needs to be added into 40mL of culture medium, stacking and culturing for 40h after uniform stirring, and the temperature during culturing is 45 ℃ to obtain the straw D; the culture medium comprises: 0.4g/L of CaCl 2 0.3g/L of MgCl 2 3g/L of peptone, 2g/L of hydrolyzed fish protein powder and the balance of disodium hydrogen phosphate-citric acid buffer solution with the pH value of 6.
Comparative example 3 similar to example 3, except that no cottonseed protein was added to the culture medium of step S2 of comparative example 3, unlike example 3.
Comparative example 4, method for performing enzymolysis on straw
A method for enzymolysis of straws comprises the following steps:
s1, pretreatment of straws:
1) Crushing straws, sieving with a 30-mesh sieve, and putting into a steamer to be steamed for 2 hours by using steam to obtain steamed straws A;
2) Adding the straw A obtained in the step 1) into a mixed solution prepared from potassium hydrogen persulfate and boric acid, wherein the added mass of the mixed solution is 3 times of that of the straw A, and standing for 1h at the temperature of 70 ℃ to obtain an acidified straw B; the preparation method of the mixed solution comprises the following steps: mixing potassium hydrogen persulfate and boric acid according to the mass ratio of 2:1, and dissolving the mixture in deionized water with the mass 5 times that of the mixture to obtain the potassium hydrogen persulfate/boric acid composite material;
3) Adding the straw B obtained in the step 2) into deionized water with the mass being 10 times that of the straw B, keeping the temperature and standing for 20min at 50 ℃ to obtain straw C, drying the straw C to constant weight in a 120 ℃ drying oven, and then performing high-pressure steam sterilization treatment at the temperature of 121 ℃, the pressure of 100kPa and the sterilization time of 20min to obtain finally pretreated straw D;
s2, compound enzymolysis culture: placing the pretreated straw D in a culture medium containing trichoderma reesei and rhizopus oryzae, wherein the dosage of the trichoderma reesei is 300 ten thousand u/kg, the dosage of the rhizopus oryzae is 100 ten thousand u/kg, the mass relation between the addition amount of the culture medium and the added straw D is that 1g of straw D needs to be added into 40mL of culture medium, stacking and culturing for 40h after uniform stirring, and the temperature during culturing is 45 ℃ to obtain the straw D; the culture medium comprises: 5.7g/L peptone, the balance disodium hydrogen phosphate-citric acid buffer pH 6.
Comparative example 4 similar to example 3, except that the hydrolyzed fish protein powder, cottonseed protein, caCl were not added to the culture medium of step S2 of comparative example 4, unlike example 3 2 And MgCl 2 。
Experiment 1 lignin removal rate
Subject: straw obtained after pretreatment in examples 1-3 and comparative example 1.
The experimental method comprises the following steps: the straws obtained after pretreatment in examples 1-3 and the straws obtained in comparative example 1 were subjected to lignin content detection kit (cat No. BC 4205) purchased from beijing soja lebao scientific and technological limited to measure the lignin content before and after pretreatment, and the lignin removal rate was measured;
the calculation formula of the lignin removal rate is as follows: (lignin content in straw before pretreatment-lignin content in straw after pretreatment)/lignin content in straw before pretreatment x 100%.
The experimental results are as follows: as shown in table 1.
Table 1: lignin removal rate results
As can be seen from the experimental results of table 1, the removal effect of lignin is significantly increased in examples 1 to 3 as compared to comparative example 1, and the removal effect of lignin by a single acid system is lower than that of examples 1 to 3 by the mixed solution of oxone and boric acid.
Experiment 2, enzymatic saccharification experiment
Subject: the filtrates obtained by the filtration of examples 1 to 3 and comparative examples 2 to 4.
The experimental method comprises the following steps: the content of reducing sugar was determined using 3,5-dinitrosalicylic acid colorimetry.
The experimental results are as follows: as shown in table 2.
