CN101988045A - Composite culture medium for algae-killing bacterium and preparation method thereof - Google Patents
Composite culture medium for algae-killing bacterium and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a composite culture medium for an algae-killing bacterium and a preparation method thereof, relates to a culture medium, and provides a composite culture medium suitable for culturing the algae-killing bacterium Alteromonas addita DH46 and capable of obviously improving the cell density of the algae-killing bacterium, and a preparation method thereof. The composite culture medium comprises the following components, namely 1L of aged seawater contains 1 to 15g of tryptone, 0.5 to 5g of yeast powder, 0.5 to 5g of soluble starch, 0.1 to 3g of NaNO3, and 0.1 to 3g of MgSO4, wherein the pH is 7.2 to 7.8, and the salinity of the aged seawater is 2 to 4 percent. The preparation method comprises the following steps of: adding the tryptone, the yeast powder, the soluble starch, the NaNO3, and the MgSO4 into the aged seawater for fixing the volume, regulating the pH to be 7.2-7.8, and sterilizing to obtain the composite culture medium for the algae-killing bacterium.
Description
Technical field
The present invention relates to a kind of substratum, especially relate to complex medium of a kind of phycomycete Alteromonas addita DH46 of killing and preparation method thereof.
Background technology
Along with the aggravation of global body eutrophication, the occurrence frequency of harmful algal, scale and harm heighten, and have caused serious ecology, resource, environmental problem and great financial loss.At present, because red tide has become a kind of global public hazards, more than 30 countries and regions such as the U.S., Japan, China, Canada, France, Sweden, Norway, Philippines, India, Indonesia, Malaysia, Korea S, Hong Kong red tide takes place all very frequent, therefore study the red-tide control method, formulate the task of top priority that relevant Preventing Countermeasures has become many countries and regions.The Marine Planktonic algae is the main biology that causes red tide, has kind more than 260 can form red tide in more than the 4000 kind of Marine Planktonic algae in the whole world, wherein has kind more than 70 can produce toxin.The control techniques of algae can reduce physical method, chemical process and biological method at present.Exist under all circumscribed situations adopting physics and chemical process to kill algae, has the primary stage (Yang Xiaoru that is in researchdevelopment before safety, efficient, the easy biological method control Cutleriales, Su Jianqiang, Zheng Tianling. meaning and the prospect [J] of allelopathy in the red tide regulation and control. ACTA Scientiae Circumstantiae, 2008,8 (2): 219-226).Biological method mainly is to utilize biological ecological relationship each other to eliminate red tide, and these biologies comprise animal, plant and microorganism, and wherein utilizing method such as molten algae bacterium that harmful algal is carried out biological control is the domestic and international research focus.
Molten algae bacterium (algicidal bacteria) is that a class suppresses algal grown in direct or indirect mode, or kills the general designation of the bacterium of algae, dissolving frustule.Current, domestic research to molten algae bacterium still is in the preliminary stage, also just just begins (Zhang Yong for the research of the molten algae active metabolite of bacterium, Xi Yu, Wu Gang. molten algae bacterium kills the progress [J] of algae material. microbiology circular, 2004,31 (1): 127-131).Kill be seen in the algae bacterium report maximum be to lean on to discharge the outer material of born of the same parents and kill algae, and the outer material of secretion born of the same parents what usually with the height correlative connections of cell density, the optimization substratum then is a kind of effective ways that improve cell density.In order to obtain a certain amount of expansion that the algae material is used for effective separation of follow-up active substance and kills the work of algae Mechanism Study of killing, the optimization work of killing phycomycete substratum and the culture condition particularly important (He Peiqing that just seems, Tian Li, Li Youguang, Deng. the physicochemical property [J] of the outer antibacterial substance of marine bacteria B-9987 Optimizing Conditions of Fermentation and born of the same parents. Chinese marine drug, 2001,2 (80): 8-12).Nutrient media components is that the people is the most important culture environment that offers microorganism growth, and being influences microorganism growth propagation and meta-bolites synthetic important factor.The factor that influences microorganism growth process comprises cultivating forms (composition, concentration) and culture condition (temperature, pH value, shaking speed and inoculum size etc.), because the fermention medium composition is numerous, and often there is interaction in each factor, so the amount of medium optimization work is big and complicated.Multiple optimization method in the mathematical statistics has begun to be widely used in the optimization work of microbiological culture media, experimental technique wherein relatively more commonly used has single factor method, the orthogonal experiment design, homogeneous design, (Xia Li such as total divisor experimental design and response surface method, Tongcheng Xu, Xiaohang Ma, et al.2008.Optimization of culture conditions for production of cis-epoxysuccinic acid hydrolase using response surface methodology.Bioresource Technology.99,5391-5396).
