CN103642722B - Nicotine-degrading bacterium and application thereof - Google Patents
Nicotine-degrading bacterium and application thereof Download PDFInfo
- Publication number
- CN103642722B CN103642722B CN201310616712.1A CN201310616712A CN103642722B CN 103642722 B CN103642722 B CN 103642722B CN 201310616712 A CN201310616712 A CN 201310616712A CN 103642722 B CN103642722 B CN 103642722B
- Authority
- CN
- China
- Prior art keywords
- nicotine
- nicotine degradation
- bacterium
- degradation bacterium
- heap body
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Abstract
The invention relates to a nicotine-degrading bacterium and an application thereof. The nicotine-degrading bacterium is characterized in that the bacterium is named as Arthrobacter histidinolovorans EA-17 according to taxonomy and was collected in China Center for Type Culture Collection on September 15, 2013, and the collection number is CCTCC No. M2013405. A strain is fermented by a 2M3 fermentation tank for production, then mixed with a self-made decomposition agent according to the mass ratio of 1:10, further inoculated into a tobacco straw compost according to the inoculation amount of 1% and fermented for 30-45d, and then straw can be decomposed; compared with traditional compost fermentation, the nicotine degradation rate is improved by 60%-78%, the total nitrogen content is increased by 7%-9%, the total phosphorus content is increased by 0.3%-0.6%, and the total potassium content is increased by 6%-8%.
Description
Technical field
The present invention relates to a kind of nicotine degradation bacterium and application thereof.
Background technology
In tobacco planting is produced, there is the generation of a large amount of these agricultural wastes of cigarette stalk.Cigarette stalk contains the malignant pollution material nicotine of high density, and carries pathogenic bacteria and worm's ovum etc., a large amount of cigarette stalk is abandoned or is burned by soil serious for generation, topsoil and the propagation increasing the weight of soil-borne disease.So rationally process cigarette stalk, making discarded cigarette stalk get utilization, is the basis of tobacco planting benign development.Cigarette stalk is rich in plant desired nutritional element and the organic matters such as N, P, K, utilizes microbiological deterioration cigarette stalk can be changed into the organic fertilizer of high-quality by During High-Temperature Composting technology.And the hot environment of During High-Temperature Composting can kill various pathogenic bacteria in cigarette stalk and parasite, reduce the also generation of disease and pest and the appearance of weeds in field ensuing crop process of growth of cigarette stalk fertilizer.So the Waste Composting of cigarette stalk utilizes has good actual application value in the discarded cigarette stalk of process.
The crop material During High-Temperature Composting comprising cigarette stalk is at present become thoroughly decomposed, and mainly utilizes the nature microorganism of energy decomposing lignocellulose maybe can promote the complex micro organism fungicide fermented.But tabacco straw has its singularity, it contains higher concentration nicotine, nicotine itself can suppress microbial growth, in addition, because nicotine itself is a kind of alkaloid, the existence of a large amount of nicotine in cigarette stalk, make the pH of cigarette stalk in alkalescence, general more than 8.5, higher pH also can affect microbial growth in cigarette stalk heap body, thus affects become thoroughly decomposed speed and the degree of humification etc. of cigarette stalk compost.
Summary of the invention
The present invention seeks in order to the problems such as microorganism suppresses by cigarette stalk middle and high concentration nicotine in cigarette stalk composting process, cigarette stalk maturity delays, quality of compost products reduction that solve prior art, utilize nicotine degradation bacterium to accelerate tabacco straw to become thoroughly decomposed, improve compost quality.
A kind of nicotine degradation bacterium of the present invention, it is characterized in that nicotine degradation bacterium, this bacterium taxonomy called after bites histidinol Arthrobacter (Arthrobacter histidinolovorans) EA-17, be preserved in Luo Jia Shan, wuchang, wuhan Wuhan University China typical culture collection center on September 10th, 2013, preserving number is CCTCCM2013405.
