CN103898012A - Thalassospira sp. strain and method for preparing agarase - Google Patents
Thalassospira sp. strain and method for preparing agarase Download PDFInfo
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Abstract
The invention relates to a Thalassospira sp. fjfst-201307 strain and a method for preparing agarase by using the strain. The strain has been registered and collected at China Center for Type Culture Collection on December 25th, 2013, and the collection number is CCTCC M2013708. In a fermentation medium composed of a carbon source, a nitrogen source and inorganic salts by using the strain as the seed, the activity of the agarase is up to 11.2 U/ml under optimized culture conditions. The method for preparing the agarase comprises the following steps: salting out the fermentation liquor by ammonium sulfate, centrifugating, collecting the precipitate, dialyzing, passing the dialysate through an ion-exchange column and a gel column, and carrying out freeze-drying to obtain the agarase dry powder.
Description
Technical field
The invention belongs to technical field of food biotechnology, relate to screening one strain sea revolve bacterium (
thalassospirasp.) and cultivation and fermentation prepare the method for agarase.
Background technology
Agar-agar (agar), having another name called agar, is a kind of Sargassum polysaccharides obtaining through hot water extraction in marine red alga, its aboundresources, be widely used in food, medicine, biotechnology field, agar-agar is mainly made up of neutral agarose (agarose) and ionic agaropectin (agar), agarose is by 1,3 β-D-the semi-lactosis that connect is connected with Isosorbide-5-Nitrae 3,6-inner ether-α-L-galactose residue straight-chain polymer of connection alternately repeatedly.
Agarase wide material sources, are mainly present in marine bacteria: alternately zygosaccharomyces, and Cytophaga, Rhodopseudomonas, pseudoalteromonas belongs to, streptomyces, micro-Pseudomonas that quivers, Vibrio.In the animal take marine alga as food belongs to, be all found, as sea hare belongs to, Littorina, is preced with extra large Zhan's genus etc.
Agaropectin oligose just refers to the oligose that agar polysaccharide is 2-10 through hydrolysis post polymerization degree (DP), claim again agar oligosaccharide, mainly be formed by connecting by the repeating unit of fine jade disaccharides, good water solubility, be conducive to absorption of human body, greatly improve its using value, it is a kind of novel oceanic functional oligose, agaropectin oligose has a wide range of applications at cosmetic field, mainly stems from its skin care moisturizing, delays senility, the function of whitening and the effect that can prevent skin infections and allergic dermatitis having as cosmetics additive.Simultaneously due to itself and the biological sugared structural similitude of endogenous, therefore to organism toxicological harmless, growing to the research of agaropectin oligose function.And in foodstuff production, because the not digested enzyme of agaropectin oligose decomposes, and empty calory produces, can do the production for beverage, bread and low calorific food of the weighting agent of high sweeting agent and dispersion agent; In addition, agaropectin oligose has stronger bacteriostatic action, is a kind of well natural antiseptic agent and antioxidant.
Fujian Province has wide marine site, has deepened in recent years the exploitation aspect oceanic resources.In ocean, abundant marine algae resource provides sufficient source for Sargassum polysaccharides.The production of agar simultaneously depends on the industrialized utilization to marine algas such as gelidium, Pterocladia tenuis Okam., fragrant plant mentioned in ancient texts, Eucheuma muricatum (Gmel.) Web． Van Bos．, lavers, and be the Preliminary Exploitation to these marine algae resources for the production of agar, the product that its follow-up deep processing obtains has very wide application prospect on market.If agar is further processed, the agaropectin oligose obtaining, is applied in the industries such as medicine, protective foods and makeup, and it is worth considerably beyond agar.Therefore the further intensive processing of marine alga is not only the making full use of of resource, and has deepened abundant understanding and the research to oceanic resources, is all very important and necessary for production application and scientific research.Although Fujian Province's marine algae resource is quite abundant, the green production of agaropectin oligose depends on the application of agarase.Because the source of agarase is relatively less, activity is low, expensive, in production application, there is certain restriction.Microbe-derived agarase can be produced in enormous quantities, but the wild strain that general screening obtains, and its inulinase-producing activity often can be very not high.Therefore, improve the product enzyme activity of bacterial strain, obtain the relatively high agarase of purity, have great importance to adopting enzymolysis process to produce agaropectin oligose.Using microbe selection by mutation technology improves the product enzyme activity of bacterial strain, and the agarase obtaining that ferments is carried out to separation and purification, and the vigor of agarase and purity are all improved greatly, is conducive to the application at industry and field of scientific study of agarase and product thereof.
