CN105925486B - The fermentation process of chlorella three-dimensional pipeline bioreactor and its application method of chlorella obtained - Google Patents
The fermentation process of chlorella three-dimensional pipeline bioreactor and its application method of chlorella obtained Download PDFInfo
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- CN105925486B CN105925486B CN201610325818.XA CN201610325818A CN105925486B CN 105925486 B CN105925486 B CN 105925486B CN 201610325818 A CN201610325818 A CN 201610325818A CN 105925486 B CN105925486 B CN 105925486B
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- chlorella
- dimensional pipeline
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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
- C12N1/12—Unicellular algae; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/02—Photobioreactors
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/48—Automatic or computerized control
Abstract
The present invention relates to technical field of bioengineering, a kind of fermentation process of chlorella three-dimensional pipeline bioreactor and its application method of chlorella obtained are disclosed.The following steps are included: biogas slurry or nutrient solution are put into three-dimensional pipeline bioreactor by (1), access chlorella vulgaris is added amino-laevulic acid or its hydrochloride, is sufficiently mixed the present invention；(2) it is passed through high-purity CO2Gas, ventilatory capacity 1.0-2.5vvm control turbulent parameters, turbulivity 1-5% by automated system, and control dissolved oxygen amount is not less than 16%, continuously ferments；(3) it ferments from next day, daily addition amino-laevulic acid or its hydrochloride are sufficiently mixed；(4) continuously ferment after 3-4d, fermentation liquid becomes bottle green from colourless, as mature bead algae solution.Fermentation period of the present invention is short, CO2High conversion rate avoids the influence of environmental condition, has effectively completely cut off the harm of miscellaneous bacteria infection, can continuously ferment.
Description
Technical field
The present invention relates to technical field of bioengineering, more particularly to it is a kind of for chlorella three-dimensional pipeline photo-biological it is anti-
Answer the fermentation process of device and its application method of chlorella obtained.
Background technique
Chlorella protein rich in, amino acid, pigment and polyunsaturated fatty acid etc., nutrition is comprehensive, has pole
Its nutritional ingredient abundant and excellent medical care effect.Chlorella protein content is up to 50%-67%, wherein containing someone
20 kinds of amino acid, multivitamin necessary to body and the ingredients such as microelement and linolenic acid, linoleic acid, carotenoid.
It is rich in chlorella growth factor (CGF) in chlorella, damage caused by cultivated animals body can be restored rapidly.Contain in chlorella
Bioactive substance glycoprotein, polysaccharide body and up to 13% the substances such as nucleic acid, there is significant tumor-inhibiting anticancer, enhancing to exempt from
The activity of epidemic disease and viral infection resisting.
Early in the 1940s, there are many institution of scientific research and livestock-raising fields in the world applies chlorella
In the health care product of people and poultry, livestock, companion animals, raising fish and shrimp, honeybee and silkworm aquaculture.Experimental results demonstrate small
Ball algae all has obvious effects on the raising of the survival rate of human health and cub, production performance and product quality.
Currently, European and American developed countries are very universal to development and utilization of the chlorella on health care product and biological feedstuff.It is international
About 10,000 tons of chlorella product dry matter of market year consumption, only this year day consumes chlorella product dry matter up to 1,000,000,000 dollars or more.
Currently used production method is mostly runway recognition pond or the photosynthetic culture of pocket type, this training method climate condition (temperature
Degree, illumination etc.) influence it is bigger, and invasion vulnerable to enemy (wheel animalcule, infusorian etc.), land occupation and water consumption are big, product
Production cost is higher, and cultured output is low and is difficult to stablize.
