CN103114046B - Aspergillus tubingensis SZX-6 and fermentation liquid, fermentation liquid extract and application thereof - Google Patents
Aspergillus tubingensis SZX-6 and fermentation liquid, fermentation liquid extract and application thereof Download PDFInfo
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- CN103114046B CN103114046B CN201310065886.3A CN201310065886A CN103114046B CN 103114046 B CN103114046 B CN 103114046B CN 201310065886 A CN201310065886 A CN 201310065886A CN 103114046 B CN103114046 B CN 103114046B
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Abstract
The invention provides aspergillus tubingensis SZX-6 with a patent number of NO: M2012540 in CCTCC (China Center For Type Culture Collection). The invention further discloses a fermentation liquid and a fermentation liquid extract of the aspergillus tubingensis SZX-6. The fermentation liquid and the fermentation liquid extract of aspergillus tubingensis have obvious inhibition effects on Vibrio anguillarum, Vibrio parahaemolyticus, Aeromonas hydrophila, Vibrio harveyi and Vibrio cholerae. Therefore, the fermentation liquid and the fermentation liquid extract can be applied to antibacterial medicines or can be used for the extraction separation of compounds with the antibacterial activity for development of the antibacterial medicine.
Description
Technical field
The invention belongs to biological medicine technology field, be specifically related to a kind of Tabin aspergillus (
aspergillus tubingensis) SZX-6, the invention still further relates to the fermented liquid of this bacterial strain and the extract of fermented liquid, the invention still further relates to the purposes of this fermented liquid and fermentation broth extract.
Background technology
Since the fifties in last century Newton etc. from thalassiomycetes
cephalosporium acremoniumtunning in find antimicrobial compounds cephalosporin C since, the research of marine microorganism antibacterial substance becomes the hot fields of drug research day by day.The totally different unique living environment in Lu Sheng fungi of thalassiomycetes (as oligotrophic, high salt, weakly alkaline etc.) makes thalassiomycetes form special metabolism and defense system, thereby can produce, is different from the special and active good secondary metabolite of the structure of Lu Sheng fungi.Up to the present, from the tunning of thalassiomycetes, found the new secondary metabolite of kind more than 1,000, structure type relates to alkaloid, polyketone, macrocyclic lactone and terpene etc., and these meta-bolitess present the good biological activity such as antibacterial, antitumor, antiviral.
In recent ten years, rapidly, output of aquatic products occupies first place in the world China's aquaculture industry development, but also constantly occurs various Animal diseases.Vibrio anguillarum (
vibrio anguillarum), Vibrio parahaemolyticus (
vibrio parahaemolyticus), Aeromonas hydrophila (
aeromonas hydrophila), Vibrio harveyi (
vibrio harveyi) and vibrio cholerae (
vibrio cholerae) etc. be the main pathogen bacterium that causes cultivated animals disease.The life-time service of antibacterials or to abuse caused pathogenic bacterium resistance, high residue and problem of environmental pollution day by day serious in aquaculture.Therefore the water of, developing new efficient, wide spectrum and low dirt has great importance with microbiotic.
Summary of the invention
Technical problem to be solved by this invention is for the deficiencies in the prior art, and a kind of new Tabin aspergillus SZX-6 bacterial strain is provided.
Another technical problem to be solved by this invention is to provide fermented liquid and the fermentation broth extract of above-mentioned Tabin aspergillus SZX-6 bacterial strain.
Another technical problem to be solved by this invention is to provide the fermented liquid of above-mentioned Tabin aspergillus SZX-6 bacterial strain and the purposes of extract thereof.
The present invention be a kind of Tabin aspergillus (
aspergillus tubingensis) SZX-6 CCTCC NO:M 2012540.Bacterial strain Tabin aspergillus involved in the present invention (
aspergillus tubingensis) SZX-6, on December 20th, 2012, being deposited in Chinese Typical Representative culture collection center C CTCC, deposit number is CCTCC NO:M 2012540, preservation address: Wuhan, China city Wuhan University, phone 027-68754052.
