CN101343649B - Method for biodegradation of wheat bran araboxylan - Google Patents
Method for biodegradation of wheat bran araboxylan Download PDFInfo
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- CN101343649B CN101343649B CN2008100713431A CN200810071343A CN101343649B CN 101343649 B CN101343649 B CN 101343649B CN 2008100713431 A CN2008100713431 A CN 2008100713431A CN 200810071343 A CN200810071343 A CN 200810071343A CN 101343649 B CN101343649 B CN 101343649B
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Abstract
The invention provides a method for biodegrading wheat bran araboxylan. The method adopts edible Agaricus blazei Murill which can be used as medicine and food as biodegradable probiotic bacteria, and a liquid culturing technique and a culture medium particularly selected are adopted to induce an extracellular compound fiber degeneration enzyme system of the biodegraded fungus to degrade the wheat bran araboxylan and to improve the transformation utilization efficiency, therefore, the nutrition converting utilization efficiency of the wheat bran when being adopted as the feed resource is enhanced.
Description
Technical field
The invention belongs to biological technical field, more specifically relate to a kind of method of biodegradation of wheat bran araboxylan.
Background technology
Wheat bran is the main sub product of wheat processing, and ratio accounts for 20% of processing total amount.On Production of Livestock and Poultry, because wheat bran contains a large amount of anti-nutritions, degradation efficiency is lower, has limited it as the utilization of animal-feed resources effective.Raise practice and show, if Wheat bran content can cause that too much livestock and poultry production performance descends degradation problem under the carcase quality in the monogastric animal daily ration.On the one hand, with the Wheat bran non-starch polysaccharide in staple solubility araboxylan content direct relation is arranged; On the other hand, also receive the influence of (comprising the insolubility araboxylan) of insolubility non-starch polysaccharide in the Wheat bran, suppressed degraded utilising efficiency the chimeric structural nutritive ingredient of wheat bran cell walls.Therefore, in the efficient feed of Testa Tritici utilizes,, be the important gordian technique that must solve to the degraded of araboxylan.
Araboxylan is present in a kind of main cell wall polysaccharides in the wheat bran, by β-D-xylopyranose residue through the xylan backbone that β-(1 → 4)-glycosidic link forms in succession, and with α-L-furans pectinose be that side chain is formed by connecting.α-L-furans arabinose side chains is to pass through 1-2 with 2 or α-L-furans pectinose monose molecule more than 2; 1-3; The 1-5 key couples together, and also contains a certain amount of ferulic simultaneously, and is covalently bound through the form and the araboxylan of esterification.The cell wall structure characteristics of this complicacy are difficult to by enteron aisle digestion and metabolism enzyme liberating it.
The complicated physicochemical characteristic of araboxylan is the effect basis of its ANFs, and it has bigger water holding capacity, causes digestive tube chyme viscosity to increase, and reduces wheat bran digestion and metabolism rate and livestock and poultry food ingestion.Reduce the anti-nutritional method of araboxylan and mainly contain chemical treatment, processing with enzyme preparation and microbiological deterioration conversion processing etc.But defectives such as the NSP enzyme source mikrobe ubiquity enzymic activity to the degraded of wheat bran araboxylan in the prior art is low, thermotolerance, resistance to acid difference; Enzyme production efficiency is low; And the research to beta-glucanase focuses mostly on genus bacillus; And the degraded araboxylan is mainly utilized fungal species such as black mold and brown root enzyme, less report is studied in the degraded utilization of edible fungus.In addition, utilize the submerged fermentation biodegradation technique of edible fungus, except the anti-nutrition of degraded wheat bran araboxylan, can improve hypha of edible fungus polysaccharide and biological activity nutritional product content thereof in the fermenting wheat bran bacterium powder simultaneously.