Agarase is mainly derived from sea mollusk and marine microorganism.The bacterium of decomposing agar-agar is mainly present in ocean environment.Since Gran in 1902 is separated to agarolytic bacteria one agar-agar pseudomonas for the first time from seawater
(Psudomonas galatica) since, people have been separated to multiple agar-agar decomposer from sea water system.As (
cytophaga (cytophaga) (Duckworth M et al., 1968),
vibrio(Vibrio) (Aoki et al., 1990),
streptomyces(streptomyces) (Bibb MJ et al., 1987),
pseudomonas-like(class Rhodopseudomonas) (Von Hofsten B et al., 1975),
alterococcus(alternately Coccus) (Shieh WY et al., 1998)
pseudomonas(Rhodopseudomonas) (Belas R et al., 1989)
pseudomanas wellantypicum(Jing Shi pseudomonas) (Wang Xuanliang, 1985),
altermonas agarlyticsgJ1B(pseudoalteromonas) (Potin et al., 1993),
thalassomonassp. cold Zymomonas mobilis is had a liking in JAMB-A33(ocean) (Ohta et al., 2005).Therefore screen new microbial method and prepare the new task that agarase is called investigator.
Summary of the invention
The object of this invention is to provide a strain sea and revolve algae, and a kind of method of utilizing this bacterial strain to prepare agarase.
Technical scheme of the present invention: a strain screening is taked the bacterium in sample liquid from the seawater of offshore sea waters, Xiamen, can be for the preparation of agarase (agarase), its Classification And Nomenclature be sea revolve bacterium (
thalassospirasp.) fjfst-2013007, is being deposited in the registration preservation of Chinese Typical Representative culture collection center on December 25th, 2013, and its deposit number is CCTCC M 2013708, and preservation address is Wuhan University.
The screening of microorganism strains and evaluation:
The present invention takes sample liquid from the seawater of offshore sea waters, Xiamen, be inoculated in enrichment medium and carry out coating in agar screening culture medium after illumination cultivation, at 25 ℃, cultivate bacterium colony that 48h selects significant depressions and enter the separation and purification of ruling in agar screening culture medium, purebred 25 ℃ of the fermention mediums of receiving, 120 rpm, shake-flask culture 24-72h, detect the agarase vigor in substratum, finishing screen is selected aimed strain, to its carry out physio-biochemical characteristics identify and molecular biology experiment guide carry out 16s rDNA sequential analysis, determine its belong to sea revolve bacterium (
thalassospirasp.), intend called after sea revolve bacterium (
thalassospirasp.) fjfst-2013007.
With described sea revolve bacterium (
thalassospirasp.) method of fjfst-2013007 production agarase comprises the steps:
(1) sea is revolved bacterium (
thalassospirasp.) fjfst-2013007 is in seed culture medium, with the bottled 20-60mL seed culture medium of 250mL taper, sterilizing according to a conventional method, cooling, and inoculates bacterium colony, after inoculation, after 20-30 ℃ of 160rpm shaking table cultivated 12-24h, obtains seed fermentation liquid;
(2) with the bottled 30mL fermention medium of 250mL taper, sterilizing according to a conventional method, cooling, and the inoculum size of pressing 6%-10%, by seed fermentation liquid access fermention medium, after inoculation, in 25-30 ℃, 160rpm shaking table obtains the fermented liquid containing agarase after cultivating 48-72h;
(3) then by fermention medium through 4 ℃, after 6000-8000g frozen centrifugation 10-20min, get supernatant liquor;
(4) supernatant liquor being added to massfraction is that the saturated ammonium sulphate of 60%-80% is saltoutd, and places refrigerator overnight, and 4 ℃, 8000-10000g is centrifugal, and 20-40min gets precipitation;
(5) with buffer A balance DEAE-Sepharose Fast Flow post (Φ 2cm × 10cm), cross 4-5 bed volume.Take 0.3g agarase powder and be dissolved in 8mL buffer A, after the centrifugal 20min of 6000g, get ion exchange column on supernatant liquor.After loading, cross 2-3 bed volume by buffer A and wash away the foreign protein not being adsorbed, the NaCl solution of the 0-0.5mol/L then preparing in order to buffer A carries out gradient elution.Flow rate control is at 0.5mL/min, and detection wavelength is 280nm.Elutriant fraction collection, 4.5mL/ pipe.In the test tube of mensuration wash-out pipe peak part, the enzyme activity of solution, merges activated part, obtains enzyme work in-process after lyophilize;
(6) Sephcryl S-100 gel permeation chromatography post (Φ 1.6cm × 80cm) is used after buffer A balance, take work in-process agarase 0.05g and be dissolved in 4mL level pad, after 6000g low-temperature centrifugation 20min, get chromatography column on supernatant liquor.Carry out wash-out by buffer A, coutroi velocity is 0.1mL/min.Elutriant is through ultraviolet detection fraction collection (4.0mL/ pipe).Determine respectively the enzyme activity of peak part test tube, activated part is merged, through fully concentrating through ultra-filtration membrane after dialysis desalting, to concentrated solution lyophilize, obtain finished product agarase.
