CN106085909A - Ripening microbial inoculum that corn straw is matrixing and application - Google Patents
Ripening microbial inoculum that corn straw is matrixing and application Download PDFInfo
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Abstract
The present invention provides the ripening microbial inoculum that a kind of corn straw is matrixing, its active component is Clostridium thermocellum (Clostridium thermocellum), solves cellulose wood polysaccharide Zymomonas mobilis (Xylanimonas cellulosilytica), bacillus cereus (Bacillus cereus) and Bacillus licheniformis (Bacillus licheniformis), the ratio of the colony forming unit number of above four kinds of bacterium is respectively 4 5:2 3:2 3:2 3, and the living bacteria count of described microbial inoculum is 109Individual/more than mL.The present invention also provide for described ripening microbial inoculum corn straw matrixing become thoroughly decomposed fermentation in application.The ripening microbial inoculum of the present invention is high to the degradation rate of corn straw, when carrying out extensive ripening fermentation, in 20 days the 40 70% of decomposable asymmetric choice net straw, provide important technical for corn straw is matrixing, have broad application prospects at organic culture substrate production field.
Description
Technical field
The present invention relates to the recycling field of agricultural wastes, specifically, relate to matrixing ripe of corn straw
Change microbial inoculum and application.
Background technology
In recent years, developing rapidly, for the demand of cultivation matrix along with industrial breeding technique and cultivation technique without soil
Amount is increased sharply day by day, the common growing nursery and culture substrate such as resource-constrained such as the peat composed of rotten mosses, Vermiculitum, and the short time is non-renewable, and rock wool etc. makes
With rear more intractable, easily to environment, in the urgent need to developing novel replacement cultivation matrix.China is large agricultural country,
The yield of annual generation substantial amounts of agricultural crop straw, especially corn straw is huge, if it is ripe that these corn straw are carried out high temperature
Change fermentation, it is achieved harmless treatment, substantial amounts of commodity cultivation matrix can be produced, really realize the resource profit of agricultural wastes
With.
Corn straw contains substantial amounts of lignocellulosic elements, decomposes difficult and speed is slow；It is coarse as substrate physical behavior,
Density is little, and hole is big；Burn root as cultivation matrix serious, suppress crop growth.Corn straw is utilized to produce organic farming base
Matter must carry out high-temperature maturing fermentation, it is intended to decomposes macromolecular substances (lignocellulose, saccharide, starch, fat, albumen), fall
Low ratio of carbon to ammonium, eliminates weeds, pathogen, to adapt to plant growth.Ripening fermentation is under the effect of microorganism, by decomposable
Organic substance all decomposes, and enters safe handling state, and Partial Elements is mineralized into the process of plant nutrient.In maturing process, first
The easy analyte of material such as saccharide, starch, fat, albumen decomposed the temperature rise period of heat production before this, divided with cellulose, hemicellulose
Solve and continue the hot fermentation stage of heat production.Wherein decomposition speed and the degree of decomposition of cellulose and hemicellulose decides ripening
Degree.In recent years, microbial inoculum applied research during the fermentation is more, but for cultivation matrix ripening microbial inoculum not
Many, in terms of the rapid aging microbial inoculum that especially corn straw is matrixing report phoenix feathers and unicorn horns especially.
Summary of the invention
It is an object of the invention to provide the ripening microbial inoculum that a kind of corn straw is matrixing.
It is a further object of the present invention to provide described ripening microbial inoculum corn straw matrixing become thoroughly decomposed fermentation in application.
In order to realize the object of the invention, a kind of ripening microbial inoculum of the present invention, its active component is Clostridium thermocellum
(Clostridium thermocellum), solution cellulose wood polysaccharide Zymomonas mobilis (Xylanimonas cellulosilytica),
Bacillus cereus (Bacillus cereus) and Bacillus licheniformis (Bacillus licheniformis).Above four kinds
The ratio of the CFU (colony forming unit number) of bacterium is respectively 4-5:2-3:2-3:2-3 (preferably 4:2:2:2), having of described microbial inoculum
Effect viable count is 109Individual/more than mL.