Summary of the invention
The method that the purpose of this invention is to provide a kind of biodegradation of wheat bran araboxylan; This method is utilized grass living edible mushrooms Agaricus blazei Murrill (Agaricus blazei Murill) Degradation and Transformation wheat bran araboxylan complex construction, makes full use of the outer conjugated fibre degrading enzyme system of Agaricus blazei Murrill born of the same parents to degraded of wheat bran araboxylan and trans-utilization efficient, and changing effect is good; The Agaricus blazei Murrill liquid strain biological activity of degraded usefulness is strong; Transformation efficiency is high, and the small molecules oligose metabolite content after degradable fermented is high, and mesostates such as its granulose possess the biological action of medicine-food two-purpose; And method is simple; Workable, be fit to scale operation, have remarkable economic efficiency.
1) method of biodegradation of wheat bran araboxylan of the present invention; It is characterized in that: said method is with Agaricus blazei; Adopt liquid culture technology and the special culture medium that screens, induce the outer conjugated fibre degrading enzyme system of born of the same parents of this biological degradation bacterium that the wheat bran araboxylan is degraded; The concrete steps of said method are: the preparation of Agaricus blazei Murrill test tube strains; Adopt conventional PDA substratum, conventional test tube strains technology of preparing and cultivation, preservation condition, carry out the Agaricus blazei Murrill test tube strains and cultivate preservation;
2) the dull and stereotyped activation culture of Agaricus blazei: under gnotobasis, test tube strains is inoculated in flat board; Carry out activation culture; Said culture condition is 25 ± 0.5 ℃, leaves standstill activation culture 10d, adopts the PDA enriched medium that the Agaricus blazei Murrill test tube strains is carried out dull and stereotyped activation culture;
3) the Agaricus blazei Murrill liquid strain is cultivated: under gnotobasis, the Agaricus blazei after the dull and stereotyped activation culture inoculated and carry out liquid strain in the culture vessel that liquid nutrient medium is housed and cultivate; Said training method is cultivated for the vibration deep layer: from the dull and stereotyped activated spawn of growth animated period; Use the 5-10 ferfas piece of punch tool cut-off footpath along flat-plate bacterial colony periphery vegetative point as 0.5-0.8cm; Insert in the culture vessel, culture vessel is the 250mL triangular flask; Said vibration deep layer culture condition is: 25 ± 0.5 ℃, and 170 ± 2r.min
-1, constant-temperature shaking culture 5-7d; Liquid nutrient medium is the 1/3-2/5 of culture vessel capacity in the said culture vessel;
4) Agaricus blazei Murrill liquid strain fermentative degradation wheat bran: adopt the Agaricus blazei Murrill liquid strain that wheat bran is carried out fermentative degradation: boil wheat bran 20-30min, matrix need not to filter, according to containing wheat bran 10g in every 100ml wheat bran fermented liquid; Semen Maydis powder 0-3g; Sucrose 2g, yeast powder 0.1g, sal epsom 0.1g; Potassium primary phosphate 0.1g, V
B10.01g mix, treat that all compositions fully dissolve, uniform mixing, the water constant volume is to volume required; The wheat bran fermented liquid is packed in the fermentative degradation container, and said Intake Quantity is the 2/5-1/2 of fermentative degradation container volume, and through 121 ℃, 0.1MPa autoclaving 20min; Treat that culture medium is cooled to room temperature, under gnotobasis, insert the Agaricus blazei Murrill liquid strain of step (3) preparation, inoculum size is the 10-20% of substratum capacity according to volume ratio, adopts the vibration deep layer to cultivate; Said vibration deep layer culture condition is 25 ± 0.5 ℃, 150-170r.min-1, constant-temperature shaking culture 7d.
Remarkable advantage of the present invention is:
(a) the present technique invention is eaten raw with fungi Agaricus blazei Murrill (Agaricus blazei Murill) as the biological degradation probiotic bacterium with the medicine-food two-purpose grass; Adopt liquid culture technology and the special culture medium that screens; The outer conjugated fibre degrading enzyme system of born of the same parents that induces this biological degradation bacterium is to degraded of wheat bran araboxylan and trans-utilization efficient, thus the nutrition conversion utilization ratio that raising wheat bran utilizes as feed resource.