Described fermention medium is: sea salt 20-30g/L, and peptone 3-7g/L, yeast powder 0.5-2.5g/L, agar 1.5-2.5g/L, adjusting pH is 7.2-8.4, deionized water preparation; Described seed culture medium: agar 2g/ L, sea salt 30 g/L, peptone 5g, yeast powder 1g, PH is 8.0, deionized water preparation.
Described buffer A is 0.02mol/L, the Tris-HCl damping fluid of pH7.5.
The detection method of the agarase of producing by the method:
Get above-mentioned steps (3) and obtain the fermented liquid supernatant liquid 1mL that contains agarase, add the PBS damping fluid (wherein containing agar-agar 0.2%) of 1mL, be placed in 40 ℃ of water-bath enzymolysis 30min, after add 1.5mLDNS, boiling water bath 15min.Be cooled to after room temperature, with distilled water polishing scale, use spectrophotometer to detect under 520nm.1U is defined as the value of 1min generation 1ug agaropectin oligose.
Beneficial effect of the present invention: filter out and a kind ofly produce the bacterial strain of agarase for microbial method, Classification And Nomenclature be sea revolve bacterium (
thalassospirasp.) fjfst-2013007.The present invention is using this bacterium as bacterial classification, domesticly yet there are no the report of producing agarase with this bacterial classification.This bacterium is with common carbon source, nitrogenous source, and the fermention medium of inorganic salt composition, with short production cycle, 10h can reach logarithmic phase, and this bacterial classification energy abduction delivering agarase can reach maximum enzyme output in 24h.Can in crude enzyme liquid, collect and obtain active high agarase by simple separation purification method.Agarase preparation used in the present invention and purifying process are simple, with low cost, and the product enzyme time is short, is suitable for industrial production or field of medicaments, the fields such as food chemistry and mariculture.
Embodiment
Be below explanation specific embodiments of the invention, but the present invention is not limited to this.
Embodiment 1
Get the seawater of offshore sea waters, Xiamen and take sample liquid, draw 1ml and dilute (10
-1, 10
-2, 10
-3, 10
-4), draw 0.5ml and coat in agar screening culture medium, at 25 ℃, cultivate bacterium colony that 48h selects significant depressions and enter the separation and purification of ruling in agar screening culture medium, by purebred 25 ℃ of the liquid nutrient mediums of receiving, 100rpm, shake-flask culture 48h, centrifuging and taking supernatant, adopts the agarase activity in spectrophotometry substratum, through primary dcreening operation, be divided into from obtaining the bacterium that 3 strains can long agarase, in conjunction with the throughput of agarase, finally select as starting strain.
Embodiment 2
Through morphology and physio-biochemical characteristics research, the feature of producing agarase bacterial strain is as follows:
Colonial morphology: bacterium colony presents circle, smooth surface, oyster white, edge has neatly.
Cellular form: this is Gram-negative bacteria, form is like vibrios shape.
Physiological and biochemical property: for strictly aerobic, can utilize starch,, glucose, sucrose, sodium-acetate, the multiple compounds such as D-semi-lactosi, as carbon source, also can utilize peptone, yeast extract paste, ammonium nitrate, ammonium sulfate, waits multiple compounds as nitrogenous source.
16S rRNA gene to bacterial strain carries out pcr amplification and sequencing, and the Gene Partial fragment length of finding its 16S rRNA is 1487bp, compares through NCBI and rrna database, be accredited as sea revolve bacterium (
thalassospirasp.), intend called after sea revolve bacterium (
thalassospirasp.) fjfst-2013007, the concrete visible sequence table of 16S rRNA sequence of bacterial strain.
