CN110272849A - The method for being remarkably improved Growth of Spirulina Platensis speed and nutrition content - Google Patents

The method for being remarkably improved Growth of Spirulina Platensis speed and nutrition content Download PDF

Info

Publication number
CN110272849A
CN110272849A CN201910625341.0A CN201910625341A CN110272849A CN 110272849 A CN110272849 A CN 110272849A CN 201910625341 A CN201910625341 A CN 201910625341A CN 110272849 A CN110272849 A CN 110272849A
Authority
CN
China
Prior art keywords
culture
spirulina
blunt top
top spirulina
blunt
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.)
Granted
Application number
CN201910625341.0A
Other languages
Chinese (zh)
Other versions
CN110272849B (en
Inventor
马玉心
崔大练
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.)
Etuoke Jinchang Spirulina Co ltd
Wuxi Xiangyuan Information Technology Co ltd
Original Assignee
Zhejiang Ocean University ZJOU
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 Zhejiang Ocean University ZJOU filed Critical Zhejiang Ocean University ZJOU
Priority to CN201910625341.0A priority Critical patent/CN110272849B/en
Publication of CN110272849A publication Critical patent/CN110272849A/en
Application granted granted Critical
Publication of CN110272849B publication Critical patent/CN110272849B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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
    • 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
    • C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
    • 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
    • C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves

Landscapes

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

Abstract

The present invention is about a kind of method for being remarkably improved Growth of Spirulina Platensis speed and nutrition content, including filtering out algae solution and obtaining algal gel 1) with the base culture base blunt top spirulina of the bacterial fermentation bacterium solution containing inactivation to the logarithmic growth phase later period；2) first stage optimization culture is carried out to above-mentioned algal gel with the Optimal Medium containing ethyl alcohol and alpha, beta-lonone, irradiation culture is combined using monochromatic light and obtains the blunt top spirulina containing swarm cell；3) it separately takes and fulvic acid is added in Optimal Medium, carry out second stage culture after the blunt top spirulina of step 2) is obtained in the form of algal gel in Optimal Medium, at least partly swarm cell is made to be converted into non motile cell.This method promotes the growth and breeding of blunt top spirulina, the enrichment of enriched nutritive substance well, and especially induction blunt top spirulina produces the nutriments such as beta carotene, polysaccharide, promotes the reproductive efficiency and economic well-being of workers and staff of blunt top spirulina.