(b) the present invention utilizes the degradation method of grass living edible mushrooms Agaricus blazei Murrill (Agaricus blazei Murill) Degradation and Transformation wheat bran araboxylan complex construction, and its advantage is: (1) utilizes the living edible mushrooms of grass to have multiple semicellulose, Mierocrystalline cellulose and lignin degradation prozyme system, wherein; Comprise β-1,4 endo-xylanase, β-1; 4 circumscribed zytases; Xylobiase, α-L-A Labaifunantangganmei, α-Pu Taotang aldehydic acid enzyme; Acetyl xylan esterase and FLA, PHCA esterase etc. are given birth to plant cell wall to the unifacial leaf grass and are had the degraded advantage; (2) being used for the living edible mushrooms of grass of the present invention is known food and medicament dual-purpose fungi; Through the fellowship of conjugated fibre degrading enzyme and xylan degrading enzyme system, in degraded wheat bran non-starch polysaccharide, utilize its degraded product; Transform metabolism, produce its biological activity Nutrition and Metabolism product.
(c) Agaricus blazei Murrill of the present invention shows the biological degradation effect of Testa Tritici fiber polysaccharide: as the substratum substrate, the content of the araboxylan in the Agaricus blazei Murrill degradation solution, total reducing sugar, protein and its meta-bolites all is higher than the filtration group of wheat bran substratum substrate with wheat bran; Compare with hydrolysis wheat bran blank control group, though araboxylan content does not have notable difference (P＞0.05) in the fermented liquid, the total reducing sugar and the protein contnt of Agaricus blazei Murrill degraded wheat bran group (AW) significantly improve P＜0.05.
(d) utilize the present invention's Testa Tritici of degrading, improved degradation effect, promote the nutrition digestion of wheat bran to absorb the water-soluble araboxylan of wheat bran macromole.Adopt the feed production performance of mouse of the wheat bran of Agaricus blazei Murrill degraded all significantly to be superior to undressed wheat bran group feeding effect with the material anharmonic ratio; P＜0.05, and obviously promote the villus length of mouse intestinal to increase, P＜0.05; The crypts degree of depth significantly reduces; P＜0.05, and improve the serum D-wood sugar absorption strength of mouse, P＜0.05.The meta-bolites of Agaricus blazei Murrill degraded Wheat bran can improve small intestine's structure, strengthens small intestine and digests and assimilates ability.
(e) adopt the wheat bran zymocyte liquid of the inventive method preparation to prepare fermenting wheat bran bacterium powder as nutritional additive: the Agaricus blazei Murrill liquid fermenting; The ANFs structure function mechanism of the water-soluble araboxylan of degraded wheat bran provides metabolic bioactive product mycelia polysaccharide of Agaricus blazei Murrill medicine edible mushrooms and thalline gp.Therefore, fermenting wheat bran bacterium powder is suitable to aid nutrition, improves the cub immunological competence; As digestive aid, improve carcass and digest and assimilate ability.
(f) adopt the wheat bran zymocyte liquid of the inventive method preparation to prepare fermenting wheat bran bacterium powder as the wheat bran feed resource: the insolubility non-starch polysaccharide in degraded wheat bran; Comprise the insolubility araboxylan, improved the digestion and metabolism of degraded wheat bran and utilized effect.Therefore, the alternative wheat bran raw material that is untreated of fermenting wheat bran is used for the wheat bran feed formulation, improves its efficiency of feed utilization.
Embodiment
The present invention adopts Agaricus blazei Murrill (Agaricus blazei Murill) bacterial classification.
