CN102077903B - Method for jointly treating stalks by steam explosion and microorganism fermentation - Google Patents
Method for jointly treating stalks by steam explosion and microorganism fermentation Download PDFInfo
- Publication number
- CN102077903B CN102077903B CN2010105574199A CN201010557419A CN102077903B CN 102077903 B CN102077903 B CN 102077903B CN 2010105574199 A CN2010105574199 A CN 2010105574199A CN 201010557419 A CN201010557419 A CN 201010557419A CN 102077903 B CN102077903 B CN 102077903B
- Authority
- CN
- China
- Prior art keywords
- stalk
- fermentation
- spore
- explosion
- stalks
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- 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
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a method for jointly treating stalks by steam explosion and microorganism fermentation, comprising the following steps of: firstly, carrying out steam explosion pretreatment on the stalks to obtain exploded stalks; then preparing spore seed liquid: inoculating aspergillus oryzae or trichoderma Koningi to a PDA (Potato Dextrose Agar) culture medium, standing and culturing for 3-5 days at the temperature of 28-32 DEG C, and preparing the spore seed liquid with the concentration of 106-108 per mL; and finally carrying out microorganism fermentation: uniformly mixing 18g of exploded stalks, 2g of bran and 30mL of mineral element nutrient solution, adjusting pH to be 7.0 with Ca(OH)2, sterilizing for 15min at the temperature of 121 DEG C and the pressure of 0.15MPa to obtain a solid fermentation medium a, inoculating the spore seed liquid according to 2-4% of inoculum size, and culturing the spore seed liquid for 5-7 days at the temperature of 28-32 DEG C. The fermentation stalks obtained by utilizing the method have low content of lignose, cellulose and hemicellulose and high activity of filter paper carbohydrase, CMC (carboxymethyl cellulose) enzyme, amylase and protease.
Description
Technical field
The present invention relates to a kind of processing method of stalk, be specifically related to the method for a kind of steam blasting and microbial fermentation Combined Treatment stalk.
Background technology
A large amount of studies confirm that both at home and abroad utilizes biotechnology to excavate fully and utilize the potential value of crop stalk, is one of effective means of alleviating global grain resource shortage.According to according to a preliminary estimate, if substituting cereals grain, the biofermentation straw with 10～20% is used for pig industry production, annual nearly 3000～5,000 ten thousand tons of the grain of just can saving in the whole nation, can solve the gap problem of the annual 3000 ten thousand tons of cereals feed grains of China fully, have particular importance society and economic implications.
The main component of stalk is crude fibre; comprise cellulose, hemicellulose and lignin; and the height polymerization of cellulose macromolecule and crystallinity and hemicellulose, lignin and its mutual protective effect of twining; so that utilize the methods such as traditional hydrolysis and biofermentation to come degrade coarse fibers, efficient is lower.At present, the preprocess method of stalk has the processing method that physics, chemistry, biology and multiple processing combine.Biologic pretreatment method is because of the utilization ratio that can improve stalk and can environment more and more be subject to people's attention, but there is also shortcoming: namely biotransformation efficiency is lower.Traditional physical treatment is adopted minimizing lignocellulosic size and crystal structure to increase specific surface area and is fallen the low-fiber degree of polymerization, generally needs more energy consumption.At present aspect physical treatment, the steam blasting preliminary treatment is a kind of the most promising preprocess method of innovation, it utilizes saturated vapor that stalk is heated to certain pressure, the high steam infiltrated fiber is inner, mode with air-flow discharges from the blind bore crack, make fiber that certain mechanical breaking occur, then sudden pressure reduction to atmospheric pressure carries out pretreated a kind of physical means to stalk.As: can in blasting process, with 160 ℃～260 ℃ saturated vapors stalk be heated to 0.69～4.83MPa; action time be several seconds to a few minutes; then sudden pressure reduction is to atmospheric pressure; cellulose is separated from the structure of complexity; shortened the length of fiber; be conducive to follow-up cellulase to cellulosic attack, make the easier effect that is subject to enzyme and microorganism of fiber, the production of scale can be saved energy consumption and cost greatly.But the research that present domestic and international blasting technique with stalk is applied to animal feed production aspect is less.The alkali preliminary treatment can be carried out at normal temperatures, and effectively reduces the content of lignin in the stalk, but can cause the loss of part fermentable sugars in the process of processing.The acid treatment hemicellulose in the lignocellulose raw material of can degrading makes that cellulose is easier to be utilized by microorganism, but the soda acid of high concentration has increased the cost that reclaims in actual production, be not suitable for industrialized large-scale production.
This comprehensive study is considered biological and physics processing method; stalk is carried out first explosion treatment carry out again microbial fermentation; filter out a kind of economical and effective, the straw pretreatment method of safe and feasible is developed the application that reaches in animal husbandry is produced for the scale of stalk resource and is laid the foundation.
Summary of the invention
The object of the invention is to provide the method for a kind of steam blasting and microbial fermentation Combined Treatment stalk, utilizes in the fermented stalk that the method obtains lignin, cellulose and hemicellulose level low, and filter paper carbohydrase, CMC enzyme, amylase and prolease activity are high.
