CN103881937A - Method for preparing photosynthetic bacteria liquid-solid dual-fermentation high-density microecological preparation - Google Patents
Method for preparing photosynthetic bacteria liquid-solid dual-fermentation high-density microecological preparation Download PDFInfo
- Publication number
- CN103881937A CN103881937A CN201210556654.3A CN201210556654A CN103881937A CN 103881937 A CN103881937 A CN 103881937A CN 201210556654 A CN201210556654 A CN 201210556654A CN 103881937 A CN103881937 A CN 103881937A
- Authority
- CN
- China
- Prior art keywords
- fermentation
- solid
- preparation
- minutes
- liquid
- 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
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method for preparing a photosynthetic bacteria liquid-solid dual-fermentation high-density microecological preparation, which comprises the steps of culturing a plating medium, culturing a test tube, culturing a conical flask, preparing second grade seed, preparing the third grade seed, and preparing the finished product. By employing the organic mediums with low concentration, such as peptone, extract yeast and NPK salt, so that the pure and bright color of the cultured photosynthetic bacteria can be endured, shallowing, greening and darkening are not generated; low cost can be guaranteed by employing nitrogen source and carbon source with low concentration, so that the method of the invention is suitable for large-scale industrial production; the photosynthetic bacteria have the advantages of fast growth speed and high bacteria liquid concentration; the preparation method is simple and easy to control, competitor is difficulty infected, and the bacterial strain concentration is high, cost of post-treatment is low, and production environment is good.
Description
Technical field
The invention belongs to technical field of bioengineering, be specifically related to the preparation method of the liquid-solid double fermentation high-density of light and bacterium probiotics.
Background technology
The current solid-state probiotics particularly formulation method of the solid-state micro-ecology of photosynthetic bacterium has two kinds of process technology schemes conventionally, and one is first to carry out liquid submerged fermentation, finally adds solid-state carrier absorption, dry forming; Though and liquid state fermentation is cultivated and can be obtained purer thalline, cost is high, fermentation is subject to product feedback control, is difficult to obtain high density bacterial strain, so that the effective bacterium of the method is counted content is lower; Another kind is the pattern of conventional solid state fermentation; But existing these technical matters method ubiquities processing step complexity, and preparation cost is high, the shortcomings such as product yield is low, miscellaneous bacteria content height.
Summary of the invention
The above-mentioned defect existing in order to overcome prior art field, the object of the invention is to, the preparation method of the liquid-solid double fermentation high-density of a kind of light and bacterium probiotics is provided, the formulation method that solves the solid-state micro-ecology of prior art photosynthetic bacterium exists cost high, be difficult to obtain high density bacterial strain, it is lower that effective bacterium is counted content, complex process, the problem that miscellaneous bacteria content is high.
The preparation method of the liquid-solid double fermentation high-density of light provided by the invention and bacterium probiotics, comprises the following steps:
One, dull and stereotyped cultivation, photosynthetic bacterium preserves bacterial classification streak inoculation to plate culture medium, and 121 DEG C of sterilizings 30 minutes, cultivate 24h for 32 DEG C;
Two, test tube inoculation, gets and grows healthy and strong, the typical inoculation 10ml of bacterium colony, and 121 DEG C of sterilizings 30 minutes, cultivate 24h for 32 DEG C;
Three, triangular flask inoculation 100ml, 121 DEG C of sterilizings 30 minutes, cultivate 24h for 32 DEG C;
Four, triangular flask inoculation 1000ml, 121 DEG C of sterilizings 30 minutes, cultivate 24h for 32 DEG C;
Five, the preparation of secondary seed, by photosynthetic bacterium, in 30 DEG C, air flow is 0.24m
3under/h condition, cultivate 24h, obtain second class inoculum 9L;
Six, the preparation of three grades of seeds, first in filling with, the liquid fermenting of 500L adds 100L tap water, by load weighted peptone 90g, yeast extract paste 90g, sodium acetate 180g, sodium bicarbonate 180g, ammonium chloride 180g, magnesium chloride 180g, dipotassium hydrogen phosphate 180g, glucose 3.6kg puts into fermentation and fills with, stir 10 minutes to all dissolving, then stop stirring, with tap water to 180L, determine after liquid level, open and stir, then open steam valve and be warming up to 100 DEG C, row's freezing air, follow-up continuous 121 DEG C of maintenances 30 minutes that are warming up to of the freezing air of draining, sterilizing finishes, cooling, for subsequent use, when substratum is cooled to 30 DEG C, the mode that photosynthetic bacterium secondary seed 9L is inoculated by pressure reduction, be inoculated in 500L fermentor tank, 30 DEG C, air flow is 9.6m
3/ h condition bottom fermentation 48h, obtains the about 200L of three-class strain,
Seven, solid substrate and culture medium raw material are joined in solid-state fermentation tank, 1.0MPa, 121 DEG C of sterilizing half an hour, while waiting substratum to be cooled to 30 DEG C, liquid spawn is mixed by 4:10 with solid fermentation matrix, the solid fermentation material mixing is contained into fermentation tray, and layer thickness 4-5cm left and right, is placed in sweathouse, make fermentation material aerobic fermentation, room temperature remains on 30 DEG C of left and right, and fermentation time is about 6 days, and between yeast phase, turn over twice every day.
