CN109355346A - A kind of method of apple pomace semisolid fermentation production biochemical fulvic acid - Google Patents
A kind of method of apple pomace semisolid fermentation production biochemical fulvic acid Download PDFInfo
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- CN109355346A CN109355346A CN201811414770.5A CN201811414770A CN109355346A CN 109355346 A CN109355346 A CN 109355346A CN 201811414770 A CN201811414770 A CN 201811414770A CN 109355346 A CN109355346 A CN 109355346A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
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Abstract
The present invention provides a kind of methods of apple pomace semisolid fermentation production biochemical fulvic acid, the present invention in discarded pomace by accessing the various fermenting microbes expanded after cultivating, semisolid fermentation is carried out to produce biochemical fulvic acid, and biochemical fulvic acid solution is made using mix bacterium agent inoculum concentration, urea additive amount, fermentation substrate water content and fermentation time as variable during the fermentation.This method can not only reduce the burden of apple relevant enterprise; protect environment; also by the cost of low production biochemical fulvic acid; use optimal zymotechnique; it can be made into multi-functional, high efficiency and extensively applicable biochemical fulvic acid solution, there is practical reference significance to semisolid fermentation production biochemical fulvic acid.
Description
Technical field
The present invention relates to field of biotechnology, the side of specifically a kind of apple pomace semisolid fermentation production biochemical fulvic acid
Method.
Background technique
Fulvic acid is water-soluble, the highest part of activity in humic acid material, and initial fulvic acid is from humic acid
Middle extraction and obtain, but the chemical substances such as a large amount of acid, alkali need to be consumed, some also needs high temperature, high pressure and special installation, consumes greatly
The energy of amount, while waste water, exhaust gas are generated, it pollutes the environment, production cost is higher.Using microbial fermentation technology from agriculture
The renewable resources such as woods waste extract yellow corruption by the rotten bacterium of biotechnology inoculation speed, composting, fermentation and chemical extraction technology etc.
Acid, this fulvic acid are also referred to as biochemical fulvic acid.Biochemical fulvic acid compared with the natural fulvic acid extracted in peat, lignite,
Molecular weight is small, good water solubility, and bioactivity is high, and acid-fast alkali-proof, anti-divalent ion does not flocculate under various water quality, with N, P, K and
Microelement cosolvency is good.
Eighties of last century the eighties latter stage, the production and application of biochemical fulvic acid are surging forward and are developed.It grinds
Study carefully report, after biochemical fulvic acid handles soil, the granule content in soil greater than 0.25mm increases 8.5%~20.0%, table
Open-birth Yellow humic acid fertilizer has the function of being obviously improved soil enzyme activity.Biochemical fulvic acid not only has stronger physiology living
Property, C, N and water metabolism have facilitation in the development and plant to root system, while by reinforcing soil microbial activities,
Improve soil chemical properties, activating soil nutrient discharges the nutrients such as N, P, K gradually with complex state, to improve fertilizer benefit
With rate.Biochemical fulvic acid can also improve the drought-resistant ability of crop, increase amonifying bacteria in soil, solid ammonia bacterium, cellulose-decomposing bacteria
Equal quantity of useful microbe and chlorophyll content of plant etc..
In terms of forestry, the biochemical fulvic acid solution of various concentration soaks root to Chinese pine, pinus sylvestris var. mongolica, Larix principis-rupprechtii and dragon spruce
Processing, can be improved the surrival rate of afforestation 11.9~16.3%；Dragon spruce, the Eurya plant surrival rate of afforestation can be respectively increased by pouring root
11.5%, 10.5%.Change alkali fertilizer in western Forest-tree culture with biochemical fulvic acid to test, the results showed that be used for Xinjiang poplar seedling
Wood, nursery stock height and ground diameter rough segmentation do not increase 49cm, 0.49cm than control, and survival rate improves 8.7%, and shoot length increases
3.7cm, is used for dragon spruce, arbor-vitae young growth, and annual plant height increases 2.9 and 4.9cm than control respectively；The fertilizer can not only promote seedling
Wood and young ebon growth, improve the surrival rate of afforestation, also have positive effect to improvement nursery and reproducing area.
Using agricultural waste material fermenting and producing biochemical fulvic acid, waste can be made to be utilized, dirt not will cause to environment
Dye, raw material is easy to get, at low cost, does not generate any pollution to environment in the production and use process, product can be produced in agriculture and forestry
In show the advantages that multi-functional, high efficiency and applicability are wide, be a kind of extremely promising sustainable development Eco-agro-forestry
New bio technology.
