CN103352016A - Method for preparing biological fertilizer by utilizing Alteromonas colwelliana A321 to ferment enteromorpha - Google Patents
Method for preparing biological fertilizer by utilizing Alteromonas colwelliana A321 to ferment enteromorpha Download PDFInfo
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- CN103352016A CN103352016A CN2013102784589A CN201310278458A CN103352016A CN 103352016 A CN103352016 A CN 103352016A CN 2013102784589 A CN2013102784589 A CN 2013102784589A CN 201310278458 A CN201310278458 A CN 201310278458A CN 103352016 A CN103352016 A CN 103352016A
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Abstract
The invention relates to a method for preparing a biological fertilizer by utilizing Alteromonas colwelliana A321 to ferment enteromorpha. The enteromorpha biological organic fertilizer is prepared through the processes of slant culture, seed culture, strain culture, preparation of microbial agents, enteromorpha fermentation and preparation. The Alteromonas colwelliana A321 provided by the invention is an autonomously directional induction screened strain and is preserved in CCTCC (China Center for Type Culture Collection) in Wuhan University, with the preservation number of CCTCC NO: M2012132. Characteristic polyose, proteins and cellulose in the intercellular substance and cell walls of the enteromorpha can be efficiently degraded by the microbial agents so as to fully release nutrient elements in the enteromorpha and increase the fermentation speed of the enteromorpha.
Description
Technical field
Biotechnology
Technical background
Enteromorpha is nutritious, and butt contains protein 9～14%, ash content 32～36%, and moisture 6.7～9%, fat 2%, all the other are carbohydrate, approximately 40%.Particularly Enteromorpha has the peculiar Sargassum polysaccharides of marine organisms, highly unsaturated fatty acids, alginic acid and spontaneous growth Auto-regulator etc., is the plant widely distributed in the ocean, that reserves are huge.Particularly because the in recent years frequent outburst of green damp Enteromorpha, annual a large amount of Enteromorpha is salvaged disembarkation and can't effectively be processed, some Enteromorpha is piled up in the waste that resource is not only caused on the bank throughout the year, and the environment around also polluting is unfavorable for the development of the industries such as tourism, fishery.In order to take full advantage of the Enteromorpha resource, solve Enteromorpha to the harm of environment, Enteromorpha to be fermented, the method for producing a kind of Enteromorpha biological organic fertilizer is practical needs.At present, Enteromorpha also is in the exploratory stage to the degraded of Enteromorpha with development and use both at home and abroad as one of plant that has exploitation value in the ocean.
Microbial fermentation microbial inoculum on the market mainly contains now: 1, the synthetic class of nutrient, such as vinelandii, photosynthetic bacterium etc.; 2, the nutrient degraded discharges class, such as cellulose-degrading bacteria, phosphorus bacteria etc.; 3, antibioticly press down sick class, Bacillus licheniformis, bacillus pumilus etc.These microbial inoculums are to research and develop for the degraded of terrestrial plant stalk and waste mostly.Compare terrestrial plant, Enteromorpha contains abundant specific polysaccharide sugar chain, has the effect of coordinate plant growth and enhancing disease resistance, and its content accounts for 10～30% of butt.The monose of sea grass polysaccharide forms and comprises wood sugar, semi-lactosi, pectinose, rhamnosyl and glucose, and these monose are connecting the Arabic croak of the β-L-saccharide residue of muttering by 1 → 4, and the Arabic croak of this β-L-is muttered and had sulfate group on the part C-3 of saccharide residue.Common Lu Sheng microorganism or the zymin sea grass polysaccharide of can't effectively degrading, and then cause existing microbiobacterial agent to be difficult to utilize by sea grass polysaccharide closely frond Mierocrystalline cellulose and the albumen of parcel, therefore can not degrade fully to Enteromorpha.Through retrieval, Zhao Ming etc. utilize the EM microbial inoculum to study Enteromorpha composting condition, and pure Enteromorpha compost temperature is less than 50 ℃.Coptis light etc. utilizes natural bacteria to study Enteromorpha composting condition, and pure Enteromorpha compost top temperature only has 54 ℃, and reaching top temperature needs 7 days.In the patent: Zhu Qiang utilizes complex micro organism fungicide that subtilis, Pseudoalteromonas, bacillus pumilus, Bacillus licheniformis and viride form that the mixture of Enteromorpha and stalk is fermented.Dan Junwei etc. have developed a kind of method of utilizing bacterium, fungi and actinomycetes to form microbiobacterial agent fermentation Enteromorpha.Not about with relevant report and the patent of this patent Related Bacteria fermentation Enteromorpha.
