CN103352016B - 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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- CN103352016B CN103352016B CN201310278458.9A CN201310278458A CN103352016B CN 103352016 B CN103352016 B CN 103352016B CN 201310278458 A CN201310278458 A CN 201310278458A CN 103352016 B CN103352016 B CN 103352016B
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Abstract
The invention relates to a method for preparing a biological fertilizer by utilizing Alteromonas cdwelliana 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 cdwelliana 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%, moisture 6.7 ~ 9%, fat 2%, and all the other are carbohydrate, and about 40%.Particularly Enteromorpha has Sargassum polysaccharides specific to marine organisms, highly unsaturated fatty acids, alginic acid and spontaneous growth Auto-regulator etc., is widely distributed in ocean, that reserves are huge plant.Particularly due to the frequent outburst of green damp Enteromorpha in recent years, the salvaged disembarkation of Enteromorphas a large amount of every year and cannot effectively processing, some Enteromorpha is piled up in the waste that resource is not only caused on bank throughout the year, also pollutes the environment of surrounding, is unfavorable for the development of the industry such as tourism, fishery.In order to make full use of Enteromorpha resource, solving Enteromorpha to the harm of environment, being fermented by Enteromorpha, a kind of method producing Enteromorpha biological organic fertilizer is practical needs.At present, Enteromorpha, as one of plant with Development volue in ocean, is also in the exploratory stage to the degraded of Enteromorpha with exploitation both at home and abroad.
Now microbial fermentation inoculum commercially mainly contains: 1, nutrient synthesis class, as vinelandii, photosynthetic bacterium etc.; 2, nutrient degraded release class, as cellulose-degrading bacteria, phosphorus bacteria etc.; 3, antibacterial press down sick class, Bacillus licheniformis, bacillus pumilus etc.These microbial inoculums are mostly degraded for terrestrial plant stalk and waste and research and develop.Compare terrestrial plant, Enteromorpha contains abundant specific polysaccharide sugar chain, and have the effect of coordinate plant growth and enhancing disease resistance, its content accounts for 10 ~ 30% of butt.The monose composition of sea grass polysaccharide comprises wood sugar, semi-lactosi, pectinose, rhamnosyl and glucose, and these monose are connected to the Arabic croak of β-L-by 1 → 4 and mutter saccharide residue, and the Arabic croak of this β-L-is muttered saccharide residue part C-3 on there is sulfate group.Common Lu Sheng microorganism or zymin cannot effectively be degraded sea grass polysaccharide, and then cause existing microbiobacterial agent to be difficult to utilize the frond Mierocrystalline cellulose and albumen closely wrapped up by sea grass polysaccharide, therefore can not degrade fully to Enteromorpha.Through retrieval, Zhao Ming etc. utilize EM microbial inoculum to have studied Enteromorpha composting condition, and pure Enteromorpha compost temperature is less than 50 DEG C.Coptis light etc. utilizes natural bacteria to have studied Enteromorpha composting condition, and pure Enteromorpha compost top temperature only has 54 DEG C, and reaching top temperature needs 7 days.In patent: the mixture of complex micro organism fungicide to Enteromorpha and stalk that Zhu Qiang utilizes subtilis, Pseudoalteromonas, bacillus pumilus, Bacillus licheniformis and viride to form ferments.Dan Junwei etc. develop a kind of method utilizing bacterium, fungi and actinomycetes to form microbiobacterial agent fermentation Enteromorpha.Not not ferment the relevant report of Enteromorpha and patent about with this patent Related Bacteria.
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 mentioned in the present invention is Alteromonas colwelliana (the examining formula Alteromonad) A321 be separated in the seawater of Qingdao sea area, this Alteromonad A321 is deposited in Wuhan, China Wuhan University China typical culture collection center on April 23rd, 2012, and deposit number is CCTCC NO:M 2012132.
The present invention's application Kao Shi Alteromonad A321 ferments Enteromorpha, and top temperature can reach 66 DEG C, within 15 days, has fermented.Utilize this kind of microorganism to carry out fermentation and there is features such as heating up rapidly, temperature is high, the cycle is short, tissue degradation is thorough, this bacterial classification Kao Shi Alteromonad A321 effect is stablized, can efficient selective degradation sea grass polysaccharide, dissociate the materials such as wrapped frond Mierocrystalline cellulose and albumen, the nutritive element of effective release Enteromorpha inside, can promote the growth of crop and the raising of quality effectively.
