CN107535675B - Inonotus obliquus solid fermentation method for wheat straw conversion feed - Google Patents
Inonotus obliquus solid fermentation method for wheat straw conversion feed Download PDFInfo
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Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
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Abstract
The invention discloses a solid fermentation method of inonotus obliquus for wheat straw conversion feed, which takes wheat straws as raw materials and adopts white rot fungus inonotus obliquus to carry out solid fermentation and degrade the wheat straws to finally obtain the wheat straw conversion feed. The invention has simple fermentation process, low cost, good flavor of the fermented product, improved crude protein and amino acid contents in different degrees, high in-vitro digestibility and rich nutrition, contains high-activity carboxymethyl cellulose, filter paper enzyme, lignin peroxidase and manganese peroxidase, and achieves the effect of producing high-quality fermented feed.
Description
Technical Field
The invention relates to the technical field of solid fermentation feed preparation, in particular to a solid fermentation method of inonotus obliquus for wheat straw conversion feed.
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. Thus, straw utilization studies have shifted from the past degradation of cellulose to the degradation of lignin. Research shows that the white rot fungi can effectively degrade lignin, and because the white rot fungi have wide adaptability to lignocellulose and can secrete high-activity ligninase and cellulase, the white rot fungi can utilize various lignocelluloses which cannot be directly utilized by livestock to synthesize own mycoprotein. White-rot fungi are increasingly valued by researchers at home and abroad because of the advantages of multiple purposes, reproducibility, short period, high nutritional value and the like in straw conversion.
The mycoprotein is the microbial protein produced by the self-reproduction of microorganisms in the fermentation process, a large amount of useful metabolites such as vitamins, lysine, organic acid and the like are produced and accumulated in the straw fermentation process, and some of the mycoprotein has the antiseptic effect such as lactic acid, acetic acid, ethanol and the like; some of them can enhance the disease resistance of animals and promote the growth and reproduction of beneficial flora. Especially in recent years, the research of producing single cell protein by using cellulose raw materials such as straws and the like through microbial fermentation and conversion has attracted attention due to the characteristics of wide raw material sources, low cost, contribution to solving the environmental pollution and the like. The method for producing the feed by microbial fermentation mainly comprises two modes of liquid fermentation and solid fermentation, and compared with the solid fermentation, the method has the remarkable advantages of high yield, short period, low energy consumption, less waste water and waste residues and the like.
Although there are many patent reports of preparing biological feed by taking substrates such as vinasse, bean pulp, leaves, cassava and the like as raw materials at home and abroad at present, the patent report of preparing the biological feed by using wheat straws is few. For example, patent 201010295193.x reports that ginkgo leaf residues and straws are added with water and sealed and then directly fermented, no any strain is added in the method for directly fermenting, the prepared fermented feed is low in protein content and nutritional value, and the method adopts liquid fermentation, generates a large amount of wastewater, is difficult to treat, is high in energy consumption and pollutes the environment. Patent 201210549464.9 reports that ginkgo leaves are used as raw materials and single strain bacillus subtilis is used for solid fermentation to prepare the biological feed additive, and patent 200910031310.9 reports that ginkgo leaves are used as raw materials and single strain candida utilis is used for solid fermentation to prepare the biological feed additive. These patents all adopt a certain strain for fermentation, and the strain does not have special capability for degrading lignocellulose, so the prepared biological feed has the serious problems of single nutrient component, low protein content, low animal digestibility, poor palatability and the like.
Disclosure of Invention
The invention aims to provide a solid fermentation method of inonotus obliquus for converting wheat straws into feed, which solves the problems of low crude protein content and high lignin content in the use process of the wheat straws as the feed and improves the nutritional value of the wheat straws as the feed.
The technical scheme adopted by the invention for solving the technical problems is as follows:
a solid fermentation method of inonotus obliquus for wheat straw conversion feed comprises the following steps:
(1) preparing a seed solution: the inonotus obliquus strain is firstly activated in a malt peptone agar culture medium and then transferred to a liquid seed culture medium for culture to obtain a seed solution.
