CN106801074A - A kind of gliocladin bacterium dregs innocent treatment method - Google Patents
A kind of gliocladin bacterium dregs innocent treatment method Download PDFInfo
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Abstract
The present invention relates to castoff regenerative recycling field, there is provided the modification Application way of a kind of gliocladin bacterium dregs innocent treatment method, wherein gliocladin bacteria residue includes：Bacteria residue residual gliocladin treatment, bacteria residue after treatment is primary raw material, and other agricultural by-products are auxiliary material, by being inoculated with selective composite flora solid fermentation, then through drying, pulverize and sieve, packaging and other steps are completed；There is simple production process using this processing method, the features such as easy, process time is short is operated.Modified product protein >=25%, can be reused for the fermented and cultured of gliocladin, there is provided the new way that environmental pollution and scrap loop in a kind of solution gliocladin production are recycled.
Description
Technical field
The present invention relates to solid waste recycling field, it is related to a kind of gliocladin bacterium dregs innocent treatment method.
Background technology
Gliocladin be table polysulfide for piperazinedione toxoid, be the secondary metabolite of the fungies such as trichoderma.It is low molecule
Measure the compound of the sulphur bridge type of polarity.Weidling and Emerson (1936) are first by this material from Trichoderma
Separated in lignorum, this material is named as gliocladin (gliotoxin) by Weidling, and 1958 by Bell etc.
People identifies its structure.
Current gliocladin mainly produces agricultural biological bactericide by Trichoderma viride liquid deep layer fermenting, is mainly used in cotton
Spend the anti-of the soil-borne diseases such as droop, rice sheath blight disease, sclerotinia rot of colza, canker of apple fruit, bitter rot or anthracnose of grape, pepper anthracnose
Control.Because it has good killing, inhibitory action to soil-borne disease pathogen, the focus as current research.But it is mould in glue
Rhzomorph production process Green trichoderma will produce substantial amounts of mycelium, after gliocladin is extracted in thalline, these mycelium and
It is minimal amount of not synthesize gliocladin fermentation bacteria residue, abbreviation gliocladin bacteria residue using complete culture medium.According to calculating, often produce
1 xanthan moldin produces 10 tons of bacteria residues, causes serious environmental pollution and the wasting of resources.Analysis shows：Gliocladin bacteria residue contains
Protein is preferable sources of nitrogen more than 20%, but cannot function as feed and use, because from safety considerations, existing
The problems such as gliocladin is remained, therefore find a kind of method of comprehensive utilization of gliocladin bacteria residue and have great importance.
The content of the invention
A kind of problems that the present invention exists for prior art, there is provided gliocladin bacterium dregs innocent treatment side
The modification Application way of method, wherein gliocladin bacteria residue includes：Bacteria residue residual gliocladin is processed, and the bacteria residue after treatment is
Primary raw material, other agricultural by-products are auxiliary material, by being inoculated with selective composite flora solid fermentation, then through drying, crushed
Sieve, packaging and other steps are completed；There is simple production process using this processing method, the features such as easy, process time is short is operated.
Product protein content >=30% after treatment, while the gliocladin, the organic solvent residual that will be remained in gliocladin bacteria residue
Pre-processed, reduced gliocladin residual quantity, increased security, the fermented and cultured of gliocladin can be reused for, there is provided
A kind of environmental pollution solved in gliocladin production and the new way of fertilizer cycling and reutilization.
The present inventor has found that gliocladin waste residue can be two in gliocladin culture medium after studying for a long period of time
It is secondary to use, replace part dregs of beans, corn pulp, yeast extract etc. as nitrogen source.Therefore, gliocladin bacteria residue is a kind of gliocladin
Ideal nutritional composition in fermentation medium, can recycle in gliocladin fermentation process, solve the waste residue to environment
Pollution and regeneration problem, realize clean manufacturing target.