Description

The method for being remarkably improved Growth of Spirulina Platensis speed and nutrition content
Technical field
The present invention is remarkably improved Growth of Spirulina Platensis speed about blunt top spirulina culture field, especially with regard to one kind The method of degree and nutrition content.
Background technique
Blunt top spirulina (Spirulina platensis) is a kind of miniature cyanobacteria of rich in nutrition content, is rich in albumen Matter, polysaccharide, unsaturated fatty acid, carotenoid (beta carotene and zeaxanthin) etc., especially beta carotene contain Amount is 15 times of content beta-carotene in carrot.Spirulina photosynthetic efficiency is high, growth and breeding is fast, it is strong to environmental suitability and Easily harvesting, is one of the main microalgae of current industrially scalable culture.Spirulina is considered as 21 century optimal health care product, Spirulina the industries such as food, health care product, cosmetics, medicine, feed have been widely used in both at home and abroad at present.
The effect of blunt top spirulina is relatively more, and general the most frequently used also most important effect is raising immunity of organisms, promotes Enterogastric peristalsis, the conventional element and microelement for supplementing body.It is worth mentioning that spirulina phycocyanin, algae indigo plant egg is a kind of Natural edible pigment, bath do not dissolve in grease and alcohols, are a kind of blue powders in water.With anticancer, promote cytothesis Function can be used as advanced natural pigment.Spirulina polysaccharide is a kind of water-soluble polysaccharide in spirulina cells, is had anti-swollen The bioactivity such as tumor, anti-radiation, anti-mutation, can improve the immune function of body cell and body fluid, resist cancer cell multiplication, mitigate The caused genetic damage etc. of radiation has broad application prospects in terms of anti-cancer, anti-aging, enhancing. Spirulina can improve Public nutrition due to after material abundance people taken in excessive high-fat, high sugar, high-protein food, cause Macroscopical overnutrition and microcosmic malnourished situation.Because undesirable eating habit have become influence national health and illness compared with High principal element.As it can be seen that research is remarkably improved the method for Growth of Spirulina Platensis speed and nutrition content with latent Application space.
The disclosure of background above technology contents is only used for auxiliary and understands inventive concept and technical solution of the invention, not The prior art for necessarily belonging to present patent application shows above content in the applying date of present patent application in no tangible proof In the case where having disclosed, above-mentioned background technique should not be taken to the novelty and creativeness of evaluation the application.
Summary of the invention
The purpose of the present invention is to provide a kind of Growth of Spirulina Platensis speed and nutrition contents of being remarkably improved Method promotes the growth and breeding of blunt top spirulina with basis culture, dual-stage optimization culture well, enriched nutritive substance Enrichment, especially induction blunt top spirulina produce beta carotene, the nutriments such as polysaccharide, so that nutriment contains in frustule Amount dramatically increases, and promotes the reproductive efficiency and economic well-being of workers and staff of blunt top spirulina.
The technical solution that the present invention is taken to achieve the above object are as follows: be remarkably improved Growth of Spirulina Platensis speed and The method of nutrition content, the method includes the following steps:
1) belonged to the red bacterium containing inactivation and the basal medium of the zymocyte liquid of Rhodopseudomonas bacterium will activate Blunt top spirulina culture afterwards filters out algae solution, obtains algal gel after centrifugation or filtration washing to the logarithmic growth phase later period；
2) first stage optimization training is carried out with algal gel of the Optimal Medium containing ethyl alcohol and alpha, beta-lonone to step 1) It supports, using orange-colored light and blue light artificial LED light source joint irradiation culture, total intensity of illumination is 2500~4500Lux, 18~ 5~10d is cultivated at a temperature of 22 DEG C, obtains the blunt top spirulina containing swarm cell；
3) it under natural optical mode condition of culture, separately takes and fulvic acid is added in Optimal Medium, step 2) is contained into trip The blunt top spirulina of dynamic cell carries out second stage culture after obtaining in the form of algal gel in Optimal Medium, makes at least partly to swim Dynamic cell transformation is non motile cell.
In the preferred embodiment of the application, the basal medium composition is as follows: 0.2~0.4g/L of sodium bicarbonate, 0.5~1.0g/L of potassium nitrate, 0.4~0.6g/L of sodium chloride, 0.02~0.03g/L of zinc sulfate, 0.05~0.08g/ of frerrous chloride L, 0.01~0.02g/L of ferric citrate, A5 component 1mL/L, remaining is water, and pH is 7.5~9.5.
In the preferred embodiment of the application, the A5 component is: Boratex 3.0g/L, tetrahydrate manganese chloride 2.5g/L, Cupric sulfate pentahydrate 0.2g/L, ammonium molybdate 0.6g/L, four water cobalt nitrate 0.1g/L, remaining is water.
In the preferred embodiment of the application, the red thin campylobacter bacteria is preferably Rhodobacter capsulatus (Rhodobacter capsulatus)。
In the preferred embodiment of the application, the Rhodopseudomonas bacterium is preferably Rhodopseudomonas palustris (Rhodopseudomonas palustris)。
In the preferred embodiment of the application, the fermentation process of the zymocyte liquid is:
Prepare fermentation medium are as follows: 0.4~1.0g/L of ferric citrate, 0.3~0.6g/L of dipotassium hydrogen phosphate, magnesium sulfate 0.2~0.4g/L, 0.4~1.0g/L of sodium chloride, 0.5~2.0g/L of yeast powder, 3.0~5.0g/L of sodium acetate, remaining is distillation Water；
After sterilizing according to number proportion 3:1 be inoculated with altogether the red thin campylobacter bacteria and the Rhodopseudomonas bacterium 1~ 1.2×107A/mL, at a temperature of 25~28 DEG C, ferment 12~96h under 5000~8000Lux continuous light, and complete inactivation is simultaneously dense It is reduced to density and is at least 1.15g/mL to obtain the final product.Utilize the basis belonged to containing red bacterium with the zymocyte liquid of Rhodopseudomonas bacterium For culture medium by blunt top spirulina culture to logarithmic growth phase later period, the algae for being conducive to significantly improve blunt top spirulina in culture solution is thin Born of the same parents' density, due to being added to mixed bacteria liquid, to promote frustule fast breeding, so that frustule concentration increases, frustule is living Jerk enhancing, splitting ability improve, further promotion and nutrients conducive to algal grown speed in further incubation The enrichment of matter.
In the preferred embodiment of the application, the red bacterium category and Rhodopseudomonas that are inactivated in the basal medium The additive amount of the zymocyte liquid of bacterium is 1.2~1.5mL/L.