The invention also discloses a kind of Tabin aspergillus SZX-6 fermented liquid, be characterized in, this fermented liquid obtains with described Tabin aspergillus SZX-6 fermentation, fermentation process is: Tabin aspergillus SZX-6 bacterial classification is inoculated in seawater Sharpe liquid nutrient medium, 160 r/min, cultivate 7 d for 28 ℃, obtain Tabin aspergillus SZX-6 tunning, after filtering mycelium, obtain fermented liquid; Consisting of of described seawater Sharpe liquid nutrient medium: glucose 40 g/L, peptone 10 g/L, Chen Haishui constant volume.
The invention also discloses a kind of Tabin aspergillus SZX-6 fermentation broth extract, be characterized in, the Tabin aspergillus SZX-6 fermented liquid that this extract be take described in technique scheme is raw material, use successively equal-volume ethyl acetate, n-butanol extraction 3 times, obtain ethyl acetate extract, n-butanol extract or extract remainder.
Tabin aspergillus SZX-6 fermented liquid of the present invention or Tabin aspergillus SZX-6 fermentation broth extract have significant bacteriostatic application, its suppressible bacterium be Vibrio anguillarum (
vibrio anguillarum), Vibrio parahaemolyticus (
vibrio parahaemolyticus), Aeromonas hydrophila (
aeromonas hydrophila), Vibrio harveyi (
vibrio harveyi) and vibrio cholerae (
vibrio cholerae).
Tabin aspergillus SZX-6 of the present invention is that separation obtains from marine cultured animal Portunus trituberculatus Miers symbiotic and epiphyte microorganism.Its separation purification method is: Portunus trituberculatus Miers is shredded with aseptic scissors, add sterilized water to make suspension, get 0.1 mL and be evenly coated with in Sharpe solid medium, 28 ℃ of thermostat containers are cultivated, after bacterium colony grows, and picking colony line purifying.Described Sharpe solid medium is by after 4% (W/V) glucose, 1% (W/V) peptone and 2% (W/V) agar heating for dissolving, adds Chen Haishui constant volume, after 121 ℃ of sterilizing 20 min, obtains.
The biology morphology of Tabin aspergillus SZX-6 of the present invention is described as: on seawater sabouraud culture medium, growth rapidly, is cultivated 7 days for 28 ℃, and colony diameter can reach 65-70 mm, quality fine hair shape, mycelia nascent is white, fades to palm fibre, black, without transudate, reverse side tawny, as shown in Figure 1; Under Optical microscope and SEM, observe, mycelia has tabula, and conidial head is spherical in shape, brown-black; Top capsule expands, the radial close two-wheeled stalklet of giving birth on the capsule of top; Conidinphore water white transparency; Conidium ovalize, brown-black, rough, there is spinule, concatenate, as shown in Figure 2,3.
The heredity feature of Tabin aspergillus SZX-6 of the present invention is as follows: belong to thalassiomycetes, its ITS sequence is in Genbank login, and accession number is JX287371.This sequence and NCBI nucleic acid database (Genbank http://www.ncbi.nlm.nih.gov/) BLAST comparison result shows, this sequence and aspergillus fungi (
aspergillussp.) ITS sequence has very high homology, wherein with bacterial strain
aspergillus tubingensisa1KA,
a. tubingensisthe ITS sequence homology of wxm77 (sequence number is respectively AM745112214443.1 and HM037959) is 99%, gathers as same cluster group take on the phylogenetic tree that N-J method builds.
Compared with prior art, the fermented liquid of Tabin aspergillus SZX-6 bacterial strain of the present invention is, the extract of fermented liquid all has the feature of anti-microbial activity.Therefore, the extract of its fermented liquid, fermented liquid can be used for antibacterials, or can be used for extracting the separated compound with anti-microbial activity and develop antibacterials.
