CN104450588A - Method for culturing rhodopseudomonas palustris - Google Patents
Method for culturing rhodopseudomonas palustris Download PDFInfo
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- CN104450588A CN104450588A CN201410816451.2A CN201410816451A CN104450588A CN 104450588 A CN104450588 A CN 104450588A CN 201410816451 A CN201410816451 A CN 201410816451A CN 104450588 A CN104450588 A CN 104450588A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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Abstract
The invention discloses a method for culturing rhodopseudomonas palustris. The method comprises the following steps: (1) preparing a culture medium; (2) disinfecting and sterilizing; (3) inoculating; (4) culturing, namely culturing an inoculated culture bottle on a culture shelf of a culture room at a temperature of 28-32 DEG C, and transferring 1-2 times after the culture bottle is cultured for 2-3 days, wherein the wall of the culture room is respectively formed by a conventional wall, a heat preserving and insulating material and a heating cable from outside to inside; every 10-12 culture bottles are closely enclosed to form a circle, and an incandescent lamp which has power of 40W is arranged right above the circle; and the distance from the culture bottles to the bulb vertically above the culture bottles is 10cm. The rhodopseudomonas palustris cultured by the method has the bacterial concentration of 5*10<9>/mL in each bottle, and has high and uniform concentration.
Description
Technical field
The invention belongs to the cultural method field of bacterium, be specifically related to a kind of cultural method of Rhodopseudomonas palustris.
Background technology
Photosynthetic bacterium (Photosynthetic bacteria) is called for short PSB, is to utilize luminous energy and CO
2maintain the general name of coloured bacterium of its autotrophy life, also be the earth occurs the earliest, the ubiquitous a kind of prokaryotic organism of occurring in nature, can under anaerobism illumination condition or under aerobic dark condition, using light as the energy, utilize the organism of occurring in nature, ammonia etc. as keeping body to carry out photosynthetic microorganism.Photosynthetic bacterial thallus is nontoxic, the very high and A wide selection of colours and designs of its protein content, in addition, also stores thing and Poly-β-hydroxybutyric acid, ubiquinone, antiviral substance and somatomedin etc. containing abundant carbon element in somatic cells.
Rhodopseudomonas palustris (Rhodop seudanonas palustris) belongs to the one of photosynthetic bacterium, belongs to Rhodospirillaceae, Rhodopseudomonas, can be used as a kind of fodder additives, have very wide application prospect in aquaculture; Also can be used for purifying water, improving plant salt endurance, resistance to lead etc.But the density unevenness one of the Rhodopseudomonas palustris that cellar culture goes out, difference is comparatively large, therefore in the utilizing of Rhodopseudomonas palustris, operates more loaded down with trivial details, increases and utilize cost.
Summary of the invention
The present invention is directed to above-mentioned weak point and the cultural method of a kind of Rhodopseudomonas palustris provided, the method is simple to operate, and the Rhodopseudomonas palustris concentration uniform quality of turning out.
For achieving the above object, the technical solution adopted for the present invention to solve the technical problems is:
A cultural method for Rhodopseudomonas palustris, comprises the following steps:
(1) substratum is prepared
Take NaCl 5g, NH respectively
4cl 5g, CH
3cOONa 15g, peptone 15g, MgSO
41g, K
2hPO
41g, 5% NaHCO
3solution 100mL, is dissolved in 5L distilled water, stirs;
(2) sterilization
The substratum that stirs loaded in stainless steel cask, to carry out disinfection sterilizing with ozonizer, sterilization 10min, the usage quantity of ozone is 1g;
(3) inoculate
Get the Rhodopseudomonas palustris mother liquor 3-5mL after activation and be placed in 500mL triangular flask, in culturing bottle, then add sterilized substratum to bottleneck, whole seeded process equipment used is all sterilized;
(4) cultivate
The culturing rack finished culturing bottle being put into culturing room is cultivated, and culture temperature is 28 DEG C-32 DEG C, cultivates and transfers 1-2 time after 2-3 days.
In order to ensure the homo(io)thermism of culturing room, further, the wall of culturing room is designed to multilayer double wall, is conventional wall, heat preserving and insulating material and heating cables from outside to inside respectively.
In order to provide balanced photoenvironment to Rhodopseudomonas palustris, further, near surrounding a circle, directly over it, 1 incandescent light is installed for the every 10-12 of the culturing bottle on culturing rack, the power of incandescent light is 40W, and the bulb distance of culturing bottle and vertical direction is 10cm.
The cultural method of a kind of Rhodopseudomonas palustris provided by the invention, has following beneficial effect:
(1) culturing room is multilayer double wall, conventional wall, heat preserving and insulating material and heating cables from outside to inside respectively, indoor are also provided with temperature regulator, vent fan, aerofoil fan etc. simultaneously, ensure that room temp is constant and evenly, the growth for Rhodopseudomonas palustris provides best growing environment.
(2) the every 10-12 of the culturing bottle on culturing rack near surrounding a circle, directly over it, 1 incandescent light is installed, the power of incandescent light is 40W, the bulb distance of culturing bottle and vertical direction is 10cm, this is designed to Rhodopseudomonas palustris and improves uniform light intensity, makes Rhodopseudomonas palustris growth concentration balanced.
