CN101363005A - Method for coculturing fine algae and photosynthetic bacteria - Google Patents
Method for coculturing fine algae and photosynthetic bacteria Download PDFInfo
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- CN101363005A CN101363005A CNA2008102223140A CN200810222314A CN101363005A CN 101363005 A CN101363005 A CN 101363005A CN A2008102223140 A CNA2008102223140 A CN A2008102223140A CN 200810222314 A CN200810222314 A CN 200810222314A CN 101363005 A CN101363005 A CN 101363005A
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Abstract
The invention provides a co-culture method of micro-algae and photosynthetic bacteria, and the co-culture method belongs to the field of biotechnology. The co-culture method is characterized in that: the co-culture method adopts a translucent glass or plastic container as the container and adopts tap water for preparing a liquid culture medium, the liquid culture medium is composed of 0.2 to 1.0g/L of MgSO47H2O, 1.0 to 3.0g/L of NaHCO3, 0.01 to 0.02g/L of CaCl2, 1.0 to 3.0g/L of NH4Cl, 1.0 to 2.0g/L of KH2PO4, 1.0 to 5.0g/L of NaCl, 0.2 to 1.0g/L of yeast extract, 2.0 to 5.0g/L of sodium acetate and 1.0ml/L of trace element liquid. The micro-algae and the photosynthetic bacteria are co-cultured at the average temperature of larger than 20 DEG C and under the solar radiation after the inoculation, the micro-algae and the photosynthetic bacteria with the high cell concentration can be obtained after 5 days, the obtained biomass of the micro-algae or the biomass of the photosynthetic bacteria can be respectively improved by 65 percent or 100 percent compared with the single culture, thereby realizing the purpose of high efficient co-culture of the micro-algae and the photosynthetic bacteria, and an algae bacteria culture which is obtained by culture can be applied as a water quality purifier and a feed additive for the aquaculture.
Description
Technical field
The invention belongs to biological technical field, a kind of fine algae and photosynthetic bacterium cultured method altogether particularly is provided.
Background technology
Fine algae belongs to lower plant, is simple photosynthesis organism.Chlorella is the unicell green alga of Chlorophyta (Chlorphyta) Chlorella (Chlorella), and nearly 10 kinds, the common chlorella vulgaris of China has chlorella ellipsoidea, Chlorella vulgaris and Chlorella pyrenoidesa etc.Chlorella rich in proteins, unsaturated fatty acids, carotenoid, xenthophylls, astaxanthin and multivitamin have high nutritive value and the function that improves immunizing power.In addition, the chlorella growth factor in the chlorella (Chlorella Growth Factor) had both had the function of bringing out the Interferon, rabbit immune cell activated, had again to promote detoxifcation and the excretory effect of body to objectionable impurities, and can promote the growth of cereuisiae fermentum.At present, chlorella is being a kind of good model animals aspect the researchs such as photosynthesis mechanism, transmembrane transport mechanism not only, and in sewage disposal, aspects such as protective foods, aquaculture bait and livestock feed additive are widely used.The scale of chlorella is cultivated the four kinds of modes that mainly contain: cultivate in open pond, cultivate in closed pond, closed illumination reaction device and unglazed according to the heterotrophic fermentation cultivation.
Photosynthetic bacterium (Photosynthetic Bacteria) is that a big class can be carried out the photosynthetic procaryotic general name of non-oxygen-production under the anaerobism illumination condition, belong to gram-negative bacteria, be divided into Rhodospirillaceae (Rhodospirillaceae), Chromatiaceae (Chromatiaceae), Chlorobacteriaceae (Chlorobiaceae) and 4 sections of green thread Cordycepps (Chloroflexaceae), mycetocyte mainly is divided into shaft-like and spherical 2 kinds of different shapes, be one of pioneer of life on earth origin, wherein not have sulphur bacterium (Rhodospirillaceae) be the more photosynthetic bacterium bacterial classification of industrialization production for rhodopseudomonas (Rhodopseudomonas) and purple.The protein content of photosynthetic bacterium is rich in Coenzyme Q10 99.0 (ubiquinone) that can be used as cellular metabolism activator and natural antioxidants and the carotenoid with antitumous effect simultaneously up to more than 60%, has important nutrition and pharmaceutical use.Photosynthetic bacterium has all been brought into play more and more important effect at aspects such as promoting growth of animal, processing high concentrated organic wastewater, aquaculture water purifying and the active bacterial manure of agricultural high-effiency as additive for farm animal feed.At present, photosynthetic bacterium mainly contains dynamically and leaves standstill 2 kinds of training methods, and culture apparatus has open and airtight 2 types.The report that the dynamic cultured continuously photosynthetic bacterium of closed bioreactor of adopting printing opacity is abroad arranged, but many at home employing glass jars and plastic tank leave standstill airtight cultivation under illumination.Dynamic airtight cultivation is carried out in the reactor of printing opacity, generally makes the photosynthetic bacterium liquid culture flow through the Glass tubing of spiral printing opacity to guarantee that culture can access sufficient illumination by pump.