1) Agaricus blazei Murrill test tube strains preparation: adopt conventional PDA culture medium culturing, the usage ratio and the culture condition of conventional test tube strains technology of preparing, inoculum size and substratum;
2) the dull and stereotyped activation culture of Agaricus blazei: under gnotobasis, test tube strains is inoculated in the plate culture medium; Carry out activation culture; Said culture condition is 25 ± 0.5 ℃, leaves standstill activation culture 10d, adopts the PDA enriched medium that the Agaricus blazei Murrill test tube strains is carried out dull and stereotyped activation culture;
3) the Agaricus blazei Murrill liquid strain is cultivated: under gnotobasis, the Agaricus blazei after the dull and stereotyped activation culture inoculated and carry out liquid strain in the culture vessel that liquid nutrient medium is housed and cultivate; Said training method is cultivated for the vibration deep layer: from the dull and stereotyped activated spawn of growth animated period; Use the 5-10 ferfas piece of punch tool cut-off footpath along flat-plate bacterial colony periphery vegetative point as 0.5-0.8cm; Insert in the culture vessel, culture vessel is the 250mL triangular flask; Said vibration deep layer culture condition is: 25 ± 0.5 ℃, and 170 ± 2r.min
-1, constant-temperature shaking culture 5-7d; Liquid nutrient medium is the 1/3-2/5 of culture vessel capacity in the said culture vessel;
4) Agaricus blazei Murrill liquid strain fermentative degradation wheat bran: adopt the Agaricus blazei Murrill liquid strain that wheat bran is carried out fermentative degradation: boil wheat bran 20-30min, matrix need not to filter, according to containing wheat bran 10g in every 100ml wheat bran fermented liquid; Semen Maydis powder 0-3g; Sucrose 2g, yeast powder 0.1g, sal epsom 0.1g; Potassium primary phosphate 0.1g, V
B10.01g mix, treat that all compositions fully dissolve, uniform mixing, the water constant volume is to volume required; The wheat bran fermented liquid is packed in the fermentative degradation container, and said Intake Quantity is the 2/5-1/2 of fermentative degradation container volume, and through 121 ℃, 0.1MPa autoclaving 20min; Treat that culture medium is cooled to room temperature, under aseptic condition, insert the Agaricus blazei Murrill liquid strain of step (3) preparation, inoculum size is the 10-20% of substratum capacity according to volume ratio, adopts the vibration deep layer to cultivate; Said vibration deep layer culture condition is 25 ± 0.5 ℃, 150-170r.min-1, constant-temperature shaking culture 7d.
The PDA enriched medium is in the step (2): in every 100ml constant volume substratum, raw materials used: yam 20g, glucose 2g, yeast powder 0.15g, potassium primary phosphate 0.3g, sal epsom 0.15g, V
B10.01g, agar 2g; Said PDA enriched medium is prepared as: take by weighing the peeling fresh potato in proportion, thinly slice and be put in the 1000mL beaker, put boil 30min on the electric furnace after, with four layers of filtered through gauze; To filtrate with culture medium prescription in other component proportional mixing, the heating, agar is melted fully; Be settled to volume requiredly, be sub-packed in while hot in the triangular flask, every bottled liquid measure is the 1/2-3/5 of container; After finishing, packing seals with tinfoil and kraft paper wrapping, in 121 ℃, and 0.1MPa autoclaving 20min.
The liquid nutrient medium warp is 121 ℃ in the step (3), 0.1MPa autoclaving 20min; Treat to insert bacterial classification after culturing bottle is cooled to room temperature; Said liquid nutrient medium is: in every 100ml constant volume substratum, raw materials used: Semen Maydis powder 3-5g, sucrose 2g, yeast powder 0.15-0.3g, potassium primary phosphate 0.1g, sal epsom 0.1g, ammonium sulfate 0.1g, V
B10.01g.
To pass through the wheat bran zymocyte liquid that Agaricus blazei Murrill liquid strain fermentative degradation handles and prepare fermenting wheat bran bacterium powder: adopt the rotation vacuum concentration, through-50 ℃, 8～15Pa, 48 hours vacuum freezedrying, pulverized 25 mesh sieves and make again.
Said fermenting wheat bran bacterium powder is used for nutritional additive or is used for the wheat bran feed resource.
Embodiment
1. application example
Instance 1: to be contrast without the water-soluble araboxylan content of the degradable fermented wheat bran of Agaricus blazei Murrill, utilize the Agaricus blazei Murrill liquid fermenting, the water-soluble araboxylan in the catabolism wheat bran substratum obviously reduces its content.
Table-1 water-soluble araboxylan of Agaricus blazei Murrill fermentative degradation wheat bran (AX)
| Contrast | Degraded wheat bran | |
| Degradation solution AX content (mg/mL) | 12.72±2.55 a | 0.37±0.02 b |
Instance 2: to be contrast without the total sugar content in the degradable fermented wheat bran liquid nutrient medium of Agaricus blazei Murrill, relatively through the Agaricus blazei Murrill liquid fermenting, the soluble non-starch polysaccharide in the degraded wheat bran, obviously the raising soluble polysaccharide that can supply utilize.