Embodiment 3
(1) sea is revolved bacterium (
thalassospirasp.) fjfst-2013007 is in seed culture medium, with the bottled 30mL seed culture medium of 250mL taper, sterilizing according to a conventional method, cooling, and inoculate bacterium colony, after inoculation, in 25 ℃, 140rpm shaking table obtains seed fermentation liquid after cultivating 24h, described seed culture medium is: sea salt 25g/L, peptone 5g/L, yeast powder 1g/L, agar 2g/L, regulating pH is 7.4, deionized water preparation.
(2) with the bottled 30mL fermention medium of 250mL taper, sterilizing according to a conventional method, cooling, and by 4% inoculum size, by seed fermentation liquid access fermention medium, after inoculation, in 25 ℃, 140rpm shaking table obtains the fermented liquid containing agarase after cultivating 48h, described seed culture medium is: sea salt 25g/L, peptone 5g/L, yeast powder 1g/L, agar 2g/L, regulating pH is 7.4, deionized water preparation.
The vigor of the agarase of this embodiment gained is 1.71U/mL
Embodiment 4
(1) sea is revolved bacterium (
thalassospirasp) fjfst-2013007 is in seed culture medium, with the bottled 30mL seed culture medium of 250mL taper, sterilizing according to a conventional method, cooling, and inoculate bacterium colony, after inoculation, in 25 ℃, 160rpm shaking table obtains seed fermentation liquid after cultivating 24h, described seed culture medium: agar 2g/ L, sea salt 30 g/L, peptone 5g, yeast powder 1g, PH is 8.0, deionized water preparation.
(2) with the bottled 30mL fermention medium of 250mL taper, sterilizing according to a conventional method, cooling, and by 6% inoculum size, by seed fermentation liquid access fermention medium, after inoculation, in 25 ℃, 160rpm shaking table obtains the fermented liquid containing agarase after cultivating 72h, described fermention medium is: sea salt 25g/L, peptone 5g/L, yeast powder 1g/L, agar 2.5g/L, regulating pH is 7.5, deionized water preparation; .
The vigor of the agarase of this embodiment gained is 11.2U/mL
Embodiment 5
(1) by above-mentioned fermention medium through 4 ℃, after 8000g frozen centrifugation 15min, get supernatant liquor.
(2) supernatant liquor being added to massfraction is that 80% saturated ammonium sulphate is saltoutd, and places refrigerator overnight, and 4 ℃, 10000g is centrifugal, and 30min gets precipitation.
(3) with buffer A (0.02mol/L, the Tris-HCl damping fluid of pH7.5) balance DEAE-Sepharose Fast Flow post (Φ 2cm × 10cm), cross 4-5 bed volume.Take 0.3g agarase powder and be dissolved in 8mL buffer A, after the centrifugal 20min of 6000g, get ion exchange column on supernatant liquor.After loading, cross 2-3 bed volume by buffer A and wash away the foreign protein not being adsorbed, the NaCl solution of the 0-0.5mol/L then preparing in order to buffer A carries out gradient elution.Flow rate control is at 0.5mL/min, and detection wavelength is 280nm, elutriant fraction collection, 4.5mL/ pipe.In the test tube of mensuration wash-out pipe peak part, the enzyme activity of solution, merges activated part, obtains enzyme work in-process after lyophilize.
(4) Sephcryl S-100 gel permeation chromatography post (Φ 1.6cm × 80cm) is used after buffer A balance, take work in-process agarase 0.05g and be dissolved in 4mL level pad, after 6000g low-temperature centrifugation 20min, get chromatography column on supernatant liquor.Carry out wash-out by buffer A, coutroi velocity is 0.1mL/min.Elutriant is through ultraviolet detection fraction collection (4.0mL/ pipe).Determine respectively the enzyme activity of peak part test tube, activated part is merged, through fully concentrating through ultra-filtration membrane after dialysis desalting, to concentrated solution lyophilize, obtain finished product agarase.