Though at the report that a large amount of culture studies are all arranged aspect photosynthetic bacterium or the fine algae, but utilize the characteristics of physiological ecological separately of photosynthetic bacterium and fine algae how, realize that the two can both be grown fast and reach and but do not see the research report aspect the higher biomass under same substratum and physico chemical factor.Fine algae and photosynthetic bacterium are cultivated altogether, can make full use of space and resource on producing, and can realize the mutual supplement with each other's advantages of bacterium algae on using, and therefore have very important using value aspect the large-scale production phycomycete.
Summary of the invention
The object of the present invention is to provide the method for a kind of fine algae and photosynthetic bacterium co-cultivation, adopt the substratum of development to cultivate, can cultivate the phycomycete culture that produces high cell concentration fine algae and photosynthetic bacterium simultaneously.
The present invention adopts the glass of printing opacity or plastic ware as container, adopts tap water obtaining liq substratum, and liquid nutrient medium consists of (g/L): MgSO
47H
2O 0.2-1.0, NaHCO
31.0-3.0, CaCl
20.01-0.02, NH
4Cl1.0-3.0, KH
2PO
41.0-2.0, NaCl 1.0-5.0, yeast extract paste 0.2-1.0, sodium acetate 2.0-5.0, liquid microelement 1.0ml/L.Cultured fine algae and photosynthetic bacterium are inoculated, are carried out co-cultivation in medial temperature greater than under 20 ℃ and the natural solar light irradiation then, can obtain the fine algae and the photosynthetic bacterium of high cell concentration after 5 days,
Liquid microelement comprises (mg/L): FeCl
36H
2O 5.0, CuSO
45H
2O 0.05, H
3BO
31.0, MnCl
24H
2O 0.5, ZnSO
47H
2O 1.0, CoCl
26H
2O 0.5.
Photosynthetic bacterium that the present invention cultivated and fine algae all belong to photosynthetic microorganism, though that they carry out photosynthetic pathways metabolism is different with mechanism, can produce synergy to the efficient utilization of substratum under the co-cultivation condition.Under aerobic and anaerobism different condition, multiple pathways metabolism is arranged as photosynthetic bacterium, product carbonic acid gas to organic metabolism can be utilized by fine algae, and fine algae can help photosynthetic bacterium to utilize organism by aerobic mode by the oxygen that photosynthesis produces simultaneously.Therefore, fine algae and photosynthetic bacterium co-cultivation not only can make the composition of substratum be fully used, and physiological ecological characteristic of being had itself also can make the two be benefited simultaneously separately, promotes syntrophism.
The invention has the advantages that: under the co-cultivation condition, can make the growth continuously and healthily simultaneously of fine algae and photosynthetic bacterium, and finally obtain higher biomass, realized that industrialization produces the phycomycete culture of high cell concentration, can be used as aquaculture bait and livestock feed additive etc. and use.
Description of drawings
Fig. 1 is under the culture medium condition of being prepared in the embodiment of the invention 1, and single culture chlorella ellipsoidea or Rhodopseudomonas palustris are the growth curve of index with dry cell weight concentration.Wherein, X-coordinate is the time, unit: day; Ordinate zou is a dry cell weight concentration.
