CN101054555A - An epiphyte capable of digesting agar - Google Patents
An epiphyte capable of digesting agar Download PDFInfo
- Publication number
- CN101054555A CN101054555A CN200710021152.XA CN200710021152A CN101054555A CN 101054555 A CN101054555 A CN 101054555A CN 200710021152 A CN200710021152 A CN 200710021152A CN 101054555 A CN101054555 A CN 101054555A
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- Prior art keywords
- agar
- cladosporium
- txj13021
- digesting
- fungi
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- 229920001817 Agar Polymers 0.000 title claims abstract description 32
- 239000008272 agar Substances 0.000 title claims abstract description 32
- 241000233866 Fungi Species 0.000 claims abstract description 14
- 244000005700 microbiome Species 0.000 claims abstract description 12
- 241001149955 Cladosporium cladosporioides Species 0.000 claims abstract description 9
- 238000003113 dilution method Methods 0.000 claims abstract description 4
- 230000001580 bacterial effect Effects 0.000 claims description 23
- 241000894006 Bacteria Species 0.000 claims description 15
- 239000006916 nutrient agar Substances 0.000 claims description 8
- 238000011081 inoculation Methods 0.000 claims description 5
- 230000004083 survival effect Effects 0.000 claims description 4
- 239000000725 suspension Substances 0.000 claims description 4
- 238000000034 method Methods 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 241000222290 Cladosporium Species 0.000 claims description 2
- 238000000926 separation method Methods 0.000 claims description 2
- 230000001954 sterilising effect Effects 0.000 claims description 2
- 238000004659 sterilization and disinfection Methods 0.000 claims description 2
- 238000012360 testing method Methods 0.000 claims description 2
- 238000005303 weighing Methods 0.000 claims description 2
- 238000012364 cultivation method Methods 0.000 claims 1
- 238000002955 isolation Methods 0.000 claims 1
- 238000002474 experimental method Methods 0.000 abstract description 5
- 239000000126 substance Substances 0.000 abstract description 4
- 239000001963 growth medium Substances 0.000 abstract description 2
- 238000012216 screening Methods 0.000 abstract description 2
- 239000002699 waste material Substances 0.000 abstract description 2
- 230000012010 growth Effects 0.000 description 6
- 108020004463 18S ribosomal RNA Proteins 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 241000282376 Panthera tigris Species 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 241000228197 Aspergillus flavus Species 0.000 description 2
- 241000453701 Galactomyces candidum Species 0.000 description 2
- 235000017388 Geotrichum candidum Nutrition 0.000 description 2
- 241000985530 Penicillium glabrum Species 0.000 description 2
- 102000035195 Peptidases Human genes 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- LCTONWCANYUPML-UHFFFAOYSA-N Pyruvic acid Chemical compound CC(=O)C(O)=O LCTONWCANYUPML-UHFFFAOYSA-N 0.000 description 2
- 241000223261 Trichoderma viride Species 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 238000012882 sequential analysis Methods 0.000 description 2
- 229920000936 Agarose Polymers 0.000 description 1
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 description 1
- 241000206572 Rhodophyta Species 0.000 description 1
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- AEMOLEFTQBMNLQ-WAXACMCWSA-N alpha-D-glucuronic acid Chemical compound O[C@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-WAXACMCWSA-N 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 235000019621 digestibility Nutrition 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000024241 parasitism Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 235000019833 protease Nutrition 0.000 description 1
- 229940107700 pyruvic acid Drugs 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- -1 sulfuric acid ester Chemical class 0.000 description 1
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention belongs to industrial microbioorganism field, more specificly a fungus capable of digesting agar. A fungus strain of Cladosporium cladosporioides which can digest agar is obtained through screening microorganism of Hainan province Jianfeng hill withered substance at pure agar culture medium by gradient dilution method. The configuration and molecular identification is carried out. For further experiment , six kinds of familiar fungi are used as contrast and proves that the six kinds of fungi can not nearly utilize the agar which prove the screened fungus can degrade or digest agar derived from industrial and experiment waste material.
Description
One, technical field
The invention belongs to the industrial microbial technology field, be specifically related to a kind of fungi of energy digesting agar.
Two, background technology
Agar is a kind of saccharan of finding in some red algae cell walls, and its main chemical compositions is the sulfuric acid ester of poly semi-lactosi, comprises D-glucuronic acid, L-semi-lactosi, D-semi-lactosi, 3,6-dehydration-L-semi-lactosi and pyruvic acid etc.Chemical structure also can be modified into agarose for special applications.Be easy to just make matrix become solid in the agar adding substratum.Early stage biologist attempts to use the gel solidification substratum, but many microorganisms can digest it, because many microorganisms can extracellular proteinase, has caused the liquefaction of gel.And the agar steady chemical structure is a kind of derivative of sugar, thereby is not subjected to the influence of proteolytic enzyme, is difficult for being decomposed utilization by mycelia, only plays peptizer in substratum, and it is melted into liquid in the time of 96 ℃, be cooled to promptly solidify again below 45 ℃.Because the solid medium with the agar preparation is Clear ﹠ Transparent, is convenient to observe the form of mycelia, so become regular bevel or plate culture medium indispensable material.Therefore it is generally acknowledged do not have microorganism can utilize agar.But this understanding may mislead us, particularly in the quantitative experiment of microorganism now.We have found one fungal strain-Cladosporium cladosporioides TXJ13021 in experiment, it can digest pure agar and grow.