The single-factor laboratory method is the common method in the optimization traditional operation of fermentation, promptly only changes the level of a factor at every turn, and other factors keep constant levels, then the optimization method investigated of factor one by one.In the routine experimentation, owing to do not need the knowledge of mathematical statistics aspect, simple to operate, therefore the result can be one of the most frequently used medium optimization method with pictorialization also.Though also there are some shortcomings in this method,, also can't show the interaction relationship between each factor as needs experiment and the long test period repeatedly.But the factor quantity that ought investigate more for a long time, single-factor laboratory method commonly used is observed the influence of each factor to microorganism growth and product generation, filter out the horizontal extent of bigger influence factor and definite factor, be used for statistical method and design multifactor level experiment, thereby reduce to test quantity greatly and analyze difficulty.
Homogeneous design be original creation such as China mathematician Fang Kaitai a kind of multifactorial experiment optimized method (Fang Kaitai, Wang Yuan. homogeneous design and uniform designs table [M]. Beijing: Science Press, 1994,21-23).This method combines the principle and the multivariate statistics of number theory, at first obtains remarkable application in China's winged missile design, is obtaining many successful examples aspect the optimization of microbiological culture media in recent years.The basic ideas of homogeneous design are exactly to make the experimental point homodisperse as far as possible, make each experimental point have better representativeness, but give up neat comparable requirement simultaneously, reducing experiment number, this itself and the maximum difference of orthogonal experimental design method; Method by multivariate statistics remedies this defective then, makes experimental result same effectively reliable.Under the identical condition of experiment number, the deviation of homogeneous design is little more than orthogonal design.Because homogeneous design is no longer considered the neat comparability of orthogonal test, so the processing of its test-results will be adopted regression analysis-linear regression or polynomial regression analysis.
Current, the report of the fermentation optimization of directed toward bacteria extracellular products is existing a lot, mainly lays particular emphasis on medium component and Optimizing Conditions of Fermentation.But rarely has report at killing the research of phycomycete with the optimization aspect of killing the algae material.Pre-stage test shows that phycomycete DH46 can secrete the outer active substance of born of the same parents extremely, the host algae is risen suppress or killing action.At present, domestic the research of killing phycomycete still is in the starting stage, but efficient, single-minded by killing the algae bacteria screening, can biodegradablely kill the new approaches that the algae material become the exploitation algicide.For its active result is applied to the practice in, the problem that at first runs into is exactly the optimization of culturing process.What of tunning are usually related with the height of cell density, and optimization substratum and culture condition thereof then are a kind of effective ways that improve cell density.
In addition, substratum is the people for offering the usefulness of thalli growth, breeding and synthetic product, for make thalli growth reach high-density certainly will to design a kind of contain must composition the balance nutritional medium.Zobell 2216E substratum is as ocean aerobic bacteria isolation medium commonly used, be by father-Zuo Beier (C.E.ZoBell) invention for marine microbiology, prescription is: peptone (Peptone) 5g, yeast extract (Yeast Extract) 1g, high ferric phosphate 0.1g, agar powder 10g (solid medium), pH7.0-7.2, Chen Haishui constant volume to 1L (referring to document: Su, J.Q., Yang, X.R., Zheng, T.L., Tian, Y., Jiao, N.Z., Cai, L.Z., Hong, H.S.Isolation and characterization of a marine algicidal bacterium against the toxic dinoflagellate Alexandrium tamarense[J] .Harmful Algae, 2007,6 (6): 799-810).It has vital role in practice as the traditional substratum that separates, cultivates marine bacteria, but different bacteriums has different nutritional needss.