Table-2 Agaricus blazei Murrill fermentative degradation wheat bran insolubility non-starch polysaccharides
| Contrast | Degraded wheat bran | |
| Degradation solution total sugar content (mg/mL) | 19.40±0.94 b | 43.54±12.28 a |
Instance 3: being contrast without the total sugar content in the degradable fermented wheat bran liquid nutrient medium of Agaricus blazei Murrill; Nitrogenous metabolite content through in the Agaricus blazei Murrill fermented liquid shows; The water-soluble araboxylan and the soluble non-starch polysaccharide of degraded wheat bran substratum; Can obviously improve Agaricus blazei Murrill thalline metabolic activity, increase the nitrogenous biologically active metabolite product in the nutrient solution.
Protein contnt in table-3 Agaricus blazei Murrill degraded wheat bran fermented liquid
| Contrast | Degraded wheat bran | |
| Protein contnt (μ g/mL) | 381.99±12.66 b | 594.53±60.75 a |
Instance 4: the degraded wheat bran bacterium powder that utilizes the preparation of Agaricus blazei Murrill degraded wheat bran substitutes the wheat bran that is untreated as feed resource, prepares raw material as the mouse daily ration.The wheat bran bacterium powder of relatively degrading preparation mouse daily ration is the feeding effect of male mice to healthy Kunming: obviously improve the mouse daily ingestion amount, promote the mouse day weight gain, reduce mouse material anharmonic ratio; Show the degraded wheat bran bacterium powder of feeding through measuring serum D-wood pool concentration, can obviously improve mouse and digest and assimilate ability; In addition, can obviously improve the growth of mouse digestive tube weave construction.
The production performance comparison that table-4 degraded wheat bran bacterium powder are fed mouse
Claims (3)
1. the method for a biodegradation of wheat bran araboxylan; It is characterized in that: said method is with Agaricus blazei; Adopt liquid culture technology and the special culture medium that screens, induce the outer conjugated fibre degrading enzyme system of born of the same parents of this biological degradation bacterium that the wheat bran araboxylan is degraded; The concrete steps of said method are:
1) Agaricus blazei Murrill test tube strains preparation; Adopt conventional PDA substratum, conventional test tube strains technology of preparing and cultivation, preservation condition, carry out the Agaricus blazei Murrill test tube strains and cultivate preservation;
2) the dull and stereotyped activation culture of Agaricus blazei: under gnotobasis, test tube strains is inoculated in flat board; Carry out activation culture; Said culture condition is 25 ± 0.5 ℃, leaves standstill activation culture 10d, adopts the PDA enriched medium that the Agaricus blazei Murrill test tube strains is carried out dull and stereotyped activation culture;
3) the Agaricus blazei Murrill liquid strain is cultivated: under gnotobasis, the Agaricus blazei after the dull and stereotyped activation culture inoculated and carry out liquid strain in the culture vessel that liquid nutrient medium is housed and cultivate; Said training method is cultivated for the vibration deep layer: from the dull and stereotyped activated spawn of growth animated period; Use the 5-10 ferfas piece of punch tool cut-off footpath along flat-plate bacterial colony periphery vegetative point as 0.5-0.8cm; Insert in the culture vessel, culture vessel is the 250mL triangular flask; Said vibration deep layer culture condition is: 25 ± 0.5 ℃, and 170 ± 2rmin
-1, constant-temperature shaking culture 5-7d; Liquid nutrient medium is the 1/3-2/5 of culture vessel capacity in the said culture vessel;
4) Agaricus blazei Murrill liquid strain fermentative degradation wheat bran: adopt the Agaricus blazei Murrill liquid strain that wheat bran is carried out fermentative degradation: boil wheat bran 20-30min, matrix need not to filter, according to containing wheat bran 10g in every 100ml wheat bran fermented liquid; Semen Maydis powder 0-3g; Sucrose 2g, yeast powder 0.1g, sal epsom 0.1g; Potassium primary phosphate 0.1g, V