In the preferred embodiment of the application, the wavelength of the orange-colored light is 590~620nm, the wave of the blue light Length is 445~475nm, and the intensity of illumination ratio of the orange-colored light and blue light is 3~5:1.In the application, with special wavelength model It encloses, the orange-colored light of particular light intensity ratio and blue light joint irradiation blunt top spirulina, Growth of Spirulina Platensis can be stimulated, induced Synthesis of the blunt top spirulina to beta carotene, this may be the orange due to above-mentioned special wavelength range, particular light intensity ratio Coloured light and blue light joint irradiation promote the synthesis of isoprene and isoprene is the precursor substance for synthesizing carotenoid, because This can be obviously improved the content of the beta carotene in frustule, accelerate the accumulation of beta carotene, promote yield.
In the preferred embodiment of the application, 10~12h gives 3000~3500Lux light when the step 1) is cultivated According to cultivation temperature is 30~32 DEG C, and air mass flow is 0.4~0.5vvm.
In the preferred embodiment of the application, the Optimal Medium: 12.6~15.5g/L of sodium bicarbonate, sodium carbonate 1.2~1.8g/L, 1.2~1.5g/L of dipotassium hydrogen phosphate, 3.5~4.5g/L of sodium nitrate, 2.2~2.5g/L of potassium sulfate, sodium chloride 2.6~3.4g/L, 0.12~0.2g/L of magnesium sulfate, 0.08~0.1g/L of calcium chloride, 0.05~0.08g/L of ferrous sulfate, A5 group Divide 1mL/L, remaining is water, pH value 8.0~9.0.
In the preferred embodiment of the application, ethyl alcohol presses concentration of volume percent in the Optimal Medium of the step 2) It is 2.0~4.2%.
In the preferred embodiment of the application, the additive amount of alpha, beta-lonone is in the Optimal Medium of the step 2) 0.2~20mg/L, preferably additive amount are 10~15mg/L.Inventor it was unexpectedly observed that during the cultivation process state add it is micro Alpha, beta-lonone is remarkably improved the content of spirulina platensis polysaccharide, and the possible process and alpha, beta-lonone help to promote algae thin Born of the same parents' amino acid and protein synthase activity is related, and the promotion of polyoses content further improves the yield and production of blunt top spirulina Rate promotes economic benefit, additionally due to the dosage of alpha, beta-lonone is few, while stimulating blunt top spirulina to accumulate polysaccharide not Cost, control cultivation investment can be obviously increased.
In the preferred embodiment of the application, in the Optimal Medium of the step 3) additive amount of fulvic acid be 35~ 45mg/L, preferably additive amount are 35~40mg/L.
In the preferred embodiment of the application, in the second stage culture of the step 3), 10~12h gives 3500~ 4000Lux illumination, cultivation temperature are 30~32 DEG C, and air mass flow is 0.5~0.6vvm, and incubation time is 8~10d.
Further to show the object of the invention, the present invention also provides above-mentioned any one to be remarkably improved blunt top spiral shell The application of the rotation algae speed of growth and the method for nutrition content in blunt top spirulina culture comprising trained using the method Support blunt top spirulina.
The method of the present invention by be added in basal medium inactivation red bacterium belong to and Rhodopseudomonas bacterium hair Ethyl alcohol and alpha, beta-lonone are added in Optimal Medium and carries out the to blunt top spirulina under orange, blue light irradiation for yeast-like fungi liquid Fulvic acid is added to blunt top spirulina progress second stage culture in the culture of one stage in Optimal Medium, can promote well Into the growth and breeding of blunt top spirulina, strengthen the enrichment of Spirulina Platensis Nutritious substance, especially induction blunt top spirulina produces β- The nutriments such as carrotene, polysaccharide promote the numerous of blunt top spirulina so that the content of nutriment dramatically increases in frustule Efficiency and economic well-being of workers and staff are grown, is that a kind of high efficiency, low cost are remarkably improved Growth of Spirulina Platensis speed and nutriment The method of content.
The invention has the benefit that
1) utilize the basal medium of the zymocyte liquid containing red bacterium category and Rhodopseudomonas bacterium by blunt top spiral Algae cultivates the algae cell density for being conducive to significantly improve blunt top spirulina in culture solution to the logarithmic growth phase later period, due to addition Mixed bacteria liquid, thus promote frustule fast breeding, so that frustule concentration increases, the enhancing of frustule liveness, break-up energy Power improves, conducive to the further promotion and the enrichment of nutriment of algal grown speed in further incubation；
2) irradiation blunt top spirulina is combined with special wavelength range, the orange-colored light of particular light intensity ratio and blue light, it can Growth of Spirulina Platensis is stimulated, induces synthesis of the blunt top spirulina to beta carotene, this may be due to above-mentioned special wavelength Range, the orange-colored light of particular light intensity ratio and blue light joint irradiation promote the synthesis of isoprene and isoprene is synthesis The precursor substance of carotenoid, therefore the content for the beta carotene that can be obviously improved in frustule accelerate beta carotene Accumulation, promoted yield；
3) state adds the content that micro alpha, beta-lonone is remarkably improved spirulina platensis polysaccharide during the cultivation process, can Can the process and alpha, beta-lonone facilitate that promotion frustule amino acid and protein synthase activity is related, and polyoses content mentions It rises and further improves the productivity and yield of blunt top spirulina, economic benefit is promoted, additionally due to the dosage pole of alpha, beta-lonone It is few, stimulating blunt top spirulina not obviously increase cost, control cultivation investment while accumulating polysaccharide；
4) the method for the present invention is belonged to and Rhodopseudomonas bacterium by the way that the red bacterium of inactivation is added in basal medium Ethyl alcohol and alpha, beta-lonone are added in Optimal Medium and carries out under orange, blue light irradiation to blunt top spirulina for zymocyte liquid First stage culture, addition fulvic acid carries out second stage culture to blunt top spirulina in Optimal Medium, can be well Promote the growth and breeding of blunt top spirulina, strengthen the enrichment of Spirulina Platensis Nutritious substance, especially induction blunt top spirulina produces The nutriments such as beta carotene, polysaccharide promote blunt top spirulina so that the content of nutriment dramatically increases in frustule Reproductive efficiency and economic well-being of workers and staff are that a kind of high efficiency, low cost are remarkably improved Growth of Spirulina Platensis speed and nutrients The method of matter content.
Present invention employs above-mentioned technical proposals to provide model essay, compensates for the deficiencies in the prior art, design is reasonable, operation side Just.
Detailed description of the invention
For above and other purpose, feature, advantage and embodiment of the invention can be clearer and more comprehensible, appended attached drawing is said It is bright as follows:
Fig. 1 is the content schematic diagram of the beta carotene of blunt top spirulina of the invention；