(2) Pretreatment of wheat straws: cutting the wheat straw to 2-2.5cm, fully soaking the wheat straw in tap water for 3 days, taking out and airing; the wheat straw is cut to 2-2.5cm in length, so that the wheat straw is convenient to degrade, and if the wheat straw is ground into powder, liquid is added subsequently to mix, so that the wheat straw is not beneficial to strain culture; the soaking with water is to make the water fully permeate the matrix and is also beneficial to the degradation of the inner part of the wheat straw by the strains; the reason why the soaked wheat straws are taken out and dried in the air is that the inonotus obliquus can not be cultured on the substrate with too large water content, so the wheat straws are dried in the air until no water obviously flows out.
(3) Solid fermentation: and (3) uniformly mixing the wheat straws treated in the step (2) with the nutrient solution, sterilizing, inoculating the seed solution, fermenting at the temperature of 24-28 ℃ for 10-15 days, taking out a fermentation product after the fermentation is finished, and then drying in a drying oven at the temperature of 50 ℃ for 3 days to obtain the product.
The white rot fungus inonotus obliquus adopted by the invention can propagate the thalli of the white rot fungus inonotus obliquus in a large quantity due to the strong capability of decomposing cellulose, thereby improving the protein content in the wheat straw; on the other hand, the wheat straw degrading agent has strong lignin degrading capacity, can improve the palatability of the wheat straw and improves the digestibility of the ruminant to the wheat straw. Based on the two aspects, the fermented wheat straws are used as the feed of ruminants, and a new way is provided for the deep utilization of wheat straw resources.
In the invention, the inonotus obliquus can decompose and utilize lignin, cellulose and hemicellulose during solid fermentation, and produce a large amount of mycoprotein and metabolite, thereby greatly improving the protein content and the nutritional quality of the wheat straw converted feed. The in vitro gas production method shows that the gas production of the wheat straws is increased from 125mL/g to 168.6mL/g after the wheat straws are treated by the white rot fungi, and the gas production is obviously increased after the treatment, which shows that the rumen degradability of the wheat straws after biological treatment is better, so the wheat straws are more suitable for feeding ruminants. The invention has simple fermentation process, low cost, good flavor of the fermented product, improved crude protein and amino acid contents in different degrees, high in-vitro digestibility and rich nutrition, contains high-activity carboxymethyl cellulose, filter paper enzyme, lignin peroxidase and manganese peroxidase, and achieves the effect of producing high-quality fermented feed.
Preferably, in the step (1), the malt peptone agar medium formula is as follows: 30g/L of malt extract powder, 3g/L of peptone, 15 g/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 ℃ and the time is 72 h.
Preferably, in the step (1), the liquid seed culture medium formula is as follows: 20 g/L glucose, 3g/L peptone, 2 g/L yeast extract, KH2PO41 g/L，MgSO41.5 g/L，CaCl20.1 g/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 ℃, the rotating speed is 150r/min, and the culture time is 48 h.
Preferably, in the step (3), the formula of the nutrient solution is as follows: corn flour 200 g/L, (NH)4）2SO410 g/L，MgSO4·7H2O 0.3 g/L，KH2PO40.11 g/L, Tween-804 g/L, pH 7.4. Aiming at the specific solid fermentation method, the inventor develops an adaptive nutrient solution formula through exploration tests, and the formula is different from the existing formula.
Preferably, in the step (3), the solid-to-liquid ratio of the wheat straws to the nutrient solution is 2g to 1mL, and the dosage of the seed solution is 0.1-0.3mL per 1g of wheat straws.
The invention has the beneficial effects that:
the invention takes the agricultural resource waste wheat straw as the raw material, and adopts the biotransformation method to prepare the feed protein, thereby not only changing waste into valuable, but also providing a new high-quality feed protein with safety and high nutritive value.