On the basis of above-mentioned technology contents, inventor further provides following technical scheme：
A kind of gliocladin bacterium dregs innocent treatment method, comprises the following steps that：
(1), bacteria residue remains the treatment of gliocladin：The bacteria residue for having extracted gliocladin is well mixed with alkaline matter, makes
The pH value of mixed material is banked up between 8-10, afterwards and is processed 5-10 days, wherein daily turning is once；
(2), the integration of material：The bacteria residue after 70-90 parts of step (1) treatment is taken by weight, and 10-20 parts of agricultural byproducts are mixed
Close uniform, between the moisture 50-60wt% of regulation material；
(3), fermentation process：With bafillus natto：Saccharomyces cerevisiae bacteria liquid accumulates ratio=1：5th, inoculum concentration 2-10vt% connects
Enter in compound, be well mixed, ferment 72-96h at 30-55 DEG C, when temperature is higher than 55 DEG C, is dropped by stirring and ventilation
Temperature；
(4), last handling process：Material to having fermented is dried, and makes moisture less than 9wt%, and 80 mesh were crushed afterwards
Sieve.
In step (1), the pH value of mixed material is controlled between 8-10, under these conditions, gliocladin is extremely not
Stabilization can be decomposed quickly, so that the gliocladin content remained in reducing mixed material, increases the security of mixed material, typically
Can complete within 5-10 days, so control time is as above, and the purpose of turning is to be well mixed material during banking up, while
Promoting the volatilization of moisture makes the material moisture for the treatment of drop to below 60wt%, facilitates the use at rear portion；
In step (2), if mixed material moisture is less than 50-60wt%, needing to add water is adjusted；
In step (3) from bafillus natto can extracellular proteinase, the insoluble protein in gliocladin thalline
Matter degraded becomes soluble albumen or oligopeptides；And high-protein yeast bacterium is mainly yeast growth process, will be inorganic in material
Nitrogen changes into organic nitrogen, improves the protein content of product.
In above-mentioned processing method, the gliocladin bacteria residue in step (1) be extracted gliocladin fresh wet bacteria slag or
The dried bacteria residue of person, bacteria residue mainly contains gliocladin residual, has alkalescence, high temperature due to gliocladin, sees photo-labile
Feature, the mode such as can be fermented by adjusting material acid-base value, aerobic bacteria, and gliocladin is decomposed, and make gliocladin in material
Content drops to below 5mg/kg, to reach condition of culture；
Described alkaline matter can be that one or more in NaOH, potassium hydroxide, calcium oxide, calcium hydroxide are mixed
Compound；
Agricultural byproducts described in step (2) can be one or more mixtures in wheat bran, corn flour, wheat flour；
The addition of above-mentioned substance can adjust the carbon-nitrogen ratio in whole mixed material, increase quick-acting nutritious, make strain quick
Breeding, for fermentation provides enough nutritional supports, while the discarded object for realizing agricultural byproducts is recycled, improves its value；
Bafillus natto and saccharomyces cerevisiae described in step (3) are bought in the management of Chinese agriculture Microbiological Culture Collection
Center, wherein bafillus natto are selected from ACCC19833 or ACCC10614；Saccharomyces cerevisiae be selected from ACCC20039 or
ACCC20042 or ACCC20107；
Described bacterium solution can be cultivated using the cultural method of its original description, it is also possible to be entered using following culture mediums
Row culture：
Both are respectively strain activation and culture base：Bafillus natto activation medium selects nutrient broth medium, will
Bafillus natto accesses beef-protein medium and carries out activation 12-18h at 30-35 DEG C in inclined-plane culture mode, then
It is inoculated into bacillus natto to ferment culture medium, 32-35 DEG C of fermentation 36-48h prepares bafillus natto bacterium solution；
Saccharomyces cerevisiae activation medium selects malt juice liquid medium, specially by saccharomyces cerevisiae at 26-30 DEG C with oblique
The mode in face accesses malt extract medium and is activated, and is then seeded into fermentation by saccharomyces cerevisiae culture medium, 28-32 DEG C of culture
48-72h is used to prepare saccharomyces cerevisiae bacterium solution；
Above-mentioned each culture medium uses prior art；
Thalline quantity >=2,000,000,000/mL, thalline in saccharomyces cerevisiae bacterium solution in final prepared bafillus natto bacterium solution
Quantity >=500,000,000/mL；
The material obtained by the above method, wherein containing protein and total reducing sugar, wherein protein content more than 25%, always
Sugar more than 50%, below gliocladin 5mg/kg, can replace part dregs of beans, corn pulp, yeast extract mould in glue as nitrogen source
Used in rhzomorph culture medium, during concrete application, consumption is the 0.5-3% of whole gliocladin culture medium weight, due to raw material master
Gliocladin thalline is come from, nutrition is more balanced, is conducive to the utilization of nutrition.