Summary of the invention
The object of the present invention is to provide and a kind ofly be suitable for cultivating extremely phycomycete Alteromonas addita DH46, can significantly improve complex medium that kills phycomycete of its cell density and preparation method thereof.
Described phycomycete extremely is Alteromonas addita DH46, is preserved in Chinese typical culture collection center on 07 14th, 2010, and the address is a Chinese Wuhan Wuhan University (postcode 430072), and preservation center deposit number is CCTCC NO:M2010177.
Consisting of of the complex medium of described phycomycete extremely, in the Chen Haishui of 1L, contain:
Yeast powder 0.5～5g;
Zulkovsky starch 0.5～5g;
NaNO
3 0.1～3g；
MgSO
4 0.1～3g。
The described pH that kills the complex medium of phycomycete can be 7.2～7.8; The salinity of described old seawater can be 20 ‰～40 ‰.
The described composition that kills the complex medium of phycomycete is preferably, and contains in the Chen Haishui of 1L:
Tryptones 14g;
Yeast powder 1.63g;
Zulkovsky starch 5.0g;
NaNO
3 1.6g；
MgSO
4 2.3g。
The described pH that kills the complex medium of phycomycete is preferably 7; The salinity of described old seawater is preferably 30 ‰.
The described preparation method who kills the complex medium of phycomycete is:
With Tryptones, yeast powder, Zulkovsky starch, NaNO
3And MgSO
4Add constant volume among the Chen Haishui, regulating pH is 7.2～7.8, after the sterilization, promptly gets the complex medium that kills phycomycete.
Described sterilization can be adopted 121 ℃ of autoclavings, 20min.
Kill phycomycete Alteromonas addita DH46 has the special physiological function as a strain microorganism, because its growth may be subjected to the influence of various ambient conditionss, as temperature, pH, salinity, dissolved oxygen amount etc., therefore be badly in need of being optimized, thereby promote to kill the generation of algae material and the development of algicide killing phycomycete substratum and culture condition.
The present invention is based on ZoBell 2216E substratum, the composition and the concentration of substratum have been investigated, and culture condition etc. is to killing the influence of phycomycete Alteromonas addita DH46 shake-flask culture, and utilize the homogeneous design experiment that substratum is optimized, provided the simple composite substratum that a kind of suitable high-density culture is killed phycomycete Alteromonas addita DH46.
Description of drawings
Fig. 1 is that differing temps is to the growth of phycomycete DH46 and the influence of algae killing effect extremely.In Fig. 1, X-coordinate be temperature (℃), left ordinate zou is for killing algae rate (%), right ordinate zou is the optical density value OD600 under the wavelength 600nm; Oblique line partly is the algae rate of killing, ● be OD600.
Fig. 2 is that different pH values are to the growth of phycomycete DH46 and the influence of algae killing effect extremely.In Fig. 2, X-coordinate is the pH value, and left ordinate zou is for killing algae rate (%), and right ordinate zou is the optical density value OD600 under the wavelength 600nm; Oblique line partly is the algae rate of killing, ● be OD600.
Fig. 3 is that different salinity is to the growth of phycomycete DH46 and the influence of algae killing effect extremely.In Fig. 3, X-coordinate is salinity (‰), and left ordinate zou is for killing algae rate (%), and right ordinate zou is the optical density value OD600 under the wavelength 600nm; Oblique line partly is the algae rate of killing, ● be OD600.