Summary of the invention
The object of the present invention is to provide a kind of marine microorganism microbial inoculum of energy high-efficiency fermenting Enteromorpha biomass and utilize this bacterial strain to prepare the method for Enteromorpha biological organic fertilizer.
The bacterial strain of mentioning among the present invention is the Alteromonas colwelliana A321 that separates in the seawater of Qingdao sea area, alternately Zymomonas mobilis A321 is deposited in Wuhan, China Wuhan University Chinese Typical Representative culture collection center on April 23rd, 2012, and deposit number is CCTCC NO:M2012132.
The present invention uses Alteromonas colwelliana A321 fermentation Enteromorpha, and top temperature can reach 66 ℃, and fermentation in 15 days is finished.Utilize this kind microorganism to ferment and have the characteristics such as intensification is rapid, temperature is high, the cycle is short, tissue degradation is thorough, this bacterial classification Alteromonas colwellianaA321 effect is stable, efficient specificity degraded sea grass polysaccharide, the materials such as wrapped frond Mierocrystalline cellulose and albumen dissociate, effectively discharge the nutritive element of Enteromorpha inside, can effectively promote the growth of crop and the raising of quality.
Embodiment
The production method of Alteromonas colwelliana A321
1, slant culture: under aseptic condition, Alteromonas colwelliana A321 original strain is inoculated on the solid medium 28 ℃ of lower cultivations 3 days.
2, seed culture: be inoculated in liquid nutrient medium under the bacterial classification aseptic condition with step 1 cultivation, 28 ℃, the 200r/min shaking table was cultivated 3 days.The bacteria suspension optical density(OD) OD600 of Alteromonas colwelliana A321 reaches 3.0.
3, spawn culture: be 8% inoculum size by the volume ratio of liquid nutrient medium, seed liquor is inoculated in the fermentor tank, 28 ℃, 200r/min cultivated 3 days.
4, the preparation of microbial inoculum: with the strain fermentating liquid that makes, add supporting agent and carry out drying by spray drying technology, make Alteromonas colwelliana A321 microbial inoculum.
Wherein, the used substratum of Alteromonas colwelliana A321 is the sea grass polysaccharide substratum in step 1～3: sea grass polysaccharide 10g, sodium-chlor 10g, calcium chloride 0.1g, dipotassium hydrogen phosphate 0.5g, sal epsom 0.5g, ammonium chloride 1g, high ferric phosphate 0.02g, ferric sulfate 0.01g, water 1000mL, pH7.0.Solid medium is to have added an amount of agar gained in liquid culture medium.
The fermenting enteromorpha method
Raw material: Enteromorpha.Alteromonas colwelliana A321 bacterium inoculum size: 0.5%.
1, the pre-treatment of microbial inoculum: use first water dissolution, fully stir.
2, batching: material water 1: 2.0, C/N value 30: 1, pH7.5, material stack height 60cm.
3, ferment: Alteromonas colwelliana A321 is evenly splashed by 0.5% inoculum size treating on the fermented product, and abundant mixing.In the fermenting process, turning in per 3 days once, continuously turning is 5 times, fermentation in 15 days is finished.
The characteristic product of Alteromonas colwelliana A321 degraded Enteromorpha detects
1, sample pretreatment: take by weighing 10g fermenting enteromorpha end product, in dissolving and the 90mL distilled water.Add the 400mL dehydrated alcohol, leave standstill 30min, centrifugal 10 minutes of 3000rpm collects supernatant liquor and rotary evaporation and is concentrated into 10mL, obtains the feature oligosaccharides concentrated solution of Alteromonas colwelliana A321 degraded Enteromorpha.