For achieving the above object, the present invention adopts following technical scheme:
The method of a kind of steam blasting and microbial fermentation Combined Treatment stalk, it comprises the steps:
1. stalk steam blasting preliminary treatment: the stalk that will pulverize 10～20 mesh sieves, moisture content≤15wt% carries out steam blasting to be processed, and then it is dried to get naturally the explosion stalk;
2. prepare aspergillus oryzae/koning trichoderma seed liquor: aspergillus oryzae or koning trichoderma are inoculated in the culture dish of being made by the PDA culture medium, 28～32 ℃ of static cultivation 3～5d, wash culture dish with sterile saline after the spore maturation, spore in the culture dish is transferred in the sterilization triangular flask compound concentration 1 * 10
6～10
8The spore suspension of individual/mL is the spore seed liquor; Described PDA culture medium prior to sterilization treatment 15min under 121 ℃, 0.15MPa pressure, is then made the culture dish that contains 15～20 mL PDA culture mediums in advance under aseptic condition;
3. microbial fermentation: get 18g explosion stalk and 2g wheat bran, 30mL mineral element nutrient solution mixes, with Ca (OH)
2Transferring to pH is 7.0, and then sterilization 15min gets the solid fermentation culture medium under 121 ℃, 0.15MPa pressure; By 2～4%(volume/mass percentage) inoculum concentration the spore seed liquor is inoculated in the solid fermentation culture medium, cultivate 5～7d for 28～32 ℃.
Test used aspergillus oryzae and koning trichoderma available from Institute of Microorganism, Academia Sinica, and effluent south agriculture university's Animal nutrition and the preservation of Biotechnology Experiment chamber.
Described stalk can be wheat straw or maize straw etc.
The step 2. component of middle PDA culture medium is: soluble starch 6 g, peptone 5 g, yeast 2 g, glucose 20 g, KH
2PO
42 g, MgSO
47H
2O 0.3 g, agar powder 20 g, distilled water 1000 mL.
The step 3. component of Mineral Elements nutrient solution is: (NH
4)
2SO
41.4 g, KH
2PO
42.0 g, MgSO
40.3 g, CaCl
20.3 g, NaCl 0.5 g, FeSO
45.0 mg, MnSO
41.6 mg, ZnCl
21.7 mg, CoCl
22.0 mg, distilled water 1000 mL.
Be compared with existing technology beneficial effect of the present invention:
The present invention at first utilizes the method for steam blasting, and (the steam blasting device explosion pulse width of employing only is 0.00875 S, the abrupt release high density energy is finished explosion to destroy the structure of stalk), make lignin in the straw and cellulose obtain loose and Partial digestion, make that microorganism is easier to play a role; Then utilize the microorganism that can secrete high activity cellulase to unite cultivation, lignin and cellulose in the straw are degraded simultaneously, thereby reach the purpose of thorough degrading straw.The straw that the processing of employing this method obtains also contains a large amount of digestive enzymes (such as cellulase, protease and amylase etc.) except containing a large amount of indigestible carbohydrates.When stalk behind explosion and aspergillus oryzae fermentation process 5～6d, the content of cellulose and hemicellulose has reduced by 27.89% and 64.80% than original stalk respectively, filter paper carbohydrase, CMC enzyme, amylase and prolease activity can reach respectively 335.10,1138.92,32.57 and 201.99 U/g, therefore, stalk after the processing is not only the good feed of herbivore, also can be used as the feed resource of pig fowl, this is for the meaning that solves food shortage that people and animals strive the grain problem and alleviate China and even the world and have particular importance.In addition, the method economical and effective, safe and feasible is for scale exploitation and the application in animal husbandry is produced of stalk resource are laid a good foundation.
Figure of description
Fig. 1 is the glucose calibration curve;
Enzyme is lived over time in Fig. 2 explosion stalk+aspergillus oryzae group sweat;
Soluble sugar content over time in Fig. 3 explosion stalk+aspergillus oryzae group sweat.
The specific embodiment
The present invention is described further by the following examples, but protection scope of the present invention is not limited to this.
Test material: maize straw comes from energy research key lab of the Ministry of Agriculture of Agricultural University Of He'nan, and after pulverizer was pulverized, it was for subsequent use to cross 10～20 mesh sieves.
Testing equipment: stalk steam blasting machine QB-200: produced by Henan Province Hebi City right way heavy-duty machine factory, vapor pressure can reach 6 MPa, and heating power is 8 kW, and effectively the blast chamber volume is 0.405 L.Quick-fried of this vapour adopts gas bullet technology, can finish earth pressure release in 0.00875 S, is different from the in the past quick-fried mode of so-called vapour (expanded and hot spray), has realized explosion truly.
