CN107868760A - A kind of cultural method of Rhodopseudomonas palustris - Google Patents

A kind of cultural method of Rhodopseudomonas palustris Download PDF

Info

Publication number
CN107868760A
CN107868760A CN201611249071.0A CN201611249071A CN107868760A CN 107868760 A CN107868760 A CN 107868760A CN 201611249071 A CN201611249071 A CN 201611249071A CN 107868760 A CN107868760 A CN 107868760A
Authority
CN
China
Prior art keywords
rhodopseudomonas palustris
culture
light
bacterium
strain
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201611249071.0A
Other languages
Chinese (zh)
Inventor
廖艳娟
黄在银
覃小芳
韦慧
李辉
钟声扬
杨悦娟
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Guangxi University for Nationalities
Original Assignee
Guangxi University for Nationalities
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Guangxi University for Nationalities filed Critical Guangxi University for Nationalities
Priority to CN201611249071.0A priority Critical patent/CN107868760A/en
Publication of CN107868760A publication Critical patent/CN107868760A/en
Pending legal-status Critical Current

Links

Classifications

    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, 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
    • C12N1/20Bacteria; Culture media therefor

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • Wood Science & Technology (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The present invention relates to one kind to use the micro hot systems of light, passes through Rhodopseudomonas palustris cultural method that is high-precision, controlling illumination wavelength, power and temperature in high sensitivity.Strain is brought back to life first and transfers 3 on behalf of strain stoste, standard curve is made using blood plate counting method, according to this experiment with computing bacterium solution bacterial population, so as to ensure bacterial population in fixed range.Then by the micro hot systems of light, 532 nm wavelength light sources are opened, progress luminous power is 4 W/m2, temperature be 30 DEG C Rhodopseudomonas palustris growth in situ heat score-curve determine.The thermokinetic parameters and the quantity of heat production of different metabolic phase of exponential phase are finally calculated, determine light λ=532 nm, 4 W/m2The photosynthetic cells effect of bacterium can be most transferred, is metabolized beneficial to bacterial growth.The Rhodopseudomonas palustris lag phase time of this method culture is short, only with 27395 s;Exponential phase is up to 48.1851 μ W, 1343.66 J for up to 45570 s, thermal power and thermal discharge;Present invention illumination reaction condition simple to operate, bacterium is accurate, and reaction is efficient.