Show that by Fig. 1 respectively (the chlorella ellipsoidea cell concn is 8.0 * 10 for 0.15g/L in the initial cell dry weight concentrations
6/ ml or Rhodopseudomonas palustris cell concn are 6.0 * 10
7/ ml) time, the dry cell weight concentration of cultivating chlorella ellipsoidea in 5 day time or Rhodopseudomonas palustris all presents ascendant trend always, and cultivating and obtaining chlorella ellipsoidea dry cell weight concentration on the 5th day respectively is that (the chlorella ellipsoidea cell concn is 1.1 * 10 to 2.0g/L
8/ ml) or Rhodopseudomonas palustris dry cell weight concentration 1.65g/L (the Rhodopseudomonas palustris cell concn is 75.0 * 10
8/ ml).
Fig. 2 is under the culture medium condition of being prepared in the embodiment of the invention 1, and co-cultivation chlorella ellipsoidea and Rhodopseudomonas palustris are the growth curve of index with dry cell weight concentration.Wherein, X-coordinate is the time, unit: day; Ordinate zou is a dry cell weight concentration.
Shown by Fig. 2: in the initial cell dry weight concentrations all is under the 0.15g/L, the dry cell weight concentration of chlorella ellipsoidea and Rhodopseudomonas palustris all presents ascendant trend always in the culturing process, cultivating the 5th day acquisition chlorella ellipsoidea dry cell weight concentration is 1.8g/L, and corresponding frustule concentration is 1.0 * 10
8/ ml; Rhodopseudomonas palustris dry cell weight concentration is 1.5g/L, and corresponding Rhodopseudomonas palustris cell concn is 62.5 * 10
8/ ml; The total dry cell weight concentration of phycomycete is 3.3g/L, and dry cell weight concentration 2.0g/L or 1.65g/L (Fig. 1) much larger than single culture chlorella ellipsoidea or photosynthetic bacterium acquisition have improved 65% or 100% culture biomass respectively.Therefore adopt phycomycete cultured method altogether, both obtained very high photosynthetic bacteria cell concentration, also obtained simultaneously a large amount of algae bio amounts, very important using value and prospect have been arranged at the industrialization producer mask of fine algae and photosynthetic bacterium mixed culture.
Fig. 3 is under the culture medium condition of being prepared in the embodiment of the invention 2, and co-cultivation chlorella ellipsoidea and Rhodopseudomonas palustris are the growth curve of index with dry cell weight concentration.Wherein, X-coordinate is the time, unit: day; Ordinate zou is a dry cell weight concentration.
Shown by Fig. 3: in the initial cell dry weight concentrations all is under the 0.15g/L, the dry cell weight concentration of chlorella ellipsoidea and Rhodopseudomonas palustris all presents ascendant trend always in the culturing process, cultivating the 5th day acquisition chlorella ellipsoidea dry cell weight concentration is 0.8g/L, and corresponding frustule concentration is 0.4 * 10
8/ ml; Rhodopseudomonas palustris dry cell weight concentration is 0.7g/L, and corresponding Rhodopseudomonas palustris cell concn is 29.2 * 10
8/ ml; The total dry cell weight concentration of phycomycete only is 1.5g/L, obviously than the total dry cell weight concentration of the phycomycete 3.3g/L low (Fig. 2) that obtains under each moiety mean value embodiment 1 condition of substratum that adopts development.
Fig. 4 is under the culture medium condition of being prepared in the embodiment of the invention 3, and co-cultivation chlorella ellipsoidea and Rhodopseudomonas palustris are the growth curve of index with dry cell weight concentration.Wherein, X-coordinate is the time, unit: day; Ordinate zou is a dry cell weight concentration.
Shown by Fig. 4: in the initial cell dry weight concentrations all is under the 0.15g/L, the dry cell weight concentration of chlorella ellipsoidea and Rhodopseudomonas palustris all presents ascendant trend always in the culturing process, cultivating the 5th day acquisition chlorella ellipsoidea dry cell weight concentration is 2.2g/L, and corresponding frustule concentration is 1.2 * 10
8/ ml; Rhodopseudomonas palustris dry cell weight concentration is 1.8g/L, and corresponding Rhodopseudomonas palustris cell concn is 75.0 * 10
8/ ml; The total dry cell weight concentration of phycomycete is 4.0g/L.Though than the total dry cell weight concentration of the phycomycete 3.3g/L height (Fig. 2) that obtains under each moiety mean value embodiment 1 condition of substratum that adopts development, the amplitude that improves only is 21.2%.