In sum, this processing method provided by the present invention has simple production process, and operation is easy, process time
Short the features such as.Product protein content >=25% after treatment, while the gliocladin, organic that will be remained in gliocladin bacteria residue
Dissolvent residual is pre-processed, and reduces gliocladin residual quantity, increases security, can be reused for the fermentation training of gliocladin
Support, there is provided the new way of a kind of environmental pollution solved in gliocladin production and fertilizer cycling and reutilization.
Specific embodiment
The present invention is further illustrated with reference to embodiment, can make those skilled in the art that this hair is more completely understood
It is bright, but do not limit the invention in any way.
Embodiment 1
Gliocladin bacteria residue 200kg is taken, water content 75% sprinkles the calcium oxide of 1kg, be well mixed, after testing material
PH value is 8.52, and once, inspection in the 6th day does not measure bacteria residue gliocladin content for daily turning；It is well mixed plus 10kg corn flour,
Between the moisture 50-60% of regulation material.
Bafillus natto ACCC19833 is accessed into beef-protein medium in inclined-plane culture mode at 35 DEG C is carried out
Fermentation 18h, is then seeded into bacillus natto to ferment culture medium,
Culture medium is constituted：Dregs of beans：18g；White sugar：18g；Calcium carbonate：9g；Sodium chloride：0.54g；Potassium chloride：0.54g；Sulfuric acid
Magnesium 0.54g；Ammonium sulfate：0.9g, manganese sulfate：0.054g；Ferrous sulfate：0.054g, water 1L；PH7.5,121 DEG C of 30mins, 35 DEG C
Culture 48h, the bafillus natto bacterium number of preparation is 5,200,000,000/mL.
Saccharomyces cerevisiae ACCC20039 is accessed into malt extract medium in the way of inclined-plane at 28 DEG C to be activated, Ran Houjie
Plant in fermentation by saccharomyces cerevisiae culture medium,
Culture medium is constituted：Glucose 10g, yeast extract 4.5g, ammonium chloride 1g, magnesium sulfate 0.50g, calcium chloride 0.2g, water 1L；
PH7.5,121 DEG C of 30min, 30 DEG C of cultures 72h, the saccharomyces cerevisiae bacterium number >=1,800,000,000/mL of preparation.
With bafillus natto：Saccharomyces cerevisiae bacteria liquid accumulates ratio=1：5th, during inoculum concentration 5vt% accesses compound, mixing is equal
It is even, in 30-55 DEG C of fermentation, when temperature is higher than 55 DEG C, lowered the temperature by stirring and ventilation.When temperature drops to less than 35 DEG C,
Stop fermentation, the material to having fermented is dried, moisture is less than 9%, the protein content of material reaches 28.6% after testing,
80 mesh sieves were crushed, was packed.
Embodiment 2：
Gliocladin bacteria residue 1000kg is taken, water content 80% sprinkles the calcium oxide of 5kg, be well mixed, after testing material
PH value is 8.52, and once, inspection in the 7th day does not measure bacteria residue gliocladin content for daily turning；It is well mixed plus 20kg corn flour,
Between the moisture 50-60% of regulation material.