Preferably, its active component is the Clostridium thermocellum of preserving number DSM No.1237, the solution of preserving number DSM No.15894
Cellulose wood polysaccharide Zymomonas mobilis, the bacillus cereus of preserving number ACCC No.03315 and the ground of preserving number ACCC No.01958
Clothing bacillus cereus.
Wherein, described Clostridium thermocellum is anaerobic bacteria, has in cellulase activity agricultural wastes high, rapid decomposable
Cellulose and hemicellulose, this culture presevation in Germany Culture Collection (DSMZ), preserving number is DSM
No.1237；Solve cellulose wood polysaccharide Zymomonas mobilis and support antibacterial, decomposable asymmetric choice net cellulose and hemicellulose components, this culture presevation preferably
In Germany's Culture Collection, preserving number is DSM No.15894；Bacillus cereus is aerobic bacteria, has generation
Cellulose enzyme viability, this culture presevation is in Chinese agriculture Microbiological Culture Collection administrative center (ACCC), and preserving number is ACCC
No.03315；Bacillus licheniformis is aerobic bacteria, has cellulose decomposition function, and this culture presevation is in Chinese agriculture microorganism
Culture presevation administrative center, preserving number is ACCC No.01958.Above four kinds of strains can be purchased by corresponding Spawn preservation organization
Buy acquisition.
Preferably, the ripening microbial inoculum of the present invention also includes the ratio of absorption carrier, described active ingredient and absorption carrier
Relation is 1CFU:(1 × 10-6～1 × 10-5) g absorption carrier, preferably 1CFU:5 × 10-6G absorption carrier.
It is highly preferred that described absorption carrier is pressed 2-4:6-8 by Maifanitum and the corn stalk powder of 40-60 mesh (preferably 40 mesh)
The weight ratio composition of (preferably 2:8).
The present invention also provides for the preparation method of described ripening microbial inoculum, comprises the following steps:
(1) preparation of seed liquor:
Clostridium thermocellum is inoculated in peptone cellulose improved culture medium, static gas wave refrigerator 2d under the conditions of 50 DEG C, collects
Fermentation liquid is as seed liquor；
Solution cellulose wood polysaccharide Zymomonas mobilis is inoculated in peptone cellulose improved culture medium, static under the conditions of 50 DEG C
Cultivate 2d, collect fermentation liquid as seed liquor；
Bacillus cereus is inoculated in peptone cellulose improved culture medium, static gas wave refrigerator 1d under the conditions of 50 DEG C,
Collect fermentation liquid as seed liquor；
Bacillus licheniformis is inoculated in peptone cellulose improved culture medium, static gas wave refrigerator 1d under the conditions of 50 DEG C,
Collect fermentation liquid as seed liquor；
(2) amplification culture of strain:
In in step (1), each seed liquor of gained is inoculated in peptone cellulose improved culture medium respectively, in cultivation temperature
Being static amplification culture step by step in the fermentation tank of 50 DEG C, expansion process is transferred, when in fermentation liquid by the volume ratio of 5%～10%
Living bacteria count reaches 109Individual/mL time terminate cultivate；
(3) preparation of ripening microbial inoculum:
The each fermentation liquid of gained in step (2) is mixed in proportion, is then added to absorption carrier mixing, obtains ripening
Microbial inoculum.
The compound method of peptone cellulose improved culture medium used in step (1) and (2) is as follows: by peptone 5g,
Yeast extract 1g, NaCl 5g, K2HPO4 1g、MgSO4·7H2O0.35g and CaCO33g is dissolved in 1L water, sterilizing, thus obtaining the product.
The present invention also provides for the application in cellulose and hemicellulose decompose of the described ripening microbial inoculum.
The present invention also provide for described ripening microbial inoculum corn straw matrixing become thoroughly decomposed fermentation in application.
The present invention also provides for the preparation method of a kind of cultivation matrix, corn straw, carbamide, calcium carbonate and water is mixed, adds
Adding appropriate described ripening microbial inoculum, after mixing, composting carries out fermentation of becoming thoroughly decomposed, and fermentation period is 15-25 days (preferably 20 days), and gained is sent out
Ferment product is i.e. as cultivation matrix.