Fig. 4 is that different rotating speeds is to the growth of phycomycete DH46 and the influence of algae killing effect extremely.In Fig. 4, X-coordinate is rotating speed (r/min), and left ordinate zou is for killing algae rate (%), and right ordinate zou is the optical density value OD600 under the wavelength 600nm; Oblique line partly is the algae rate of killing, ● be OD600.
Fig. 5 is that the different composite nutritive ingredient is to the growth of phycomycete DH46 and the influence of algae killing effect extremely.In Fig. 5, X-coordinate is the compound nutritional composition, is extractum carnis, yeast powder, Tryptones, peptone from left to right, and left ordinate zou is for killing algae rate (%), and right ordinate zou is the optical density value OD600 under the wavelength 600nm; Oblique line partly is the algae rate of killing, ● be OD600.
Fig. 6 is that different added ingredientss are to the growth of phycomycete DH46 and the influence of algae killing effect extremely.In Fig. 6, X-coordinate is the compound nutritional composition, from left to right is contrast 1 (Control1), contrast 2 (Control2), sucrose, glucose, Trisodium Citrate, Zulkovsky starch, soy peptone, acid hydrolysis casein peptone, glycine, NaNO
3, MgSO
4, left ordinate zou is the optical density value OD600 under the wavelength 600nm, right ordinate zou is for killing algae rate (%); A is for killing the algae rate, and b is OD600.
Embodiment
Following embodiment further specifies of the present invention, but the invention is not restricted to following embodiment.
1. bacterial classification and algae kind
Bacterial classification: DH46 derives from resource and environment institute of microbiology of Xiamen University to be separated from the seawater sample that red tide district in the East Sea collects and screens in red tide 973 project MC2003-2 flight numbers in 2003, is the alternately Zymomonas mobilis that a strain has algae effect extremely.
The algae kind: the no bacterial strain of Alexandrium tamarense (AT), algae kind system is provided by hydrobiont institute of Ji'nan University, and the Alexandrium tamarense that obtains through the degerming of the aseptic algae technology of the applicant does not have bacterial strain.The used nutrient solution of algae is the f/2 nutrient solution.Algae places indoor triangular flask to cultivate, and temperature is 20 ± 1 ℃, and illumination condition is 12h illumination: the 12h dark.
2. measuring method
Tuurbidimetry (optical density, OD): get the thalline fermented liquid of suitable stoste or dilution, do contrast, measure optical density value OD down in wavelength 600nm with honourable photometer with blank substratum.
Dry cell weight (DCW): draw fermented liquid 10ml, the centrifugal 10min of 8000r/min uses deionized water wash thalline 3 times after the supernatant discarded, and 80 ℃ of constant temperature dry to constant weight, and weigh and calculate DCW (g/L).
Kill the algae rate: get the cell-free filtrate after the fermentation culture, the ratio according to 5% is added in the test algae, fixes with Compound Iodine Solutlon behind the effect 24h, counts under opticmicroscope.Algicidal activity (%)=(N
C-N
T)/N
C* 100, in the formula, N
CViable count in the expression control group, N
TViable count in the expression experimental group.
Below provide the described optimizing process that kills the complex medium of phycomycete.
From the factor of influence of substratum, find out the remarkably influenced factor, adopt uniform design to be optimized.Data Processing System (DPS) 3.01 data processing softwares are adopted in experimental design among the present invention, data analysis.
At first adopting Zobell 2216E is initial medium, carries out the exploration that single factor method is optimized culture condition.
Temperature Influence: setting culture temperature respectively is 20,25,28,30 and 32 ℃, it is 100ml that pH7.0～7.2,1% inoculum sizes, 250ml triangle shake bottled liquid measure, after the 150r/min shaking table is cultivated 24h, get bacterium liquid and measure optical density value (being OD600) down and kill the algae rate in wavelength 600nm; The result as shown in Figure 1.