Embodiment 1(is elaborated to the inventive method take maize straw as example)
The method of a kind of steam blasting and microbial fermentation Combined Treatment stalk, it comprises the steps:
1. stalk steam blasting preliminary treatment: will pulverize the maize straw of 20 mesh sieves, moisture content 10wt% under steam pressure 2.5 Mpa conditions, release pressure behind pressurize 200 S, carry out steam blasting and process, the stalk natural drying after the processing gets the explosion stalk to moisture content≤10wt%, and is for subsequent use;
2. prepare the aspergillus oryzae seed liquor: aspergillus oryzae is inoculated in the culture dish of being made by the PDA culture medium, 30 ℃ of static cultivation 3d wash culture dish with sterile saline after the spore maturation, the spore in the culture dish are transferred in the sterilization triangular flask compound concentration 1 * 10
6The spore suspension of individual/mL is the spore seed liquor; Described PDA culture medium prior to sterilization treatment 15min under 121 ℃, 0.15MPa pressure, is then made the culture dish that contains 20mL PDA culture medium in advance under aseptic condition; Described PDA nutrient media components is: soluble starch 6 g, peptone 5 g, yeast 2 g, glucose 20 g, KH
2PO
42 g, MgSO
47H
2O 0.3 g, agar powder 20 g, distilled water 1000 mL;
3. aspergillus oryzae fermentation: (component is: (NH to add 18g explosion stalk, 2 g wheat brans and 30 mL mineral element nutrient solutions in the triangular flask of 500 mL
4)
2SO
41.4 g, KH
2PO
42.0 g, MgSO
40.3 g, CaCl
20.3 g, NaCl 0.5 g, FeSO
45.0 mg, MnSO
41.6 mg, ZnCl
21.7 mg, CoCl
22.0 mg, distilled water 1000 mL), after mixing, with Ca (OH)
2Transferring to pH is 7.0, and then sterilization 15min gets the solid fermentation culture medium under 121 ℃, 0.15MPa pressure; By 4%(volume/mass percentage) inoculum concentration the spore seed liquor is inoculated in the solid fermentation culture medium, in 30 ℃ constant incubator, cultivate.Interval 24h for the first time, every interval 48h maize straw sample of getting fermentation from incubator carries out the mensuration of index of correlation afterwards.By the index determining result as can be known, generally cultivate 5～7d and just can obtain preferably treatment effect.
With reference to embodiment 1, difference is carried out fermentation test for substituting aspergillus oryzae with koning trichoderma.
The method of a kind of steam blasting and microbial fermentation Combined Treatment stalk, it comprises the steps:
1. stalk steam blasting preliminary treatment: will pulverize the stalk of 10 mesh sieves, moisture content 15wt% under the condition of steam pressure 1.8MPa, release pressure behind the pressurize 240s, carrying out steam blasting and process, is that 15wt% gets the explosion stalk with its natural drying to moisture content then, for subsequent use;
2. prepare the koning trichoderma seed liquor: koning trichoderma is inoculated in the culture dish of being made by the PDA culture medium, 28 ℃ of static cultivation 5d, wash culture dish with sterile saline after the spore maturation, the spore in the culture dish is transferred in the sterilization triangular flask compound concentration 1 * 10
8The spore suspension of individual/mL is the spore seed liquor; Described PDA culture medium prior to sterilization treatment 15min under 121 ℃, 0.15MPa pressure, is then made the culture dish that contains 15 mL PDA culture mediums in advance under aseptic condition; Described PDA nutrient media components is: soluble starch 6 g, peptone 5 g, yeast 2 g, glucose 20 g, KH
2PO
42 g, MgSO
47H
2O 0.3 g, agar powder 20 g, distilled water 1000 mL;
3. koning trichoderma fermentation: get 18g explosion stalk and 2g wheat bran, 30mL mineral element nutrient solution mixes, usefulness Ca (OH)
2Transferring to pH is 7.0, and then sterilization 15min gets the solid fermentation culture medium under 121 ℃, 0.15MPa pressure; By 2%(volume/mass percentage) inoculum concentration the spore seed liquor is inoculated in the solid fermentation culture medium, cultivate 5～7d for 28 ℃ and get final product.
Use the inventive method to process the performance measurement test of gained fermented maize stalk.
Experimental design and grouping
It is 6 groups that test is divided into, i.e. common stalk group, explosion stalk group, common stalk+aspergillus oryzae group, common stalk+koning trichoderma group, explosion stalk+aspergillus oryzae group and explosion stalk+koning trichoderma group.Every group of 6 repetitions are cultivated in 30 ℃ constant incubator after each winding kind, interval 24h for the first time, and every interval 48h gets the mensuration that part fermented stalk sample carries out index of correlation from incubator afterwards.
The physical and chemical index of measuring in the process of the test
2.1 the mensuration of cellulose and content of lignin
Employing Vansoest method (Feng Jihua, etc. use the comparison [J] that Van Soest method and conventional method are measured cellulose and lignin.
Southwest Nationalities College's journal.1994,20 (1): 55-56.) measure respectively each processed group in the variation of different fermentations time inner cellulose and content of lignin.
2.2 the mensuration of enzyme activity and soluble sugar
Get the fermented maize straw sample of 4.0 g processing gained, add 46 mL physiological saline, soak 2 h after, centrifugal 10 min of 3000 r/min, get supernatant as crude enzyme liquid, measure wherein filter paper carbohydrase, carboxymethylcelluloenzyme enzyme, protease and amylase activity and soluble sugar content.
