CN112931687A - Method for converting wheat straw into feed cooperatively through Fenton reaction pretreatment and inonotus obliquus solid fermentation - Google Patents
Method for converting wheat straw into feed cooperatively through Fenton reaction pretreatment and inonotus obliquus solid fermentation Download PDFInfo
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
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- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
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Abstract
The invention discloses a method for converting wheat straw into feed by cooperation of Fenton reaction pretreatment and inonotus obliquus solid fermentation, which takes agricultural resource waste wheat straw as a raw material and adopts the Fenton reaction pretreatment and inonotus obliquus solid fermentation methods to prepare feed protein. The wheat straw feed protein prepared by the invention can effectively improve the production performance and feed conversion rate of ruminants, and has good market prospect.
Description
Technical Field
The invention belongs to the technical field of fermentation engineering, and particularly relates to a method for converting wheat straw into a feed by combining Fenton reaction pretreatment and inonotus obliquus solid fermentation.
Background
In recent years, the best way for treating crop straws by biological means is always searched at home and abroad, and the problem has the greatest difficulty that water-insoluble lignocellulose in the straws is difficult to degrade by acid, alkali and enzyme, mainly caused by the crystallinity and the polymerization degree of the cellulose and lignin wrapping the cellulose and hemicellulose. Lignin and hemicellulose are combined in a covalent bond mode, cellulose molecules are embedded in the lignin and the hemicellulose to form a natural barrier, so that enzymes are not easily contacted with the cellulose molecules, and the water insolubility and the chemical structure complexity of the lignin cause the difficult degradation of straws. Therefore, to completely degrade cellulose, the problem of lignin degradation must be solved.
Some common pretreatment methods of crop straws include physical treatment, chemical treatment and the like. Physical treatments such as grinding, soaking, granulating, etc., all of which reduce the crystallinity of the cellulose, ultimately resulting in better degradation of the cellulose by hydrolytic enzymes. However, the physical treatment method requires machinery and processing procedures, so that the application in agriculture is limited, and although the reduction of the particle size is really beneficial to the subsequent enzyme treatment process, the crushed crop straws are not beneficial to the later-stage feeding operation. Although the acid-base treatment method can reduce the crystallinity of cellulose and increase the yield of saccharides, the application of sulfuric acid and sodium hydroxide is not only expensive, but also causes serious environmental pollution. Meanwhile, the method does not have the function of increasing the protein content, and the single microbial solid fermentation method cannot obviously increase the protein content.
The Fenton reaction is characterized in that a chain reaction between divalent iron ions (Fe2+) and hydrogen peroxide catalyzes a hydroxyl radical to generate stronger oxidation capacity, the oxidation potential of the hydroxyl radical is second to that of fluorine, in addition, the hydroxyl radical has high electronegativity or electrophilicity, the electron affinity of the hydroxyl radical is as high as 569.3kJ, and the hydroxyl radical has strong addition reaction characteristics, so that the Fenton reagent can indiscriminately oxidize most organic matters in water, and is particularly suitable for oxidation treatment of organic wastewater which is difficult to biodegrade or generally difficult to effectively oxidize chemically.
Disclosure of Invention
The invention aims to provide a method for converting wheat straw into a feed by combining Fenton reaction pretreatment and inonotus obliquus solid fermentation, solves the problems of low crude protein content and high lignin content in the process of using the wheat straw as the feed, and improves the nutritional value of the wheat straw as the feed.
The technical scheme adopted by the invention for solving the technical problems is as follows:
a method for transforming wheat straw into feed by cooperation of Fenton reaction pretreatment and inonotus obliquus solid fermentation comprises the following steps:
(1) preparing a seed solution: activating the inonotus obliquus strain in a malt peptone agar culture medium, and transferring to a liquid seed culture medium for culture to obtain a seed solution;
(2) straw Fenton pretreatment: cutting the wheat straws to wheat straw sections with the length of 1.0-1.5cm, carrying out Fenton pretreatment, washing cleanly after the treatment is finished, and airing;
(3) solid fermentation: and (3) uniformly mixing the wheat straws treated in the step (2) with the nutrient solution, sterilizing, inoculating seed solution, fermenting for 25-30 days at the humidity of 75 +/-3% and the temperature of 26-28 ℃, taking out a fermentation product after the fermentation is finished, and drying to obtain the product.