A kind of nicotine degradation bacterium of the present invention, it is characterized in that nicotine degradation bacterium EA-17 industrial production is second order fermentation, its production method is: activated spawn is connected to LB liquid nutrient medium, cultivates 10-18h, is prepared into first order seed for 30-37 DEG C; Again one-level kind is inoculated in the 2M that LB liquid medium is housed with the inoculum size of 0.1%-0.5%
3in fermentor tank, fermentor tank liquid amount is 60%, 30-37 DEG C, mixing speed 250rpm, ventilation ratio 0.2VVM-1VVM, and fermentation 16-24h, biomass reaches 3 × 10
9cFU/mL.
The application of a kind of nicotine degradation bacterium of the present invention in cigarette stalk heap body.
The application of a kind of nicotine degradation bacterium of the present invention in cigarette stalk heap body, it is characterized in that described nicotine degradation bacterium EA-17 use-pattern is: after nicotine degradation fermented liquid mixes with the mass ratio of 1:10 with self-control decomposing agent, by the inoculum size access cigarette stalk heap body of 1%, keep heap body moisture 60%-65%, fermentation 30-45d, can become thoroughly decomposed, described self-control decomposing agent contains aspergillus niger spore 10
7individual/g, subtilis 10
8cFU/g.
The application of a kind of nicotine degradation bacterium of the present invention in cigarette stalk heap body, it is characterized in that described nicotine degradation bacterium EA-17, after 30-45d fermentation, in cigarette stalk heap body, nicotine degradation rate can reach 43%-50%, compared with not inoculating nicotine degradation bacterium and only add the Traditional compost zymotechnique of decomposing agent, nicotine degradation rate improves 60%-78%, total nitrogen accretion is 7%-9%, total phosphorus increasing amount is 0.3%-0.6%, total potassium increasing amount is 6%-8%, cellulose degradation rate adds 15%-20%, and hemicellulose degradation rate adds 25%-30%.
The application of a kind of nicotine degradation bacterium of the present invention in cigarette stalk heap body:
Nicotine degradation bacterium EA-17 bacterium solution preparation
The present invention nicotine degradation bacterium used is for biting histidinol Arthrobacter (Arthrobacterhistidinolovorans), and deposit number is CCTCCM2013405, bacterial classification code name EA-17.
LB substratum: peptone 10g, yeast powder 5g, sodium-chlor 10g, agar 20g, distilled water 1000ml, pH7.0-pH7.2,121 DEG C, sterilizing 30min.
Strain fermentation
Nicotine degradation bacterium EA-17 is inoculated on LB substratum and (LB medium base adds 2% agar), cultivate 18-24h for 30 DEG C-37 DEG C.
Transfer in LB liquid medium by above-mentioned cultured nicotine degradation bacterium EA-17,30 DEG C-37 DEG C, 180r/min cultivates 10-18h.
Above-mentioned cultured liquid spawn is transferred to the 2M containing LB liquid medium
3in fermentor tank, fermentation parameter is: temperature 30 DEG C-37 DEG C, fermentation 16-24h, liquid amount 60%, mixing speed 250rpm, ventilation ratio 0.2VVM ~ 1VVM.
After fermentation cylinder for fermentation 16-24h, fermented liquid answers biomass to reach 3 × 10
9cFU/mL.
The use of nicotine degradation bacterium, EA-17 fermented liquid and from tobacco curing stalk decomposing agent (aspergillus niger spore: 10
7individual/g, subtilis 10
8cFU/g) mix with the mass ratio of 1:10, by the inoculum size access tabacco straw heap body of 1%, keep heap body moisture 60%-65%, fermentation 30 ~ 45d, can become thoroughly decomposed.
The present invention utilizes biotechnology to produce cigarette stalk fertilizer, promotes that tabacco straw becomes thoroughly decomposed with high-efficiency tobacco alkaline degradation bacterium EA-17.