Table 2: determination of reducing sugar content results
| Number of groups | Example 1 | Example 2 | Example 3 | Comparative example 2 | Comparative example 3 | Comparative example 4 |
| Reducing sugar content | 272mg/g | 312mg/g | 328mg/g | 233mg/g | 254mg/g | 196mg/g |
As can be seen from the experimental results in Table 2, examples 1-3 have higher enzymolysis effect, comparative example 2 has obviously reduced reducing sugar content when reducing hydrolyzed fish protein powder and cottonseed protein, and comparative example 4 reduces hydrolyzed fish protein powder, cottonseed protein, caCl 2 And MgCl 2 The enzymolysis effect is further reduced, while the comparative example 3 only adds the hydrolyzed fish protein powder and CaCl 2 And MgCl 2 While no cottonseed protein was added, no better enzymatic hydrolysis effect than in examples 1-3 could be obtained, and it can be seen that hydrolyzed fish protein powder, cottonseed protein, caCl were added to the culture medium 2 And MgCl 2 The effect of obtaining reducing sugar by performing enzymolysis on the straws by trichoderma reesei and rhizopus oryzae can be effectively improved, and particularly, more remarkable effect can be obtained by using the synergistic effect of the hydrolyzed fish protein powder and the cottonseed protein.
Claims (6)
1. A method for efficiently performing enzymolysis on straws is characterized by comprising the following steps:
s1, pretreatment of straws, wherein the pretreatment comprises the following steps:
1) Crushing the straws, sieving the crushed straws with a 30-mesh sieve, and putting the crushed straws into a steamer to be steamed for 2 to 3 hours by using water vapor to obtain steamed straws A;
2) Adding the straw A obtained in the step 1) into a mixed solution prepared from potassium hydrogen persulfate and boric acid, wherein the added mass of the mixed solution is 2-3 times of that of the straw A, and standing for 1-2h at the temperature of 65-80 ℃ to obtain an acidified straw B;
3) Adding the straw B obtained in the step 2) into deionized water, preserving heat and standing for 10-30min at 40-60 ℃ to obtain straw C, drying the straw C to constant weight in a drying oven at 100-150 ℃, and then performing high-pressure steam sterilization treatment to obtain finally pretreated straw D;
s2, compound enzymolysis culture: placing the pretreated straws in a culture medium containing trichoderma reesei and rhizopus oryzae, uniformly stirring, and performing stacking culture to obtain the straw-containing culture medium; the culture medium comprises: 0.3-0.5g/L CaCl2, 0.1-0.3g/L MgCl2, 0.5-2g/L peptone, 1-2g/L hydrolyzed fish protein powder, 1-2g/L cottonseed protein, and the balance buffer solution.
2. The method for efficiently hydrolyzing straws in an enzymolysis manner as claimed in claim 1, wherein the preparation method of the mixed solution in the step 2) comprises the following steps: mixing potassium hydrogen persulfate and boric acid according to the mass ratio of 1-3:1, and dissolving the mixture in deionized water to obtain the potassium hydrogen persulfate/boric acid composite material.
3. The method for efficiently hydrolyzing straws by using enzymes according to claim 1, wherein the dosage of trichoderma reesei in the step S2 is 200-300 ten thousand u/kg, and the dosage of rhizopus oryzae is 80-120 ten thousand u/kg.
4. The method for efficiently hydrolyzing straws as claimed in claim 1, wherein the buffer solution is disodium hydrogen phosphate-citric acid buffer solution, and the pH value of the buffer solution is 5-6.
5. The method for efficiently hydrolyzing straws with enzymes as claimed in claim 1, wherein the mass relationship between the added amount of the culture medium and the added straws D is that 30-45mL of the culture medium is required to be added for 1g of straws D.
6. The method for efficiently hydrolyzing straws in an enzymolysis manner as claimed in claim 1, wherein the cultivation time in the step S2 is 36-48h, and the cultivation temperature is 35-45 ℃.
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| CN102077903A (en) * | 2010-11-25 | 2011-06-01 | 河南农业大学 | Method for jointly treating stalks by steam explosion and microorganism fermentation |
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| CN109112172A (en) * | 2018-09-25 | 2019-01-01 | 中国农业科学院麻类研究所 | A kind of method of microorganism enzymatic saccharification stalk |
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