2.2.1 the mensuration of filter paper carbohydrase (FPU) vigor (Jiang Jiala, etc. the bacterial strain screening of cellulose degradation and utilization thereof [J].
The modern biomedical progress.2009,18 (9): 3451-3454.)
Get crude enzyme liquid and boil each 0.5 mL of crude enzyme liquid of 10min deactivation, the citrate buffer solution and the 50 mg filter paper bars that add respectively 1.5 mL concentration, 0.05 mol/L, pH4.5, in 50 ℃ of water bath heat preservation l h, then add respectively 1.5 mL 3,5-dinitrosalicylic acid (DNS), boil 5 min, take the crude enzyme liquid group of deactivation as contrast, measure absorbance at wavelength 550 nm places.Enzyme activity unit definition: take hydrolysis per hour the catalytic substrate hydrolysis form the enzyme amount of 1 μ mol glucose as a unit.
2.2.2 the mensuration of carboxymethylcelluloenzyme enzyme (CMCase) vigor (Jiang Jiala, etc. the bacterial strain screening of cellulose degradation and utilization thereof [J].
The modern biomedical progress.2009,18 (9): 3451-3454.)
Get crude enzyme liquid and boil each 0.5 mL of crude enzyme liquid of 10min deactivation, add respectively 1.5 mL, 0.51%CMC citrate buffer solution, in 50 ℃ of thermostat water baths behind enzymolysis 30 min, from water-bath, take out, then add respectively 1.5 mL DNS nitrite ion cessation reactions, behind the abundant mixing, boiling water bath 15 min, take the crude enzyme liquid group of deactivation as contrast, measure absorbance at wavelength 550nm place.Enzyme activity unit definition: take hydrolysis per hour the catalytic substrate hydrolysis form the enzyme amount of 1 μ mol glucose as a unit.
2.2.3 the mensuration of amylase activity (Yasser B, et al. Isolation and identification of a new fungal strain for amylase biosynthesis[J].
Polish Journal of Microbiology. 2009,58 (3): 269-273.)
Get the starch of 1.25 mL 1%, 0.25 mL acetate buffer solution (0.1mol/L, pH5.0), 0.25 mL deionized water, 0.25 the crude enzyme liquid of mL adds 1.5 mL DNS, boiling water bath 15 min behind 50 ℃ of insulation 10 min, simultaneously take the crude enzyme liquid group of deactivation as contrast, in wavelength 550 nm places mensuration absorbance.Enzyme activity unit definition: form the enzyme amount of 1 μ mol glucose as a unit take the hydrolysis of hydrolysis per minute catalytic substrate.
2.2.4 the mensuration of prolease activity:
Employing Folin one phenol method mensuration albumen enzyme activity (Wang Pengpeng, etc. protease and amylase produce screening and zymologic property analysis and research [J].
China's herding magazine. 2009, (21): 48-51.).
2.2.5 the mensuration of soluble sugar:
Content of reducing sugar in employing 3, the 5-dinitrosalicylic Acid Colorimetry mensuration hydrolyzate (Wang Ping, etc. the research [J] that reaches the maize straw degradation effect is identified in the separation of aspergillus oryzae in the bovine rumen.
Agricultural University Of He'nan's journal, 2010, (44) 3:295-299.).
Accurately take by weighing DEXTROSE ANHYDROUS 1.000 g of drying to constant weight through 105 ℃, add 100 mL water, be made into the standard glucose liquid of 10 mg/mL concentration.Draw respectively the standard glucose liquid 1.0,2.0,3.0,4.0 of 10 mg/mL concentration, 5.0,6.0 mL are settled to 50 mL with distilled water in the volumetric flask of 50 mL, are made into 200,400,600,800, the standard liquid of 1000,1200 μ g/mL.Get variable concentrations standard liquid 1mL in test tube, add 3 mL DNS reagent and in boiling water, react 15 min, cooling, under 550 nm, with spectrophotometric instrumentation absorbance, take absorbance as ordinate, glucose amount is abscissa, the drawing standard curve.The blank standard glucose that replaces 1 mL with 1 mL distilled water.The mark curve is seen Fig. 1.
Data statistics and analysis
All (mean ± represent that SD) result carries out variance analysis with One-way ANOVA process in 16.0 editions statistical softwares of SPSS, and carries out the DuncanShi multiple ratio, P＜0.05 is significant difference to test data with " mean+SD ".
