CN114317275A - Iron-deficiency culture medium for improving hydrogen yield of microalgae and preparation method and application thereof - Google Patents
Iron-deficiency culture medium for improving hydrogen yield of microalgae and preparation method and application thereof Download PDFInfo
- Publication number
- CN114317275A CN114317275A CN202111438479.3A CN202111438479A CN114317275A CN 114317275 A CN114317275 A CN 114317275A CN 202111438479 A CN202111438479 A CN 202111438479A CN 114317275 A CN114317275 A CN 114317275A
- Authority
- CN
- China
- Prior art keywords
- culture medium
- final concentration
- medium
- iron
- culture
- 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
Images
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses an iron-deficiency culture medium for improving hydrogen yield of microalgae, which is used for removing FeSO in TAP culture solution4·7H2O, and adding NaHSO3. The invention adopts the formula (adding NaHSO) of the existing hydrogen production culture medium3TAP culture solution) was optimized, and it was found that NaHSO was added3The hydrogen yield of Chlamydomonas reinhardtii 124 in the TAP culture solution is higher than that of the existing culture solution added with NaHSO3The system is improved by more than 2 times, one culture medium composition is reduced, and the cost is reduced.
Description
Technical Field
The invention relates to the technical field of biology, in particular to an iron-deficiency culture medium for improving hydrogen production of microalgae and a preparation method and application thereof.
Background
The traditional biomass energy source generates serious environmental pollution after being burnt, and the reserve amount is seriously insufficient. Therefore, with the increasing problems of global environmental pollution and energy crisis, people are urgently required to find a clean and renewable life-based energy source. Compared with the traditional biomass energy, the life-based energy is a non-carbon-based green and environment-friendly energy which is emphasized to be developed, and has the characteristics of emphasizing direct energy conversion and supply in the life process and the like. The photosynthetic hydrogen production of algae happens to accord with the energy characteristic. The algae can release oxygen and perform photosynthesis, and meanwhile, hydrogenase can capture electrons in water to generate hydrogen. However, hydrogenases are particularly sensitive to oxygen released by photosynthesis.
For twenty years, many methods have been developed to activate hydrogenase and induce continuous, efficient photosynthetic hydrogen production, which mainly include increasing the tolerance of hydrogenase to oxygen and reducing the amount of oxygen released during photosynthesis.
Chlamydomonas is a common algae species for people to study the hydrogen production of algae. Ten years ago, the university of Shanghai university, Ma As the professor topic group developed a method of adding NaHSO3Important method for removing oxygen, activating hydrogenase, inducing continuous and efficient photosynthetic hydrogen production (Ma W, Chen M, Wang L, et al3 greatly enhances photobiological H2production in the green alga Chlamydomonas reinhardtii. Bioresource Technol,2011,102: 8635-. The method indirectly eliminates oxygen released by photosynthesis (Wei L, Yi J, Wang L, et al. light interaction is important for hydrogen production in NaHSO) by reaction of bisulfite ions with superoxide anions on the photosystem I acceptor side3Plant Cell Physiol,2017,58(3): 451-. Despite the addition of NaHSO3Under the condition, more hydrogen can be obtained, but the requirement of practical application can not be met. In addition, the medicines required by the culture medium preparation account for a certain proportion of the chlamydomonas hydrogen production cost.
Patent CN102925490A discloses a culture medium for increasing hydrogen production of chlamydomonas and a preparation method thereof, wherein the culture medium is composed of the following 13 substances: NH (NH)4Cl、MgCl2·6H2O、K2HPO4、KH2PO4Tris base, H3BO4、ZnCl2、MnCl2·4H2O、CoCl2·6H2O、CuCl2·2H2O、(NH4)6Mo7O24·4H2O、FeCl2·4H2O and glacial acetic acid. According to the invention, after the existing hydrogen-releasing culture medium formula (the TAP culture solution lacking sulfur) is optimized, the hydrogen yield of Chlamydomonas CC400 in the TAP culture solution lacking sulfur and calcium is improved by about two thirds compared with that of the existing TAP culture solution lacking sulfur. However, the hydrogen yield of the chlamydomonas CC400 improved by the culture medium is not obvious, and the culture medium has the defects of long hydrogen induction time (usually after 24 hours) and cell death, and the like, and is difficult to meet the requirements of practical application. Compared with the culture medium, the culture medium for improving the hydrogen yield of the chlamydomonas completely avoids the defects and is expected to meet the requirements of practical application.