Embodiment
At first adopting volume is 5 liters vial, adopts tap water according to 4.5 liters of liquid nutrient mediums of each moiety mean value preparation of substratum.The substratum that embodiment 1 adopts consists of (g/L): MgSO
40.6, NaHCO
32.0, CaCl
20.015, NH
4Cl 2.0, KH
2PO
41.5 NaCl 3.0, yeast extract paste 0.6, sodium acetate 3.5, liquid microelement 1.0ml/L.The substratum that embodiment 2 adopts consists of (g/L): MgSO
47H
2O 0.2, NaHCO
31.0, CaCl
20.01, NH
4Cl 1.0, KH
2PO
41.0 NaCl 1.0, yeast extract paste 0.2, sodium acetate 2.0, liquid microelement 1.0ml/L.The substratum that embodiment 3 adopts consists of (g/L): MgSO
47H
2O 1.0, NaHCO
33.0, CaCl
20.02, NH
4Cl 3.0, KH
2PO
42.0 NaCl 5.0, yeast extract paste 1.0, sodium acetate 5.0, liquid microelement 1.0ml/L.Liquid microelement comprises (mg/L): FeCl
36H
2O 5.0, CuSO
45H
2O 0.05, H
3BO
31.0, MnCl
24H
2O 0.5, ZnSO
47H
2O 1.0, CoCl
26H
2O 0.5.Cultured chlorella ellipsoidea and Rhodopseudomonas palustris seed are inoculated, carried out cultured continuously 5 days in medial temperature under greater than 20 ℃ natural solar irradiation condition, it is 1.8g/L that embodiment 1 has obtained chlorella ellipsoidea dry cell weight concentration, Rhodopseudomonas palustris dry cell weight concentration is 1.5g/L, the total dry cell weight concentration of phycomycete is 3.3g/L, dry cell weight concentration 2.0g/L or 1.65g/L (Fig. 1) much larger than single culture chlorella ellipsoidea or photosynthetic bacterium acquisition have improved 65% or 100% culture biomass respectively.The Rhodopseudomonas palustris of embodiment 1 high cell concentration and chlorella ellipsoidea (Fig. 2), the biomass (Fig. 1) that obtains than single culture chlorella ellipsoidea or Rhodopseudomonas palustris has improved 65% or 100% respectively.Embodiment 2 is under each moiety lower bound condition of substratum, and cultivating acquisition chlorella ellipsoidea dry cell weight concentration through 5 days is 0.8g/L, and Rhodopseudomonas palustris dry cell weight concentration is 0.7g/L, and the total dry cell weight concentration of phycomycete only is 1.5g/L.Embodiment 3 is under the high limit condition of each moiety of substratum, and cultivating acquisition chlorella ellipsoidea dry cell weight concentration through 5 days is 2.2g/L, and Rhodopseudomonas palustris dry cell weight concentration is 1.8g/L, and the total dry cell weight concentration of phycomycete is 4.0g/L.Therefore, fine algae and photosynthetic bacterium co-cultivation not only can make the composition of substratum be fully used, and physiological ecological characteristic of being had itself also can make the two be benefited simultaneously separately, reaches the promotion syntrophism, has important industrialization production application and is worth.
Claims (2)
1, the method for a kind of fine algae and photosynthetic bacterium co-cultivation is characterized in that, the present invention adopts the glass of printing opacity or plastic ware as container, adopts tap water obtaining liq substratum, and liquid nutrient medium consists of: MgSO
47H
2O 0.2-1.0g/L, NaHCO
31.0-3.0g/L, CaCl
20.01-0.02g/L, NH
4Cl 1.0-3.0g/L, KH
2PO
41.0-2.0g/L, NaCl 1.0-5.0g/L, yeast extract paste 0.2-1.0g/L, sodium acetate 2.0-5.0g/L, liquid microelement 1.0ml/L, cultured fine algae and photosynthetic bacterium are inoculated, carried out co-cultivation in medial temperature greater than under 20 ℃ and the natural solar light irradiation then, obtain the fine algae and the photosynthetic bacterium of high cell concentration after 5 days.
2, in accordance with the method for claim 1, it is characterized in that liquid microelement comprises: FeCl
36H
2O 5.0mg/L, CuSO
45H
2O 0.05mg/L, H
3BO
31.0mg/L, MnCl
24H
2O 0.5mg/L, ZnSO
47H
2O 1.0mg/L, CoCl
26H
2O 0.5mg/L.
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