Aforesaid method, presses corn straw, carbamide and calcium carbonate the mass ratio mixing of 98:1:0.1, is added to jade
Rice straw, carbamide and the water of calcium carbonate gross weight 55-60% (preferably 60%), gained ripening fermentation raw material and ripening microbial inoculum are by 100:
The mass ratio mixing of 0.5-1 (preferably 100:0.5), piles wide 1-3m, high 1-2m, long 2-100m (the widest 3m, high 1.3m, length
Cuboid 5m) carries out fermentation of becoming thoroughly decomposed, and fermentation period is 20 days, in sweat when heap temperature is more than 50 DEG C, turns over every day
Heap once, until having fermented.
The present invention also provides for the cultivation matrix prepared by said method.
The present invention further provides the application in plant soil-less culturing of the described cultivation matrix.Described plant includes Oryza sativa L..
The invention have the advantages that
(1) the matrixing ripening microbial inoculum of corn straw that the present invention provides, cellulose and hemicellulase are lived height, can be
Initial stage effective decomposition of cellulose of corn straw ripening fermentation and hemicellulose components, thus the wood fibre that rapid damage is hard
Element structure, reaches the effect of loose straw structure, enters the synchronous high-efficiency maturation stage of cellulose, hemicellulose afterwards.
(2) the ripening microbial inoculum of the present invention is the micro-aerobic composite microbial system of antibacterial, can in micro-aerobic ripening sweat,
The most aerobic bacterial strain solution cellulose wood polysaccharide Zymomonas mobilis, bacillus cereus and the leading decomposition of cellulose of Bacillus licheniformis and
Consume partial oxygen while hemicellulose, create local anaerobic, create for Anaerobic cellulose decomposing bacteria-Clostridium thermocellum and decompose ring
Border, further speeds up the decomposition of cellulose and hemicellulose, promotes the ripening progress that raw material is overall.
(3) the ripening microbial inoculum of the present invention is high to the degradation rate of corn straw, when carrying out extensive ripening fermentation, and 20 days
The 40-70% of interior decomposable asymmetric choice net straw.
(4) breach cannot efficient-decomposition native cellulose by purification bacterium and catabolic enzyme thereof for the ripening microbial inoculum of the present invention
Limitation, provides important technical for corn straw is matrixing, has wide application at organic culture substrate production field
Prospect.
Detailed description of the invention
Following example are used for illustrating the present invention, but are not limited to the scope of the present invention.If not specializing, embodiment
In the conventional means that is well known to those skilled in the art of technological means used, raw materials used be commercial goods.
In following example, the compound method of peptone cellulose (PCS) improved culture medium used is as follows: by peptone 5g,
Yeast extract 1g, NaCl 5g, K2HPO4 1g、MgSO4·7H2O0.35g and CaCO33g is dissolved in 1L water.121 DEG C of steam
Sterilizing 15 minutes.
The preparation of the ripening microbial inoculum that embodiment 1-3 corn straw is matrixing
The ripening microbial inoculum of preparation in embodiment 1-3, its active component is Clostridium thermocellum (Clostridium
Thermocellum) preserving number DSM No.1237, solution cellulose wood polysaccharide Zymomonas mobilis (Xylanimonas
Cellulosilytica) preserving number DSM No.15894, bacillus cereus (Bacillus cereus) preserving number ACCC
No.03315 and Bacillus licheniformis (Bacillus licheniformis) preserving number ACCC No.01958.
1, the preparation of seed liquor:
Clostridium thermocellum preserving number DSM No.1237 is inoculated in peptone cellulose (PCS) improved culture medium, at 50 DEG C
Under the conditions of static gas wave refrigerator 2d, collect fermentation liquid as seed liquor；
Solution cellulose wood polysaccharide Zymomonas mobilis preserving number DSM No.15894 is inoculated in peptone cellulose improved culture medium
In, static gas wave refrigerator 2d under the conditions of 50 DEG C, collects fermentation liquid as seed liquor；
Bacillus cereus preserving number ACCC No.03315 is inoculated in peptone cellulose improved culture medium, 50
Static gas wave refrigerator 1d under the conditions of DEG C, collects fermentation liquid as seed liquor；
Bacillus licheniformis preserving number ACCC No.01958 is inoculated in peptone cellulose improved culture medium, 50
Static gas wave refrigerator 1d under the conditions of DEG C, collects fermentation liquid as seed liquor.