Cladosporium cladosporioides is a kind of fungi of the commonplace a kind of saprophytic double parasitism that distributes, and the research of this bacterium is mainly concentrated on pathogenic bacteria (Moore et al., 2001.Mycopathologia, 154:25-28; Richards and Beuchat, 2005.International Journal of Food Microbiology is 103:1-10) with the biological absorption of metal (Pethkar et al., 2001.Bioresource Technology, 80:211-215; Buszman et al., 2006.Science of the Total Environment, aspect such as 363:195-205).And do not appear in the newspapers aspect utilizing at agar for this bacterium.Research for this bacterium can be understood, be not do not have microorganism can utilize with digesting agar as nutrition source.
Three, summary of the invention
The problem that the present invention need solve is to utilize the microorganism digesting agar, allow the investigator understand, be not that agar can not be utilized and digestion by Institute of Micro-biology, so that microbe research afterwards, particularly relate in the microorganism quantitative examination of agar, provide the help of usefulness, the while also provides certain research basis for the degraded of agar.
The fungi that the present invention adopts is dendritic cladosporium Cladosporium cladosporioides TXJ13021 (being designated hereinafter simply as the TXJ13021 bacterial strain), be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, preservation date is on March 23rd, 2007, and preserving number is CGMCC1987.
Production bacterial strain TXJ13021 bacterial strain of the present invention divides the litter from spike mountain range, China Hainan mountane rain forest.Separate preparation according to conventional gradient dilution method.Be about to litter water wash-out, then elutriant carried out gradient dilution, diluent is seeded on the pure nutrient agar of solid, the superincumbent bacterium that will survive is after a while cultivated again, obtains bacterial strain.For determining further whether bacterial strain has utilized pure agar, and six kinds of common bacterial classifications are connected to pure nutrient agar, find almost all not growths, and the bacterial strain that is obtained can smooth growth then.
The present invention has carried out the observation of the form of bacterium colony and spore, mycelia, and growth characteristic, and 18S rDNA sequential analysis are the new bacterial strain (TXJ13021) of Cladosporium cladosporioides with this identification of strains.This bacterial strain has following feature: 1. bacterium colony is irregular subcircular, and bacterium colony is smooth, and aerial hyphae is undeveloped, and bacterium colony is a grey black; 2. bacterium colony is that growth is vigorous under the condition of 28 ℃ and pH 7.0 in temperature; 3. compare with other common bacteria, bacterial strain can be grown in above the pure agar, need not to add any other nutrition; 4. the 18SrDNA The sequencing results is carried out the maximum homology comparison in GenBank, found that the sequencing result of 18S rDNA and Cladosporium cladosporioides 18S rDNA sequence homology reach 100%.
The invention has the beneficial effects as follows: dendritic cladosporium Cladosporium cladosporioidesTXJ13021 has stable natural matter source, and bacterial classification derives from litter, and screening method is simple; Constant product quality, aerobic type is easily preserved, and light, temperature, soda acid are all had stronger stability.Dendritic cladosporium Cladosporium cladosporioides TXJ13021 can digesting agar, with the agar waste material that is produced in degraded microbiological industry or the experiment cultivation, the mistaken ideas of thinking that traditionally microorganism can not digestibility and utilization agar have been corrected, for the microorganism quantitative examination of agar provides useful help.
Four, embodiment
1.TXJ13021 the separation of bacterial strain, evaluation
Litter is from spike mountain range, Hainan, and litter is cut into fritter about 2mm * 2mm with the scissors of sterilization in the aseptic technique platform, takes by weighing then about 1 gram, add in the test tube, add the sterilized water of 9ml again, vibration 1min, then, use the gradient dilution method, bacteria suspension is diluted to 10
-1, 10
-2, 10
-3, 10
-4, four concentration get 10
-3, 10
-4, the bacterial suspension inoculation of two concentration to pure nutrient agar (2% agar, pH7.0) on, the culture dish with inoculation places incubator to cultivate the bacterial strain of observing survival in 20 days under 28 ℃ then.The bacterium colony of survival is a grey black, has at first carried out identification of morphology, is defined as with a kind of, carries out 18S rDNA sequential analysis then, is accredited as Cladosporium cladosporioides TXJ13021.