Accompanying drawing explanation
Fig. 1 is the colonial morphology figure of Tabin aspergillus SZX-6 bacterial strain;
Fig. 2 is the conidial fructification figure of Tabin aspergillus SZX-6 bacterial strain;
Fig. 3 is the conidium figure of Tabin aspergillus SZX-6 bacterial strain;
Fig. 4 is that fermented liquid and the fermentation broth extract of Tabin aspergillus SZX-6 bacterial strain schemed the inhibition of main aquatic pathogenic bacterium is active.
Embodiment
Below further describe concrete technical scheme of the present invention, so that those skilled in the art understands the present invention further, and do not form the restriction to its right.
Embodiment 1, a kind of Tabin aspergillus (
aspergillus tubingensis) SZX-6 CCTCC NO:M 2012540.
Embodiment 2, a kind of Tabin aspergillus SZX-6 fermented liquid, this fermented liquid obtains with the Tabin aspergillus SZX-6 fermentation described in embodiment 1, fermentation process is: Tabin aspergillus SZX-6 bacterial classification is inoculated in seawater Sharpe liquid nutrient medium, 160 r/min, cultivate 7 d for 28 ℃, obtain Tabin aspergillus SZX-6 tunning, after filtering mycelium, obtain fermented liquid; Consisting of of described seawater Sharpe liquid nutrient medium: glucose 40 g/L, peptone 10 g/L, Chen Haishui constant volume.
Embodiment 3, a kind of Tabin aspergillus SZX-6 fermentation broth extract, the Tabin aspergillus SZX-6 fermented liquid that this extract be take described in embodiment 2 is raw material, uses successively equal-volume ethyl acetate, n-butanol extraction 3 times, obtains ethyl acetate extract, n-butanol extract or extract remainder.
Embodiment 4, the Tabin aspergillus SZX-6 fermented liquid described in embodiment 2 and the antibacterial experiment of the Tabin aspergillus SZX-6 fermentation broth extract described in embodiment 3.
1. anti-microbial activity indication bacterium solution preparation: by Vibrio anguillarum (
vibrio anguillarum), Vibrio parahaemolyticus (
vibrio parahaemolyticus), Aeromonas hydrophila (
aeromonas hydrophila), Vibrio harveyi (
vibrio harveyi) and vibrio cholerae (
vibrio cholerae) with sterilized water, make spore suspension (10 respectively
7individual/mL).Spore suspension is inoculated in the sterilized 500 mL triangular flasks that include 200 mL beef-protein mediums by 5% inoculum size, and in the constant-temperature shaking incubator of 37 ℃ of temperature, rotating speed 160 r/min, shaking culture 1 day, obtains indicator liquid.
2. bacteriostatic activity test: the beef extract-peptone solid medium that is cooled to 45-50 ℃ after fusing is poured in 90.00 mm flat boards, and every plate falls about 20 mL, places to be cooled to and solidifies.Respectively at smearing 100 μ L indicator liquid on each flat board, place 30 min.Then on flat board, put Oxford cup well, 100 μ L fermented liquids or fermentation broth extract are added in hand-hole.In 37 ℃ of incubators, cultivate after 24 h, use vernier caliper measurement antibacterial circle diameter, active size represents with antibacterial circle diameter (mm).The results are shown in Figure 4.
Experiment shows: Tabin aspergillus SZX-6 fermented liquid and fermentation broth extract (acetic acid ethyl ester extract, n-butyl alcohol extract and extract remainder) to 5 kinds of main aquatic pathogenic bacterium Vibrio anguillarums (
vibrio anguillarum), Vibrio parahaemolyticus (
vibrio parahaemolyticus), Aeromonas hydrophila (
aeromonas hydrophila), Vibrio harveyi (
vibrio harveyi) and vibrio cholerae (
vibrio cholerae) all there is significant inhibition activity, by the sequence of bacteriostatic activity size, be fermented liquid > extract remainder > acetic acid ethyl ester extract > n-butyl alcohol extract.Therefore, the extract of Tabin aspergillus SZX-6 fermented liquid, fermented liquid can be used for antibacterials, or can be used for extracting the separated compound with anti-microbial activity and develop antibacterials.