Results and analysis
4.1 embodiment 1 method is processed gained maize straw chemical composition with the variation of fermentation time
[0032]As shown in Table 2, adopt embodiment 1 method to process the maize straw of (being explosion stalk+aspergillus oryzae group) along with the prolongation of fermentation time, the content of the neutral detergent fiber in the fermented stalk, acid detergent fiber, cellulose and hemicellulose is downward trend.At the 1st d of aspergillus oryzae fermentation, the cellulose in the maize straw and hemicellulose level namely have obvious reduction (p＜0.05); At the 5th d of fermentation, the hemicellulose level in the sample significantly is lower than the 1st d and the 3rd d, reduces 39.11%(p＜0.05 during than 0 d); And content of cellulose and the 3rd d difference not significantly (p＞0.05), but reduce 17.35%(p＜0.05 during than 0 d).Consider that from stalk composition degraded situation the effect of 5 d that ferment is better than front 3 d.In addition, most of indexs of the 5th d that ferments and fermentation the 7th d are without significant difference, thereby from the economic angle consideration, selecting the suitable fermentation time of aspergillus oryzae is 5 d.
The aspergillus oryzae fermentation can not reduce the content of lignin in the explosion stalk during the fermentation, make on the contrary content of lignin rise to some extent (p＞0.05), mainly be because microbial fermentation increases the Soluble matter content in the fermented stalk, in measuring the content of lignin process, because the loss of solable matter, the gross weight of the final dry of fermented stalk is reduced, cause the relative rising (table 3 has also obtained similar result) of content of lignin.
From the above, when fermentation 5 d, each performance indications of maize straw namely obtain preferably effect.Therefore, when below getting fermentation time and being 5 d, the chemical composition of maize straw respectively organized in record.
4.2 the different disposal method is in fermentation impact on maize straw degradation rate during 5 d
The chemical composition (take dry as the basis) of different disposal group maize straw during table 3 fermentation 5d
| Group forms | Cellulose % | Neutral detergent fiber % | Acid detergent fiber % | Hemicellulose % | Lignin % |
| Common stalk group | 41.30±2.31A | 79.26±1.31A | 59.23±2.00A | 20.02±1.18A | 12.58±1.07A |
| Explosion stalk group | 37.80±0.40B | 60.94±0.34C | 51.12±1.12B | 9.92±0.86C | 7.97±1.21B |
| Common stalk+aspergillus oryzae group | 33.77±1.23C | 71.78±1.27B | 52.19±0.60B | 19.59±1.02A | 14.58±2.18A |
| Common stalk+koning trichoderma group | 34.02±1.74C | 72.66±0.24B | 53.62±1.69B | 14.76±2.40B | 12.44±0.90A |
| Explosion stalk+koning trichoderma group | 33.33±1.10C | 60.42±1.41C | 52.31±0.89B | 7.30±0.54C | 10.07±0.22B |
| Explosion stalk+aspergillus oryzae group | 29.78±2.92D | 52.76±2.36D | 47.28±3.75C | 7.06±1.03C | 9.54±0.97B |
---------as shown in Table 3, explosion and various microbial fermentation are processed and have all been reduced significantly the content of cellulose in the stalk (p＜0.05), and common stalk and explosion stalk are after aspergillus oryzae and koning trichoderma fermentation, its content of cellulose all significantly is lower than explosion stalk group (p＜0.05), illustrates that the sweat of microorganism has utilized the cellulose components in the maize straw.Wherein, stalk is behind the inoculation aspergillus oryzae (being explosion stalk+aspergillus oryzae group) after the explosion, its content of cellulose significantly is lower than explosion stalk+koning trichoderma group (p＜0.05), illustrated that aspergillus oryzae utilizes the efficient aspect the explosion stalk cellulose to be higher than koning trichoderma in decomposition.Therefore, when next step measures the variation that various enzymes are lived in the fermented stalk sample, take " explosion stalk+aspergillus oryzae group " as main reference object.Explosion treatment has significantly reduced the content of lignin in the stalk (p＜0.05), but all without significant impact (p＞0.05), this does not secrete the cause of lignin-degrading enzymes mainly due to aspergillus oryzae and koning trichoderma on the lignin in common stalk and the explosion stalk for aspergillus oryzae and koning trichoderma fermentation.
In the preprocessing process of stalk resource, the most important thing is to reduce the ratio of lignin in the stalk, and avoid cellulosic degraded as far as possible.Under these trial shots condition, blasting process has significantly reduced the content (p＜0.05) of the cellulose in the stalk, hemicellulose and lignin, make that cellulose degradation rate is that 8.47%(significantly is lower than common stalk+aspergillus oryzae group p＜0.05 in the stalk), the degradation rate of hemicellulose is 50.45%, and the degradation rate of lignin reaches 36.65%.This explanation explosion treatment has a significant effect aspect cellulose, hemicellulose and the lignin in degrading straw.Lignin is the barrier that straw biological utilizes, and is accompanied by the degraded of hemicellulose and lignin in the blasting process of stalk, and the structural lignin in the stalk is damaged, thereby more is conducive to the effect of microorganism and enzyme.Studies show that of Moyson (Moyson E, Verachtert H. Growth of higher fungi on wheat straw and their impact on the digestibility of the substrate[J].
Appl. Microbial. Biotechnol. 1991,36:421-424.), animal increases along with the reduction of lignin in the stalk the digestibility of maize straw, and this illustrates simultaneously that also this explosion treatment has great importance to the utilization of straw feed resource.
