CN1724656A - A kind of preparation method who is used for the high vigor compound enzymic preparation of caragana microphylla degraded - Google Patents
A kind of preparation method who is used for the high vigor compound enzymic preparation of caragana microphylla degraded Download PDFInfo
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- CN1724656A CN1724656A CNA2004100215442A CN200410021544A CN1724656A CN 1724656 A CN1724656 A CN 1724656A CN A2004100215442 A CNA2004100215442 A CN A2004100215442A CN 200410021544 A CN200410021544 A CN 200410021544A CN 1724656 A CN1724656 A CN 1724656A
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Abstract
The present invention relates to Woody feed, particularly contain xylogen and high-content robust fibre and produce feed, specifically a kind of preparation method who is used for the high vigor compound enzymic preparation of Woody feed caragana microphylla degraded, with the bacterial suspension inoculation of whiterot fungi on solid medium, then adopt two warm culture methods, at ambient moisture 20～50%, under the natural ventilation oxygenating condition, can make compound enzymic preparation; Solid medium consists of, 8～15% corn steep liquors, 20～35% wheat bran, 5～15% (NH
4)
2SO
4, surplus is a water; It is two stages of branch that two temperature is cultivated, previous stage: 50～80 hours, and 32～38 ℃; The latter half: 350～420 hours, between 28～32 ℃; Cultivated altogether 400～500 hours.Advantage of the present invention is: it is higher to produce enzyme activity, and enzyme system is balanced, and to the good degrading effect of caragana microphylla, cost is low, and it is all good that the growth of livestock is had a security feeding effect.
Description
Technical field
The present invention relates to Woody feed, particularly contain xylogen and high-content robust fibre and produce feed, specifically a kind of preparation method who is used for the high vigor compound enzymic preparation of Woody feed caragana microphylla degraded.
Background technology
One of key constraints of China's animal husbandry development is the shortage of feed, and particularly the western arid of China, semiarid pastoral area are even more serious.Woody feed is a kind of feed resource, occupies an important position in livestock feed.Compare with the draft feed, Woody feed contains higher nutritive ingredient, particularly high-protein nutrient and calcium contents, therefore mixes use with the draft feed, can remedy proteic deficiency.Woody feed is more obvious as the effect that livestock feed is used for lacking in winter and early spring grass its performance in season in addition, can alleviate the contradiction that lacks feed during this well.
Caragana microphylla is Inner Mongol Erdos area a kind of Woody feed distributed more widely, is the animal feed resource of potentialization.But, influenced nutritive value and palatability and failed to obtain a large amount of developments and utilizations because of its xylogen and crude fibre too high levels.
Xylogen is a class organic polymer, extensively in distribution and the higher plant, be only second to content of cellulose, the lignin structure complexity, Mierocrystalline cellulose and hemicellulose " deadlocked " are in the same place, its bag is applied serious obstruction cellulose degradation, so lignin degradation is a key of effectively utilizing lignocellulose raw material.Cellulosic structure is also complicated, and water insoluble, Mierocrystalline cellulose must directly contacts with enzyme during enzymolysis, can only rely on enzyme in the contact process to be diffused in the cellulose net structure of complexity and could realize hydrolysis.In plant feed, pectin is the cell walls of plant, and it forms gelatinous mass in animal digestive tract, influences the absorption of livestock to nutritive substance, also is one of problem of being utilized of restriction nutrition.
The enzyme class that participates in lignin degradation is a lot, and wherein laccase is one of main enzyme.Cellulase can be converted into carbohydrate with the intravital Mierocrystalline cellulose of plant, hemicellulose, simultaneously by decomposing vegetable cell, contents such as soluble protein in the cell, non-protein nitrogen(NPN) thing, mineral substance is released.Polygalacturonase can decompose pectin substance, thereby eliminates the inhibition that livestock is digested and assimilated nutritive substance.
Caragana microphylla to be developed to the main feed resource of Erdos, at first will produce a kind of compound enzymic preparation of high vigor.That is that all right is ripe for the suitability for industrialized production of China's compound enzymic preparation, and the zymin industry remains to be further developed, and still there are situations such as enzymic activity is low, cost is high, utilization ratio is lower in enzyme preparation product, and especially strength is comparatively weak aspect exploitation.
Summary of the invention
For the caragana microphylla feed is extensively utilized, solve the existing problem of above-mentioned zymin, the object of the present invention is to provide the preparation method of the low compound enzymic preparation that is used for the caragana microphylla degraded of a kind of enzymic activity height, cost.