In described step 1 or two or three or four, used medium is all by 0.5g peptone, 0.5g yeast extract paste, 1.0g sodium acetate, 1.0g sodium bicarbonate, 1.0g ammonium chloride, 0.5g magnesium chloride, 1.0g dipotassium hydrogen phosphate, 15g agar, the configuration of 1000ml distilled water forms, and pH value is 7; In described step 7, solid substrate is by 500kg Pericarppium arachidis hypogaeae or cotton seed hulls, 150kg Semen Maydis powder, and 200kg bean cake powder, 100kg wheat bran, 5kg glucose, the configuration of 200kg water forms.
The preparation method of the liquid-solid double fermentation high-density of light provided by the invention and bacterium probiotics, its beneficial effect is, in the present invention, Medium's PH Value does not raise along with the prolongation of incubation time, remains between 5.5-7.5, can not suppress the growth of photosynthetic bacterium; Adopt organic substratum such as peptone, yeast extract paste and the nitrogen phosphorus sylvite of lower concentration to ensure that the photosynthetic bacterium color of turning out is pure, beautiful, can be not thin out, turn green, blackening etc.; Adopt nitrogenous source and the carbon source of lower concentration to ensure low cost, more can be suitable for large-scale industrialization and produce; Photosynthetic bacterium growth speed is fast, and bacterial concentration is high; Its preparation technology is simple and be easy to control, is difficult for dying miscellaneous bacteria, and bacterial strain concentration is high, and post-treatment cost is low, production environment close friend.
Embodiment
Below in conjunction with an embodiment, the preparation method of light provided by the invention and the liquid-solid double fermentation high-density of bacterium probiotics is described in detail.
Embodiment
The preparation method of the liquid-solid double fermentation high-density of the light of the present embodiment and bacterium probiotics, comprises the following steps:
One, dull and stereotyped cultivation, photosynthetic bacterium preserves bacterial classification streak inoculation to plate culture medium, and 121 DEG C of sterilizings 30 minutes, cultivate 24h for 32 DEG C;
Two, test tube inoculation, gets and grows healthy and strong, the typical inoculation 10ml of bacterium colony, and 121 DEG C of sterilizings 30 minutes, cultivate 24h for 32 DEG C;
Three, triangular flask inoculation 100ml, 121 DEG C of sterilizings 30 minutes, cultivate 24h for 32 DEG C;
Four, triangular flask inoculation 1000ml, 121 DEG C of sterilizings 30 minutes, cultivate 24h for 32 DEG C;
Five, the preparation of secondary seed, by photosynthetic bacterium, in 30 DEG C, air flow is 0.24m
3under/h condition, cultivate 24h, obtain second class inoculum 9L;
Six, the preparation of three grades of seeds, first in filling with, the liquid fermenting of 500L adds 100L tap water, by load weighted peptone 90g, yeast extract paste 90g, sodium acetate 180g, sodium bicarbonate 180g, ammonium chloride 180g, magnesium chloride 180g, dipotassium hydrogen phosphate 180g, glucose 3.6kg puts into fermentation and fills with, stir 10 minutes to all dissolving, then stop stirring, with tap water to 180L, determine after liquid level, open and stir, then open steam valve and be warming up to 100 DEG C, row's freezing air, follow-up continuous 121 DEG C of maintenances 30 minutes that are warming up to of the freezing air of draining, sterilizing finishes, cooling, for subsequent use, when substratum is cooled to 30 DEG C, the mode that photosynthetic bacterium secondary seed 9L is inoculated by pressure reduction, be inoculated in 500L fermentor tank, 30 DEG C, air flow is 9.6m
3/ h condition bottom fermentation 48h, obtains the about 200L of three-class strain, and total bacteria containing amount reaches 1.3 × 10
9cfu/mL.