It is disclosed in Chinese patent CN103060199B " semicontinuous fermentation method of heterophytic chlorella " a kind of to culture medium
It is inoculated with heterophytic chlorella, using the semicontinuous fermentation method for the method fermentation that spreads cultivation, bigger culture medium/seed rate is met and wants
Ask, reduce the inadaptability of cell growth, make cell growth faster, reduce deadtime, fermentation period in 10-15d or so,
Fermentation period is longer, and fermentation process is semicontinuous, inconvenient, and there are still the threats of miscellaneous bacteria infection.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of fermentation period is short, CO2High conversion rate avoids environmental condition
Influence, effectively completely cut off miscellaneous bacteria infection harm, the fermentation for the chlorella three-dimensional pipeline bioreactor that can continuously ferment
Method.
In order to solve the above technical problems, the present invention adopts the following technical scheme:
The fermentation process of chlorella three-dimensional pipeline bioreactor of the present invention, comprising the following steps:
(1) biogas slurry or nutrient solution are put into three-dimensional pipeline bioreactor, access chlorella vulgaris, glycyl is added
Propionic acid or its hydrochloride, are sufficiently mixed；
(2) it is passed through high-purity CO2Gas, ventilatory capacity 1.0-2.5vvm control turbulent parameters, turbulent flow by automated system
Degree is 1-5%, and control dissolved oxygen amount is not less than 16%, continuously ferments；
(3) it ferments from next day, daily addition amino-laevulic acid or its hydrochloride are sufficiently mixed；
(4) continuously ferment after 3-4d, fermentation liquid becomes bottle green from colourless, as mature bead algae solution.
The fermentation process of chlorella three-dimensional pipeline bioreactor of the present invention further includes further step (5), will
The mature bead algae solution centrifugal concentrating that step (4) obtains obtains chlorella concentrate, packs after cooling, storage.
The fermentation process of chlorella three-dimensional pipeline bioreactor of the present invention, further, step (1) neutral body pipeline
The three-dimensional pipeline bioreactor that bioreactor selects the high light conversion agent containing rare earth to be fabricated.
The fermentation process of chlorella three-dimensional pipeline bioreactor of the present invention, further, that accesses in step (1) is small
The concentration of ball algae is the 5-10% (volume ratio) of biogas slurry or nutrient solution, and the amount of the amino-laevulic acid of addition or its hydrochloride is
0.1-2.0ppb (volume ratio).
The fermentation process of chlorella three-dimensional pipeline bioreactor of the present invention, further, CO in step (2)2Gas
Purity be not less than 99.9%.
The fermentation process of chlorella three-dimensional pipeline bioreactor of the present invention, further, control fermentation in step (2)
Temperature is 10-35 DEG C, illuminance 600-2500Lux, pH 3.0-8.0.
The fermentation process of chlorella three-dimensional pipeline bioreactor of the present invention, further, step are added daily in (3)
Amino-laevulic acid or its hydrochloride amount be 0.1-2.0ppb (volume ratio).
The fermentation process of chlorella three-dimensional pipeline bioreactor of the present invention, further, step (1) and step (3)
In amino-laevulic acid or its hydrochloride are added under the conditions of being protected from light.
The fermentation process of chlorella three-dimensional pipeline bioreactor of the present invention, further, in step (4) every milliliter at
Dry matter content is 1.5-2.5g in ripe bead algae solution.
The mature bead algae solution that the fermentation process of chlorella three-dimensional pipeline bioreactor of the present invention is prepared makes
With method, mature bead algae solution can be mixed in feed or water source and be used to poultry, can also be dried to powder and health care product or food is added
In eat.
The fermentation process of chlorella three-dimensional pipeline bioreactor of the present invention compared with prior art, has following beneficial
Effect:
The fermentation process of chlorella three-dimensional pipeline bioreactor of the present invention is built using bioreactor to be closed, is steady
Fixed environment realizes continuously fermenting for chlorella, substantially increases light by the best turbulent parameters of automated system control fermentation
The efficiency of cooperation；In maturation bead algae solution made from the method for the present invention, amount of dry matter 1.5-2.5g/mL, much higher than existing
Chlorella stoste made from fermentation process, and shearing force is small in bioreactor, it is structurally reasonable conducive to the growth of chlorella,
It is easy to operate, there is excellent frustule growing environment, power consumption is small, and reactor occupies little space, and equipment cost is low.