Bafillus natto ACCC10614 is accessed into beef-protein medium in inclined-plane culture mode at 35 DEG C is carried out
Fermentation 18h, is then seeded into bacillus natto to ferment culture medium,
Culture medium is constituted：Dregs of beans：18g；White sugar：18g；Calcium carbonate：9g；Sodium chloride：0.54g；Potassium chloride：0.54g；Sulfuric acid
Magnesium 0.54g；Ammonium sulfate：0.9g, manganese sulfate：0.054g；Ferrous sulfate：0.054g, water 1L；PH7.5,121 DEG C of 30mins, 35 DEG C
Culture 48h, the bafillus natto bacterium number of preparation is 4,500,000,000/mL.
Saccharomyces cerevisiae ACCC20042 is accessed into malt extract medium in the way of inclined-plane at 28 DEG C to be activated, Ran Houjie
Plant in fermentation by saccharomyces cerevisiae culture medium,
Culture medium is constituted：Glucose 10g, yeast extract 4.5g, ammonium chloride 1g, magnesium sulfate 0.50g, calcium chloride 0.2g, water 1L；
PH7.5,121 DEG C of 30mins, 30 DEG C of cultures 72h, the saccharomyces cerevisiae bacterium number >=1,500,000,000/mL of preparation.
With bafillus natto：Saccharomyces cerevisiae bacteria liquid accumulates ratio=1：5th, during inoculum concentration 5% accesses compound, mixing is equal
It is even, in 30-55 DEG C of fermentation, when temperature is higher than 55 DEG C, lowered the temperature by stirring and ventilation.When temperature drops to less than 35 DEG C,
Stop fermentation, the material to having fermented is dried, moisture is less than 9%, the protein content of material reaches 30.2% after testing,
80 mesh sieves were crushed, was packed.
Test example
The gliocladin strain test tube slant strain that will be stored in PDA culture medium under the conditions of 4 DEG C is moved under room temperature condition
(20 DEG C -25 DEG C) activation 4h；On aseptic operating platform, the distilled water after being sterilized with 10mL is by through the test tube slant strain of overactivation
Thallus suspension liquid is made, aseptic condition undershoot is washed in the triangular flask equipped with sterilizing seed culture medium, 1 test tube strains inoculation 1
Individual triangular flask, pH value nature, 24 DEG C of cultivation temperature, shaking speed 200r/min, incubation time is 48h；
Seed culture medium constitutes (w/w)：(w/w) is by weight：Glucose 2%, peptone 0.2%, magnesium sulfate
0.05%, potassium dihydrogen phosphate 0.025% adds water to 1000mL；Sterilising conditions are 121 DEG C, 0.15Mpa sterilizings 20min；
It is used for the fermentation of gliocladin, specific culture medium with fermentation materials Substitution for Soybean Meal：
Control group：Glucose sugar 0.2%, dregs of beans 1%, corn flour:2%；Soya-bean oil 2%；Vitamin K 0.005%, lipoic acid
0.003%, magnesium sulfate 0.1%, potassium dihydrogen phosphate 0.05%, zinc sulfate 0.005%, ferrous sulfate 0.005% is added water to
1000mL；Sterilising conditions are 121 DEG C, 0.15Mpa sterilizings 20min；
Experimental group：Glucose sugar 0.2%, fermentation materials 1%, corn flour:2%；Soya-bean oil 2%；Vitamin K 0.005%, sulphur is pungent
Acid 0.003%, magnesium sulfate 0.1%, potassium dihydrogen phosphate 0.05%, zinc sulfate 0.005%, ferrous sulfate 0.005% is added water to
1000mL；Sterilising conditions are 121 DEG C, 0.15Mpa sterilizings 20min；
Liquid seeds are connected in the above-mentioned fermentation mediums of 2L with the inoculum concentration of 10% (v/v), speed of agitator 200r/
Min, 26 DEG C of culture 48h, detects the fermentation level of gliocladin, compares 860mg/L, experimental group 894mg/L, it is seen that use this hair
The fermentation level fermented after bright bacteria residue treatment is higher, and yield is preferable, while realizing the innoxious of gliocladin bacteria residue
Treatment, utilizes for its is secondary and provides better way.