Fig. 2 is the polyoses content schematic diagram of blunt top spirulina of the invention.
Specific embodiment
Unless otherwise defined, technical and scientific term used herein has ordinary skill of the art The identical meaning that personnel are generally understood.The present invention uses method described herein and material；But as is generally known in the art Other suitable methods and material can also be used.Material, method and example described herein are merely illustrative, It is not intended for limiting.All publications, patent application case, Patent Case, Provisional Application, data base entries and herein Other bibliography referred to etc., entirety is incorporated herein as reference.It include being defined as with this specification if there is conflict It is quasi-.
Embodiment 1:
The method for being remarkably improved Growth of Spirulina Platensis speed and nutrition content, the method includes following steps It is rapid:
1) belonged to the red bacterium containing inactivation and the basal medium of the zymocyte liquid of Rhodopseudomonas bacterium will activate Blunt top spirulina culture afterwards filters out algae solution, obtains algal gel after centrifugation or filtration washing to the logarithmic growth phase later period；
2) first stage optimization training is carried out with algal gel of the Optimal Medium containing ethyl alcohol and alpha, beta-lonone to step 1) It supports, using orange-colored light and the artificial LED light source joint irradiation culture of blue light, total intensity of illumination is 4000Lux, at a temperature of 20 DEG C 7d is cultivated, the blunt top spirulina containing swarm cell is obtained；
3) it under natural optical mode condition of culture, separately takes and fulvic acid is added in Optimal Medium, step 2) is contained into trip The blunt top spirulina of dynamic cell carries out second stage culture after obtaining in the form of algal gel in Optimal Medium, makes at least partly to swim Dynamic cell transformation is non motile cell.
In a presently preferred embodiment, the basal medium composition is as follows: sodium bicarbonate 0.3g/L, potassium nitrate 0.6g/ L, sodium chloride 0.5g/L, zinc sulfate 0.02g/L, frerrous chloride 0.06g/L, ferric citrate 0.02g/L, A5 component 1mL/L, Remaining is water, and pH is 8.2.
In a presently preferred embodiment, the A5 component is: Boratex 3.0g/L, tetrahydrate manganese chloride 2.5g/L, five water sulphur Sour copper 0.2g/L, ammonium molybdate 0.6g/L, four water cobalt nitrate 0.1g/L, remaining is water.
In a presently preferred embodiment, the red thin campylobacter bacteria is Rhodobacter capsulatus.
In a presently preferred embodiment, the Rhodopseudomonas bacterium is Rhodopseudomonas palustris.
In a presently preferred embodiment, the fermentation process of the zymocyte liquid is: preparing fermentation medium are as follows: ironic citrate Ammonium 0.6g/L, dipotassium hydrogen phosphate 0.4g/L, magnesium sulfate 0.3g/L, sodium chloride 0.5g/L, yeast powder 1.0g/L, sodium acetate 3.0g/ L, remaining is distilled water, is inoculated with the red thin campylobacter bacteria and the Rhodopseudomonas altogether according to number proportion 3:1 after sterilizing Bacterium 1.2 × 107A/mL, at a temperature of 28 DEG C, ferment 48h under 7000Lux continuous light, and complete inactivation is simultaneously concentrated into density 1.15g/mL to obtain the final product.Using belonging to containing red bacterium and the basal medium of the zymocyte liquid of Rhodopseudomonas bacterium is by blunt top spiral shell Rotation algae cultivates the algae cell density for being conducive to significantly improve blunt top spirulina in culture solution to the logarithmic growth phase later period, due to adding Add mixed bacteria liquid, thus promote frustule fast breeding, so that frustule concentration increases, the enhancing of frustule liveness, division Ability improves, conducive to the further promotion and the enrichment of nutriment of algal grown speed in further incubation.
In a presently preferred embodiment, the red bacterium inactivated in the basal medium belongs to and Rhodopseudomonas bacterium The additive amount of zymocyte liquid is 1.5mL/L.
In a presently preferred embodiment, the wavelength of the orange-colored light is 620nm, and the wavelength of the blue light is 460nm, institute The intensity of illumination ratio for stating orange-colored light and blue light is 3:1.In the application, with special wavelength range, the orange of particular light intensity ratio Coloured light and blue light joint irradiation blunt top spirulina, can stimulate Growth of Spirulina Platensis, induce blunt top spirulina to β-carrot The synthesis of element, this may be to combine to irradiate due to above-mentioned special wavelength range, the orange-colored light of particular light intensity ratio and blue light Promote the synthesis of isoprene and isoprene is the precursor substance for synthesizing carotenoid, therefore can be obviously improved in frustule Beta carotene content, accelerate the accumulation of beta carotene, promote yield.
In a presently preferred embodiment, 12h gives 3200Lux illumination when the step 1) is cultivated, and cultivation temperature is 30 DEG C, Air mass flow is 0.4vvm.
In a presently preferred embodiment, the Optimal Medium: sodium bicarbonate 13.0g/L, sodium carbonate 1.6g/L, phosphoric acid hydrogen Dipotassium 1.4g/L, sodium nitrate 4.0g/L, potassium sulfate 2.4g/L, sodium chloride 3.0g/L, magnesium sulfate 0.15g/L, calcium chloride 0.1g/L, Ferrous sulfate 0.06g/L, A5 component 1mL/L, remaining is water, pH value 8.8.
In a presently preferred embodiment, ethyl alcohol by concentration of volume percent is in the Optimal Medium of the step 2) 2.5%.
In a presently preferred embodiment, the additive amount of alpha, beta-lonone is 10mg/ in the Optimal Medium of the step 2) L.Inventor is it was unexpectedly observed that state adds micro alpha, beta-lonone and is remarkably improved spirulina platensis polysaccharide during the cultivation process Content, it is related that the possible process and alpha, beta-lonone help to be promoted frustule amino acid and protein synthase activity, more The promotion of sugared content further improves the productivity and yield of blunt top spirulina, economic benefit is promoted, additionally due to β-violet The dosage of ketone is few, and blunt top spirulina is being stimulated not obviously increase cost, control cultivation investment while accumulating polysaccharide.
In preferred embodiments, the additive amount of fulvic acid is 38mg/L in the Optimal Medium of the step 3).
In preferred embodiments, in the second stage culture of the step 3), 10h gives 3500Lux illumination, culture temperature Degree is 32 DEG C, and air mass flow is 0.6vvm, and incubation time is 9d.
The present invention also provides above-mentioned any one to be remarkably improved Growth of Spirulina Platensis speed and nutrition content Application of the method in blunt top spirulina culture comprising utilize the method culture blunt top spirulina.
Comparative example D2~D4: influence of the zymocyte liquid to culture blunt top spirulina:
Comparative example D2:
Comparative example D2 is substantially the same manner as Example 1, the difference is that in the step 1) of comparative example D2, zymocyte liquid is only 1.2×107The Rhodobacter capsulatus of a/mL fermented after zymocyte liquid.