Common stalk and explosion stalk are after the aspergillus oryzae fermentation, and its cellulosic content all significantly is lower than explosion stalk group (p＜0.05), illustrates that the process of microbial fermentation has been utilized the cellulose components in the stalk.Explosion stalk wherein neutral detergent fiber and acid detergent fiber content after aspergillus oryzae fermentation all significantly reduces (p＜0.05), illustrate that aspergillus oryzae has obvious facilitation to the conversion of stalk insoluble fibre composition, stalk after the explosion can promote nutrition and the part active material that it is converted into thalline self through the fermentation process of aspergillus oryzae, animal is produced have certain facilitation.
4.3 explosion stalk+aspergillus oryzae group maize straw is the variation of enzyme activity and soluble sugar during the fermentation
In the aspergillus oryzae sweat, Fig. 2 and Fig. 3 are seen respectively in the variation of filter paper carbohydrase, CMC enzyme, amylase, protease and soluble sugar in the maize straw sample.
The filter paper carbohydrase is with the index utilizing cellulose total enzyme alive height of filter paper as reaction substrate.As shown in Figure 2, in aspergillus oryzae fermentation explosion stalk process, the enzyme in early stage is lived lower, raises gradually behind the 2nd d, and the 6th d is significantly higher than At All Other Times point (p＜0.05).
The work of CMC enzyme is an index of reacting cellulase restriction endonuclease height, the CMC enzyme of aspergillus oryzae fermentation explosion stalk is lived, namely reach the highest at 1d, this may with the stalk blasting process in to produce the carbohydrate of more short chain fiber and short chain relevant, these materials can impel aspergillus oryzae to secrete more restriction endonuclease and promote its utilization.The 2nd d reduces gradually afterwards, and 4d rises to again the highest, and the 7th d enzyme of fermentation is lived and significantly reduced (p＜0.05).Many studies show that, the explosion treatment of stalk helps to weaken the structure of lignocellulosic, makes it be easy to be subject to the attack of enzyme and microorganism.This result illustrates that also the structure of the straw lignocellulose after the explosion is damaged, and makes that wherein short chain cellulose is easier to be utilized by microorganism.
The remarkable rising from the 2d of fermentation of the diastatic activity of explosion stalk all keeps higher level in several days after the fermentation.The protease activity of explosion stalk presents the variation tendency that raises first and descend afterwards, and the enzyme work of ferment the 3rd d and the 4th d reaches the highest, and the 7th d enzyme work of fermenting is down to minimum.
Because of the nitrogenous source that contains in the fermentation medium relatively less, main cellulolytic filter paper carbohydrase reaches at the 6th d and is up to 335.10 U/g, the CMC of the 6th d and diastatic activity are relatively high, are respectively 1138.92 U/g and 32.57 U/g, and protease activity is 201.99 U/g.6 d that can consider to ferment in explosion stalk microbe trans-utilization process effectively bring into play the effect that it utilizes stalk.
As shown in Figure 3, aspergillus oryzae in fermentation explosion stalk process along with the increase of fermentation time, soluble sugar in the maize straw sample reduces gradually, be that explosion stalk soluble sugar content when fermentation the 1st～2 d is the highest, significantly descend subsequently, maintain a lower level, this is relevant with the soluble sugar that microorganism has utilized explosion to produce during the fermentation.
Conclusion
The explosion preliminary treatment of this research (pressurize 200s under the 2.5MPa pressure) makes the content of cellulose in the maize straw, hemicellulose and lignin reduce respectively 8.47%, 50.45% and 36.65%.Behind aspergillus oryzae fermentation 5～6 d, cellulose wherein and the content of hemicellulose have reduced by 27.89% and 64.80% than original stalk respectively to the explosion stalk, have reduced by 21.17% and 28.28% than steam blasting stalk again.Filter paper carbohydrase in the fermented stalk, CMC enzyme, amylase and prolease activity reach respectively 335.10,1138.92,32.57 and 201.99 U/g.This proves absolutely maize straw after explosion and microbial fermentation Combined Treatment, and its degradation rate and nutritive value have all obtained increasing substantially, for the development and use of agricultural crop straw are laid a good foundation.
Claims (4)
1. the method for a steam blasting and microbial fermentation Combined Treatment stalk is characterized in that, comprises the steps:
1. stalk steam blasting preliminary treatment: the stalk that will pulverize 10～20 mesh sieves, moisture content≤15wt% carries out steam blasting to be processed, and then it is dried to get naturally the explosion stalk;
2. prepare aspergillus oryzae/koning trichoderma seed liquor: aspergillus oryzae or koning trichoderma are inoculated in the culture dish of being made by the PDA culture medium, 28～32 ℃ of static cultivation 3～5d, wash culture dish with sterile saline after the spore maturation, spore in the culture dish is transferred in the sterilization triangular flask compound concentration 1 * 10
6～10
8The spore suspension of individual/mL is the spore seed liquor;
3. microbial fermentation: get 18g explosion stalk and 2g wheat bran, 30mL mineral element nutrient solution mixes, with Ca (OH)
2Transferring to pH is 7.0, and then sterilization 15min gets the solid fermentation culture medium under 121 ℃, 0.15MPa pressure; Inoculum concentration by 2～4% is inoculated in the spore seed liquor in the solid fermentation culture medium, cultivates 5～7d for 28～32 ℃;
Described step 1. middle steam blasting is treated to: under the condition of steam pressure 1.8～2.5MPa, and release pressure behind pressurize 200～240s.