For achieving the above object, the technical solution used in the present invention is: the microorganism of employing is a whiterot fungi, carry out solid by two temperature culture methods and produce enzymic fermentation, via slant culture, shake bottle enlarged culturing and novel solid substratum and produce the enzyme solid ferment stage, make the compound enzymic preparation of high vigor degraded caragana microphylla, the concrete operations step is as follows:
(1) slant culture:
In sucrose potato sloped tube, insert Corilus versicolor Quel., put in 28～35 ℃ of incubators, cultivated 8～15 days, get slant pore suspension;
(2) shake a bottle enlarged culturing:
The 100 milliliters of sucrose potato nutrient solutions of packing in 500 milliliters of triangular flasks insert spore suspension, and inoculum size is 2～5% (V/V) of nutrient solution volume, carry out enlarged culturing, culture condition is at 25～35 ℃, and shaking speed is 120～150r/min, cultivated 5～10 days, and must shake phialosporae suspension;
(3) solid fermentation:
With solid medium, behind autoclaving, by with 5～10% amount (V/W that shake phialosporae suspension, be suspension vol/solid medium weight) be inoculated in solid medium, then adopt two warm culture methods, ambient moisture is controlled between 20～50%, and the natural ventilation oxygenating can make compound enzymic preparation;
Wherein: described microorganism whiterot fungi is Corilus versicolor Quel. (Coriolus versicolor); The Corilus versicolor Quel. spore is to wash and make with sterilized water on the slant medium; The prescription of described solid medium by weight percentage, by 8～15% corn steep liquors, 20～35% wheat bran, 5～15% (NH
4)
2SO
4And the moisture of surplus, make through uniform mixing; Described two temperature culture method is meant that culture temperature divides two stages, previous stage: temperature is controlled at 32～38 ℃, the latter half in during 50～80 hours: temperature is controlled between 28～32 ℃ in during 350～420 hours, cultivates altogether 400～500 hours; Described two temperature culture method is meant that culture temperature divides two stages, previous stage: temperature is controlled at 35～37 ℃ for good in during 60～70 hours, the latter half: temperature is controlled between 28～30 ℃ in during 380～400 hours, cultivates 440～470 hours altogether for good.
The present invention has following advantage:
1. it is higher to produce enzyme activity, and enzyme system is balanced, to the good degrading effect of caragana microphylla.The prozyme that carries out solid fermentation production by prescription of the present invention has very high laccase, cellulase and pectinase activity, and (its laccase work can reach 9.58U/g through actual measurement, filter paper enzyme activity actual measurement can reach 35.9mg glucose/(gDM.h), pectinase activity actual measurement can reach 29.55mg glucose/(gDM.h), this prozyme agent have stronger degradation capability to the xylophyta fiber, is fit to the production of feeds such as higher plant of caragana microphylla class xylogen and crop straw, timber tankage fully.In addition, adopt the two temperature culture method, the high-temperature cultivation in early stage helps mycelial growth, and the suitable cooling in later stage is cultivated, and has controlled the mycelial speed of growth, and the enzymatic production process is prolonged, and is beneficial to the generation that promotes enzyme.
2. cost is low.The present invention is an animal feed with the caragana microphylla, develops and utilizes to be target, pays attention to its price in the selection of mixed enzyme fermentation raw material, selecting cheap wheat bran is compound N, C source, is inorganic N source with ammonium sulfate, and corn steep liquor is inductor C source, have wide material sources, cheap advantage.The inventive method has realized optimization of fermentation condition, and fermentation production process is simple, is easy to advantages such as grasp, thereby has reduced the cost that produces enzyme, makes the operation of caragana microphylla feed have the stronger market competitiveness.
3. the growth to livestock has security, and the employed solid fermentation thing of compound enzymic preparation that the present invention produces is nontoxic to livestock, when having security, nutritive substance is had supplementary function.Environment is not directly or indirectly polluted.The present invention is used for the caragana microphylla feed fermentation and produces (external enzymolysis) or directly mix and stir feed (enzymolysis in the body) that grind fodder at caragana microphylla, and effect is all good.
Embodiment
Below by embodiment the present invention is described in further detail.
Embodiment 1
(1) slant culture:
In the test tube of sucrose potato solid medium (agar 2% remaining is water for potato 20%, sucrose 2%), insert Corilus versicolor Quel., put in 28 ℃ of incubators, cultivated 10 days;
(2) spore suspension preparation
Corilus versicolor Quel. spore on the slant medium is washed with sterilized water, make spore suspension;
(3) shake a bottle enlarged culturing
100 milliliters of sucrose potato nutrient solutions (potato 20%, sucrose 2%, yeast extract pastes 1% are being housed, remainder is a water) insert the spore suspension of a slant tube in 500 milliliters of triangular flasks, carry out enlarged culturing, put 28 ℃, rotating speed is the 130r/min shaking table, cultivates 8 days, must shake phialosporae suspension;
(4) produce enzyme solid ferment
Get above-mentioned solid medium 100g, behind autoclaving, inoculation 4ml shakes phialosporae suspension, then adopt two warm culture methods, fermented 440 hours, constant temperature was at 35 ℃ in preceding 60 hours, constant temperature was 30 ℃ in back 380 hours, ambient moisture is controlled at 30%, and the natural ventilation oxygenating can make the compound enzymic preparation of high vigor.Wherein: the prescription of solid medium is: corn steep liquor 10.8%: wheat bran: 27.7%, and (NH
4)
2SO
4: 9.0%, moisture: 52.5%, pH value: nature, about 6.4.