The present invention is directed to tabacco straw, utilize the high-efficiency tobacco alkaline degradation bacterium filtered out from physical environment, exploitation promotes the novel process that tabacco straw becomes thoroughly decomposed.
Accompanying drawing illustrates:
Fig. 1 is heap temperature variation diagram in nicotine degradation bacterium composting process of the present invention;
Fig. 2 is nicotine degradation bacteria biomass variation diagram in nicotine degradation bacterium T process composting process of the present invention;
Fig. 3 piles body nicotine content variation diagram in composting process;
Fig. 4 piles body pH value variation diagram in nicotine degradation bacterium composting process of the present invention;
Fig. 5 piles body organic content variation diagram in Split-gill composting process of the present invention;
Fig. 6 piles the total N content variation diagram of body in Split-gill composting process of the present invention;
Fig. 7 is the variation diagram piling body ammonium nitrogen and nitrate nitrogen content in nicotine degradation bacterium composting process of the present invention.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail, but working of an invention mode is not limited thereto.
Embodiment 1: a kind of nicotine degradation bacterium of the present invention, this bacterium taxonomy called after bites histidinol Arthrobacter (Arthrobacter histidinolovorans) EA-17, be preserved in China typical culture collection center on September 15th, 2013, preserving number is CCTCCM2013405.Nicotine degradation bacterium EA-17 industrial production is second order fermentation, and its production method is: activated spawn is connected to LB liquid nutrient medium, cultivates 10-18h, is prepared into first order seed for 30-37 DEG C; Again one-level kind is inoculated in the 2M that LB liquid medium is housed with the inoculum size of 0.1%-0.5%
3in fermentor tank, fermentor tank liquid amount is 60%, 30-37 DEG C, mixing speed 250rpm, ventilation ratio 0.2VVM-1VVM, and fermentation 16-24h, biomass reaches 3 × 10
9cFU/mL.
The application of a kind of nicotine degradation bacterium in cigarette stalk heap body, nicotine degradation bacterium EA-17 use-pattern is: after nicotine degradation fermented liquid mixes with the mass ratio of 1:10 with self-control decomposing agent, by the inoculum size access cigarette stalk heap body of 1%, keep heap body moisture 60%-65%, fermentation 30-45d, can become thoroughly decomposed, described self-control decomposing agent contains aspergillus niger spore 10
7individual/g, subtilis 10
8cFU/g.Nicotine degradation bacterium EA-17, after 30-45d fermentation, in cigarette stalk heap body, nicotine degradation rate can reach 43%-50%, compared with not inoculating nicotine degradation bacterium and only add the Traditional compost zymotechnique of decomposing agent, nicotine degradation rate improves 60%-78%, total nitrogen accretion is 7%-9%, total phosphorus increasing amount is 0.3%-0.6%, total potassium increasing amount is 6%-8%, and cellulose degradation rate adds 15%-20%, and hemicellulose degradation rate adds 25%-30%.
Embodiment 2: a kind of nicotine degradation bacterium of the present invention EA-17 bacterium solution preparation
1 substratum
LB substratum: peptone 10g, yeast powder 5g, sodium-chlor 10g, agar 20g, distilled water 1000ml, pH7.0-pH7.2,121 DEG C, sterilizing 30min.
2 microbial inoculum preparations
2.1 actication of culture: nicotine degradation bacterium EA-17 is inoculated on LB substratum and (LB medium base adds 2% agar), cultivate 18-24h for 30 DEG C.
2.2 bacterial strain one-level kinds are cultivated: transfer in LB liquid medium by above-mentioned cultured nicotine degradation bacterium EA-17, liquid amount 50ml/500ml triangular flask, culture temperature 30 DEG C, shaking speed 180r/min, and 12h cultivated by shaking table.
2.3 fermentor cultivation: fermenter volume 2M
3, 1200L substratum, above-mentioned first order seed inoculum size is 2.4L, mixing speed 250rpm, ventilation ratio 0.5VVM, leavening temperature 30 DEG C, fermentation 20h.