Be in traditional open fermentation process and the existing fermentation process using bioreactor use purity for
The CO of 0-50%2Gas is the photosynthetic carbon source of chlorella, all there is CO2What purity or excessive concentration can not convert completely asks
Topic, and the present invention is added to micro amino-laevulic acid or its hydrochloride, additive amount 0.1- in bioreactor
2.0ppb matches with the turbulence reaction condition of continuous-stable although additive amount is few, can greatly improve CO2Conversion
Rate, the present invention are not less than 99.9% high-purity CO using purity2, bioreactor is passed through with ventilatory capacity 1.0-2.5vvm, is passed
It send kinetic energy to consume small, energy conservation and environmental protection, reduces production run cost, CO2Utilization rate is up to 99% or more, solves existing method
In existing CO2The low problem of conversion ratio.
The present invention adds micro amino-laevulic acid or its hydrochloride daily, avoid a flash amino-laevulic acid or its
It is decomposed caused by hydrochloride concentration is excessively high and the low problem of utilization rate, and it is vigorous with metastable content to maintain chlorella
Metabolic capabilities, improve the photosynthetic efficiency of chlorella.
The present invention is selected in no solar radiation or closes under the conditions of being protected from light of LED light, and amino-laevulic acid or its hydrochloric acid is added
Salt is extremely easy in decomposition under illumination condition because it is light sensitive species and loses physiological activity, therefore is added under the conditions of being protected from light,
Chlorella can be made quickly to absorb amino-laevulic acid or its hydrochloride without improving amino-laevulic acid by illumination decomposition and consumption
Or the utilization rate of its hydrochloride, it is the CO of Efficient Conversion high-purity high-concentration2It lays a good foundation.
The fermentation period of traditional open fermentation process is generally 10-20d, and emerging is general existing anti-using photo-biological
The fermentation period of the fermentation process of device is answered to be generally 7-10d, and the present invention is due to improving CO2Conversion ratio substantially reduces hair
The time required to ferment, the method for the present invention fermentation period is 3-4d, substantially increases production efficiency, and fermentation process is continuous, easy to operate, together
When suitable for Laboratory Production and, the production of large-scale yield.
The present invention can select the culture medium that remaining biogas slurry is chlorella fermentation that ferments, and complete the benefit of biological waste liquid
With reducing the processing cost of waste liquid, alleviate environmental pressure, reduce production cost.Three-dimensional pipeline photoproduction in the present invention
Object reactor selects the three-dimensional pipeline bioreactor of the high light conversion agent manufacture containing rare earth, and light transmittance is improved, into
And photosynthetic efficiency is improved, and using the turbulivity of 1-5%, chlorella is made to be in best suspended state, it can sufficiently and CO2
Contact is completed to absorb and be converted.
The present invention selects three-dimensional pipeline bioreactor, and the dosage of equipment space occupied and water is greatly saved, and improves
Fermentation total output, water consumption are 30% or so of the production technology of traditional raceway pond, and space occupied is traditional raceway pond
40% or less.And be the structure of closed circulation, the microbiological contamination problem avoided can save 60% or more production cost.
Specific embodiment
Prepare embodiment
The fermentation process of chlorella three-dimensional pipeline bioreactor of the present invention the following steps are included:
(1) biogas slurry or nutrient solution are put into three-dimensional pipeline bioreactor, access chlorella vulgaris, inoculum density such as table
Shown in 1, amino-laevulic acid or its hydrochloride are then added under the conditions of being protected from light, additional amount is as shown in table 1, is sufficiently mixed
30min；
(2) it is passed through high-purity CO that purity is not less than 99.9%2Gas controls turbulent parameters by automated system, so that
Dissolved oxygen amount is not less than 16% in fermentation liquid, and design parameter is as shown in table 2, continuously ferments；
(3) ferment next day from, daily under the conditions of being protected from light be added 0.1-2.0ppb (volume ratio) amino-laevulic acid or
Its hydrochloride, is sufficiently mixed 30min；
(4) continuously ferment after 3-4d, when fermentation liquid becomes bottle green from colourless, as mature bead algae solution, every milliliter at
Dry matter content is 1.5-2.5g in ripe bead algae solution；
(5) the mature bead algae solution centrifugal concentrating for obtaining step (4), obtains chlorella concentrate, chlorella after concentration
The water content of concentrate is not higher than 60%, packs after cooling, storage.