2.TXJ13021 bacterial strain is to the utilization of agar
For further determining the utilization of TXJ13021 bacterial strain to agar, Penicillium frequentans, Geotrichum candidum, Trichoderma viride, Aspergillus flavus, Penicillumsimplicissimum, six kinds of fungies of and Oosporium sp. are in contrast.TXJ13021 bacterial strain and this six kinds of fungies are inoculated on the pure nutrient agar, and the culture dish with inoculation places incubator to cultivate under 28 ℃ 20 days then.Found that six kinds of fungies in contrast almost can not grow on pure nutrient agar, and the TXJ13021 bacterial strain was grown colony diameter for being about 3.6cm at 20 days.In addition, also be provided with the substratum contrast, the speed of growth great majority of these six kinds of fungies on the red substratum of the competent tiger of nutrient want fast than the TXJ13021 bacterial strain.TXJ13021 bacterial strain and the growth fraction of six kinds of common bacteria on red substratum of tiger and pure nutrient agar are
| Bacterial classification | Tiger red substratum (5 days) | Pure nutrient agar (20 days) |
| Penicillium frequentans Geotrichum candidum Trichoderma viride Aspergillus flavus Penicillum simplicissimum Oosporium sp. TXJ13021 | 1.4±0.4c 1.9±0.3b 3.7±0.6a 3.9±0.6a 1.5±0.3c 0.5±0.1d 1.5±0.4c | 0.0±0.0c 0.0±0.0c 0.0±0.0c 0.3±0.1b 0.0±0.0c 0.0±0.0c 3.6±0.4a |
Lowercase after the data in the table in the row represents to exist each other significant difference (p＜0.05).
Claims (3)
1. the fungi of an energy digesting agar is characterized in that dendritic cladosporium Cladosporium cladosporioidesTXJ13021, and strains separation is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center from spike mountain range, Hainan.
2. according to the isolation cultivation method of the described dendritic cladosporium Cladosporium cladosporioides TXJ13021 of claim 1, it is characterized in that: litter is from spike mountain range, Hainan, litter is cut into the fritter of 2mm * 2mm with the scissors of sterilization in the aseptic technique platform, take by weighing 1 gram then, add in the test tube, add the sterilized water of 9ml again, vibration 1min, then, use the gradient dilution method, bacteria suspension is diluted to 10
-1, 10
-2, 10
-3, 10
-4, four concentration get 10
-3, 10
-4, the bacterial suspension inoculation of two concentration is to 2% agar, and on the pure nutrient agar of pH7.0, the culture dish with inoculation places incubator to cultivate the bacterial strain of observing survival in 20 days under 28 ℃ then, and the bacterium colony of survival is a grey black.
3. according to the application of the described dendritic cladosporium Cladosporium cladosporioides TXJ13021 of claim 1 in digesting agar.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB200710021152XA CN100532534C (en) | 2007-03-30 | 2007-03-30 | An epiphyte capable of digesting agar |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB200710021152XA CN100532534C (en) | 2007-03-30 | 2007-03-30 | An epiphyte capable of digesting agar |
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| Publication Number | Publication Date |
|---|---|
| CN101054555A true CN101054555A (en) | 2007-10-17 |
| CN100532534C CN100532534C (en) | 2009-08-26 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB200710021152XA Expired - Fee Related CN100532534C (en) | 2007-03-30 | 2007-03-30 | An epiphyte capable of digesting agar |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101851586A (en) * | 2010-05-11 | 2010-10-06 | 林永慧 | Fungus for efficiently decomposing synthetic dye and application thereof |
| CN102703327A (en) * | 2012-02-04 | 2012-10-03 | 西北农林科技大学 | Cladosporium Sp. XJ-AC03 and application thereof |
| CN105238703A (en) * | 2015-10-27 | 2016-01-13 | 江南大学 | Cladosporiumc cladosporioides for high yield of feruloyl esterase and application of Cladosporiumc cladosporioides in edible vinegar brewing |
-
2007
- 2007-03-30 CN CNB200710021152XA patent/CN100532534C/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101851586A (en) * | 2010-05-11 | 2010-10-06 | 林永慧 | Fungus for efficiently decomposing synthetic dye and application thereof |
| CN102703327A (en) * | 2012-02-04 | 2012-10-03 | 西北农林科技大学 | Cladosporium Sp. XJ-AC03 and application thereof |
| CN105238703A (en) * | 2015-10-27 | 2016-01-13 | 江南大学 | Cladosporiumc cladosporioides for high yield of feruloyl esterase and application of Cladosporiumc cladosporioides in edible vinegar brewing |
| CN105238703B (en) * | 2015-10-27 | 2018-05-25 | 江南大学 | The dendritic cladosporium of one plant height production feruloyl esterase and the application in vinegar brewing |
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| Publication number | Publication date |
|---|---|
| CN100532534C (en) | 2009-08-26 |
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Granted publication date: 20090826 Termination date: 20120330 |