Seven, solid substrate and culture medium raw material are joined in solid-state fermentation tank, 1.0MPa, 121 DEG C of sterilizing half an hour, while waiting substratum to be cooled to 30 DEG C, liquid spawn is mixed by 4:10 with solid fermentation matrix, the solid fermentation material mixing is contained into fermentation tray, and layer thickness 4-5cm left and right, is placed in sweathouse, make fermentation material aerobic fermentation, room temperature remains on 30 DEG C of left and right, and fermentation time is about 6 days, and between yeast phase, turn over twice every day.
In described step 1 or two or three or four, used medium is all by 0.5g peptone, 0.5g yeast extract paste, 1.0g sodium acetate, 1.0g sodium bicarbonate, 1.0g ammonium chloride, 0.5g magnesium chloride, 1.0g dipotassium hydrogen phosphate, 15g agar, the configuration of 1000ml distilled water forms, and pH value is 7; In described step 7, solid substrate is by 500kg Pericarppium arachidis hypogaeae or cotton seed hulls, 150kg Semen Maydis powder, and 200kg bean cake powder, 100kg wheat bran, 5kg glucose, the configuration of 200kg water forms.
Claims (3)
1. a preparation method for the liquid-solid double fermentation high-density of light and bacterium probiotics, is characterized in that: comprise the following steps:
One, dull and stereotyped cultivation, photosynthetic bacterium preserves bacterial classification streak inoculation to plate culture medium, and 121 DEG C of sterilizings 30 minutes, cultivate 24h for 32 DEG C;
Two, test tube inoculation, gets and grows healthy and strong, the typical inoculation 10ml of bacterium colony, and 121 DEG C of sterilizings 30 minutes, cultivate 24h for 32 DEG C;
Three, triangular flask inoculation 100ml, 121 DEG C of sterilizings 30 minutes, cultivate 24h for 32 DEG C;
Four, triangular flask inoculation 1000ml, 121 DEG C of sterilizings 30 minutes, cultivate 24h for 32 DEG C;
Five, the preparation of secondary seed, by photosynthetic bacterium, in 30 DEG C, air flow is 0.24m
3under/h condition, cultivate 24h, obtain second class inoculum 9L;
Six, the preparation of three grades of seeds, first in filling with, the liquid fermenting of 500L adds 100L tap water, by load weighted peptone 90g, yeast extract paste 90g, sodium acetate 180g, sodium bicarbonate 180g, ammonium chloride 180g, magnesium chloride 180g, dipotassium hydrogen phosphate 180g, glucose 3.6kg puts into fermentation and fills with, stir 10 minutes to all dissolving, then stop stirring, with tap water to 180L, determine after liquid level, open and stir, then open steam valve and be warming up to 100 DEG C, row's freezing air, follow-up continuous 121 DEG C of maintenances 30 minutes that are warming up to of the freezing air of draining, sterilizing finishes, cooling, for subsequent use, when substratum is cooled to 30 DEG C, the mode that photosynthetic bacterium secondary seed 9L is inoculated by pressure reduction, be inoculated in 500L fermentor tank, 30 DEG C, air flow is 9.6m
3/ h condition bottom fermentation 48h, obtains the about 200L of three-class strain,
Seven, solid substrate and culture medium raw material are joined in solid-state fermentation tank, 1.0MPa, 121 DEG C of sterilizing half an hour, while waiting substratum to be cooled to 30 DEG C, liquid spawn is mixed by 4:10 with solid fermentation matrix, the solid fermentation material mixing is contained into fermentation tray, and layer thickness 4-5cm left and right, is placed in sweathouse, make fermentation material aerobic fermentation, room temperature remains on 30 DEG C of left and right, and fermentation time is about 6 days, and between yeast phase, turn over twice every day.