SEQUENCE LISTING
<110> University Of Agriculture and Forestry In Fujian
<120> revolves bacterium in mono-strain sea and prepares the method for agarase
<130> 1
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1487
<212> DNA
<213> 16S rRNA
<400> 1
cagagtttga tcctggctca gaacgaacgc tggcggcagg cctaacacat gcaagtcgga 60
cgagaaggtt ccttcgggaa ctggagagtg gcgcacgggt gagtaacgcg tggggaccta 120
cctcttagtg ggggataacg gttggaaacg accgctaata ccgcatacgc ccttcggggg 180
aaagatttat cgctaagaga tggacccgcg ttggattaga tagttggtga ggtaacggct 240
caccaagtca gcgatccata gctggtttga gaggatgatc agccacactg ggactgagac 300
acggcccaga ctcctacggg aggcagcagt ggggaatatt ggacaatggg ggcaaccctg 360
atccagccat gccgcgtgag tgaagaaggc cttcgggttg taaagctctt tcagatgcga 420
agatgatgac ggtaacatca gaagaagccc cggctaattt cgtgccagca gccgcggtaa 480
tacgaaaggg gctagcgttg ttcggattta ctgggcgtaa agggcacgca ggcggtcttg 540
ccagtcaggg gtgaaagccc ggggctcaac cccggaactg cctctgatac tgcaagacta 600
gagactagga gagggtggtg gaattcccag tgtagaggtg aaattcgtag atattgggag 660
gaacaccaga ggcgaaggcg gccacctgga ctagatctga cgctcaggtg cgaaagcgtg 720
gggagcaaac aggattagat accctggtag tccacgccgt aaacgatgag tgctagttgt 780
cgggacttcg gtttcggtga cgcagctaac gcattaagca ctccgcctgg ggagtacggt 840
cgcaagatta aaactcaaag gaattgacgg gggcccgcac aagcggtgga gcatgtggtt 900
taattcgaag caacgcgcag aaccttacca acccttgaca tccctatcgc gattaccaga 960
gatggttttc atcagttcgg ctggataggt gacaggtgct gcatggctgt cgtcagctcg 1020
tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac ccctgttccc agttgccagc 1080
atttagttgg gcactctggg gagactgccg gtgacaagcc ggaggaaggc ggggatgacg 1140
tcaagtcctc atggccctta cgggttgggc tacacacgtg ctacaatggt aactacagag 1200
ggcagcgact tagcgataag gagccaatcc caaaaagtta tctcagttcg gattgcactc 1260
tgcaactcga gtgcatgaag ttggaatcgc tagtaatcgt ggatcagcat gccacggtga 1320
atacgttccc gggccttata cacaccgccc gtcacaccat gggagttggt tttacccgaa 1380
gacggtgggc taacctttta ggaggcagcc ggccacggta aggtcagcga ctggggtgaa 1440
gtcgtaacaa ggtagccgta ggggaacctg cggctggatc acctcct 1487
Claims (6)
1. bacterium is revolved in a strain sea, it is characterized in that: described sea revolve bacterium be sea revolve bacterium (
thalassospirasp.) fjfst-2013007, is being deposited in the registration preservation of Chinese Typical Representative culture collection center on December 25th, 2013, and its deposit number is CCTCC M 2013708.
2. bacterium is revolved in a strain sea according to claim 1, it is characterized in that: described sea revolve bacterium (
thalassospirasp.) sequence of the 16S rRNA of fjfst-2013007 is as shown in SEQ ID NO.1.
3. a purposes for bacterium is revolved in a strain sea as claimed in claim 1, it is characterized in that: for the preparation of agarase.