3 fermented liquid biomasss
Contained by fermented liquid, the biomass of nicotine degradation bacterium EA-17 is 3 × 10
9cFU/mL.
The application of nicotine degradation bacterium EA-17 in tabacco straw becomes thoroughly decomposed:
2.1 nicotine degradation bacterium EA-17 application methodes
EA-17 fermented liquid and self-control decomposing agent (aspergillus niger spore: 10
7individual/g, subtilis 10
8cFU/g) mix with the mass ratio of 1:10, by the inoculum size access tabacco straw heap body of 1%, keep heap body moisture 60%-65%, fermentation 30-45d, can become thoroughly decomposed.Control experiment is the cigarette stalk heap body of inoculation 1% decomposing agent.
2.2 nicotine degradation bacterium accelerate the effect that cigarette stalk becomes thoroughly decomposed
Nicotine degradation bacterium of the present invention, can nicotine effectively in degrading tobacco stalk for biting histidinol Arthrobacter EA-17, accelerates cigarette stalk compost maturity, improve the degradation rate of organic matter, Mierocrystalline cellulose etc., thus the content of the elements such as raising compost total N, K.
The become thoroughly decomposed effect of the present invention to heap body access microbial inoculum carries out confirmatory experiment, and arrange two different treatment, be respectively inoculation from tobacco curing stalk decomposing agent (CK), access from tobacco curing stalk decomposing agent and nicotine degradation bacterium (T), experimental result is as follows.
2.2.1 temperature
As seen from Figure 1, after adding microbiobacterial agent, heap temperature rises rapidly, and in 5d, heap body CK, T process all reaches more than 50 DEG C.CK reaches top temperature 56 DEG C of used time 14d, and maintaining more than 50 DEG C temperature is 25d.And T process reaches top temperature 58 DEG C of used time 10d, maintaining more than 50 DEG C temperature is 32d.Single temperature variation analysis from heap body, with the addition of nicotine degradation bacterium, improve other microbial growth environment, microbial activities is relatively active, makes heap body reach top temperature faster and top temperature is relatively high.
2.2.2 nicotine degradation bacteria biomass
By Fig. 2, nicotine degradation bacterium slowly increases at front 8d, and then ramp breeding, reaches maximum 1.28 × 10 to biomass during 26d
8cFU/g, and the initial addition of bacterial strain is about 10
6cFU/g, proves that nicotine degradation bacterium can well adapt to compost environment, effectively utilizes cigarette stalk to grow.
2.2.3 nicotine content
In heap body, nicotine content change is as Fig. 3.In two heap body process, nicotine content is all the trend constantly reduced, but the nicotine degradation of T process comparatively CK is higher.At the end of compost, in CK process, nicotine degradation rate is that 28.27%, T is treated to 49.33%.The nicotine that the interpolation of nicotine degradation bacterium can effectively be degraded in cigarette stalk is described.
2.2.4pH value
As seen from Figure 4, pH is respectively 7.87,7.17 CK and the T process compost later stage.Nicotine degradation bacterium effectively degrades the alkaline matter nicotine discharged in heap body, thus reduces heap body pH value.Cellulose-degrading bacteria suitable growth pH environment is neutral and slant acidity, and thus T process can provide the pH environment of cellulose degradation bacteria growing preferably, thus is conducive to degrading tobacco stalk.
2.2.5 organic content
By Fig. 5, when compost starts, heap body organic content is 52%, and at the end of compost, CK process organic content is down to 37.17%, T process and is down to 34.65%, reduce respectively 28.93% and 33.49%, T process comparatively CK process organic matter degradation rate improve 15.76%.