Disclosure of Invention
The inventors have surprisingly found that addition of NaHSO3The hydrogen yield of Chlamydomonas reinhardtii 124 in the TAP culture solution is higher than that of the existing culture solution added with NaHSO3The system is improved by more than 2 times, one culture medium composition is reduced, and the cost is reduced, thereby completing the invention.
The technical scheme of the invention is as follows:
the culture medium provided by the invention consists of solute and solvent, wherein the solute consists of the following 15 substances: NH (NH)4Cl、MgSO4·7H2O、CaCl2·2H2O、K2HPO4、KH2PO4Tris base, Na2EDTA·2H2O、ZnSO4·7H2O、H3BO3、MnCl2·4H2O、CoCl2·6H2O、CuSO4·5H2O、(NH4)6Mo7O24Glacial acetic acid and NaHSO3The components of the composition are as follows,
the NH4The final concentration of Cl in the medium was 0.375 g/L;
the MgSO4·7H2The final concentration of O in the medium was 0.0925 g/L;
the CaCl is2·2H2O in the culture mediumThe final concentration is 0.05 g/L;
said K2HPO4The final concentration in the medium was 0.288 g/L;
the KH2PO4The final concentration in the medium was 0.144 g/L;
the final concentration of the Tris base in the culture medium is 2.42 g/L;
the Na is2EDTA·2H2The final concentration of O in the culture medium is 0.05 g/L;
the ZnSO4·7H2The final concentration of O in the culture medium is 0.022 g/L;
said H3BO3The final concentration in the culture medium is 0.0114 g/L;
the MnCl2·4H2The final concentration of O in the medium was 0.00506 g/L;
the CoCl2·6H2The final concentration of O in the medium was 0.00161 g/L;
the CuSO4·5H2The final concentration of O in the medium was 0.00157 g/L;
said (NH)4)6Mo7O24The final concentration in the medium was 0.0011 g/L;
the final concentration of the glacial acetic acid in the culture medium is 1 ml/L;
the NaHSO3The final concentration in the medium was 1.352 g/L;
the solvent is water.
The pH of the medium was 7.3.
The microalgae is Chlamydomonas reinhardtii (Chlamydomonas reinhardtii).
The invention provides a preparation method of a culture medium, which comprises the following steps: dissolving 15 substances in the solutes in water to the final concentration to obtain the culture medium.
The culture medium can promote microalgae to produce hydrogen; the microalgae is Chlamydomonas reinhardtii (Chlamydomonas reinhardtii).
The invention provides a method for preparing hydrogen by utilizing microalgae, which comprises the following steps: culturing Chlamydomonas reinhardtii (Chlamydomonas reinhardtii) in the culture medium, and collecting the culture product to obtain hydrogen;
the culture temperature is 25 ℃ and the culture time is 3 days (about 72 hours); the illumination intensity of the culture is 200 mu mol photons m-2s-1。
The Chlamydomonas reinhardtii (Chlamydomonas reinhardtii) mentioned above are Chlamydomonas reinhardtii (Chlamydomonas reinhardtii) 124.
Compared with the prior art, the invention has the following beneficial effects:
the invention adopts the formula (adding NaHSO) of the existing hydrogen production culture medium3TAP culture solution) was optimized, and it was found that NaHSO was added3The hydrogen yield of Chlamydomonas reinhardtii 124 in the TAP culture solution is higher than that of the existing culture solution added with NaHSO3The system is improved by more than 2 times, one culture medium composition is reduced, and the cost is reduced. At the same time, due to the addition of NaHSO3In the formula of the TAP (ferrum-deficient) culture solution, only FeSO exists in the iron element4·7H2In O, the compound can be directly removed to achieve the environment of iron deficiency, and the operation is convenient.
In a word, compared with the original chlamydomonas hydrogen production culture system, the improved system not only can save the drug dosage, but also can obviously improve the hydrogen release amount, which advances a step forward towards the goal of low-cost hydrogen production of algae.