Comparative example D3:
Comparative example D3 is substantially the same manner as Example 1, the difference is that in the step 1) of comparative example D3, zymocyte liquid is only 1.2×107The Rhodopseudomonas palustris of a/mL fermented after zymocyte liquid.
Comparative example D4:
Comparative example D4 is substantially the same manner as Example 1, the difference is that in the step 1) of comparative example D4, in basal medium It is not added with any zymocyte liquid.
Comparative example D5: influence of the alpha, beta-lonone to culture blunt top spirulina:
Comparative example D5 is substantially the same manner as Example 1, the difference is that in the step 2) of comparative example D5, first stage optimization Alpha, beta-lonone is not added in the Optimal Medium of culture.
Comparative example D6~D8: monochromatic light ray is according to the influence to culture blunt top spirulina:
Comparative example D6:
Comparative example D6 is substantially the same manner as Example 1, the difference is that the first stage in the step 2) of comparative example D6 is excellent Change the orange-colored light irradiation culture that the 620nm of 4000Lux is used alone in culture.
Comparative example D7:
Comparative example D7 is substantially the same manner as Example 1, the difference is that the first stage in the step 2) of comparative example D7 is excellent Change the blue light irradiation culture that the 420nm of 4000Lux is used alone in culture.
Comparative example D8:
Comparative example D8 is substantially the same manner as Example 1, the difference is that the first stage in the step 2) of comparative example D8 is excellent Change and uses the culture of 4000Lux white light in culture.
Comparative example D9: influence of the fulvic acid to culture blunt top spirulina:
Comparative example D9 is substantially the same manner as Example 1, the difference is that the second stage in the step 2) of comparative example D9 is excellent The Optimal Medium for changing culture is not added with fulvic acid.
Experimental example 1: the detection of influence of the cultural method to Growth of Spirulina Platensis speed:
Detection method are as follows: with spectrophotometer detecting step 1) in the blunt top spirulina cell population density that is inoculated with after activation (OD560), it is calculated as 1 (a reference value), respectively detecting step 3) blunt top spirulina cell population density in Optimal Medium；Foundation Above-mentioned detection method detects the incubation in embodiment 1, comparative example D2~D9, and statistical result is as shown in table 1.
Table 1, Growth of Spirulina Platensis speed statistics
Growth speed of the blunt top spirulina in second stage optimization it is found that in the preferred embodiment of the present invention 1 is analyzed by table 1 Degree is very fast, comparative example is significantly faster than, particularly it can further be seen that in different fermentations bacterium solution, alpha, beta-lonone, illumination and richness Acid has different degrees of influence to the growth and breeding of blunt top spirulina.
Experimental example 2: the detection of influence of the cultural method to blunt top spirulina content beta-carotene:
Detection method: taking algae solution to filter, and is centrifuged to obtain algal gel, carries out vacuum freeze drying, and acetone is added, and ice-bath ultrasonic is broken 20min takes supernatant is standby to survey；High performance liquid chromatography surveys content beta-carotene in blunt top spirulina, and mobile phase is VMethanol:VAcetone= 95:5。
The blunt top spirulina that in the above way each method culture in embodiment 1, comparative example D2~D9 is obtained respectively into Row detection, it is as shown in Figure 1 to count content beta-carotene in the blunt top spirulina of each example method.By the diagram of Fig. 1 it is found that originally The content beta-carotene of blunt top spirulina in the preferred embodiment 1 of application is higher, illustrates that the present processes can be mentioned significantly Rise the content beta-carotene and output of blunt top spirulina.
Experimental example 3: the detection of influence of the cultural method to spirulina platensis polysaccharide content:
Detection method: taking algae solution to be centrifuged through 6000r/min, takes supernatant, adjusts pH to 7, is gradually added into 20% lead acetate To precipitating completely to remove the substances such as protein, pigment, colloid, and 10% sulfuric acid isometric with lead acetate liquid is added in liquid Then sodium solution is added 95% ethanol precipitation and obtains EPS of Spirulina until no longer generating precipitating；Centrifugation gained algal gel water Add deionized water after washing；Ultrasonic disruption adjusts pH to 10, spirulina intracellular polyse is extracted with hot-water extraction method, with three chloroethenes Sour (TCA) method deproteinized matter and cytochromes；Sulfuric acid-phynol method detects polyoses content.
The blunt top spirulina that in the above way each method culture in embodiment 1, comparative example D2~D9 is obtained respectively into Row detection, counts the content (with dry cell weight basis, %) of the exocellular polysaccharide and intracellular polyse in the blunt top spirulina of each example method, Statistical result is as shown in Figure 2.
As shown in Figure 2, the exocellular polysaccharide of the blunt top spirulina in preferred embodiment of the present application 1 and intracellular polyse content are equal In higher level, illustrate that the present processes can be obviously improved the polyoses content and output of blunt top spirulina, in conjunction with the above reality The present processes known to testing example may make the content of nutriment in frustule to dramatically increase, and promote the breeding of blunt top spirulina Efficiency and economic well-being of workers and staff, be a kind of high efficiency, low cost be remarkably improved Growth of Spirulina Platensis speed and nutriment contains The method of amount.
The prior art of routine techniques dawn known to those skilled in the art in above-described embodiment, therefore herein no longer in detail It repeats.
Although above-mentioned specific embodiment has shown that, is described and pointed out novel feature applied to various embodiments, It, can form and details to illustrated device or method it should be understood that under the premise of not departing from the spirit of present disclosure Carry out various omissions, substitutions and changes.In addition, above-mentioned various features and method can be used independently of each other, or can be with various sides Formula combination.All possible combination and sub-portfolio are intended to and fall within the scope of the present disclosure.Above-mentioned many embodiment packets Similar component is included, and therefore, these similar components are interchangeable in different implementation scenarios.Although in certain realities Apply scheme and embodiment it is disclosed in the context that the present invention, it is understood by one skilled in the art that the present invention can be beyond specific Disclosed embodiment extends to other alternate embodiments and/or application and its apparent modification and equivalent.Therefore, The present invention is not intended to be limited by the specific disclosure of this paper preferred embodiment.