2.2.6 total nitrogen and water-soluble ammonium nitrogen, nitrate nitrogen content
As seen from Figure 6, when compost terminal, T process total nitrogen content slightly exceeds CK8.05%.By Fig. 7, constantly reduce the trend be on the increase with nitric nitrogen in ammonium nitrogen on the whole, when fermentation termination, CK process and T process ammonia-nitrogen content are down to 120mg/Kg and 59mg/Kg respectively, are less than 400mg/Kg, reach During High-Temperature Composting and to become thoroughly decomposed standard.Experimental result confirms the transforming relationship between ammonium nitrogen and nitric nitrogen, well indicates compost maturity process.And T process nitrate nitrogen content is higher than CK process, exceeds 27.78%.Illustrate that adding nicotine degradation bacterium effectively reduces scattering and disappearing of nitrogen, increases cigarette stalk compost total nitrogen and nitrate nitrogen content.
2.2.7 total phosphorus potassium content
By table 1, before and after compost, it is little that total phosphorous increases change difference in two process, and total potassium content increasing amount of CK process and T process is respectively 10.36%, 17.67%, and the K content namely added in the tabacco straw fertilizer of nicotine degradation bacterium fermentation is higher.
2.2.8 wood fibre cellulose content
By table 2, in CK process, Mierocrystalline cellulose, hemicellulose degradation rate are respectively 50.81% and 52.27%, add in the T process of nicotine degradation bacterium, Mierocrystalline cellulose, hemicellulose degradation rate are 59.71% and 66.01%, improve 17.52% and 26.29% compared with CK, what nicotine degradation bacterium was described adds the ligocellulose degradation effectively can improving cigarette stalk.
Claims (5)
1. a nicotine degradation bacterium, it is characterized in that nicotine degradation bacterium, this bacterium taxonomy called after bites histidinol Arthrobacter (Arthrobacter histidinolovorans) EA-17, be preserved in China typical culture collection center on September 10th, 2013, preserving number is CCTCC NO:M2013405.
2. according to a kind of nicotine degradation bacterium according to claim 1, it is characterized in that nicotine degradation bacterium EA-17 industrial production is second order fermentation, its production method is: activated spawn is connected to LB liquid nutrient medium, cultivates 10-18h, is prepared into first order seed for 30-37 DEG C; Again one-level kind is inoculated in the 2M that LB liquid medium is housed with the inoculum size of 0.1%-0.5%
3in fermentor tank, fermentor tank liquid amount is 60%, 30-37 DEG C, mixing speed 250rpm, ventilation ratio 0.2VVM-1VVM, and fermentation 16-24h, biomass reaches 3 × 10
9cFU/mL.
3. according to the application of a kind of nicotine degradation bacterium according to claim 1 in cigarette stalk heap body.
4. according to the application of a kind of nicotine degradation bacterium according to claim 3 in cigarette stalk heap body, it is characterized in that described nicotine degradation bacterium EA-17 use-pattern is: after nicotine degradation fermented liquid mixes with the mass ratio of 1:10 with self-control decomposing agent, by the inoculum size access cigarette stalk heap body of 1%, keep heap body moisture 60%-65%, fermentation 30-45d, can become thoroughly decomposed, described self-control decomposing agent contains aspergillus niger spore 10
7individual/g, subtilis 10
8cFU/g.