Table 1 respectively prepares fermentation raw material component and proportion in embodiment
Table 2 respectively prepares basic fermentation condition in embodiment
| Preparation example 1 | Preparation example 2 | Preparation example 3 | Preparation example 4 | Preparation example 5 | Preparation example 6 | |
| CO2Ventilatory capacity vvm | 1.0 | 1.2 | 1.5 | 2.5 | 2.3 | 2.0 |
| Temperature DEG C | 10 | 15 | 35 | 20 | 25 | 30 |
| Illuminance Lux | 2500 | 1500 | 600 | 2000 | 2300 | 1000 |
| pH | 5.4 | 6.9 | 7.8 | 3.1 | 4.5 | 6.2 |
| Turbulent velocity (%) | 1 | 5 | 2 | 3 | 5 | 4 |
Product made from above-mentioned preparation embodiment is provided to carry out quality testing according to concerned countries, specific testing result is such as
Described in table 3.
Table 3 respectively prepares the quality measurements that product is made in embodiment
| Testing index | Detection method | Preparation 1 | Preparation 2 | Preparation 3 | Preparation 4 | Preparation 5 | Preparation 6 |
| Dry matter g/mL | GB/T 6435-2006 | 1.5 | 2.5 | 1.7 | 2 | 2.3 | 2.1 |
| Crude protein % | GB/T 6432-1994 | 55.61 | 60.69 | 57.2 | 61.08 | 62.47 | 58.4 |
| EPA and DHA% | GB/T5009.168-2003 | 6.38 | 8.27 | 6.72 | 7.41 | 7.97 | 7.04 |
| Vitamin A IU/kg | GB/T 17817-2010 | 3217 | 3549 | 3304 | 3460 | 3597 | 3401 |
| Vitamin E mg/kg | GB/T 17812-2008 | 580.16 | 610.2 | 592.45 | 604.23 | 609.4 | 585.9 |
| Chlorophyll mg/kg | GB/T 22182-2008 | 13070 | 13459 | 13108 | 13243 | 13398 | 13327 |
As shown in table 3, in maturation bead algae solution made from the method for the present invention, amount of dry matter 1.5-2.5g/mL is much higher than
The preparation of existing method is horizontal, and for crude protein quality high percentage up between 55-63%, the mass percent of EPA and DHA are up to 6-
Between 9%, retinol equivalent is between 3200-3600IU/kg, and content of vitamin E is between 580-610mg/kg, chlorophyll
Content is between 13000-13500mg/kg, and each nutrient composition content is much higher than chlorella stoste made from existing fermentation process.
Application Example
Maturation bead algae solution made from the fermentation process of chlorella three-dimensional pipeline bioreactor of the present invention can directly be made
It is drunk for drinking-water water source to poultry.
Maturation bead algae solution made from the fermentation process of chlorella three-dimensional pipeline bioreactor of the present invention, can be with quality
It is more edible to poultry than in 10-20% incorporation poultry and livestock feed.
Maturation bead algae solution made from the fermentation process of chlorella three-dimensional pipeline bioreactor of the present invention, can be with volume
It is incorporated than 5-10% edible to poultry in tap water.
Maturation bead algae solution made from the fermentation process of chlorella three-dimensional pipeline bioreactor of the present invention, can be further
It is lyophilized into powder, is added in the health care products such as instant bean, albumen powder, rice flour or food with certain proportion.