2, the amplification culture of strain:
Each for gained in step 1 seed liquor is inoculated in peptone cellulose improved culture medium respectively, in cultivation temperature is
Static amplification culture step by step in the fermentation tank of 50 DEG C, expansion process is transferred, when having in fermentation liquid by the volume ratio of 5%～10%
Effect viable count reaches 109Individual/mL time terminate cultivate.
3, the preparation of ripening microbial inoculum:
By the Clostridium thermocellum fermentation liquid of gained, solution cellulose wood polysaccharide Zymomonas mobilis fermentation liquid, bacillus cereus in step 2
Fermentation liquid and the lichen bacillus ferments liquid press the ratio 4:2:2:2 of CFU, and (embodiment 1, in embodiment 2, the ratio of the CFU of four kinds of bacterium is
5:2:3:2, in embodiment 3, the ratio of the CFU of four kinds of bacterium is 4:3:2:3) mixing, then it is added to absorption carrier mixing, i.e.
Obtain ripening microbial inoculum.Wherein, active ingredient is 1CFU:5 × 10 with the proportionate relationship of absorption carrier-6G absorption carrier.Described absorption carries
Body is made up of by the weight ratio of 2:8 Maifanitum and the corn stalk powder of 40 mesh.
Embodiment 4 ripening microbial inoculum corn straw matrixing become thoroughly decomposed fermentation in application
Experimental group 1-experimental group 3: corn straw, carbamide and calcium carbonate are pressed the mass ratio mixing of 98:1:0.1, wherein
Add corn straw, carbamide and the water of calcium carbonate gross weight 60%, gained ripening fermentation raw material respectively with embodiment 1-3 (embodiment
1, embodiment 2 and embodiment 3 correspond respectively to experimental group 1, experimental group 2 and experimental group 3) the ripening microbial inoculum prepared is by 100:0.5
Mass ratio mixing, pile wide 3m, high 1.3m, the cuboid of long 5m carry out fermentation of becoming thoroughly decomposed, fermentation period is 20 days, sweat
In when heap temperature is more than 50 DEG C, turning every day once, until having fermented.
Matched group: the ripening microbial inoculum prepared without embodiment 1-3, directly piles wide 3m, height by ripening fermentation raw material
1.3m, the cuboid of long 5m carry out fermentation of becoming thoroughly decomposed.Fermentation condition is ibid.
Experimental result, uses after microbial inoculum 7 days, and the cellulose decomposition rate of corn straw reaches 10.5%, and hemicellulose decomposes
Rate reaches 13.0%, and structure has loose sign, enters cellulose and the vigorous catabolic phase of hemicellulose afterwards.Use 20 days
After, the resolution ratio of corn straw reaches more than 40%.
After fermenting 20 days, experimental group and matched group tunning major parameter comparative result are shown in Table 1.
Table 1 tunning major parameter comparative result
| C/N | Gross mass loss of weight | Cellulose loss of weight | Hemicellulose loss of weight | Unit weight | Total porosity | |
| Matched group | 16.3 | 29.3% | 26.5% | 33.7% | 0.19g/cm<sup>3</sup> | 60.3% |
| Experimental group 1 | 14.2 | 40.9% | 38.4% | 49.2% | 0.25g/cm<sup>3</sup> | 70.6% |
| Experimental group 2 | 14.5 | 40.1% | 37.7% | 50.1% | 0.25g/cm<sup>3</sup> | 72.1% |
| Experimental group 3 | 14.4 | 41.6% | 39.1% | 49.7% | 0.26g/cm<sup>3</sup> | 71.1% |
The application in seedling cultivation of rice of embodiment 5 soilless culture substrate
Using in embodiment 4, the tunning of acquisition is as soilless culture substrate, for seedling cultivation of rice.