Claims (4)
- A Tabin aspergillus ( aspergillus tubingensis) SZX-6 CCTCC NO:M 2012540.
- 2. a Tabin aspergillus SZX-6 fermented liquid, it is characterized in that, this fermented liquid obtains with Tabin aspergillus SZX-6 fermentation claimed in claim 1, fermentation process is: Tabin aspergillus SZX-6 bacterial classification is inoculated in seawater Sharpe liquid nutrient medium, 160 r/min, cultivate 7 d for 28 ℃, obtain Tabin aspergillus SZX-6 tunning, after filtering mycelium, obtain fermented liquid; Consisting of of described seawater Sharpe liquid nutrient medium: glucose 40 g/L, peptone 10 g/L, Chen Haishui constant volume.
- 3. a Tabin aspergillus SZX-6 fermentation broth extract, it is characterized in that, it is raw material that this extract be take Tabin aspergillus SZX-6 fermented liquid claimed in claim 2, uses successively equal-volume ethyl acetate, n-butanol extraction 3 times, obtains ethyl acetate extract, n-butanol extract or extract remainder.
- 4. the bacteriostatic application of Tabin aspergillus SZX-6 fermented liquid claimed in claim 2 or Tabin aspergillus SZX-6 fermentation broth extract claimed in claim 3, described bacterium be Vibrio anguillarum ( vibrio anguillarum), Vibrio parahaemolyticus ( vibrio parahaemolyticus), Aeromonas hydrophila ( aeromonas hydrophila), Vibrio harveyi ( vibrio harveyi) and vibrio cholerae ( vibrio cholerae).
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| CN105483027A (en) * | 2016-01-04 | 2016-04-13 | 山东星光生物科技有限公司 | Aspergillus tubingensis and method for producing fructooligosaccharides through whole-cell catalysis of aspergillus tubingensis |
| CN110292051B (en) * | 2017-02-23 | 2021-04-23 | 北京市农林科学院 | Fruit and vegetable disease inhibitor and application thereof |
| CN109207374B (en) * | 2018-10-10 | 2021-08-03 | 江苏省海洋资源开发研究院(连云港) | Corbicula fluminea symbiotic fungus aspergillus fumigatus, preparation method thereof, extract and application thereof |
| CN109706083B (en) * | 2018-12-10 | 2023-05-30 | 贵州省中国科学院天然产物化学重点实验室(贵州医科大学天然产物化学重点实验室) | Eucommia ulmoides endophytic fungus with remarkably increased antibacterial activity after metabolic regulation |
| CN109626598A (en) * | 2019-01-23 | 2019-04-16 | 南华大学 | A method of utilizing Tabin aspergillus and phytate in-situ immobilization hexavalent uranium polluted surface water |
| CN110468058B (en) * | 2019-09-10 | 2020-12-25 | 福州大学 | Eurotium cristatum fermentation broth crude peptide extract and application thereof in preservation of meat products |
| CN116515646B (en) * | 2023-06-25 | 2023-09-19 | 华南农业大学 | Aspergillus tubingensis and application thereof in preparing tannase and/or degrading tannin |
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Inventor after: Guo Lei Inventor after: Zhu Wencheng Inventor after: Xu Fuquan Inventor after: Liu Weiwei Inventor after: Chen Li Inventor before: Guo Lei Inventor before: Zhu Chengwen Inventor before: Xu Fuquan Inventor before: Liu Weiwei Inventor before: Chen Li |
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Free format text: CORRECT: INVENTOR; FROM: GUO LEI ZHU CHENGWEN XU FUQUAN LIU WEIWEI CHEN LI TO: GUO LEI ZHU WENCHENGXU FUQUAN LIU WEIWEI CHEN LI |
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