2. the method for Treating straw as claimed in claim 1 is characterized in that described stalk is wheat straw or maize straw.
3. the method for Treating straw as claimed in claim 1 is characterized in that, described step 2. in the PDA nutrient media components be: soluble starch 6g, peptone 5g, yeast 2g, glucose 20g, KH
2PO
42g, MgSO
47H
2O 0.3g, agar powder 20g, distilled water 1000mL.
4. the method for Treating straw as claimed in claim 1 is characterized in that described step 3. Mineral Elements nutrient solution prescription is: (NH
4)
2SO
41.4g, KH
2PO
42.0g, MgSO
40.3g, CaCl
20.3g, NaCl 0.5g, FeSO
45.0mg, MnSO
41.6mg, ZnCl
21.7mg, CoCl
22.0mg, distilled water 1000mL.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010105574199A CN102077903B (en) | 2010-11-25 | 2010-11-25 | Method for jointly treating stalks by steam explosion and microorganism fermentation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010105574199A CN102077903B (en) | 2010-11-25 | 2010-11-25 | Method for jointly treating stalks by steam explosion and microorganism fermentation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102077903A CN102077903A (en) | 2011-06-01 |
| CN102077903B true CN102077903B (en) | 2013-01-23 |
Family
ID=44084255
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010105574199A Active CN102077903B (en) | 2010-11-25 | 2010-11-25 | Method for jointly treating stalks by steam explosion and microorganism fermentation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102077903B (en) |
Families Citing this family (22)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102653477B (en) * | 2012-05-11 | 2013-12-25 | 贵州大学 | Preparation method of vinasse-type bio-organic fertilizer |
| CN102864671B (en) * | 2012-09-03 | 2016-01-13 | 潍坊天健源新农业科技有限公司 | The reuse method of Rhizomatic traditional Chinese medicine slag and application |
| CN103704501B (en) * | 2013-12-09 | 2015-04-08 | 平顶山中宏华龙饲料有限公司 | Method for producing high-efficiency broiler feed by utilization of corn straw fermentation |
| CN103881945A (en) * | 2014-01-08 | 2014-06-25 | 北京中科百瑞能工程技术有限责任公司 | Method for preparing culture medium for industrial fermentation by using agricultural raw materials subjected to steam explosion treatment |
| CN103892033B (en) * | 2014-02-21 | 2016-06-01 | 苏州承泰能生物质工程技术有限公司 | The preparation method of a kind of feed |
| CN104186925A (en) * | 2014-08-07 | 2014-12-10 | 成都巨星农牧科技有限公司 | Method for preparing feed through complex probiotic fermentation |
| CN104206828A (en) * | 2014-08-12 | 2014-12-17 | 成都巨星农牧科技有限公司 | Fermentation method for preparation of pig feed |
| CN104472855A (en) * | 2014-11-21 | 2015-04-01 | 河南农业大学 | Method for producing mycoprotein feed by utilizing sweet sorghum straws and/or related waste residues |
| CN104336414A (en) * | 2014-11-28 | 2015-02-11 | 张汉桥 | Fermentation treatment method for beef feed |
| CN104782883A (en) * | 2015-04-10 | 2015-07-22 | 黄飚 | Method for efficient and comprehensive utilization of cornstalks |
| CN104839429B (en) * | 2015-05-28 | 2019-03-15 | 河南双成生物科技有限公司 | The method that the combined solid fermented preparation of steam blasting is rich in fatty acid feedstuff |
| CN105146067B (en) * | 2015-07-15 | 2018-05-15 | 河南农业大学 | A kind of technique that agricultural crop straw is converted into pig chicken feed resource |
| CN105519800A (en) * | 2015-12-25 | 2016-04-27 | 广西农垦金光乳业有限公司 | Preparation method of easy-digestion sugarcane byproduct feed |
| CN105941825A (en) * | 2016-05-06 | 2016-09-21 | 江苏盐城源耀生物科技有限公司 | Corn stalk biologically fermented feed and preparation method thereof |
| CN108450935A (en) * | 2017-02-17 | 2018-08-28 | 北京中科百瑞能工程技术有限责任公司 | A kind of method that medicine-food two-purpose class raw material makes red yeast rice |
| CN108477401A (en) * | 2018-02-06 | 2018-09-04 | 安徽省天长市周氏羊业有限公司 | A kind of method that steam blasting-microbial fermentation processing stalk prepares high protein sheep feed |
| CN108532000B (en) * | 2018-04-17 | 2020-07-10 | 中国农业科学院麻类研究所 | Degumming method of apocynum venetum bast |
| CN108740300B (en) * | 2018-04-19 | 2021-12-21 | 河南农业大学 | Method for preparing monogastric animal energy feed by using crop straws |
| CN108813100B (en) * | 2018-06-15 | 2021-09-21 | 河南农业大学 | Method for preparing straw micro-storage feed by combined treatment of blasting, calcium oxide and probiotics |
| CN108850552B (en) * | 2018-07-09 | 2022-05-03 | 康桥投资控股有限公司 | Preparation method of corn straw feed fermentation |
| CN112522342B (en) * | 2020-12-17 | 2023-02-24 | 新疆希普生物科技股份有限公司 | Method for efficiently performing enzymolysis on straws |