B10.01g, treat that all compositions fully dissolve, uniform mixing, the water constant volume is to volume required; The wheat bran fermented liquid is packed in the fermentative degradation container, and said Intake Quantity is the 2/5-1/2 of fermentative degradation container volume, and through 121 ℃, 0.1MPa autoclaving 20min; Treat that culture medium is cooled to room temperature, under gnotobasis, insert the Agaricus blazei Murrill liquid strain of step (3) preparation, inoculum size is the 10-20% of substratum capacity according to volume ratio, adopts the vibration deep layer to cultivate; Said vibration deep layer culture condition is 25 ± 0.5 ℃, 150-170rmin
-1, constant-temperature shaking culture 7d;
The PDA enriched medium is in the said step (2): in every 100ml constant volume substratum, raw materials used: yam 20g, glucose 2g, yeast powder 0.15g, potassium primary phosphate 0.3g, sal epsom 0.15g, VB10.01g, agar 2g; Said PDA enriched medium is prepared as: take by weighing the peeling fresh potato in proportion, thinly slice and be put in the 1000mL beaker, put boil 30min on the electric furnace after, with four layers of filtered through gauze; To filtrate with culture medium prescription in other component proportional mixing, the heating, agar is melted fully; Be settled to volume requiredly, be sub-packed in while hot in the triangular flask, every bottled liquid measure is the 1/2-3/5 of container; After finishing, packing seals with tinfoil and kraft paper wrapping, in 121 ℃, and 0.1MPa autoclaving 20min;
121 ℃ on the liquid nutrient medium warp of Agaricus blazei Murrill liquid spawn in the said step (3), 0.1MPa autoclaving 20min; Treat to insert bacterial classification after culturing bottle is cooled to room temperature; Said liquid nutrient medium is: in every 100ml constant volume substratum, raw materials used: Semen Maydis powder 3-5g, sucrose 2g, yeast powder 0.15-0.3g, potassium primary phosphate 0.1g, sal epsom 0.1g, ammonium sulfate 0.1g, VB10.01g.
2. the method for biodegradation of wheat bran araboxylan according to claim 1; It is characterized in that: the wheat bran zymocyte liquid that will pass through the processing of Agaricus blazei Murrill liquid strain fermentative degradation prepares fermenting wheat bran bacterium powder: adopt the rotation vacuum concentration; Through-50 ℃, 8～15Pa, 48 hours vacuum freezedrying, pulverized 25 mesh sieves and made again.
3. the purposes of the fermenting wheat bran bacterium powder of the method preparation with biodegradation of wheat bran araboxylan as claimed in claim 2; It is characterized in that: in the said fermenting wheat bran bacterium powder; Contain hypha of edible fungus polysaccharide and biological activity Nutrition and Metabolism product thereof simultaneously, be used for nutritional additive or be used for the wheat bran feed resource.
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| CN101543285B (en) * | 2009-04-30 | 2012-06-27 | 福建省农业科学院农业工程技术研究所 | Wheat-bran dietary fiber composite functional fungus powder product and preparation process thereof |
| CN107937318B (en) * | 2017-12-27 | 2020-10-30 | 齐鲁工业大学 | Bacillus subtilis MXT-1 for degrading wheat pentosan and application thereof |
| CN116287335B (en) * | 2023-02-21 | 2024-01-30 | 浙江大学 | Method for evaluating intestinal microecological regulation effect of arabinoxylans and application thereof |
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| CN1303923A (en) * | 2000-01-07 | 2001-07-18 | 中国科学院微生物研究所 | Medicinal mycelial fungus Agaricus blazei liquid culture |
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| CN1303923A (en) * | 2000-01-07 | 2001-07-18 | 中国科学院微生物研究所 | Medicinal mycelial fungus Agaricus blazei liquid culture |
| CN101194674A (en) * | 2007-12-29 | 2008-06-11 | 福建省农业科学院畜牧兽医研究所 | Feed additive for improving pigling intestinal canal function and immunity function, and use method thereof |
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