4. the method for bacterium for the preparation of agarase revolved in a strain sea as claimed in claim 1, it is characterized in that: described method comprises the steps:
(1) with the bottled 20-60mL seed culture medium of 250mL taper, sterilizing according to a conventional method, cooling, and inoculate bacterium colony, after inoculation, in 20-30 ℃, 160rpm shaking table obtains seed fermentation liquid after cultivating 12-24h;
(2) with the bottled 20-60mL fermention medium of 250mL taper, sterilizing according to a conventional method, cooling, and the inoculum size of pressing 2%-10%, by seed fermentation liquid access fermention medium, after 20-30 ℃ of cultivation 24-60h, obtain the fermented liquid containing agarase;
(3) then by fermention medium through 4 ℃, after 8000-10000g frozen centrifugation 10-20min, get supernatant liquor;
(4) supernatant liquor being added to massfraction is that the saturated ammonium sulphate of 60%-80% is saltoutd, and places refrigerator overnight, and 4 ℃, 8000-10000g is centrifugal, and 20-40min gets precipitation;
(5) with buffer A balance DEAE-Sepharose Fast Flow post, cross 4-5 bed volume, taking 0.3g agarase powder is dissolved in 8mL buffer A, after the centrifugal 20min of 6000g, get ion exchange column on supernatant liquor, after loading, cross 2-3 bed volume by buffer A and wash away the foreign protein not being adsorbed, the NaCl solution of the 0-0.5mol/L then preparing in order to buffer A carries out gradient elution; Flow rate control is at 0.5mL/min, and detection wavelength is 280nm, elutriant fraction collection, and 4.5mL/ pipe, in the test tube of mensuration wash-out pipe peak part, the enzyme activity of solution, merges activated part, obtains enzyme work in-process after lyophilize;
(6) Sephcryl S-100 gel permeation chromatography post is used after buffer A balance, taking work in-process agarase 0.05g is dissolved in 4mL level pad, after 6000g low-temperature centrifugation 20min, get chromatography column on supernatant liquor, carry out wash-out by buffer A, coutroi velocity is 0.1mL/min, elutriant is through ultraviolet detection fraction collection, 4.0mL/ pipe, determine respectively the enzyme activity of peak part test tube, activated part is merged, through fully concentrating through ultra-filtration membrane after dialysis desalting, to concentrated solution lyophilize, obtain finished product agarase.
5. the method for bacterium for the preparation of agarase revolved in a strain sea according to claim 4, it is characterized in that: described fermention medium is: sea salt 20-30g/L, peptone 3-7g/L, yeast powder 0.5-2.5g/L, agar 1.5-2.5g/L, adjusting pH is 7.2-8.4, deionized water preparation; Described seed culture medium: agar 2g/ L, sea salt 30 g/L, peptone 5g, yeast powder 1g, PH is 8.0, deionized water preparation.
6. the method for bacterium for the preparation of agarase revolved in a strain sea according to claim 4, it is characterized in that: described buffer A is 0.02mol/L, the Tris-HCl damping fluid of pH7.5.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105349461A (en) * | 2015-11-26 | 2016-02-24 | 福建农林大学 | Agarase generating vibrio alginolyticus and application thereof |
| CN106754538A (en) * | 2017-01-03 | 2017-05-31 | 青岛双瑞海洋环境工程股份有限公司 | Oil degradation bacteria Oil 23, the isolation and purification methods of oil degradation bacteria Oil 23 and its application |
| CN111826297A (en) * | 2019-04-18 | 2020-10-27 | 新乡医学院 | Breeding method of agarase-producing strain |
| CN113817710A (en) * | 2021-11-09 | 2021-12-21 | 蓝脑科技(厦门)有限公司 | Agarase freeze-drying protective agent and agarase preservation method |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101691556A (en) * | 2009-09-25 | 2010-04-07 | 中国科学院南海海洋研究所 | Deep sea thalassospira and application thereof |
| CN101838617A (en) * | 2009-03-17 | 2010-09-22 | 清华大学 | Thalassospira capable of degrading polyaromatic hydrocarbon and application thereof |
-
2014
- 2014-03-14 CN CN201410093126.8A patent/CN103898012B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101838617A (en) * | 2009-03-17 | 2010-09-22 | 清华大学 | Thalassospira capable of degrading polyaromatic hydrocarbon and application thereof |
| CN101691556A (en) * | 2009-09-25 | 2010-04-07 | 中国科学院南海海洋研究所 | Deep sea thalassospira and application thereof |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105349461A (en) * | 2015-11-26 | 2016-02-24 | 福建农林大学 | Agarase generating vibrio alginolyticus and application thereof |
| CN105349461B (en) * | 2015-11-26 | 2019-03-08 | 福建农林大学 | One plant of production agarase vibrio alginolyticus and application |
| CN106754538A (en) * | 2017-01-03 | 2017-05-31 | 青岛双瑞海洋环境工程股份有限公司 | Oil degradation bacteria Oil 23, the isolation and purification methods of oil degradation bacteria Oil 23 and its application |
| CN111826297A (en) * | 2019-04-18 | 2020-10-27 | 新乡医学院 | Breeding method of agarase-producing strain |
| CN113817710A (en) * | 2021-11-09 | 2021-12-21 | 蓝脑科技(厦门)有限公司 | Agarase freeze-drying protective agent and agarase preservation method |
| CN113817710B (en) * | 2021-11-09 | 2023-11-24 | 蓝脑科技(厦门)有限公司 | Agarase freeze-drying protective agent and agarase preservation method |
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