The wheat straw is cut to 1.0-1.5cm in length to facilitate degradation, and if the wheat straw is ground into powder, liquid is added subsequently to be mixed, so that the wheat straw is not beneficial to strain culture; meanwhile, the water is soaked to ensure that the water fully permeates the matrix, which is also beneficial to the degradation of the inner part of the wheat straw by the strains; the air drying of the wheat straws is that the inonotus obliquus can not be cultured on a substrate with too large water content, so that the wheat straws are air-dried until no water obviously flows out.
The Fenton pretreatment has mild reaction conditions, low cost and environmental friendliness, can reduce the molecular weight of lignin and promote the enzymatic saccharification of lignocellulose biomass, and meanwhile, the treated wheat straw has no reagent residue, so that the method is beneficial to the solid fermentation of the inonotus obliquus at the later stage, and the protein content is improved.
The Fenton reaction can not change or improve the nutritive value of crop straws, but can destroy the lignin structure, promote the degradation capability of the inonotus obliquus on lignocellulose and further improve the protein production capability of white rot fungi. The invention adopts the Fenton reaction pretreatment and the inonotus obliquus solid fermentation synergistic treatment, can obviously increase the protein content of the fermentation product, simultaneously reduces the lignin content, and improves the nutritional value of the wheat straw as the feed.
On one hand, the Fenton reagent adopted by the invention can randomly attack cell walls due to high-activity hydroxyl free radicals (. OH) generated by the Fenton reagent, so that the lignocellulose structure is damaged, the treated wheat straw is beneficial to the growth and degradation of the inonotus obliquus, so that the protein content in the wheat straw is improved, meanwhile, the treated wheat straw is more crisp and fragile, and the palatability of the straw feed is improved; on the other hand, the Fenton reaction has low cost, low consumption, mild reaction and environmental protection, and saves a large amount of resources. Based on the two aspects, the straw wheat straw which is pretreated by Fenton and subjected to solid fermentation by the inonotus obliquus is used as the feed of the ruminant, so that a new way is provided for deep utilization of the wheat straw resource.
The straw pretreated by Fenton in the invention is crisp and fragile, and lignin, cellulose and hemicellulose can be decomposed and utilized when the straw is subjected to solid fermentation by inonotus obliquus, and a large amount of mycoprotein is generated, so that the protein content and the nutritional quality of the straw converted feed are greatly improved. The protein content is detected to be increased from 6.45 percent to 20.59 percent, so the feed is suitable for feeding ruminants. The invention has simple fermentation process, low cost, good flavor of the fermented product, and improved crude protein content to different degrees, thereby achieving the effect of producing high-quality fermented feed.
Preferably, in step (2), the Fenton reagent for Fenton pretreatment is prepared from FeSO4Solution and H2O2Composition of, wherein FeSO4Final concentration of 0.015-0.025mol/L, H2O2The final concentration is 1.5-2.5 mol/L.
Preferably, in the step (2), the Fenton pretreatment reaction parameters are as follows: the solid-liquid ratio of the wheat straw section to the Fenton reagent is 1g:15-18mL, the temperature is 50 +/-2 ℃, the rotating speed of a shaking table is 200 +/-10 r/min, and the time is 2.5-3.5 h.
Preferably, in the step (3), the formula of the nutrient solution is as follows: corn flour 200g/L, (NH)4)2SO4 10g/L，MgSO4·7H2O 0.3g/L，KH2PO4 0.11g/L，Tween-80 4g/L，pH 5.0。
Preferably, in the step (3), the solid-to-liquid ratio of the wheat straws to the nutrient solution is 1g:2-3mL, and the dosage of the seed solution is 0.5-1.0mL per 1g of wheat straws.