| CN113072599A (en) * | 2021-03-10 | 2021-07-06 | 武汉市华甜生物科技有限公司 | Method for preparing natural hesperidin by biological fermentation and extraction |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1294861A (en) * | 1999-11-03 | 2001-05-16 | 中国科学院化工冶金研究所 | Mixed solid fermentation and straw puffing process to prepare protein feed |
| CN1451755A (en) * | 2002-04-12 | 2003-10-29 | 中国农业大学 | Method for producing alcohol by solid fermentation of stalks |
| CN1706270A (en) * | 2004-06-05 | 2005-12-14 | 宁夏大学 | Stalks fermenting fungus prepn and production process and biologically fermented stalks feed |
-
2010
- 2010-11-25 CN CN2010105574199A patent/CN102077903B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1294861A (en) * | 1999-11-03 | 2001-05-16 | 中国科学院化工冶金研究所 | Mixed solid fermentation and straw puffing process to prepare protein feed |
| CN1451755A (en) * | 2002-04-12 | 2003-10-29 | 中国农业大学 | Method for producing alcohol by solid fermentation of stalks |
| CN1706270A (en) * | 2004-06-05 | 2005-12-14 | 宁夏大学 | Stalks fermenting fungus prepn and production process and biologically fermented stalks feed |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102077903A (en) | 2011-06-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102077903B (en) | Method for jointly treating stalks by steam explosion and microorganism fermentation | |
| Yang et al. | Bioconversion of corn straw by coupling ensiling and solid-state fermentation | |
| Montoya et al. | Growth, fruiting and lignocellulolytic enzyme production by the edible mushroom Grifola frondosa (maitake) | |
| US20070099282A1 (en) | Enzyme and methodology for the treatment of a biomass | |
| CN101460614A (en) | Talaromyces emersonii enzyme systems | |
| CN103958689A (en) | C1-C2 organic acid treatment of lignocellulosic biomass to produce acylated cellulose pulp, hemicellulose, lignin and sugars and fermentation of the sugars | |
| KR101771960B1 (en) | Paenibacillus jamilae BRC15-1 strain producing cellulase and use thereof | |
| CN108486017A (en) | A kind of garden waste degradation bacterial agent | |
| CN104789492B (en) | Bacillus megaterium bacterial strain and its application | |
| Fadel et al. | Cellulases and animal feed production by solid-state fermentation by Aspergillus fumigatus NRCF-122 mutant | |
| CN101624584A (en) | Preparation method of internal cutting type xylanase | |
| CN108048473A (en) | A kind of feruloyl esterase gene, engineering strain and preparation method and purposes | |
| Iram et al. | Salt and nitrogen amendment and optimization for cellulase and xylanase production using dilute acid hydrolysate of distillers’ dried grains with solubles (DDGS) as the feedstock | |
| CN104789491B (en) | Lichem bacillus strain and its application | |
| CN112725314B (en) | Method for producing endoglucanase by fermenting coarse feed through natural symbiotic mixed culture | |
| Ximenes et al. | Enzyme production by industrially relevant fungi cultured on coproduct from corn dry grind ethanol plants | |
| JP2009183264A (en) | Saccharification method of rice straw | |
| Kapoor et al. | Effect of supplementation of wheat straw on growth and lignocellulolytic enzyme potential of Lentinus edodes | |
| CN104860401A (en) | Application of Rhizopus oryzae strain JHSW01 to the fermentation of organic waste liquid and forestry and agricultural waste | |
| Singh et al. | Evaluation of chemical pre-treatment for biodegradation of agricultural lignocellulosic wastes by Aspergillus niger | |
| CN101638645A (en) | Method for producing xylanase by solid mechanical fermentation | |
| CN101067129A (en) | Industrial process of producing xylanase with Aspergillus usamii | |
| Cassa-Barbosa et al. | Isolation and characterization of yeasts capable of efficient utilization of hemicellulosic hydrolyzate as the carbon source | |
| CN104893988A (en) | Trichoderma atroviride strain capable of producing high-temperature-resistant feruloyl esterase and high-temperature-resistant cellulase and application thereof | |
| CN104845952B (en) | A kind of method that Trichoderma atroviride triangular flask liquid fermentation prepares heat-resisting feruloyl esterase and cellulase complex enzyme formulation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20160826 Address after: Taihang Road demonstration zone in Xinxiang city of Henan province and 453004 plain Tonghui River Road intersection 200 meters to the East Road Patentee after: Henan Delin Biological Products Co., Ltd. Address before: 450002 Zhengzhou Cultural Road, Henan, No. 95 Patentee before: He'nan Agricultural University |