Summary of the invention
The present invention provides a kind of low costs, the side of the apple pomace semisolid fermentation production biochemical fulvic acid of scientific matching
Method.
The present invention provides a kind of method of apple pomace semisolid fermentation production biochemical fulvic acid, step includes:
(1) fermentation medium of 40%-80% water content is made using apple pomace as fermentation raw material；
(2) it takes 20g glucose, 200g potato, 15-20g agar, 1000ml water to mix and be sufficiently stirred and nature pH is made
Potato dextrose medium, take 20g glucose, 100g potato, 15g agar, 1000ml water to mix and system be sufficiently stirred
At the bean sprouts medium of 7.2-7.4pH, take 10g peptone, 3g beef extract, 5g calcium chloride, 15-20g agar, 1000ml water mixed
The beef-protein medium that 7.0-7.2pH is made is sufficiently stirred in merging；
(3) it at 28 DEG C, with potato dextrose medium culture aspergillus niger 48h, is made wine with bean sprouts medium culture
Yeast and candida utili for 24 hours, under the conditions of 30 DEG C, with beef-protein medium culture bacillus subtilis and vacation list
Born of the same parents bacillus is for 24 hours, stand-by after overactivation by above-mentioned five kinds of strains；
(4) by aspergillus niger, candida utili, saccharomyces cerevisiae, bacillus subtilis and the vacation list after activation in step (3)
Born of the same parents bacillus, which is compounded with the inoculative proportion of 1:1:1:2:2 and carries out culture with expansion culture medium, is made mixed microorganism microbial inoculum,
Wherein, expand the incubation of culture medium are as follows: mould is cultivated for 24 hours for 30 DEG C of constant temperature in shaking table with revolving speed 220rpm, and yeast is shaking
12h is cultivated with revolving speed 180rpm 30 DEG C of constant temperature in bed, bacterium cultivates 12h in shaking table with revolving speed 180rpm for 30 DEG C of constant temperature, above
This five kinds of strains obtain corresponding seed liquor after cultivation, take 10% seed liquor to expand culture 48h in fermentation flask and are fermented
Fermentation liquid and sterilizing wheat bran are carried out mixed fermentation with 1:1 ratio, dry 1d-3d, mixed microorganism microbial inoculum is made by liquid；
(5) urea of the mixed microorganism microbial inoculum, certain additive amount that take certain inoculum concentration is linked into the fermentation in step (1)
Culture medium fermentation a period of time, screenings separates after fermentation, and fermentation liquid is biochemical fulvic acid solution.
Particularly, above-mentioned expansion culture medium refers to the remainder potato dextrose medium after removing agar, bean sprouts
Juice culture medium and beef-protein medium.
Particularly, the inoculum concentration of above-mentioned mixed microorganism microbial inoculum is 3%-7%.
Particularly, the additive amount of above-mentioned urea is 1%-3%.
Particularly, the fermentation time in above-mentioned fermentation medium is 20d-35d.
The present invention accesses the various fermenting microbes expanded after culture in discarded pomace, carries out semisolid fermentation to produce
Biochemical fulvic acid, and during the fermentation with mix bacterium agent inoculum concentration, urea additive amount, fermentation substrate water content and fermentation time
For variable, biochemical fulvic acid solution is made.This method can not only reduce the burden of apple relevant enterprise, protect environment, also
The cost of low production biochemical fulvic acid be can be made into multi-functional, high efficiency and extensively applicable biochemistry using optimal zymotechnique
Fulvic acid solution has practical reference significance to semisolid fermentation production biochemical fulvic acid.
Specific embodiment
For a further understanding of the present invention, the preferred embodiment of the invention is described below with reference to embodiment, still
It should be appreciated that these descriptions are only further explanation the features and advantages of the present invention, rather than to the claims in the present invention
Limitation.