5. according to the application of a kind of nicotine degradation bacterium according to claim 4 in cigarette stalk heap body, it is characterized in that described nicotine degradation bacterium EA-17, after 30-45d fermentation, in cigarette stalk heap body, nicotine degradation rate can reach 43%-50%, compared with not inoculating nicotine degradation bacterium and only add the Traditional compost zymotechnique of decomposing agent, nicotine degradation rate improves 60%-78%, total nitrogen accretion is 7%-9%, total phosphorus increasing amount is 0.3%-0.6%, total potassium increasing amount is 6%-8%, cellulose degradation rate adds 15%-20%, and hemicellulose degradation rate adds 25%-30%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310616712.1A CN103642722B (en) | 2013-11-27 | 2013-11-27 | Nicotine-degrading bacterium and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310616712.1A CN103642722B (en) | 2013-11-27 | 2013-11-27 | Nicotine-degrading bacterium and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103642722A CN103642722A (en) | 2014-03-19 |
| CN103642722B true CN103642722B (en) | 2015-06-24 |
Family
ID=50248009
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310616712.1A Expired - Fee Related CN103642722B (en) | 2013-11-27 | 2013-11-27 | Nicotine-degrading bacterium and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103642722B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103789238A (en) * | 2014-01-27 | 2014-05-14 | 华中农业大学 | Nicotine efficient degrading bacterium and culture method |
| CN107083343A (en) * | 2017-05-26 | 2017-08-22 | 中南民族大学 | Bacterial strains of arthrobacterium HW 17 of one plant of degraded cellulose and application thereof |
| CN107673830A (en) * | 2017-10-10 | 2018-02-09 | 长沙爱扬医药科技有限公司 | A kind of organic fertilizer products for promoting nicotine degradation |
| CN115305226B (en) * | 2022-09-22 | 2023-06-16 | 郑州轻工业大学 | Acinetobacter radioresistant ZJ-22 for degrading nicotine and producing hydrogen and application thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102643772B (en) * | 2012-05-10 | 2014-03-12 | 广西中烟工业有限责任公司 | Microbe strain and application thereof |
| CN103789238A (en) * | 2014-01-27 | 2014-05-14 | 华中农业大学 | Nicotine efficient degrading bacterium and culture method |
-
2013
- 2013-11-27 CN CN201310616712.1A patent/CN103642722B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN103642722A (en) | 2014-03-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102174398B (en) | Composite microbiological bacterial agent used for returning maize straws to field and preparation method and applications thereof | |
| CN101759494B (en) | Diseases-prevention organic fertilizer prepared by edible fungi residue | |
| CN101941851B (en) | Technology and process for preparing biochemical humic acid by using kitchen waste | |
| CN104609995B (en) | Plant growth promoting bio-organic fertilizer for saline-alkali land | |
| CN103710282B (en) | Straw decomposing agent for quick start of decomposition at low temperature and its preparation method | |
| CN102442849A (en) | Disease-preventing bio-organic fertilizer prepared from straw and its preparation method | |
| EP2479253A1 (en) | Antagonistic bacteria for preventing and treating panama wilt disease of continuously planted banana and microorganism organic fertilizer thereof | |
| CN103740693B (en) | A kind of straw decomposing inoculant and preparation method thereof | |
| CN103194392B (en) | Complex microbial inoculant for degrading straw and method for degrading straw by using same | |
| CN101759493B (en) | Biological disease-preventing organic fertilizer and preparation method thereof | |
| CN103484396B (en) | New strain of streptomyces thermocarboxydus and application thereof | |
| CN111362751B (en) | Special compound microbial disease-resistant growth-promoting bacterial fertilizer for ginseng and preparation method thereof | |
| CN103820366B (en) | The agent of a kind of tabacco straw quick composting | |
| CN104694410A (en) | Preparation method of straw decomposition agent | |
| CN103304285A (en) | Microbial agent and preparation method as well as application thereof | |
| CN104692866A (en) | Biological organic fertilizer | |
| CN104692868A (en) | Application of bioorganic fertilizer to soil improvement | |
| CN104692844A (en) | Preparation method of bioorganic fertilizer | |
| CN107502572A (en) | Application of the bacillus licheniformis in straw degradative, the microbial bacterial agent comprising the bacterium and its application | |
| CN103642722B (en) | Nicotine-degrading bacterium and application thereof | |
| CN104099281A (en) | Environment-friendly compound microbial agent | |
| CN108203352A (en) | A kind of production method of kitchen garbage organic fertilizer | |
| CN102942420B (en) | Special microbial agent and bio-organic fertilizer for sweet cherry and preparation methods of special microbial agent | |
| CN105802888A (en) | Bacillus subtilis for degrading edible mushroom dreg cellulose and application of bacillus subtilis | |
| CN101928161B (en) | Method for preparing direct seeding compression nutrition pot raw material |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20150624 Termination date: 20171127 |