Embodiment
The production method of Kao Shi Alteromonad A321
1, slant culture: aseptically, is inoculated on solid medium by Kao Shi Alteromonad A321 original strain, cultivates 3 days at 28 DEG C.
2, seed culture: inoculate sub-liquid nutrient medium under bacterial classification aseptic condition step 1 cultivated, 28 DEG C, 200r/min shaking table cultivates 3 days.The bacteria suspension optical density(OD) OD600 of Kao Shi Alteromonad A321 reaches 3.0.
3, spawn culture: be the inoculum size of 8% by the volume ratio of liquid nutrient medium, seed liquor be inoculated in fermentor tank, 28 DEG C, 200r/min cultivates 3 days.
4, the preparation of microbial inoculum: by obtained strain fermentating liquid, adds supporting agent and carries out drying by spray drying technology, obtained Kao Shi Alteromonad A321 microbial inoculum.
Wherein, the substratum that in step 1 ~ 3, Kao Shi Alteromonad A321 is used is sea grass polysaccharide substratum: sea grass polysaccharide 10g, sodium-chlor 10g, calcium chloride 0.1g, dipotassium hydrogen phosphate 0.5g, magnesium sulfate 0.5g, ammonium chloride 1g, high ferric phosphate 0.02g, ferric sulfate 0.01g, water 1000mL, pH7.0.Solid medium is in liquid culture medium, with the addition of appropriate agar gained.
Fermenting enteromorpha method
Raw material: Enteromorpha.Kao Shi Alteromonad A321 bacterium inoculum size: 0.5%.
1, the pre-treatment of microbial inoculum: first use water dissolution, fully stir.
2, prepare burden: material water 1: 2.0, C/N value 30: 1, pH 7.5, material stack height 60cm.
3, ferment: by Kao Shi Alteromonad A321 by 0.5% inoculum size evenly splash and treating on fermented product, and fully to mix.In fermenting process, once, continuous turning 5 times, has fermented for 15 days in turning in every 3 days.
The distinctive products of Kao Shi Alteromonad A321 degraded Enteromorpha detects
1, sample pretreatment: take 10g fermenting enteromorpha end product, dissolves with 90mL distilled water.Add 400mL dehydrated alcohol, centrifugal 10 minutes of standing 30min, 3000rpm, collect supernatant liquor and rotary evaporation is concentrated into 10ml, obtains the feature oligosaccharides concentrated solution of Kao Shi Alteromonad A321 degraded Enteromorpha.
2, get this concentrated solution and carry out GC and ESI-MS analysis, obtain specific product---the sulfation rhamnosyl of Kao Shi Alteromonad A321 degraded Enteromorpha, its molecular weight is 244Da.This product can be used as the characteristics index of this strain fermentation Enteromorpha of qualification.
Claims (1)
1. prepare a method for bio-fertilizer, it is characterized in that, the Alteromonascolwelliana A321 fermentation Enteromorpha utilizing preserving number to be M2012132 produces bio-fertilizer.
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| CN201310278458.9A CN103352016B (en) | 2013-06-27 | 2013-06-27 | Method for preparing biological fertilizer by utilizing Alteromonas colwelliana A321 to ferment enteromorpha |
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| CN201310278458.9A CN103352016B (en) | 2013-06-27 | 2013-06-27 | Method for preparing biological fertilizer by utilizing Alteromonas colwelliana A321 to ferment enteromorpha |
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| CN103352016B true CN103352016B (en) | 2015-03-11 |
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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| 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 |
| 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 |
| 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 |
| 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 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1281732C (en) * | 2004-03-02 | 2006-10-25 | 中国海洋大学 | Production of agar-agar enzyme by Alteromonas sp.nov.SY 37-12 strain |
| CN101486612A (en) * | 2009-03-02 | 2009-07-22 | 中国海洋大学生物工程开发有限公司 | Method producing seaweed fertilizer by processing Entermorpha prolifera by microbial fermentation method |
| CN102604868B (en) * | 2012-03-21 | 2013-05-29 | 清华大学深圳研究生院 | Alteromonas and application thereof in inhibiting growth of red-tide algae |
| CN103451119B (en) * | 2012-06-03 | 2015-06-17 | 中国海洋大学 | Alteromonas and method thereby for producing gel-type enteromorpha polysaccharide degrading enzyme by using Alteromonas |
| CN102813057B (en) * | 2012-09-17 | 2014-11-26 | 中国水产科学研究院渔业机械仪器研究所 | Method for producing fermented feed through utilizing enteromorpha leaching liquor |
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