Claims (10)

1. the method for being remarkably improved Growth of Spirulina Platensis speed and nutrition content, it is characterised in that the method includes Following step:
1) belonged to the red bacterium containing inactivation and the basal medium of the zymocyte liquid of Rhodopseudomonas bacterium is by the blunt of activation Top spirulina culture filters out algae solution and obtains algal gel after centrifugation or filtration washing to the logarithmic growth phase later period；
2) first stage optimization culture is carried out with algal gel of the Optimal Medium containing ethyl alcohol and alpha, beta-lonone to step 1), adopted With orange-colored light and the artificial LED light source joint irradiation culture of blue light, total intensity of illumination is 2500~4500Lux, at 18~22 DEG C At a temperature of cultivate 5~10d, obtain the blunt top spirulina containing swarm cell；
(3) it under natural optical mode condition of culture, separately takes and fulvic acid is added in Optimal Medium, by the thin containing travelling of step 2) The blunt top spirulina of born of the same parents carries out second stage culture after obtaining in the form of algal gel in Optimal Medium, keeps at least partly travelling thin Dysuria with lower abdominal colic turns to non motile cell.
2. according to the method described in claim 1, it is characterized by:
The red thin campylobacter bacteria is Rhodobacter capsulatus, and/or
The Rhodopseudomonas bacterium is Rhodopseudomonas palustris.
3. method according to claim 1 or 2, it is characterised in that: the fermentation process of the zymocyte liquid is:
Prepare fermentation medium are as follows: 0.4~1.0g/L of ferric citrate, 0.3~0.6g/L of dipotassium hydrogen phosphate, magnesium sulfate 0.2~ 0.4g/L, 0.4~1.0g/L of sodium chloride, 0.5~2.0g/L of yeast powder, 3.0~5.0g/L of sodium acetate, remaining is distilled water；
After sterilizing according to number proportion 3:1 be inoculated with altogether the red thin campylobacter bacteria and the Rhodopseudomonas bacterium 1~1.2 × 107A/mL, at a temperature of 25~28 DEG C, ferment 12~96h under 5000~8000Lux continuous light, and complete inactivation is simultaneously concentrated into close Degree is at least 1.15g/mL to obtain the final product.
4. method according to claim 1 or 2, it is characterised in that: the red bacterium that is inactivated in the basal medium belong to and The additive amount of the zymocyte liquid of Rhodopseudomonas bacterium is 1.2~1.5mL/L.
5. according to the method described in claim 1, it is characterized by: basal medium composition is as follows: sodium bicarbonate 0.2~ 0.4g/L, 0.5~1.0g/L of potassium nitrate, 0.4~0.6g/L of sodium chloride, 0.02~0.03g/L of zinc sulfate, frerrous chloride 0.05~ 0.08g/L, 0.01~0.02g/L of ferric citrate, A5 component 1mL/L, remaining is water, and pH is 7.5~9.5.
6. according to the method described in claim 1, it is characterized by: the wavelength of the orange-colored light is 590~620nm, the indigo plant The wavelength of coloured light is 445~475nm, and the intensity of illumination ratio of the orange-colored light and blue light is 3~5:1.
7. according to the method described in claim 1, it is characterized by:
Ethyl alcohol is 2.0~4.2% by concentration of volume percent in the Optimal Medium of the step 2)；
The additive amount of alpha, beta-lonone is 0.2~20mg/L in the Optimal Medium of the step 2).
8. according to the method described in claim 1, it is characterized by: in the Optimal Medium of the step 3) fulvic acid addition Amount is 35~45mg/L.
9. according to the method described in claim 1, it is characterized by: 10~12h gives in the second stage culture of the step 3) 3500~4000Lux illumination is given, cultivation temperature is 30~32 DEG C, and air mass flow is 0.5~0.6vvm, incubation time is 8~ 10d。
10. being remarkably improved the method for Growth of Spirulina Platensis speed and nutrition content described in any one of claim 1~9 Application in blunt top spirulina culture, it is characterised in that utilize the method culture blunt top spirulina.
CN201910625341.0A 2019-07-11 2019-07-11 Method for remarkably improving growth speed and nutrient content of spirulina platensis Active CN110272849B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201910625341.0A CN110272849B (en) 2019-07-11 2019-07-11 Method for remarkably improving growth speed and nutrient content of spirulina platensis