Experimental group 1-3: by the substrate of the experimental group of acquisition in embodiment 4, take gross dry weight about 300g, be uniformly laid on length, width and height
In the special thin vinyl disc of 60 × 30 × 4cm, stromal thickness about 3cm, irrigate the sulphuric acid water of pH5.5 after slight suppression, often dish is broadcast
Plant 100g (dry weight meter) Germinated rice seeds, cover table soil 0.5cm thick, nurse young plants in hothouses 30 days, temperature simulation northeast condition in spring, in vain
They 25 DEG C, night 10-15 DEG C.Setup Experiments 3 repetition, and after 30 days, measure the upgrowth situation of rice seedling.
Matched group: with reference to the operational approach of above-mentioned experimental group, only substrate is replaced to the base obtained in embodiment 4 matched group
Matter.
The upgrowth situation of experimental group 1-3 and matched group rice seedling is shown in Table 2.
The upgrowth situation of table 2 rice seedling
| Leaf age | Plant height | Individual plant radical | Herb hundred strain dry weight | Overground part hundred strain dry weight | Root hundred strain dry weight | |
| Matched group | 2.69 leaf | 12.6cm | 9 | 3.02g | 2.27g | 0.75g |
| Experimental group 1 | 3.07 leaf | 13.4cm | 11 | 3.59g | 2.76g | 0.84g |
| Experimental group 2 | 2.98 leaf | 13.1cm | 10 | 3.44g | 2.57g | 0.79g |
| Experimental group 3 | 3.10 leaf | 13.6cm | 11 | 3.51g | 2.79g | 0.86g |
Although, the present invention is described in detail the most with a general description of the specific embodiments, but
On the basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to the scope of protection of present invention.
Claims (10)
1. a ripening microbial inoculum, it is characterised in that its active component be Clostridium thermocellum (Clostridium thermocellum),
Solve cellulose wood polysaccharide Zymomonas mobilis (Xylanimonas cellulosilytica), bacillus cereus (Bacillus
Cereus) and Bacillus licheniformis (Bacillus licheniformis), the ratio of the CFU of above four kinds of bacterium is respectively 4-5:2-
3:2-3:2-3, the living bacteria count of described microbial inoculum is 109Individual/more than mL.
Ripening microbial inoculum the most according to claim 1, it is characterised in that its active component is preserving number DSM No.1237
Clostridium thermocellum, solution cellulose wood polysaccharide Zymomonas mobilis, the waxy bud of preserving number ACCC No.03315 of preserving number DSM No.15894
Spore bacillus and the Bacillus licheniformis of preserving number ACCC No.01958, the ratio of the CFU of preferably above four kinds of bacterium is respectively 4:2:2:
2。
Ripening microbial inoculum the most according to claim 1 and 2, it is characterised in that also include absorption carrier in described microbial inoculum, described
Active ingredient is 1CFU:(1 × 10 with the proportionate relationship of absorption carrier-6～1 × 10-5) g absorption carrier, preferably 1CFU:5 × 10- 6G absorption carrier.
4. according to the ripening microbial inoculum described in any one of claim 1-3, it is characterised in that described absorption carrier is by 40-60 mesh wheat
Meal stone and corn stalk powder press the weight ratio composition of 2-4:6-8.