Embodiment one
A kind of method of apple pomace semisolid fermentation production biochemical fulvic acid, step include:
(1) fermentation medium of 40%-80% water content is made using apple pomace as fermentation raw material；
(2) it takes 20g glucose, 200g potato, 15-20g agar, 1000ml water to mix and be sufficiently stirred and nature pH is made
Potato dextrose medium, take 20g glucose, 100g potato, 15g agar, 1000ml water to mix and system be sufficiently stirred
At the bean sprouts medium of 7.2-7.4pH, take 10g peptone, 3g beef extract, 5g calcium chloride, 15-20g agar, 1000ml water mixed
The beef-protein medium that 7.0-7.2pH is made is sufficiently stirred in merging；
(3) it at 28 DEG C, with potato dextrose medium culture aspergillus niger 48h, is made wine with bean sprouts medium culture
Yeast and candida utili for 24 hours, under the conditions of 30 DEG C, with beef-protein medium culture bacillus subtilis and vacation list
Born of the same parents bacillus is for 24 hours, stand-by after overactivation by above-mentioned five kinds of strains；
(4) by aspergillus niger, candida utili, saccharomyces cerevisiae, bacillus subtilis and the vacation list after activation in step (3)
Born of the same parents bacillus, which is compounded with the inoculative proportion of 1:1:1:2:2 and carries out culture with expansion culture medium, is made mixed microorganism microbial inoculum,
Wherein, expand the incubation of culture medium are as follows: mould is cultivated for 24 hours for 30 DEG C of constant temperature in shaking table with revolving speed 220rpm, and yeast is shaking
12h is cultivated with revolving speed 180rpm 30 DEG C of constant temperature in bed, bacterium cultivates 12h in shaking table with revolving speed 180rpm for 30 DEG C of constant temperature, above
This five kinds of strains obtain corresponding seed liquor after cultivation, take 10% seed liquor to expand culture 48h in fermentation flask and are fermented
Fermentation liquid and sterilizing wheat bran are carried out mixed fermentation with 1:1 ratio, dry 1d-3d, mixed microorganism microbial inoculum is made by liquid；
(5) urea of the mixed microorganism microbial inoculum, 1%-3% additive amount that take 3%-7% inoculum concentration is linked into step (1)
Fermentation medium ferment 20d-35d, after fermentation screenings separate, fermentation liquid is biochemical fulvic acid solution.
According to different mixed microorganism microbial inoculum inoculum concentrations, different urea additive amounts, different fermentation medium fermentations
The measurement of the progress fulvic acid content of biochemical fulvic acid solution made of time.Measuring method is potassium bichromate titrimetric method, operation
Method is as described below:
It takes 0.2g sample to be placed in 250ml conical flask, adds distilled water, small glass funnel is inserted on bottleneck, in boiling water
Ceaselessly agitation heating 30min pours into all solution and residue in 100ml volumetric flask after taking out cooling, and distilled water is added extremely to carve
It spends and shakes up, then with Medium speed filter paper filtering residue to obtain filtrate.It takes in the above-mentioned filtrate merging 250ml conical flask of 5ml, adds
Enter 4.8mol/L potassium bichromate solution 5ml, concentrated sulfuric acid 15ml is added, 30min is heated in boiling water makes its oxidation.Finally, from boiling
The solution that oxidation obtains is taken out in water-bath, is cooled to room temperature, and distilled water 70ml and adjacent Féraud beautiful jade indicator 3 is added to this solution
Drop, is titrated with ferrous sulfate standard solution, when solution stops titration from orange through green and when becoming brick-red.It calculates yellow
The formula of rotten acid content are as follows:
In formula: V0Refer to the ferrous sulfate volume consumed in titration blank tube；V refers to the sulfuric acid consumed in titration sample cell
Ferrous volume；N refers to the molar concentration of ferrous sulfate standard solution；C refers to (generally 0.5) containing carbon ratio of fulvic acid；G refers to
The sample quality being added when preparing sample solution；A refers to the volume of sample solution titration；B refers to that the sample drawn when titration is molten
Liquid product.
Obtained result such as table 1, it can be seen that 7% inoculum concentration, 50% fermentation substrate water content, 2% urea add
Dosage, 30d fermentation time be optimum process, the content of fulvic acid is more than 20% in tunning.
Table 1
Claims (5)
1. a kind of method of apple pomace semisolid fermentation production biochemical fulvic acid, step include:
(1) fermentation medium of 40%-80% water content is made using apple pomace as fermentation raw material；
(2) it takes 20g glucose, 200g potato, 15-20g agar, 1000ml water to mix and the horse that nature pH is made is sufficiently stirred
Bell potato dextrose culture-medium takes 20g glucose, 100g potato, 15g agar, 1000ml water to mix and be sufficiently stirred and be made
The bean sprouts medium of 7.2-7.4pH takes 10g peptone, 3g beef extract, 5g calcium chloride, 15-20g agar, the mixing of 1000ml water
And the beef-protein medium that 7.0-7.2pH is made is sufficiently stirred；
(3) at 28 DEG C, with potato dextrose medium culture aspergillus niger 48h, with bean sprouts medium culture saccharomyces cerevisiae
For 24 hours with candida utili, under the conditions of 30 DEG C, with beef-protein medium culture bacillus subtilis and false unit cell bar
Bacterium is for 24 hours, stand-by after overactivation by above-mentioned five kinds of strains；
(4) by aspergillus niger, candida utili, saccharomyces cerevisiae, bacillus subtilis and the false unit cell bar after activation in step (3)
Bacterium is carried out compounding and is carried out culture with expansion culture medium that mixed microorganism microbial inoculum is made with the inoculative proportion of 1:1:1:2:2, wherein
Expand the incubation of culture medium are as follows: mould is cultivated for 24 hours for 30 DEG C of constant temperature in shaking table with revolving speed 220rpm, and yeast is permanent in shaking table
30 DEG C of temperature cultivates 12h with revolving speed 180rpm, and bacterium 30 DEG C of constant temperature in shaking table with revolving speed 180rpm cultivates 12h, and above this five kinds
Strain obtains corresponding seed liquor after cultivation, takes 10% seed liquor to expand culture 48h in fermentation flask and obtains fermentation liquid, with 1:
Fermentation liquid and sterilizing wheat bran are carried out mixed fermentation by 1 ratio, dry 1d-3d, mixed microorganism microbial inoculum is made；
(5) urea of the mixed microorganism microbial inoculum, certain additive amount that take certain inoculum concentration is linked into the fermented and cultured in step (1)
Base fermentation a period of time, screenings separates after fermentation, and fermentation liquid is biochemical fulvic acid solution.