Compared with the prior art, the prepared wheat straw feed protein not only has obviously improved protein content, but also contains various high-activity cellulase, ligninase, various amino acids and trace elements, has comprehensive nutrient components and good palatability, and has high ruminant digestibility.
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.
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 solid fermentation method of inonotus obliquus for wheat straw conversion feed 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, 15 g/L of agar and 5.4-5.8 of pHs.
The formula of the liquid seed culture medium is as follows: 20 g/L glucose, 3g/L peptone, 2 g/L yeast extract, KH2PO41g/L，MgSO41.5 g/L，CaCl20.1 g/L, pH is natural.
(2) Pretreatment of wheat straws: cutting the wheat straw to 2cm length, fully soaking the wheat straw in tap water for 3 days, taking out and airing;
(3) solid fermentation: and (3) uniformly mixing 10g of wheat straw treated in the step (2) with 5mL of nutrient solution, sterilizing at 121 ℃ for 30min, inoculating 1mL of seed solution, fermenting at 28 ℃ for 10 days, taking out a fermentation product after the fermentation is finished, and then drying in a 50 ℃ oven for 3 days to obtain the product.
The nutrient solution formula is as follows: corn flour 200 g/L, (NH)4）2SO410 g/L，MgSO4·7H2O 0.3 g/L，KH2PO40.11 g/L，Tween-80 4 g/L，pH 7.4。
The detection result shows that the product contains 9.1 percent of crude protein, 35.3 percent of cellulose, 24.3 percent of hemicellulose, 16.7 percent of lignin and 168.6mL/g of in vitro gas production. The wheat straw which is not treated by the method has the crude protein content of 4.4 percent, the cellulose content of 30.3 percent, the hemicellulose content of 21.2 percent, the lignin content of 15.1 percent and the in vitro gas production rate of 125 mL/g.
After fermentation, the activity of the lignocellulose degrading enzyme is 10 th day: the carboxymethyl cellulose activity is 58.4 IU/g, the filter paper enzyme activity is 14.3 IU/g, the lignin peroxidase activity is 832.1 IU/g, the manganese peroxidase activity is 493.8 IU/g, and the blank group enzyme activity before treatment is 0 IU/g.
Example 2:
a solid fermentation method of inonotus obliquus for wheat straw conversion feed comprises the following steps:
(1) preparing a seed solution: firstly, activating the strain of the inonotus obliquus in a malt peptone agar culture medium, wherein 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, 15 g/L of agar and 5.4-5.8 of pHs.
The formula of the liquid seed culture medium is as follows: 20 g/L glucose, 3g/L peptone, 2 g/L yeast extract, KH2PO41g/L，MgSO41.5 g/L，CaCl20.1 g/L, pH is natural.
(2) Pretreatment of wheat straws: cutting the wheat straw to 2.5cm, fully soaking the wheat straw in tap water for 3 days, taking out and airing;
(3) solid fermentation: and (3) uniformly mixing 50g of the wheat straw treated in the step (2) with 25mL of nutrient solution, sterilizing at 121 ℃ for 30min, inoculating 5mL of seed solution, fermenting at 24 ℃ for 15 days, taking out a fermentation product after the fermentation is finished, and then drying in an oven at 50 ℃ for 3 days to obtain the product.
The nutrient solution formula is as follows: corn flour 200 g/L, (NH)4）2SO410 g/L，MgSO4·7H2O 0.3 g/L，KH2PO40.11 g/L，Tween-80 4 g/L，pH 7.4。
The detection result shows that the product contains 9.8 percent of crude protein, 36.2 percent of cellulose, 25.6 percent of hemicellulose, 17.2 percent of lignin and 172.3mL/g of in vitro gas production. The wheat straw which is not treated by the method has the crude protein content of 4.4 percent, the cellulose content of 30.3 percent, the hemicellulose content of 21.2 percent, the lignin content of 15.1 percent and the in vitro gas production rate of 125 mL/g.