Preferably, in the step (1), the liquid seed culture medium formula is as follows: 20g/L glucose, 3g/L peptone, 2g/L yeast extract, KH2PO4 1g/L，MgSO4 1.5g/L，CaCl20.1g/L, pH is natural.
Preferably, in the step (1), the parameters of the liquid seed culture medium culture are as follows: the temperature is 28 +/-1 ℃, the rotating speed is 150 +/-10 r/min, and the culture time is 48 +/-1 h.
Preferably, in the step (1), the malt peptone agar medium formula is as follows: 30g/L of malt extract powder, 3g/L of peptone, 15g/L of agar and 5.4-5.8 of pH.
Preferably, in step (1), the parameters of activation in malt peptone agar medium are: the temperature is 28 +/-1 ℃, and the time is 72 +/-1 h.
The invention has the beneficial effects that: the invention takes agricultural resource waste wheat straws as raw materials, adopts Fenton reaction pretreatment and inonotus obliquus solid fermentation methods to prepare the feed protein, provides a safe, low-cost and environment-friendly pretreatment method, can change waste into valuable, and provides a new high-quality feed protein with safety and high nutritional value.
The wheat straw feed protein prepared by the invention can effectively improve the production performance and feed conversion rate of ruminants, and has good market prospect.
Drawings
FIG. 1 is the change in cellulose content during fermentation.
FIG. 2 is the change in hemicellulose content during fermentation.
FIG. 3 is the change in lignin content during fermentation.
Detailed Description
The technical solution of the present invention will be further specifically described below by way of specific examples.
In the present invention, the raw materials and equipment used are commercially available or commonly used in the art, unless otherwise specified. The methods in the following examples are conventional in the art unless otherwise specified.
Example 1:
a method for transforming wheat straw into feed by cooperation of Fenton reaction pretreatment and inonotus obliquus solid fermentation comprises the following steps:
(1) preparing a seed solution: the inonotus obliquus strain (commercially available) is firstly activated in a malt peptone agar medium, and the activation parameters are as follows: the temperature is 27 ℃, and the time is 73 h; then transferring to a liquid seed culture medium for culture, wherein the culture parameters are as follows: the temperature is 27 ℃, the rotating speed is 160r/min, and the culture time is 49h, thus obtaining the seed liquid.
The malt peptone agar medium formula is: 30g/L of malt extract powder, 3g/L of peptone, 15g/L of agar and 5.4-5.8 of pH.
The formula of the liquid seed culture medium is as follows: 20g/L glucose, 3g/L peptone, 2g/L yeast extract, KH2PO4 1g/L，MgSO4 1.5g/L，CaCl20.1g/L, pH is natural.
(2) Pretreatment of wheat straws: cutting wheat straw to 1cm length, and adding equal volume of 0.015mol/L FeSO according to the solid-liquid ratio of 1:164The solution was mixed with 1.5mol/L of H2O2Taking out the mixture after 2.5 hours at the temperature of 52 ℃ and the rotating speed of 210r/min, and cleaning and airing the mixture; (3) solid fermentation: and (3) uniformly mixing 3g of wheat straw treated in the step (2) with 9mL of nutrient solution, sterilizing at 121 ℃ for 30min, inoculating 1.5mL of seed solution, fermenting at the humidity of 72% and the temperature of 27 ℃ for 30 days, taking out a fermentation product after the fermentation is finished, and then drying in an oven at 55 ℃ for 1 day to obtain the product.
The nutrient solution formula is as follows: corn flour 200g/L, (NH)4)2SO4 10g/L，MgSO4·7H2O 0.3g/L，KH2PO40.11g/L，Tween-80 4g/L，pH 5.0。
Example 2:
a method for transforming wheat straw into feed by cooperation of Fenton reaction pretreatment and inonotus obliquus solid fermentation comprises the following steps:
(1) preparing a seed solution: the inonotus obliquus strain (commercially available) is firstly activated in a malt peptone agar medium, and the activation parameters are as follows: the temperature is 29 ℃, and the time is 71 h; then transferring to a liquid seed culture medium for culture, wherein the culture parameters are as follows: the temperature is 29 ℃, the rotating speed is 140r/min, and the culture time is 47h, thus obtaining the seed liquid.