Through the composite solid zymin that above-mentioned steps makes, its laccase work can reach 8.72U/g through actual measurement, and filter paper enzyme activity actual measurement can reach 34.9mg glucose/(gDM.h), and the pectinase activity actual measurement can reach 25.3mg glucose/(gDM.h).
Embodiment 2
Difference from Example 1 is:
In the test tube of sucrose potato solid medium, insert Corilus versicolor Quel., put in 30 ℃ of incubators, cultivated 12 days; 100 milliliters of sucrose potato nutrient solutions access spore suspensions are being housed, and enlarged culturing is put 32 ℃, and rotating speed is the 110r/min shaking table, cultivates 10 days, must shake phialosporae suspension; When producing enzyme solid ferment, get solid medium 50g, inoculation 2ml shakes phialosporae suspension, then adopt two warm culture methods, fermented 470 hours, constant temperature was at 37 ℃ in preceding 70 hours, constant temperature was 28 ℃ in back 400 hours, and ambient moisture is controlled at 50%, can make the compound enzymic preparation of high vigor.Wherein: the prescription of solid medium is: corn steep liquor 15.0%: wheat bran: 23.5%, and (NH
4)
2SO
4: 12.0%, moisture: 49.5%, pH value: nature, about 6.4.
The composite solid zymin that makes after the fermentation, its laccase work can reach 1.73U/g through actual measurement, and filter paper enzyme activity actual measurement can reach 36.9mg glucose/(gDM.h), and the pectinase activity actual measurement can reach 33.8mg glucose/(gDM.h).
Claims (4)
1. preparation method who is used for the high vigor compound enzymic preparation of caragana microphylla degraded is characterized in that:
The bacterial suspension inoculation of whiterot fungi on solid medium, is then adopted two warm culture methods, ambient moisture is controlled between 20～50%, the natural ventilation oxygenating can make compound enzymic preparation;
Consisting of of solid medium by weight percentage, 8～15% corn steep liquors, 20～35% wheat bran, 5～15% (NH
4)
2SO
4, surplus is a water;
The two temperature culture method is meant that culture temperature divides two stages, previous stage: temperature is controlled at 32～38 ℃, the latter half in during 53～80 hours: temperature is controlled between 28～32 ℃ in during 350～420 hours, cultivates altogether 400～500 hours.
2. according to the described preparation method of claim 1, it is characterized in that: described two temperature culture method is meant that culture temperature divides two stages, previous stage: temperature is controlled at 35～37 ℃ for good in during 60～70 hours, the latter half: temperature is controlled between 28～30 ℃ in during 380～400 hours, cultivates altogether 440～470 hours.
3. according to the described preparation method of claim 1, it is characterized in that: the bacteria suspension of described whiterot fungi is via slant culture, shakes a bottle enlarged culturing acquisition, and detailed process is as follows,
(1) slant culture:
In sucrose potato solid medium test tube, insert the whiterot fungi spore, put in 28～35 ℃ of incubators, cultivated 8～15 days, the whiterot fungi spore on the slant medium is washed with sterilized water, make spore suspension;
(2) shake a bottle enlarged culturing:
The sucrose potato nutrient solution of packing in triangular flask inserts spore suspension, and inoculum size is 2～5%, carry out enlarged culturing, culture condition is at 25～35 ℃, and shaking speed is 120～150r/min, cultivated 5～10 days, and must shake phialosporae suspension, i.e. the bacteria suspension of whiterot fungi.
4. according to claim 1,2 or 3 described preparation methods, it is characterized in that: described whiterot fungi is a Corilus versicolor Quel..
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|---|---|---|---|
| CNA2004100215442A CN1724656A (en) | 2004-07-23 | 2004-07-23 | A kind of preparation method who is used for the high vigor compound enzymic preparation of caragana microphylla degraded |
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| CNA2004100215442A CN1724656A (en) | 2004-07-23 | 2004-07-23 | A kind of preparation method who is used for the high vigor compound enzymic preparation of caragana microphylla degraded |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101811781A (en) * | 2010-03-04 | 2010-08-25 | 南京信息工程大学 | White rot fungus rot agent and method for producing organic pollutant rot agent by using white rot fungi |
| CN105862498A (en) * | 2016-03-29 | 2016-08-17 | 山东贵和显星纸业有限公司 | Preparation method and application for recycled fiber improving enzyme used for papermaking |
| CN106638081A (en) * | 2016-10-28 | 2017-05-10 | 河南华禹环保科技有限公司 | Method for preparing filter paper fiber carrier for white-rot fungus culture |
-
2004
- 2004-07-23 CN CNA2004100215442A patent/CN1724656A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101811781A (en) * | 2010-03-04 | 2010-08-25 | 南京信息工程大学 | White rot fungus rot agent and method for producing organic pollutant rot agent by using white rot fungi |
| CN105862498A (en) * | 2016-03-29 | 2016-08-17 | 山东贵和显星纸业有限公司 | Preparation method and application for recycled fiber improving enzyme used for papermaking |
| CN106638081A (en) * | 2016-10-28 | 2017-05-10 | 河南华禹环保科技有限公司 | Method for preparing filter paper fiber carrier for white-rot fungus culture |
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