2, get this concentrated solution and carry out GC and ESI-MS analysis, obtain specificity product---the sulfation rhamnosyl of Alteromonas colwelliana A321 degraded Enteromorpha, its molecular weight is 244Da.This product can be used as the characteristics index of identifying this strain fermentation Enteromorpha.
Claims (3)
1. a strain bacterium Alteromonas colwellianaA321, this culture presevation unit: Chinese Typical Representative culture collection center.Address: Wuhan, China, Wuhan University.Preservation date is on 04 23rd, 2012, and preserving number is M2012132.
2. utilize Alteromonas colwelliana A321 fermentation Enteromorpha to produce bio-fertilizer.
3. fermentation Enteromorpha claimed in claim 2, the property material that its product contains is the sulfation rhamnosyl, molecular weight is 244Da.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105602995A (en) * | 2016-01-21 | 2016-05-25 | 青岛海洋生物医药研究院股份有限公司 | Method for preparing Enteromorpha bio-fertilizer through liquid deep quick fermentation |
| CN108239176A (en) * | 2016-12-27 | 2018-07-03 | 中国海洋大学 | Low molecular weight sea grass polysaccharide and preparation method thereof, sulphation low molecular weight sea grass polysaccharide and preparation method thereof and application |
| CN113429227A (en) * | 2021-08-24 | 2021-09-24 | 烟台上水医药科技有限公司 | Application of bacillus subtilis in biological fermentation |
| CN114410542A (en) * | 2022-02-15 | 2022-04-29 | 青岛中尚奇生物科技有限公司 | Vibrio sky blue and application thereof in enteromorpha degradation |
| CN114480201A (en) * | 2022-02-15 | 2022-05-13 | 青岛中尚奇生物科技有限公司 | Vibrio natriegens capable of strongly degrading enteromorpha protein and application of vibrio natriegens in preparation of organic fertilizer |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105602995A (en) * | 2016-01-21 | 2016-05-25 | 青岛海洋生物医药研究院股份有限公司 | Method for preparing Enteromorpha bio-fertilizer through liquid deep quick fermentation |
| CN105602995B (en) * | 2016-01-21 | 2019-05-17 | 青岛海洋生物医药研究院股份有限公司 | A kind of method that deep liquid Rapid Fermentation prepares Enteromorpha bio-fertilizer |
| CN108239176A (en) * | 2016-12-27 | 2018-07-03 | 中国海洋大学 | Low molecular weight sea grass polysaccharide and preparation method thereof, sulphation low molecular weight sea grass polysaccharide and preparation method thereof and application |
| CN108239176B (en) * | 2016-12-27 | 2021-08-17 | 中国海洋大学 | Low-molecular-weight enteromorpha polysaccharide and preparation method thereof, sulfated low-molecular-weight enteromorpha polysaccharide and preparation method and application thereof |
| CN113429227A (en) * | 2021-08-24 | 2021-09-24 | 烟台上水医药科技有限公司 | Application of bacillus subtilis in biological fermentation |
| CN114410542A (en) * | 2022-02-15 | 2022-04-29 | 青岛中尚奇生物科技有限公司 | Vibrio sky blue and application thereof in enteromorpha degradation |
| CN114480201A (en) * | 2022-02-15 | 2022-05-13 | 青岛中尚奇生物科技有限公司 | Vibrio natriegens capable of strongly degrading enteromorpha protein and application of vibrio natriegens in preparation of organic fertilizer |
| CN114410542B (en) * | 2022-02-15 | 2023-08-11 | 青岛中尚奇生物科技有限公司 | Vibrio paradise and application thereof in degradation of enteromorpha |
| CN114480201B (en) * | 2022-02-15 | 2023-08-11 | 青岛中尚奇生物科技有限公司 | Vibrio natriurus capable of strongly degrading enteromorpha protein and application of vibrio natriurus in preparation of organic fertilizer |
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| CN103352016B (en) | 2015-03-11 |
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