The malt peptone agar medium formula is: 30g/L of malt extract powder, 3g/L of peptone, 15g/L of agar and 5.4-5.8 of pH.
The formula of the liquid seed culture medium is as follows: 20g/L glucose, 3g/L peptone, 2g/L yeast extract, KH2PO4 1g/L，MgSO4 1.5g/L，CaCl20.1g/L, pH is natural.
(2) Pretreatment of wheat straws: cutting wheat straw to 1.2cm length, adding equal volume of 0.025mol/L FeSO according to solid-liquid ratio of 1:184The solution was mixed with 2.5mol/L H2O2Taking out, cleaning and airing at the temperature of 48 ℃ and the rotating speed of 190r/min for 3.5 hours; (3) solid fermentation: and (3) uniformly mixing 3g of wheat straw treated in the step (2) with 7mL of nutrient solution, sterilizing at 121 ℃ for 30min, inoculating 3mL of seed solution, fermenting at the humidity of 78% and the temperature of 28 ℃ for 25 days, taking out a fermentation product after the fermentation is finished, and then drying in an oven at 55 ℃ for 1 day to obtain the product.
The nutrient solution formula is as follows: corn flour 200g/L, (NH)4)2SO4 10g/L，MgSO4·7H2O 0.3g/L，KH2PO40.11g/L，Tween-80 4g/L，pH 5.0。
Example 3:
a method for transforming wheat straw into feed by cooperation of Fenton reaction pretreatment and inonotus obliquus solid fermentation comprises the following steps:
(1) preparing a seed solution: the inonotus obliquus strain (commercially available) is firstly activated in a malt peptone agar medium, and the activation parameters are as follows: the temperature is 28 ℃, and the time is 72 h; then transferring to a liquid seed culture medium for culture, wherein the culture parameters are as follows: the temperature is 28 ℃, the rotating speed is 150r/min, and the culture time is 48h, thus obtaining the seed liquid.
The malt peptone agar medium formula is: 30g/L of malt extract powder, 3g/L of peptone, 15g/L of agar and 5.4-5.8 of pH.
Liquid, method for producing the same and use thereofThe formula of the seed culture medium is as follows: 20g/L glucose, 3g/L peptone, 2g/L yeast extract, KH2PO4 1g/L，MgSO4 1.5g/L，CaCl20.1g/L, pH is natural.
(2) Pretreatment of wheat straws: cutting wheat straw to 1.5cm length, and adding equal volume of 0.02mol/L FeSO according to the solid-to-liquid ratio of 1:154The solution was mixed with 2.0mol/L H2O2Taking out, cleaning and airing at the temperature of 50 ℃ and the rotating speed of 200r/min after 3 hours; (3) solid fermentation: and (3) uniformly mixing 3g of wheat straw treated in the step (2) with 6mL of nutrient solution, sterilizing at 121 ℃ for 30min, inoculating 2mL of seed solution, fermenting at the humidity of 75% and the temperature of 26 ℃ for 30 days, taking out a fermentation product after the fermentation is finished, and then drying in an oven at 55 ℃ for 1 day to obtain the product.
The nutrient solution formula is as follows: corn flour 200g/L, (NH)4)2SO4 10g/L，MgSO4·7H2O 0.3g/L，KH2PO40.11g/L，Tween-80 4g/L，pH 5.0。
The product of example 3 was tested to have a mycelium content of 149.91g/kg (Table 1), a crude protein content of 20.6%, an increase of 221.9% (Table 2), a cellulose content of 39.9% (FIG. 1), a hemicellulose content of 15.4% (FIG. 2), and a lignin content of 18.3% (FIG. 3). The amount of mycelia produced from wheat straws which were not treated by the method of the present invention was 139.3g/kg (Table 1), the crude protein content was 6.7%, the increase was 4.7% (Table 2), the cellulose content was 33.79% (FIG. 1), the hemicellulose content was 23.16% (FIG. 2), and the lignin content was 22.23% (FIG. 3). Although the reduced protein content (4.5%) of straw after Fenton pretreatment (Table 2) was observed, it has a significant positive effect on the subsequent solid fermentation of Inonotus obliquus.