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201910625341.0A CN110272849B (en) 2019-07-11 2019-07-11 Method for remarkably improving growth speed and nutrient content of spirulina platensis

Publications (2)

Publication Number Publication Date
CN110272849A true CN110272849A (en) 2019-09-24
CN110272849B CN110272849B (en) 2023-01-03

Family

ID=67964278

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201910625341.0A Active CN110272849B (en) 2019-07-11 2019-07-11 Method for remarkably improving growth speed and nutrient content of spirulina platensis

Country Status (1)

Country Link
CN (1) CN110272849B (en)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107287125A (en) * 2017-08-24 2017-10-24 黑龙江科技大学 A kind of cultural method of chlorella pyrenoidosa
CN115074279A (en) * 2022-06-27 2022-09-20 李航 Method for increasing beta-carotenoid content in spirulina and spirulina thereof
CN116925983A (en) * 2023-09-14 2023-10-24 烟台泓源生物肥料有限公司 Culture method of spirulina platensis

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0746993A (en) * 1993-05-31 1995-02-21 Pentel Kk Production of phycocyanin
JP2002315569A (en) * 2001-04-24 2002-10-29 Tokai Sangyo Kk Method for culturing algae
CN105940112A (en) * 2014-01-27 2016-09-14 泰恩河畔纽卡斯尔大学 Improvements in the synthesis of phycocyanins
CN109777754A (en) * 2019-01-30 2019-05-21 北京林业大学 A method of improving blunt top spirulina beta carotene and zeaxanthin accumulation