Embodiment
Embodiment 1
A cultural method for Rhodopseudomonas palustris, comprises the following steps:
(1) substratum is prepared
Take NaCl 5g, NH respectively
4cl 5g, CH
3cOONa 15g, peptone 15g, MgSO
41g, K
2hPO
41g, 5% NaHCO
3solution 100mL, is dissolved in 5L distilled water, stirs;
(2) sterilization
The substratum that stirs loaded in stainless steel cask, to carry out disinfection sterilizing with ozonizer, sterilization 10min, the usage quantity of ozone is 1g, and this process can make the complete sterilizing of substratum, in case Rhodopseudomonas palustris is contaminated;
(3) inoculate
Get the Rhodopseudomonas palustris mother liquor 3-5mL after activation and be placed in 500mL triangular flask, in culturing bottle, then add sterilized substratum to bottleneck, whole seeded process equipment used is all sterilized;
(4) cultivate
The culturing rack finished culturing bottle being put into culturing room is cultivated, and culture temperature is 28 DEG C-32 DEG C, and cultivate after 2-3 days, bacterium liquid reddens, and then transfers 1-2 time, wants temperature and light photograph situation in frequent observation ward between Rhodopseudomonas palustris incubation period.
In order to ensure the homo(io)thermism of culturing room, culturing room's wall is designed to multilayer double wall, conventional wall, heat preserving and insulating material and heating cables from outside to inside respectively, indoor are also provided with temperature regulator, vent fan, aerofoil fan etc. simultaneously, ensure that room temp is constant and evenly, the growth for Rhodopseudomonas palustris provides best growing environment.
In order to provide balanced photoenvironment to Rhodopseudomonas palustris, the every 10-12 of the culturing bottle on culturing rack near surrounding a circle, directly over it, install 1 incandescent light, the power of incandescent light is 40W, and the bulb distance of culturing bottle and vertical direction is 10cm.
The condition that the present invention cultivates Rhodopseudomonas palustris is constant temperature, illumination anaerobism, and incubation time is 3-6 days, and cultivate after terminating, random detection 200 bottles of bacterium liquid, wherein have 1 bottle of pollution microbes, and on average every bottle of bacterial concentration is 5 × 10
9individual/mL, every bottle of bacterial concentration difference is within limit of error, and as can be seen here, the Rhodopseudomonas palustris concentration using the present invention to cultivate is high and very even.
Claims (3)
1. a cultural method for Rhodopseudomonas palustris, is characterized in that, comprises the following steps:
(1) substratum is prepared
Take NaCl 5g, NH respectively
4cl 5g, CH
3cOONa 15g, peptone 15g, MgSO
41g, K
2hPO
41g, 5% NaHCO
3solution 100mL, is dissolved in 5L distilled water, stirs;
(2) sterilization
The substratum that stirs loaded in stainless steel cask, to carry out disinfection sterilizing with ozonizer, sterilization 10min, the usage quantity of ozone is 1g;
(3) inoculate
Get the Rhodopseudomonas palustris mother liquor 3-5mL after activation and be placed in 500mL triangular flask, in culturing bottle, then add sterilized substratum to bottleneck, whole seeded process equipment used is all sterilized;
(4) cultivate
The culturing rack finished culturing bottle being put into culturing room is cultivated, and culture temperature is 28 DEG C-32 DEG C, cultivates and transfers 1-2 time after 2-3 days.
2. the cultural method of a kind of Rhodopseudomonas palustris according to claim 1, is characterized in that, the wall of culturing room is conventional wall, heat preserving and insulating material and heating cables from outside to inside respectively.
3. the cultural method of a kind of Rhodopseudomonas palustris according to claim 1, it is characterized in that, every 10-12 culturing bottle, near surrounding a circle, installs 1 incandescent light directly over it, the power of incandescent light is 40W, and the bulb distance of culturing bottle and vertical direction is 10cm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410816451.2A CN104450588A (en) | 2014-12-24 | 2014-12-24 | Method for culturing rhodopseudomonas palustris |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410816451.2A CN104450588A (en) | 2014-12-24 | 2014-12-24 | Method for culturing rhodopseudomonas palustris |
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| CN104450588A true CN104450588A (en) | 2015-03-25 |
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| CN201410816451.2A Pending CN104450588A (en) | 2014-12-24 | 2014-12-24 | Method for culturing rhodopseudomonas palustris |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107868760A (en) * | 2016-12-29 | 2018-04-03 | 广西民族大学 | A kind of cultural method of Rhodopseudomonas palustris |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101113419A (en) * | 2007-06-21 | 2008-01-30 | 侯建明 | Intensive production method for sulfur-free rhodospirillum |
| CN201396042Y (en) * | 2009-05-08 | 2010-02-03 | 上海力申科学仪器有限公司 | FRP (fiber reinforce plastic) door structure for incubator |
| CN103881934A (en) * | 2012-12-19 | 2014-06-25 | 青岛中仁药业有限公司 | Preparation method for liquid and microecological preparation of photosynthetic bacteria |
| CN203960198U (en) * | 2014-07-14 | 2014-11-26 | 安徽大学 | Photosynthetic bacterium incubator |
-
2014
- 2014-12-24 CN CN201410816451.2A patent/CN104450588A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101113419A (en) * | 2007-06-21 | 2008-01-30 | 侯建明 | Intensive production method for sulfur-free rhodospirillum |
| CN201396042Y (en) * | 2009-05-08 | 2010-02-03 | 上海力申科学仪器有限公司 | FRP (fiber reinforce plastic) door structure for incubator |
| CN103881934A (en) * | 2012-12-19 | 2014-06-25 | 青岛中仁药业有限公司 | Preparation method for liquid and microecological preparation of photosynthetic bacteria |
| CN203960198U (en) * | 2014-07-14 | 2014-11-26 | 安徽大学 | Photosynthetic bacterium incubator |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107868760A (en) * | 2016-12-29 | 2018-04-03 | 广西民族大学 | A kind of cultural method of Rhodopseudomonas palustris |
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Application publication date: 20150325 |
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