Description

A kind of cultural method of Rhodopseudomonas palustris
Technical field
The present invention relates to a kind of cultural method of Rhodopseudomonas palustris, using light-micro hot systems, by high accuracy, Wavelength, power and the temperature of illumination are controlled in high sensitivity, and bacterium is cultivated.Belong to field of microbial culture technology, have Body is related to a kind of cultural method of Rhodopseudomonas palustris.
Background technology
The factor of photosynthetic bacterium growth is influenceed in addition to own physiological condition, also has some external factor, such as temperature, soda acid Degree, the initial concentration of substrate and illumination can all limit the growth of bacterium.The wherein just energy source of photosynthetic bacterium growth, it is The basic premise of photosynthetic bacteria existence.Photosynthetic pigments are pigments specific to photosynthetic organism, are to convert light energy into chemistry The key substance of energy.Different photosynthetic bacteria strains contains different photosynthetic pigments, and the type and quantity of photosynthetic pigments are to light Capture produces material impact, therefore growth of the light to photosynthetic bacteria is most important.
Rhodopseudomonas(Rhodopseudomonas)It is most commonly seen in photosynthetic bacteria and that there is Practical significance one Individual category, Rhodopseudomonas palustris are its representative strain, Industrial Wastewater Treatment, hydrogen manufacturing, aquaculture, planting industry, cosmetics and The fields such as health products have a wide range of applications, but are concentrated mainly on culture for Rhodopseudomonas palustris cultural method at present In the improvement of base, such as cultivation temperature, acid-base value, substrate formula and concentration, for Rhodopseudomonas palustris culture illumination condition The document report seen of research be the effect grown using complex light to it, and in single wavelength, certain power and temperature Growth yet there are no document report under illumination.
The content of the invention
It is an object of the invention to the vacancy for existing area research, proposes a kind of culture side of Rhodopseudomonas palustris Method.Present invention illumination reaction condition simple to operate, bacterium is accurate, and reaction is efficient.
To realize this purpose, the present invention uses light-micro hot systems, by controlling the wavelength of illumination and the marsh of power Red pseudomonas cultural method.Strain is brought back to life first and 3 generations of transferring are as strain stoste, and standard is made using blood plate counting method Curve determination, experiment with computing bacterium solution bacterial population according to this, so as to ensure bacterial population in fixed range.Then using light-micro heat system System, 532 nm wavelength light sources are opened, progress luminous power is 4 W/m2Rhodopseudomonas palustris growth in situ heat score-curve survey It is fixed.Finally by the thermokinetic parameters and the quantity of heat production of different metabolic phase for calculating exponential phase, monochromatic light λ=532 are determined Nm, 4 W/m2The photosynthetic cells effect that power can most transfer bacterium makes bacterium keep good growth situation, beneficial to bacterial growth generation Thank.The Rhodopseudomonas palustris lag phase time that this method is turned out is short, only with 27395 s;Exponential phase for up to 45570 s, thermal power and thermal discharge are up to 48.1851 μ W, 1343.66 J.
The method of the present invention includes step in detail below:
1st, the preparation of culture medium:Medium component is weighed respectively:The g of tryptone 15.0, the g of soya peptone 5.0, sodium chloride 5.0 G, distilled water is dissolved in, 1.0 L are settled to distilled water after stirring, it is 7.3 ± 0.2 to survey solution PH, is sub-packed in 250 mL cones Shape bottle, after sealing, 121 DEG C of 30 min of sterilizing.
2nd, Rhodopseudomonas palustris actication of culture:Freeze-dried vaccine is forwarded to from slant medium 4-5 mL Liquid Cultures Base, bring back to life strain.Transferred for 3 generations again as strain stoste.
3rd, Rhodopseudomonas palustris bacterial concentration:Counted using counting method of blood cell.Put down with the strain after resurrection Plate culture, 30 DEG C are placed in, intensity of illumination is to be counted after cultivating 24-48 hours in 2500 lux illumination incubators, makes standard Curve.Rhodopseudomonas palustris bacterial concentration measure measures its its OD value at 660 nm using spectrophotometric, passes through mark Directrix curve fit equation determines bacterial population so that bacterium solution bacterial population used is about 10 every time8Individual/mL.
With this experiment with computing bacterium solution bacterial population.
4th, Rhodopseudomonas palustris culture:In light-micro hot systems, 1.5 mL culture mediums and 1.5 mL are separately added into The fluid nutrient medium of Rhodopseudomonas palustris is inoculated with into sample cell and reference cell, then it is 15 mL's to be respectively charged into volume In stainless steel sample cell and reference tube, constant temperature is to 30 DEG C, after baseline stability, opens 532 nm wavelength light sources, carries out light work( Rate is 4 W/m2Rhodopseudomonas palustris growth in situ experiment, time 345600s.
The present invention has following features:
1st, the illumination of Rhodopseudomonas palustris bacterium of the present invention culture is specific wavelength monochromatic light, i.e. the nm of λ=532, and certain power is 4W/m2, specified temp be 30 DEG C be controlled by light-micro hot systems.
2nd, the condition of Rhodopseudomonas palustris of the present invention is constant temperature, and facultative aerobic culture can be by thin in incubation " heat score-curve " of bacteria growing is monitored to its growth course state.And calculated after experiment terminates according to heat score-curve The Rhodopseudomonas palustris lag phase time that this method is turned out is 27395 s;The exponential phase time is 45570 s, hot merit Rate and thermal discharge are up to 48.1851 μ W, 1343.66 J respectively, in view of in production, come often through the time for extending logarithmic phase The fermentation efficiency of raising bacterium is visible, and the Rhodopseudomonas palustris cultivated using the present invention has low cost, high efficiency, operation letter Single, the accurate feature of illumination reaction condition of bacterium.