Example 3:
a solid fermentation method of inonotus obliquus for wheat straw conversion feed comprises the following steps:
(1) preparing a seed solution: firstly, activating the strain of the inonotus obliquus in a malt peptone agar culture medium, wherein 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, 15 g/L of agar and 5.4-5.8 of pHs.
The formula of the liquid seed culture medium is as follows: 20 g/L glucose, 3g/L peptone, 2 g/L yeast extract, KH2PO41g/L，MgSO41.5 g/L，CaCl20.1 g/L, pH is natural.
(2) Pretreatment of wheat straws: cutting the wheat straw to 2.2cm, fully soaking the wheat straw in tap water for 3 days, taking out and airing;
(3) solid fermentation: and (3) uniformly mixing 100g of the wheat straw treated in the step (2) with 50mL of nutrient solution, sterilizing at 121 ℃ for 30min, inoculating 10mL of seed solution, fermenting at 26 ℃ for 12 days, taking out a fermentation product after the fermentation is finished, and then drying in an oven at 50 ℃ for 3 days to obtain the product.
The nutrient solution formula is as follows: corn flour 200 g/L, (NH)4）2SO410 g/L，MgSO4·7H2O 0.3 g/L，KH2PO40.11 g/L，Tween-80 4 g/L，pH 7.4。
The detection result shows that the product contains 9.5 percent of crude protein, 35.8 percent of cellulose, 25.2 percent of hemicellulose, 16.9 percent of lignin and 170.4mL/g of in vitro gas production. The wheat straw which is not treated by the method has the crude protein content of 4.4 percent, the cellulose content of 30.3 percent, the hemicellulose content of 21.2 percent, the lignin content of 15.1 percent and the in vitro gas production rate of 125 mL/g.
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 (5)
1. A solid fermentation method of inonotus obliquus of a wheat straw conversion feed 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) pretreatment of wheat straws: cutting the wheat straw to 2-2.5cm, fully soaking the wheat straw in tap water for 3 days, taking out and airing;
(3) solid fermentation: uniformly mixing the wheat straws treated in the step (2) with the nutrient solution, sterilizing, inoculating seed solution, fermenting at 24-28 ℃ for 10-15 days, taking out a fermentation product after the fermentation is finished, and then drying in a drying oven at 50 ℃ for 3 days to obtain a product;
in the step (3), the nutrient solution formula is as follows: corn flour 200 g/L, (NH)4）2SO410 g/L，MgSO4·7H2O 0.3 g/L，KH2PO40.11 g/L，Tween-80 4 g/L，pH 7.4；
In the step (3), the solid-to-liquid ratio of the wheat straw to the nutrient solution is 2g to 1mL, and the dosage of the seed solution is 0.1-0.3mL per 1g of wheat straw.
2. The method for the solid fermentation of inonotus obliquus in a straw-converted feed according to claim 1, wherein in the step (1), the formulation of the malt peptone agar medium is as follows: 30g/L of malt extract powder, 3g/L of peptone, 15 g/L of agar and 5.4-5.8 of pHs.
3. The method for the solid fermentation of inonotus obliquus in wheat straw transformed feed according to claim 2, wherein the parameters of the activation in the malt peptone agar medium in the step (1) are as follows: the temperature is 28 ℃ and the time is 72 h.
4. The method for solid fermentation of Inonotus obliquus in wheat straw-converted feedstuff as claimed in claim 1, wherein in step (1), liquid seed culture is performedThe base formula is as follows: 20 g/L glucose, 3g/L peptone, 2 g/L yeast extract, KH2PO41g/L，MgSO41.5 g/L，CaCl20.1 g/L, pH is natural.
5. The solid fermentation method of Inonotus obliquus for wheat straw conversion feed according to claim 4, wherein in the step (1), the parameters of the liquid seed culture medium culture are as follows: the temperature is 28 ℃, the rotating speed is 150r/min, and the culture time is 48 h.
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