TABLE 1 variation of mycelium amount during fermentation (g mycelium/kg straw)
| Time (sky) | Without Fenton | Fenton treatment | |
| 5 | 54.45 | 57.98 | |
| 10 | 68.6 | 82.73 | |
| 15 | 82.73 | 111.02 | |
| 20 | 96.87 | 128.69 | |
| 20 | 125.16 | 139.3 | |
| 30 | 139.3 | 149.91 |
TABLE 2 Change in protein content during fermentation
Therefore, the straw pretreated by Fenton is subjected to solid fermentation, lignin can be degraded under the condition of maximally preserving the cellulose content, the crude protein content of the straw can be obviously increased, and the method is a relatively excellent pretreatment method.
The above-described embodiments are only preferred embodiments of the present invention, and are not intended to limit the present invention in any way, and other variations and modifications may be made without departing from the spirit of the invention as set forth in the claims.
Claims (9)
1. A method for transforming wheat straw into feed by cooperation of Fenton reaction pretreatment and inonotus obliquus solid fermentation is characterized by comprising the following steps:
(1) preparing a seed solution: activating the inonotus obliquus strain in a malt peptone agar culture medium, and transferring to a liquid seed culture medium for culture to obtain a seed solution;
(2) straw Fenton pretreatment: cutting the wheat straws to wheat straw sections with the length of 1.0-1.5cm, carrying out Fenton pretreatment, washing cleanly after the treatment is finished, and airing;
(3) solid fermentation: and (3) uniformly mixing the wheat straws treated in the step (2) with the nutrient solution, sterilizing, inoculating seed solution, fermenting for 25-30 days at the humidity of 75 +/-3% and the temperature of 26-28 ℃, taking out a fermentation product after the fermentation is finished, and drying to obtain the product.
2. The method of claim 1, wherein in step (2), the Fenton reagent for Fenton pretreatment is prepared from FeSO4Solution and H2O2Composition of, wherein FeSO4Final concentration of 0.015-0.025mol/L, H2O2The final concentration is 1.5-2.5 mol/L.
3. The method according to claim 1 or 2, wherein in the step (2), the Fenton pretreatment reaction parameters are as follows: the solid-liquid ratio of the wheat straw section to the Fenton reagent is 1g:15-18mL, the temperature is 50 +/-2 ℃, the rotating speed of a shaking table is 200 +/-10 r/min, and the time is 2.5-3.5 h.
4. The method of claim 1, wherein the method is performed in a batch modeCharacterized in that in the step (3), the formula of the nutrient solution is as follows: corn flour 200g/L, (NH)4)2SO4 10g/L，MgSO4·7H2O 0.3g/L，KH2PO4 0.11g/L，Tween-80 4g/L，pH 5.0。
5. The method according to claim 1 or 4, wherein in the step (3), the solid-to-liquid ratio of the wheat straw to the nutrient solution is 1g:2-3mL, and the amount of the seed solution is 0.5-1.0mL per 1g of the wheat straw.
6. The method of claim 1, wherein in step (1), the liquid seed medium is formulated as: 20g/L glucose, 3g/L peptone, 2g/L yeast extract, KH2PO4 1g/L，MgSO4 1.5g/L，CaCl20.1g/L, pH is natural.
7. The method according to claim 1 or 6, wherein in step (1), the parameters of the liquid seed culture medium culture are: the temperature is 28 +/-1 ℃, the rotating speed is 150 +/-10 r/min, and the culture time is 48 +/-1 h.
8. The method according to claim 1, wherein in step (1), the malt peptone agar medium is formulated as: 30g/L of malt extract powder, 3g/L of peptone, 15g/L of agar and 5.4-5.8 of pH.
9. The method according to claim 1 or 8, wherein in step (1), the parameters of activation in malt peptone agar medium are: the temperature is 28 +/-1 ℃, and the time is 72 +/-1 h.
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