Brief description of the drawings
Fig. 1 is the type light of RD 496-CK 2000-micro hot systems reality that the present invention is used for Rhodopseudomonas palustris culture Thing figure.
Fig. 2 is the heat production P- of the gained of the embodiment of the present invention 1tThe time of curve map and lower different growing stages, thermal change Figure.
Fig. 3 is the heat production P- of the gained of the embodiment of the present invention 2tThe time of curve map and lower different growing stages, thermal change Figure.
Fig. 4 is the heat production P- of the gained of the embodiment of the present invention 3tThe time of curve map and lower different growing stages, thermal change figure.
Embodiment
Technical scheme is further described below by way of specific embodiment.Following examples are to this hair Bright further explanation, rather than limitation the scope of the present invention.
Embodiment 1
1)The preparation of culture medium:Weigh the g of tryptone 15.0, the g of soya peptone 5.0, the g of sodium chloride 5.0, distilled water 1.0 L, after stirring, its PH is surveyed as 7.3 ± 0.2,121 DEG C of 30 min of sterilizing.
2)Actication of culture:Bring back to life strain and 3 generations of transferring are as strain stoste.
3)Bacterial concentration:Counted using counting method of blood cell.With after resurrection strain carry out flat board culture, culture 24 ~ Count within 48 hours, making standard curve, experiment with computing bacterium solution bacterial population,.
4)Bacteria Culture:In light-Calorimetry system, be separately added into 1.5 mL culture mediums and 1.5 mL to be inoculated with marsh red The fluid nutrient medium of pseudomonad is into sample cell and reference cell, then is respectively charged into the stainless steel sample cell that volume is 15 mL In reference tube, constant temperature is to 30 DEG C, after baseline stability, opens 532 nm wavelength light sources, progress luminous power is 4 W/m2's Rhodopseudomonas palustris growth in situ is tested, and the time is 345600 s.
Experiment terminate after according to heat score-curve, as shown in figure 1, A-B is lag phase, B-C exponential phases, C-D is Stationary phase, D-E are decline phase.It is 27395 s to calculate the Rhodopseudomonas palustris lag phase time that this method is turned out;It is right Number growth period is up to 48.1851 μ W, 1343.66 J respectively for up to 45570 s, thermal power and thermal discharge.
Embodiment 2
1)The preparation of culture medium:Weigh the g of tryptone 15.0, the g of soya peptone 5.0, the g of sodium chloride 5.0, distilled water 1.0 L, after stirring, its PH is surveyed as 7.3 ± 0.2,121 DEG C of 30 min of sterilizing.
2)Actication of culture:Bring back to life strain and 3 generations of transferring are as strain stoste.
3)Bacterial concentration:Counted using counting method of blood cell.With after resurrection strain carry out flat board culture, culture 24 ~ Count within 48 hours, make standard curve determination, experiment with computing bacterium solution bacterial population.
4)Bacteria Culture:In light-Calorimetry system, be separately added into 1.5 mL culture mediums and 1.5 mL to be inoculated with marsh red The fluid nutrient medium of pseudomonad(Measure bacterium solution OD values before measure every time, bacterial population is determined by standard curve fit equation, So that bacterium solution bacterial population used is about 10 every time8Individual/mL)Into sample cell and reference cell, then it is 15 mL to be respectively charged into volume Stainless steel sample cell and reference tube in, set relevant parameter, constant temperature is to 30 DEG C, after baseline stability, open 532 nm Wavelength light source, progress luminous power are 2 W/m2Rhodopseudomonas palustris growth in situ experiment, the time is 345600 s(96 h).
Test after terminating according to heat score-curve, as shown in Fig. 2 A'B' is lag phase, B'-C' exponential phases, C'- D' is stationary phase, and D'-E' is decline phase.Calculating the Rhodopseudomonas palustris lag phase time that this method is turned out is 161200 s;The exponential phase time is 11362 s, and thermal power and thermal discharge are respectively 19.687 μ W, 135.00 J.Explanation , illumination power is too small, and the growth metabolism of Rhodopseudomonas palustris is slack-off, and growth performance declines.
Embodiment 3
1)The preparation of culture medium:Weigh the g of tryptone 15.0, the g of soya peptone 5.0, the g of sodium chloride 5.0, distilled water 1.0 L, after stirring, its PH is surveyed as 7.3 ± 0.2,121 DEG C of 30 min of sterilizing.
2)Actication of culture:Bring back to life strain and 3 generations of transferring are as strain stoste.
3)Bacterial concentration:Counted using counting method of blood cell.With after resurrection strain carry out flat board culture, culture 24 ~ Count within 48 hours, make standard curve determination, experiment with computing bacterium solution bacterial population.
4)Bacteria Culture:In light-Calorimetry system, be separately added into 1.5 mL culture mediums and 1.5 mL to be inoculated with marsh red The fluid nutrient medium of pseudomonad(Measure bacterium solution OD values before measure every time, bacterial population is determined by standard curve fit equation, So that bacterium solution bacterial population used is about 10 every time8Individual/mL)Into sample cell and reference cell, then it is 15 mL to be respectively charged into volume Stainless steel sample cell and reference tube in, set relevant parameter, constant temperature is to 30 DEG C, after baseline stability, open 532 nm Wavelength light source, progress luminous power are 6 W/m2Rhodopseudomonas palustris growth in situ experiment, the time is 345600 s(96 h).
Test after terminating according to heat score-curve, as shown in figure 3, A 〞-B 〞 are lag phase, B 〞-C 〞 exponential phases, C 〞-D 〞 is stationary phase, and D 〞-E 〞 are decline phase.It is 142838 to calculate the Rhodopseudomonas palustris lag phase time that this method is turned out s;The exponential phase time is 29549 s, and thermal power and thermal discharge are respectively 38.561 μ W, 842.66 J.Illustrate, illumination Power is excessive, and the growth metabolism of Rhodopseudomonas palustris is equally slack-off, and growth performance declines.