The influence of initial pH: the initial pH value of setting substratum respectively is 3～11, and culture temperature is 28 ℃, 1% inoculum size, and it is 100ml that the 250ml triangle shakes bottled liquid measure, after the 150r/min shaking table is cultivated 24h, gets bacterium liquid and measures OD600 and kill the algae rate; The result as shown in Figure 2.
The influence of salinity: the salinity of setting substratum with artificial seawater respectively is 0,10,20,30,40 and 68 ‰, initial pH7.0～7.2, and all the other conditions are same as described above, get fermented liquid then and measure OD600 and kill the algae rate; The result as shown in Figure 3.
The influence of rotating speed: set shaking table concussion speed respectively and be 120,150,180 and 210r/min, all the other conditions are same as described above, cultivate to get fermented liquid behind the 24h and measure OD600 and kill the algae rate; The result as shown in Figure 4.
Experimental result shows, this bacterium under 28～30 ℃ of culture condition, the ability that its biomass and kill the algae rate and all can reach high value, thalline produce active substance between 20～30 ℃ in temperature influence less, but cell density increases along with the rising of temperature earlier, descends subsequently.In the temperature change process, thalli growth is consistent with the generation rule of killing the algae material.Consider and save the energy so select 28 ℃ to be culture condition from now on.Its suitableeest growth pH is separately arranged for microbial fermentation, DH46 the pH value be higher than 9 and be lower than 6 meta-acid or partially the growth of alkali environment hypothallus be subjected to press down, when pH is 7, obtains biomass and kill the maximum of algae rate.Salinity has significance for the growth increment of marine microorganism, and by the result as can be known, the generation of DH46 growth and algicdal activity material thereof all is subjected to the remarkably influenced that salt concn changes.When the substratum salinity was 0, thalline was grown hardly, and killing the algae rate only is 14.1%; Along with salinity increase thalli growth is rapid, algae killing effect obviously strengthens; When salinity greater than 30 ‰, increase though the algae rate of killing of this bacterium has slightly, thalli growth obviously slows down.Therefore 30 ‰ be the salinity range that is fit to kill phycomycete DH46.What the change of rotating speed directly influenced is exactly the dissolved oxygen amount of microorganism, too high or too low dissolved oxygen speed all is unfavorable for thalli growth, when rotating speed is 180r/min, and biomass and kill the algae rate and be respectively 1.085,100%, this value is the shaking speed of suitable thalli growth and product active substance.
So the optimal culture condition of DH46 is: culture temperature is 28 ℃, and the initial pH of suitable culture base is 7, and salinity is 30 ‰, and rotating speed is 180r/min.
Substratum mainly comprises several big compositions of carbon source, nitrogenous source and inorganic salt, because the fermention medium composition is numerous, and often there is interaction in each factor, is difficult to the model that theorizes; In addition, because take off data often comprises bigger error, also influenced the accurate assessment of medium optimization process, so the amount of medium optimization work is big and complicated.At first select several complex nutrients sources in the present invention, be that those not only can but also can make the nutritive substance (as peptone, yeast powder etc.) of nitrogenous source as carbon source, on their basis, add different trace ingredientss then, investigate each composition growth of killing phycomycete and the influence of producing active substance.
With Zobell 2216E substratum is the basis (no carbon source, nitrogenous source) of screening and optimizing, and with the 5g/L extractum carnis, yeast powder, Tryptones and peptone are complex nutrients sources, and all the other component concentrations are same as basic medium.Culture condition is: inoculum size is 1%, 28 ℃, and 180r/min, pH are 7, and salinity is 30 ‰, measures OD600 behind the shake-flask culture 24h and kills the algae rate.Result such as Fig. 5 show that DH46 grows in the nutrient solution that with the Tryptones is nutrition source, its thalline biomass is with the algae rate is all good extremely, and its value is respectively 1.18 and 96.9%, so select it as main nutrient matter, continue follow-up experiment.