Claims (5)
1. a kind of gliocladin bacterium dregs innocent treatment method, it is characterised in that：Comprise the following steps that：
(1), bacteria residue remains the treatment of gliocladin：The bacteria residue for having extracted gliocladin is well mixed with alkaline matter, makes mixing
The pH value of material is banked up between 8-10, afterwards and is processed 5-10 days, wherein daily turning is once；
(2), the integration of material：The bacteria residue after 70-90 parts for the treatment of is taken by weight, and 10-20 parts of agricultural byproducts are well mixed, regulation
Between the moisture 50-60wt% of material；
(3), fermentation process：With bafillus natto liquid：Saccharomyces cerevisiae bacteria liquid accumulates ratio=1：5th, inoculum concentration 2-10vt% is accessed
In compound, it is well mixed, ferment 72-96h at 30-55 DEG C, when temperature is higher than 55 DEG C, is dropped by stirring and ventilation
Temperature；
(4), last handling process：Material to having fermented is dried, and makes moisture less than 9wt%, and 80 mesh sieves were crushed afterwards i.e.
Can.
2. gliocladin bacterium dregs innocent treatment method according to claim 1, it is characterized in that：Glue in step (1) is mould
Rhzomorph bacteria residue is the fresh wet bacteria slag or dried bacteria residue for having extracted gliocladin；
Described alkaline matter can be one or more mixing in NaOH, potassium hydroxide, calcium oxide, calcium hydroxide
Thing.
3. gliocladin bacterium dregs innocent treatment method according to claim 1, it is characterized in that：Agriculture described in step (2)
Byproduct is one or more mixtures in wheat bran, corn flour, wheat flour.
4. gliocladin bacterium dregs innocent treatment method according to claim 1, it is characterized in that：Receiving described in step (3)
Beans bacillus and saccharomyces cerevisiae are bought in Chinese agriculture Microbiological Culture Collection administrative center, wherein bafillus natto choosing
From ACCC19833 or ACCC10614；Saccharomyces cerevisiae is selected from ACCC20039 or ACCC20042 or ACCC20107.
5. gliocladin bacterium dregs innocent treatment method according to claim 1, it is characterized in that：Receiving described in step (3)
Thalline quantity >=2,000,000,000/mL in beans bacillus bacterium solution, thalline quantity >=500,000,000/mL in saccharomyces cerevisiae bacterium solution.
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Cited By (2)
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| CN107089994A (en) * | 2017-06-08 | 2017-08-25 | 黄河三角洲京博化工研究院有限公司 | A kind of method that gliocladin is reclaimed in the ointment from gliocladin |
| CN112725385A (en) * | 2019-10-28 | 2021-04-30 | 中国石油化工股份有限公司 | Method for preparing long-chain dicarboxylic acid by fermentation |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107089994A (en) * | 2017-06-08 | 2017-08-25 | 黄河三角洲京博化工研究院有限公司 | A kind of method that gliocladin is reclaimed in the ointment from gliocladin |
| CN107089994B (en) * | 2017-06-08 | 2019-04-05 | 黄河三角洲京博化工研究院有限公司 | A method of recycling gliocladin from gliocladin ointment |
| CN112725385A (en) * | 2019-10-28 | 2021-04-30 | 中国石油化工股份有限公司 | Method for preparing long-chain dicarboxylic acid by fermentation |
| CN112725385B (en) * | 2019-10-28 | 2022-09-09 | 中国石油化工股份有限公司 | Method for preparing long-chain dicarboxylic acid by fermentation |
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