Claims (2)

1. using the method for light-micro hot systems culture Rhodopseudomonas palustris, it is characterised in that high-precision, highly sensitive Light specific wavelength is λ=532 nm, certain power 4W/m2, specified temp is 30 DEG C.
2. according to claim 1, a kind of cultural method of Rhodopseudomonas palustris, comprise the following steps:
1) preparation of culture medium:Medium component is the g of tryptone 15.0, the g of soya peptone 5.0, the g of sodium chloride 5.0, is distilled The L of water 1.0, after stirring, its pH is surveyed as 7.3 ± 0.2,121 DEG C of 30 min of sterilizing;
2) actication of culture:Bring back to life strain and 3 generations of transferring are as strain stoste;
3) Bacteria Culture:In light-Calorimetry system, it is separately added into 1.5 mL culture mediums and 1.5 mL is inoculated with the red false list in marsh The fluid nutrient medium of born of the same parents bacterium is into sample cell and reference cell, then is respectively charged into the stainless steel sample cell and ginseng that volume is 15 mL Than in pipe, bacterium solution bacterial population used is about 108Individual/mL, constant temperature is to 30 DEG C, after baseline stability, opens 532 nm wavelength lights Source, row luminous power are 4 W/m2Rhodopseudomonas palustris growth in situ experiment, the time is 345600 s.
CN201611249071.0A 2016-12-29 2016-12-29 A kind of cultural method of Rhodopseudomonas palustris Pending CN107868760A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201611249071.0A CN107868760A (en) 2016-12-29 2016-12-29 A kind of cultural method of Rhodopseudomonas palustris

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201611249071.0A CN107868760A (en) 2016-12-29 2016-12-29 A kind of cultural method of Rhodopseudomonas palustris

Publications (1)

Publication Number Publication Date
CN107868760A true CN107868760A (en) 2018-04-03

Family

ID=61761455

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201611249071.0A Pending CN107868760A (en) 2016-12-29 2016-12-29 A kind of cultural method of Rhodopseudomonas palustris

Country Status (1)

Country Link
CN (1) CN107868760A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112877242A (en) * 2021-02-05 2021-06-01 广东工业大学 Bacterial source photosensitizer for wastewater treatment and preparation method and application thereof