Prescription with reference to classical substratum Zobell 2216E substratum adds various material compositions respectively on the basis of Tryptones, study their nutritional supplementation effects to Tryptones, for the screening and optimizing substratum is done further preparation.Be specially: the glucose, sucrose, Trisodium Citrate, Zulkovsky starch, yeast powder, soy peptone, acid hydrolyzed casein peptone, glycine, the NaNO that on the basis of 5g/L Tryptones, add 1g/L respectively
3And MgSO
4, all the other component concentrations are same as basic medium.Culture condition is: inoculum size is 1%, 28 ℃, and 180r/min, pH are 7, and salinity is 30 ‰, measures OD600 behind the shake-flask culture 24h and kills the algae rate.Whether the component in conjunction with increment and algae rate investigation adding extremely exists promoter action to killing phycomycete.Wherein, only add a kind of composition of Tryptones in the contrast 1, contrast 2 is for adding yeast powder (being Zobell 2216E substratum) on the Tryptones basis.The result as shown in Figure 6, the fermented liquid biomass difference that shows different tests group substratum DH46 is little, but algae killing effect difference is clearly, may since medium component and proportioning Different Effects DH46 produce active ability of killing the algae material, wherein principle remains further research.
The contrast 1 that only is added with Tryptones is an increment or killing the algae rate all is starkly lower than contrast 2, promptly former basic medium; Carbon source is added in the component, it only is 23.6% that the somatic cells filtrate of the substratum results of interpolation sucrose is killed the algae rate, glucose is 68.9%, Trisodium Citrate is 71.6%, all be lower than 89.3% the algae rate of killing of former basic medium, show that these carbon source compositions do not have promoter action to killing phycomycete DH46 secretion activity material; Nitrogenous source adds the soy peptone in the component, acid hydrolyzed casein peptone, the thalline biomass of glycine and kill the algae rate also not as good as former basic medium effect.Add two kinds of inorganic salt NaNO
3, MgSO
4The algae rate of killing of substratum all reach more than 95%, improved thalline effectively and produced the ability of killing the algae material.Consider the huge contribution (its OD600 be 1.47) of Zulkovsky starch, contract Tryptones, yeast powder, NaNO to the thalline biomass
3, MgSO
4All list factor of influence in, participate in the medium optimization process.
Screening and optimizing kills the phycomycete complex medium, adopts uniform design to determine this substratum optimum formula composition.
Comprehensive The above results is selected Tryptones, starch, yeast powder, NaNO
3, MgSO
4For factor of influence carries out the homogeneous design of 5 factors, 15 levels, all the other components are same as initial medium.5 factors are respectively independent variable(s) X1, X2, X3, X4 and X5, and dry cell weight is dependent variable Y.Culture condition is selected: inoculum size is 1%, 28 ℃, and 180r/min, pH are 7, and salinity is 30 ‰.Tryptones is provided with 15 levels, and all the other all are provided with 5 levels.Experimental result through DPS software data treatment system quadratic polynomial stepwise regression analysis, and is carried out test of significance to this model, experimental design and the results are shown in Table 1.
Table 15 factors 15 horizontal uniform designs table U15 (5
4* 15) and test-results
Multiple regression equation is as follows:
Y＝3.219+0.0969X1+0.484X2-0.543X3-0.0542X5+0.00174X1*X1-0.0325
Coefficient R=1, F value=38461.5, conspicuous level p=0.004, residual standard deviation S=0.00015, adjusted coefficient R a=0.99999, Durbin-Watson statistic d=1.78.Illustrate that this equation is believable to the simulation and forecast of dry cell weight.
By the experiment of above homogeneous design, each factor all has certain influence to killing algae rate and thalli growth amount as can be known, should kill phycomycete DH46 the optimal medium scheme be (g/L): Tryptones 14.0, yeast powder 1.63, starch 5.0, NaNO
31.6, MgSO
42.3.
Below provide the present invention and prepare the described embodiment that kills the complex medium of phycomycete.