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0746993A (en) * 1993-05-31 1995-02-21 Pentel Kk Production of phycocyanin
JP2002315569A (en) * 2001-04-24 2002-10-29 Tokai Sangyo Kk Method for culturing algae
CN105940112A (en) * 2014-01-27 2016-09-14 泰恩河畔纽卡斯尔大学 Improvements in the synthesis of phycocyanins
CN109777754A (en) * 2019-01-30 2019-05-21 北京林业大学 A method of improving blunt top spirulina beta carotene and zeaxanthin accumulation

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107287125A (en) * 2017-08-24 2017-10-24 黑龙江科技大学 A kind of cultural method of chlorella pyrenoidosa
CN107287125B (en) * 2017-08-24 2020-08-14 黑龙江科技大学 Method for culturing chlorella pyrenoidosa
CN115074279A (en) * 2022-06-27 2022-09-20 李航 Method for increasing beta-carotenoid content in spirulina and spirulina thereof
CN115074279B (en) * 2022-06-27 2024-07-09 李航 Method for increasing content of beta-carotenoid in spirulina and spirulina
CN116925983A (en) * 2023-09-14 2023-10-24 烟台泓源生物肥料有限公司 Culture method of spirulina platensis
CN116925983B (en) * 2023-09-14 2023-12-01 烟台泓源生物肥料有限公司 Culture method of spirulina platensis

Also Published As

Publication number Publication date
CN110272849B (en) 2023-01-03

Similar Documents

Publication Publication Date Title
Soni et al. Spirulina–From growth to nutritional product: A review
Iwamoto Industrial production of microalgal cell‐mass and secondary products‐major industrial species: chlorella
Barghbani et al. Investigating the effects of several parameters on the growth of Chlorella vulgaris using Taguchi's experimental approach
Borowitzka Commercial production of microalgae: ponds, tanks, tubes and fermenters
Chacón‐Lee et al. Microalgae for “healthy” foods—possibilities and challenges
WO2012100583A1 (en) Culturing method for microalgae with high yields
CN110272849A (en) The method for being remarkably improved Growth of Spirulina Platensis speed and nutrition content
CN105755088B (en) The method for inducing Haematococcus pluvialis production astaxanthin
WO2012071983A1 (en) Heterotrophic culturing method for microalgae with high yields
CN108410737B (en) A kind of two-steps tissue culture method of purple ball algae
CN106804285A (en) Culture method for improving triterpenoid components of antrodia camphorata
CN102094061B (en) Method for producing lutein from microalgae
CN106190853B (en) A kind of red algae cultural method of high yield phycocyanin
CN103627639B (en) A kind of method utilizing spirulina breeding liquid residual liquid cultivation Dunaliella salina
CN103460994A (en) Cordyceps militaris nano-selenium and preparation method thereof
CN104212865A (en) Production process for producing astaxanthin by micro-alga culture
Pan-utai et al. Photoautotrophic cultivation of Arthrospira maxima for protein accumulation under minimum nutrient availability
CN106399108A (en) Simple high-efficiency haematococcus pluvialis nutritive cell culturing and harvesting method
Borowitzka Algae as food
CN106868085A (en) A kind of method for promoting haematococcus pluvialis rapid conversion to accumulate astaxanthin
CN104711203A (en) Selenium-rich germanium yeast powder
CN109777754A (en) A method of improving blunt top spirulina beta carotene and zeaxanthin accumulation
CN103396953B (en) Batch feeding method for cultivating chlorella
CN103299823A (en) Method for improving yield and trace element enrichment of liquid fermentation mycelium of lucid ganoderma
WO2012065545A1 (en) Microalgae culturing method for oil and lutein rapid accumulation

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
GR01 Patent grant
GR01 Patent grant
TR01 Transfer of patent right
TR01 Transfer of patent right

Effective date of registration: 20230613

Address after: 016199 Spirulina Industrial Park, Otog Banner, Wulan Town, Otog Banner, Ordos City, Inner Mongolia

Patentee after: Etuoke Jinchang Spirulina Co.,Ltd.

Address before: 2081, building a, 88 Jianghai West Road, Liangxi District, Wuxi City, Jiangsu Province, 214000

Patentee before: Wuxi Xiangyuan Information Technology Co.,Ltd.

Effective date of registration: 20230613

Address after: 2081, building a, 88 Jianghai West Road, Liangxi District, Wuxi City, Jiangsu Province, 214000

Patentee after: Wuxi Xiangyuan Information Technology Co.,Ltd.

Address before: 316100 plot C2-10, Xiaohui Industrial Zone, Putuo exhibition, Putuo marine science and Technology Industrial Park, Putuo District, Zhoushan City, Zhejiang Province

Patentee before: Zhejiang Ocean University