Embodiment described above only describe the preferred embodiments of the invention, not to model of the invention
It encloses and is defined, without departing from the spirit of the design of the present invention, those of ordinary skill in the art are to technical side of the invention
The various changes and improvements that case is made should all be fallen into the protection scope that claims of the present invention determines.
Claims (7)
1. the fermentation process of chlorella three-dimensional pipeline bioreactor, it is characterised in that: the following steps are included:
(1) biogas slurry or nutrient solution are put into three-dimensional pipeline bioreactor, access chlorella vulgaris, amino-laevulic acid is added
Or its hydrochloride, it is sufficiently mixed；
(2) it is passed through high-purity CO2Gas, ventilatory capacity 1.0-2.5vvm control turbulent parameters by automated system, and turbulivity is
1-5%, control dissolved oxygen amount are not less than 16%, continuously ferment；
(3) it ferments from next day, daily addition amino-laevulic acid or its hydrochloride are sufficiently mixed；
(4) continuously ferment after 3-4d, fermentation liquid becomes bottle green from colourless, as mature bead algae solution；
The concentration of the chlorella vulgaris accessed in step (1) is the volume ratio 5-10% of biogas slurry or nutrient solution, the glycyl of addition
The amount of propionic acid or its hydrochloride is 0.1-2.0ppb.
2. the fermentation process of chlorella three-dimensional pipeline bioreactor according to claim 1, it is characterised in that: also wrap
Step (5) are included, the mature bead algae solution centrifugal concentrating that step (4) is obtained obtains chlorella concentrate, packs after cooling, storage
It deposits.
3. the fermentation process of chlorella three-dimensional pipeline bioreactor according to claim 1, it is characterised in that: step
(1) the three-dimensional pipeline optical-biological reaction that neutral body pipeline bioreactor selects the high light conversion agent containing rare earth to be fabricated
Device.
4. the fermentation process of chlorella three-dimensional pipeline bioreactor according to claim 1, it is characterised in that: step
(2) CO in2The purity of gas is not less than 99.9%.
5. the fermentation process of chlorella three-dimensional pipeline bioreactor according to claim 1, it is characterised in that: step
(2) control fermentation temperature is 10-35 DEG C in, illuminance 600-2500Lux, pH 3.0-8.0.
6. the fermentation process of chlorella three-dimensional pipeline bioreactor according to claim 1, it is characterised in that: step
(1) and in step (3) amino-laevulic acid or its hydrochloride are added under the conditions of being protected from light.
7. the fermentation process of chlorella three-dimensional pipeline bioreactor according to claim 1, it is characterised in that: step
(4) dry matter content is 1.5-2.5g in every milliliter of mature bead algae solution in.
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| CN115449485B (en) * | 2022-11-14 | 2023-03-31 | 天津长芦汉沽盐场有限责任公司 | Method for culturing marine chlorella |
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| CN1837351A (en) * | 2006-04-12 | 2006-09-27 | 华东理工大学 | Method for culturing chlorella with high-density and high-quality |
| CN101712929A (en) * | 2008-10-08 | 2010-05-26 | 青岛生物能源与过程研究所 | Method for carrying out microalgae photobiological culture by utilizing luminous particle |
| CN103114041A (en) * | 2012-12-19 | 2013-05-22 | 盐城工学院 | Method for rapidly cultivating chlorella |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1837351A (en) * | 2006-04-12 | 2006-09-27 | 华东理工大学 | Method for culturing chlorella with high-density and high-quality |
| CN101712929A (en) * | 2008-10-08 | 2010-05-26 | 青岛生物能源与过程研究所 | Method for carrying out microalgae photobiological culture by utilizing luminous particle |
| CN103114041A (en) * | 2012-12-19 | 2013-05-22 | 盐城工学院 | Method for rapidly cultivating chlorella |
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