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0065574B1 (en) * 1980-12-04 1986-05-21 Nippon Carbide Kogyo Kabushiki Kaisha Photosynthetic bacteria culture
JPH0471696A (en) * 1990-07-10 1992-03-06 Yoshida Sogo Kenkyusho:Kk Method for purifying water
CN101650324B (en) * 2009-08-21 2011-04-06 重庆大学 Device for testing heat map spectra of microbial biochemical reaction
CN104450588A (en) * 2014-12-24 2015-03-25 四川省星川生物科技有限公司 Method for culturing rhodopseudomonas palustris
CN105087430A (en) * 2015-07-27 2015-11-25 厦门市科环海洋生物科技有限公司 Culture medium and culture method of high-concentration photosynthetic bacteria

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0065574B1 (en) * 1980-12-04 1986-05-21 Nippon Carbide Kogyo Kabushiki Kaisha Photosynthetic bacteria culture
JPH0471696A (en) * 1990-07-10 1992-03-06 Yoshida Sogo Kenkyusho:Kk Method for purifying water
CN101650324B (en) * 2009-08-21 2011-04-06 重庆大学 Device for testing heat map spectra of microbial biochemical reaction
CN104450588A (en) * 2014-12-24 2015-03-25 四川省星川生物科技有限公司 Method for culturing rhodopseudomonas palustris
CN105087430A (en) * 2015-07-27 2015-11-25 厦门市科环海洋生物科技有限公司 Culture medium and culture method of high-concentration photosynthetic bacteria

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
CHUAN ZHANG等: "Enhancing continuous photo-H2 production using optical fiber for biofilm formation", 《ENVIRONMENTAL PROGRESS AND SUSTAINABLE ENERGY》 *
刘德海等: "沼泽红假单胞菌发酵工艺研究", 《中国饲料》 *
孙明星: "光合细菌Rhodopseudomonas Palustris PB-Z产氢性能的研究", 《中国优秀硕士学位论文全文数据库·工程科技I辑》 *
王永忠等: "序批式培养沼泽红假单胞菌光照产氢的能量分析", 《太阳能学报》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112877242A (en) * 2021-02-05 2021-06-01 广东工业大学 Bacterial source photosensitizer for wastewater treatment and preparation method and application thereof

Similar Documents

Publication Publication Date Title
Henkanatte-Gedera et al. Removal of dissolved organic carbon and nutrients from urban wastewaters by Galdieria sulphuraria: Laboratory to field scale demonstration
Costa et al. Improving Spirulina platensis biomass yield using a fed-batch process
CN102994367B (en) High-efficiency phototroph reaction system for pure culture of photosynthetic bacteria and sterilizing method thereof
CN101580870A (en) Method for fast and continuously detecting faecal coliform and colon bacillus group
Palamae et al. Production of renewable biohydrogen by Rhodobacter sphaeroides S10: a comparison of photobioreactors
CN100408669C (en) Oral biological film dynamic model device and its oral biological film forming method
CN110184193A (en) It is a kind of efficiently to expand numerous continuous gradient feed process and device applied to haematococcus pluvialis
CN106556599B (en) A kind of method for rapid inspecting animalcule
Ross et al. Investigating and modeling the effect of light intensity on Rhodopseudomonas palustris growth
CN102115776B (en) Microalgae screening method and system thereof
CN107868760A (en) A kind of cultural method of Rhodopseudomonas palustris
CN104313182B (en) Detection method of phage
CN207958369U (en) A kind of microbial detection device
CN106479898A (en) A kind of culture medium of Haematocoocus Pluvialls and its application
Huang et al. Screening of flocculant-producing strains by NTG mutagenesis
CN200952006Y (en) Oral cavity biological membrane dynamic model device
CN109929898A (en) A method of promoting the energy-efficient production hydrogen of HAU-M1 photosynthetic bacteria group
CN102250764A (en) Micro holographic biological sensing reactor system
CN107488718A (en) Detect C60Nanoparticles generate the method and device influenceed to Microcystin
JPH11346760A (en) Cultivation of microalga
CN110261267A (en) A kind of detection method of fishing photosynthetic bacteria preparation product
CN106635782A (en) Oleaginous microalgae integrated culture reactor and culture method
CN101948745A (en) Artificial mouth simulating device
Ogbonna et al. Production of pure photosynthetic cell biomass for environmental biosensors
CN204981878U (en) Little algae screening and culture apparatus

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20180403