5. the preparation method of ripening microbial inoculum described in claim 3 or 4, it is characterised in that comprise the following steps:
(1) preparation of seed liquor:
Clostridium thermocellum is inoculated in peptone cellulose improved culture medium, static gas wave refrigerator 2d under the conditions of 50 DEG C, collects fermentation
Liquid is as seed liquor；
Solution cellulose wood polysaccharide Zymomonas mobilis is inoculated in peptone cellulose improved culture medium, static gas wave refrigerator under the conditions of 50 DEG C
2d, collects fermentation liquid as seed liquor；
Bacillus cereus is inoculated in peptone cellulose improved culture medium, static gas wave refrigerator 1d under the conditions of 50 DEG C, collects
Fermentation liquid is as seed liquor；
Bacillus licheniformis is inoculated in peptone cellulose improved culture medium, static gas wave refrigerator 1d under the conditions of 50 DEG C, collects
Fermentation liquid is as seed liquor；
(2) amplification culture of strain:
In in step (1), each seed liquor of gained is inoculated in peptone cellulose improved culture medium respectively, it is 50 in cultivation temperature
DEG C fermentation tank in static amplification culture step by step, expansion process by 5%～10% volume ratio switching, effective when in fermentation liquid
Viable count reaches 109Individual/mL time terminate cultivate；
(3) preparation of ripening microbial inoculum:
The each fermentation liquid of gained in step (2) is mixed in proportion, is then added to absorption carrier mixing, obtains ripening bacterium
Agent；
The compound method of peptone cellulose improved culture medium used in step (1) and (2) is as follows: by peptone 5g, yeast
Extract 1g, NaCl 5g, K2HPO4 1g、MgSO4·7H2O 0.35g and CaCO33g is dissolved in 1L water, sterilizing, thus obtaining the product.
6. the application in cellulose and hemicellulose decompose of the ripening microbial inoculum described in any one of claim 1-4.
7. ripening microbial inoculum described in any one of claim 1-4 corn straw matrixing become thoroughly decomposed fermentation in application.
8. the preparation method of a cultivation matrix, it is characterised in that corn straw, carbamide, calcium carbonate and water are mixed, adds suitable
Ripening microbial inoculum described in amount any one of claim 1-4, after mixing, composting carries out fermentation of becoming thoroughly decomposed, and fermentation period is 15-25 days, gained
Tunning is i.e. as cultivation matrix.
Method the most according to claim 8, it is characterised in that corn straw, carbamide and calcium carbonate are pressed 98:1:0.1's
Mass ratio mixes, and is added to corn straw, carbamide and the water of calcium carbonate gross weight 55-60%, gained ripening fermentation raw material with
The mass ratio mixing of 100:0.5-1 pressed by ripening microbial inoculum, piles wide 1-3m, high 1-2m, the cuboid of long 2-100m carry out becoming thoroughly decomposed and send out
Ferment, fermentation period is 20 days, in sweat when heap temperature is more than 50 DEG C, turning every day once, until having fermented.
The cultivation matrix that the most according to claim 8 or claim 9, prepared by method.
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| CN115011648A (en) * | 2022-07-07 | 2022-09-06 | 东北农业大学 | Fermentation method for producing sugar by using clostridium thermocellum to strengthen corn straws |
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| CN107805615A (en) * | 2017-11-13 | 2018-03-16 | 上海创博生态工程有限公司 | A kind of maize straw decomposing agent and preparation method thereof |
| CN107805615B (en) * | 2017-11-13 | 2021-06-08 | 上海创博生态工程有限公司 | Corn straw decomposing inoculant and preparation method thereof |
| CN107986908A (en) * | 2017-12-06 | 2018-05-04 | 云南云天化股份有限公司 | A kind of microbial manure matrix of suitable bacillus growth |
| CN110093300A (en) * | 2019-05-22 | 2019-08-06 | 北京师范大学 | A kind of method of bacillus composite bacteria agent of degrading maize straws and preparation method thereof, degrading straw |
| CN110923174A (en) * | 2019-12-26 | 2020-03-27 | 宁夏嘉日康诚农业科技有限公司 | Rapid decomposition microbial inoculum for corn straw basification and application thereof |
| CN114045234A (en) * | 2021-11-03 | 2022-02-15 | 哈尔滨工业大学 | Bacterial liquid for promoting aerobic fermentation of corn straw to produce heat, preparation method and application |
| CN115011648A (en) * | 2022-07-07 | 2022-09-06 | 东北农业大学 | Fermentation method for producing sugar by using clostridium thermocellum to strengthen corn straws |
| CN115011648B (en) * | 2022-07-07 | 2024-03-15 | 东北农业大学 | Fermentation method for enhancing corn straw sugar production by clostridium thermocellum |
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