2. the preparation method of the liquid-solid double fermentation high-density of light according to claim 1 and bacterium probiotics, it is characterized in that: in described step 1 or two or three or four, used medium is all by 0.5g peptone, 0.5g yeast extract paste, 1.0g sodium acetate, 1.0g sodium bicarbonate, 1.0g ammonium chloride, 0.5g magnesium chloride, 1.0g dipotassium hydrogen phosphate, 15g agar, the configuration of 1000ml distilled water forms, and pH value is 7.
3. the preparation method of the liquid-solid double fermentation high-density of light according to claim 1 and bacterium probiotics, it is characterized in that: in described step 7, solid substrate is by 500kg Pericarppium arachidis hypogaeae or cotton seed hulls, 150kg Semen Maydis powder, 200kg bean cake powder, 100kg wheat bran, 5kg glucose, the configuration of 200kg water forms.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210556654.3A CN103881937A (en) | 2012-12-20 | 2012-12-20 | Method for preparing photosynthetic bacteria liquid-solid dual-fermentation high-density microecological preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210556654.3A CN103881937A (en) | 2012-12-20 | 2012-12-20 | Method for preparing photosynthetic bacteria liquid-solid dual-fermentation high-density microecological preparation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103881937A true CN103881937A (en) | 2014-06-25 |
Family
ID=50951063
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210556654.3A Pending CN103881937A (en) | 2012-12-20 | 2012-12-20 | Method for preparing photosynthetic bacteria liquid-solid dual-fermentation high-density microecological preparation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103881937A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109880779A (en) * | 2019-04-24 | 2019-06-14 | 安徽黄河水处理科技股份有限公司 | A kind of preparation method of photosynthetic bacteria liquid |
-
2012
- 2012-12-20 CN CN201210556654.3A patent/CN103881937A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109880779A (en) * | 2019-04-24 | 2019-06-14 | 安徽黄河水处理科技股份有限公司 | A kind of preparation method of photosynthetic bacteria liquid |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105462872B (en) | A kind of compound micro-ecological preparation and preparation method thereof | |
| CN103168613B (en) | Fast manufacture method for liquid strain | |
| CN103484421B (en) | A kind of chlamydosporic method of liquid fermenting scale up test Gliocladium roseum | |
| CN102550293B (en) | Method for liquid fermentation cultivation of Agaricus bisporus strain | |
| CN105316255B (en) | A kind of method of soil beneficial microbe mixed fermentation | |
| CN105039194A (en) | Yeast fused strain mixed bacterial agent and method of fermenting organic waste water therewith to prepare liquid fertilizer | |
| CN104403953A (en) | High-density fermentation culture medium formula for saccharomyces cerevisiae for feed and applications thereof | |
| CN103300209A (en) | Marsh rhodopseudomonas activation preparation and preparation method thereof | |
| CN102242075B (en) | Semi-continuous fermentation method for bacillus used for microbial fertilizer | |
| CN103695315B (en) | A kind of fermentable produces the method for chitin oligosaccharide | |
| CN102851330A (en) | Method for preparing citric acid through fermentation | |
| CN102533570B (en) | Aspergillus niger, application of Aspergillus niger and method for preparing citric acid by fermentation | |
| CN110205250A (en) | One plant of cellulase high-yield and its screening technique and application | |
| CN102550294B (en) | Method for liquid fermentation cultivation of Pleurotus cornucopiae strain | |
| CN102399702A (en) | Aspergillus niger and application thereof as well as citric acid preparation method through fermentation | |
| CN103881934A (en) | Preparation method for liquid and microecological preparation of photosynthetic bacteria | |
| CN105385608A (en) | Lentinus edodes liquid strain submerged fermentation technology | |
| CN103099162B (en) | Making method of L-lactic acid pickled vegetable | |
| CN102061279A (en) | Method for producing rhodopseudomonas palustris fermentation liquor by high-density fermentation | |
| CN103881937A (en) | Method for preparing photosynthetic bacteria liquid-solid dual-fermentation high-density microecological preparation | |
| CN105420130A (en) | Liquid-solid two-phase fermentation method for saccharomyces cerevisiae used for feed | |
| CN111621436B (en) | Culture solution suitable for producing prodigiosin by Serratia marcescens XD1-B-1 | |
| CN104152365B (en) | One strain produces bacterial strain and the production method thereof of KGA | |
| CN103305437B (en) | L-ammonium lactate tolerant bacterium and application thereof | |
| CN105296407A (en) | Method for culturing avibacterium paragallinarum bacterial solution |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140625 |