Consisting of of the complex medium of described phycomycete extremely, in the Chen Haishui of 1L, contain:
Tryptones 14g;
Yeast powder 1.63g;
Zulkovsky starch 5.0g;
NaNO
3 1.6g；
MgSO
4 2.3g；
Regulating pH is 7, and the salinity of described old seawater is 30 ‰, and its preparation method is: with Tryptones, yeast powder, Zulkovsky starch, NaNO
3And MgSO
4Add constant volume among the Chen Haishui, regulating pH is 7,121 ℃ of autoclavings, 20min.Promptly get the complex medium that kills phycomycete.
By above-mentioned optimization experiment, obtained killing optimum medium prescription and the optimal culture condition of phycomycete DH46.With initial medium with optimize the shake-flask culture that substratum is respectively applied for DH46, culture condition adopts optimizes culture condition.Thalline fermentation obvious effect of increasing production after the optimization is as table 2.Cultivate 24h in shaking bottle after, the dry cell weight value of its stationary phase can be up to 7.36g/L, surpasses the 6.37g/L with the predictor of regression model, and has improved nearly one times than the basic medium before optimizing.Illustrate and utilize not only feasibility of homogeneous design optimization experiment, still effectively.
Culture effect contrast before and after table 2 is optimized
Similar to Example 1, its difference is that the component of substratum is for to contain in the Chen Haishui of 1L:
Tryptones 1g;
Yeast powder 1g;
Zulkovsky starch 0.5g;
NaNO
3 1g；
MgSO
4 0.1g；
Regulating pH is 7.5, and the salinity of described old seawater is 20 ‰.
Similar to Example 1, its difference is that the component of substratum is for to contain in the Chen Haishui of 1L:
Tryptones 15g;
Yeast powder 5g;
Zulkovsky starch 5g;
NaNO
3 3g；
MgSO
4 3g；
Regulating pH is 7.8, and the salinity of described old seawater is 40 ‰.
Embodiment 4
Similar to Example 1, its difference is that the component of substratum is for to contain in the Chen Haishui of 1L:
Tryptones 5g;
Yeast powder 1g;
Zulkovsky starch 1g;
NaNO
3 1g；
MgSO
4 1g；
Regulating pH is 7.2, and the salinity of described old seawater is 35 ‰.
Embodiment 5
Similar to Example 1, its difference is that the component of substratum is for to contain in the Chen Haishui of 1L:
Tryptones 10g;
Yeast powder 3g;
Zulkovsky starch 3g;
NaNO
3 1.5g；
MgSO
4 1.5g；
Regulating pH is 7.4, and the salinity of described old seawater is 25 ‰.
Claims (8)
1. a complex medium that kills phycomycete is characterized in that described phycomycete extremely is Alteromonas addita DH46, is preserved in Chinese typical culture collection center on 07 14th, 2010, and preservation center deposit number is CCTCC NO:M2010177;
Consisting of of the complex medium of described phycomycete extremely, in the Chen Haishui of 1L, contain:
Tryptones 1～15g;
Yeast powder 0.5～5g;
Zulkovsky starch 0.5～5g;
NaNO
3 0.1～3g；
MgSO
4 0.1～3g。
2. a kind of complex medium that kills phycomycete as claimed in claim 1 is characterized in that the described pH that kills the complex medium of phycomycete is 7.2～7.8.
3. a kind of complex medium that kills phycomycete as claimed in claim 1, the salinity that it is characterized in that described old seawater is 20 ‰～40 ‰.
4. a kind of complex medium that kills phycomycete as claimed in claim 1 is characterized in that consisting of of the described complex medium that kills phycomycete, contains in the Chen Haishui of 1L:
Tryptones 14g;
Yeast powder 1.63g;
Zulkovsky starch 5.0g;
NaNO
3 1.6g；
MgSO
4 2.3g。
5. a kind of complex medium that kills phycomycete as claimed in claim 2 is characterized in that the described pH that kills the complex medium of phycomycete is 7.