2. a kind of method of apple pomace semisolid fermentation production biochemical fulvic acid according to claim 1, it is characterised in that:
The culture medium that expands refers to remainder potato dextrose medium, bean sprouts medium and beef extract after removing agar
Peptone culture medium.
3. a kind of method of apple pomace semisolid fermentation production biochemical fulvic acid according to claim 1, it is characterised in that:
The inoculum concentration of the mixed microorganism microbial inoculum is 3%-7%.
4. a kind of method of apple pomace semisolid fermentation production biochemical fulvic acid according to claim 1, it is characterised in that:
The additive amount of the urea is 1%-3%.
5. a kind of method of apple pomace semisolid fermentation production biochemical fulvic acid according to claim 1, it is characterised in that:
Fermentation time in fermentation medium is 20d-35d.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110616246A (en) * | 2019-10-11 | 2019-12-27 | 北京大伟嘉生物技术股份有限公司 | Fermentation method for increasing content of sodium fulvate and microbial agent thereof |
| CN113331211A (en) * | 2021-06-04 | 2021-09-03 | 广西壮族自治区亚热带作物研究所（广西亚热带农产品加工研究所） | Medicament for preventing and treating passion fruit stem basal rot and preparation method thereof |
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| CN1974490A (en) * | 2006-12-11 | 2007-06-06 | 陈五岭 | Process of fermenting waste from farm and sideline product processing to produce fulvic acid bacterial manure |
| CN107574136A (en) * | 2017-10-30 | 2018-01-12 | 中国水产科学研究院淡水渔业研究中心 | A kind of preparation method of semi-solid probiotics used for aquiculture |
| CN108587944A (en) * | 2018-04-04 | 2018-09-28 | 北京航天恒丰科技股份有限公司 | A kind of composite bacteria agent and preparation method thereof for the fermentation of agricultural waste gurry |
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- 2018-11-26 CN CN201811414770.5A patent/CN109355346B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1974490A (en) * | 2006-12-11 | 2007-06-06 | 陈五岭 | Process of fermenting waste from farm and sideline product processing to produce fulvic acid bacterial manure |
| CN107574136A (en) * | 2017-10-30 | 2018-01-12 | 中国水产科学研究院淡水渔业研究中心 | A kind of preparation method of semi-solid probiotics used for aquiculture |
| CN108587944A (en) * | 2018-04-04 | 2018-09-28 | 北京航天恒丰科技股份有限公司 | A kind of composite bacteria agent and preparation method thereof for the fermentation of agricultural waste gurry |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110616246A (en) * | 2019-10-11 | 2019-12-27 | 北京大伟嘉生物技术股份有限公司 | Fermentation method for increasing content of sodium fulvate and microbial agent thereof |
| CN113331211A (en) * | 2021-06-04 | 2021-09-03 | 广西壮族自治区亚热带作物研究所（广西亚热带农产品加工研究所） | Medicament for preventing and treating passion fruit stem basal rot and preparation method thereof |
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Effective date of registration: 20221130 Address after: 337000 Ma Shan Zhen Xiao Qiao Cun, Xiangdong District, Pingxiang City, Jiangxi Province Patentee after: Jiangxi Sanhui Technology Co.,Ltd. Address before: 337000 Ma Shan Zhen Xiao Qiao Cun, Xiangdong District, Pingxiang City, Jiangxi Province Patentee before: PINGXIANG LELE HUMIC ACID FACTORY |