6. a kind of complex medium that kills phycomycete as claimed in claim 3, the salinity that it is characterized in that described old seawater is 30 ‰.
7. a kind of preparation method who kills the complex medium of phycomycete as claimed in claim 1 is:
With Tryptones, yeast powder, Zulkovsky starch, NaNO
3And MgSO
4Add constant volume among the Chen Haishui, regulating pH is 7.2～7.8, after the sterilization, promptly gets the complex medium that kills phycomycete.
8. a kind of preparation method who kills the complex medium of phycomycete as claimed in claim 7 is characterized in that described sterilization, is to adopt 121 ℃ of autoclavings, 20min.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102604868A (en) * | 2012-03-21 | 2012-07-25 | 清华大学深圳研究生院 | Alteromonas and application thereof in inhibiting growth of red-tide algae |
| CN103451140A (en) * | 2013-09-27 | 2013-12-18 | 厦门大学 | Immobilized method of algal inhibiting bacteria DH46 and application thereof |
| CN103468622A (en) * | 2013-09-27 | 2013-12-25 | 厦门大学 | Method for fixing algae inhibiting bacteria DH46 through sodium alginate-microporous starch |
| CN114698652A (en) * | 2022-01-17 | 2022-07-05 | 中国科学院生态环境研究中心 | Application of composition in preparation of algaecide, algaecide and in-situ algaecide method |
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| JPH0528597B2 (en) * | 1987-03-03 | 1993-04-26 | Meiji Seika Co | |
| JPH05103641A (en) * | 1991-10-15 | 1993-04-27 | Marino Forum 21 | New food using degradation product of seaweed of brown algae |
| US5418156A (en) * | 1991-04-15 | 1995-05-23 | University Of Maryland | Agarase enzyme system from alteromonas strain 2-40 |
| US7927604B2 (en) * | 2006-12-25 | 2011-04-19 | Sugiyo Co., Ltd. | Method of preparing a seaweed degradation product and a composition for preparing a seaweed degradation product |
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| JPH0528597B2 (en) * | 1987-03-03 | 1993-04-26 | Meiji Seika Co | |
| US5418156A (en) * | 1991-04-15 | 1995-05-23 | University Of Maryland | Agarase enzyme system from alteromonas strain 2-40 |
| JPH05103641A (en) * | 1991-10-15 | 1993-04-27 | Marino Forum 21 | New food using degradation product of seaweed of brown algae |
| US7927604B2 (en) * | 2006-12-25 | 2011-04-19 | Sugiyo Co., Ltd. | Method of preparing a seaweed degradation product and a composition for preparing a seaweed degradation product |
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| 《FEMS Microbiol Lett》 20100528 Yan Wang,et al Aestuariibacter aggregatus sp.nov., a moderately halophilic bacterium isolated from seawater of the Yellow Sea 48-54 1-8 第309卷, 2 * |
| 《厦门大学学报(自然科学版)》 20061231 郑天凌等 海洋微生物研究的回顾与展望 150－157 1－8 第45卷, 2 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102604868A (en) * | 2012-03-21 | 2012-07-25 | 清华大学深圳研究生院 | Alteromonas and application thereof in inhibiting growth of red-tide algae |
| 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 |
| CN103451140A (en) * | 2013-09-27 | 2013-12-18 | 厦门大学 | Immobilized method of algal inhibiting bacteria DH46 and application thereof |
| CN103468622A (en) * | 2013-09-27 | 2013-12-25 | 厦门大学 | Method for fixing algae inhibiting bacteria DH46 through sodium alginate-microporous starch |
| CN114698652A (en) * | 2022-01-17 | 2022-07-05 | 中国科学院生态环境研究中心 | Application of composition in preparation of algaecide, algaecide and in-situ algaecide method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101988045B (en) | 2012-08-22 |
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