Drawings
FIG. 1 is a graph showing the results of hydrogen production by Chlamydomonas reinhardtii 124.
Detailed Description
The invention is described in detail below with reference to the figures and specific embodiments.
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Example 1
Hydrogen production by Chlamydomonas in Normal TAP culture fluid
One, Normal TAP culturePreparation of the base: NH (NH)4Cl 0.375g/L；MgSO4·7H2O 0.0925g/L；CaCl2·2H2O 0.05g/L；K2HPO4 0.288g/L；KH2PO40.144 g/L; tris alkali 2.42 g/L; hunter's trace elements (1 ml/L), glacial acetic acid (1 ml/L), and water (the rest).
Hunter trace elements (Hunter's trace elements): na (Na)2EDTA·2H2O 50g/L；ZnSO4·7H2O 22.0g/L；H3BO3 11.4g/L；MnCl2·4H2O 5.06g/L；FeSO4·7H2O 4.99g/L；CoCl2·6H2O 1.61g/L；CuSO4·5H2O 1.57g/L；(NH4)6Mo7O24·4H2O1.10 g/L, and the balance of water.
The pH of the medium was 7.3.
Second, Chlamydomonas pre-culture
2.43X 107Inoculating Chlamydomonas cells into a reagent bottle (500ml) containing 300ml of normal TAP culture solution, and culturing in a constant-temperature light incubator (25 deg.C, 40 μmol photons m)-2s-1)。
Third, detecting hydrogen production of Chlamydomonas
To be OD750When reaching about 1.0, the mixture is subpackaged into 60ml hydrogen bottles (30ml overhead; 30ml algal cells, chlorophyll 300 mu g).
The hydrogen-releasing bottle was placed at 25 ℃ and 200. mu. mol phosns m-2s-1Continuously culturing for 4.5 days in an illumination incubator, and detecting the hydrogen amount at different time points.
100 μ l of gas was withdrawn from above in the closed vessel with a micro-syringe and a sample of the withdrawn gas was injected into a gas phase system equipped with a thermal conductivity detector (GC, 7890A, Agilent, USA) for hydrogen detection.
The experiment was repeated three times and the results averaged.
As a result, as shown in FIG. 1, no hydrogen production was detected within 4.5 days.
Example 2
In the presence of NaHSO3Normal hydrogen production of Chlamydomonas in TAP culture solution
Preparation of Normal TAP Medium
See example 1.
Second, Chlamydomonas pre-culture
See example 1.
Third, detecting hydrogen production of Chlamydomonas
Chlamydomonas cell OD to be grown in Normal TAP culture solution750When reaching about 1.0, the mixture is subpackaged into 60ml hydrogen bottles (30ml overhead; 30ml algal cells, chlorophyll 300 mu g).
The hydrogen-releasing bottle was placed at 25 ℃ and 200. mu. mol phosns m-2s-1After continuous light culture for 1.5 days in a light incubator, 1.352g/L NaHSO is added by a syringe3To each hydrogen-evolving bottle; and then respectively taking different time points to detect the hydrogen quantity.
100 μ l of gas was withdrawn from above in the closed vessel with a micro-syringe and a sample of the withdrawn gas was injected into a gas phase system equipped with a thermal conductivity detector (GC, 7890A, Agilent, USA) for hydrogen detection.
The experiment was repeated three times and the results averaged.
As a result, as shown in FIG. 1, the amount of hydrogen produced was 73.25. + -. 5.72ml/L in 3 days.
Example 3
In the presence of NaHSO3The hydrogen yield of Chlamydomonas in the TAP culture solution
Preparation of Normal TAP Medium
See example 1.
Second, Chlamydomonas pre-culture
See example 1.
Preparation of iron-deficiency TAP culture medium
Removing FeSO from normal TAP culture solution4·7H2O; the pH of the medium was 7.3.
Chlamydomonas cell OD to be grown in Normal TAP culture solution750When the temperature reached about 1.0 ℃, the mixture was centrifuged at 2500rpm for 5 minutes at 25 ℃.
Pouring off the supernatant, and rinsing the obtained cells with an iron-deficient TAP culture solution for three times; then, the mixture was dispensed into 60ml hydrogen bottles (30ml overhead; 30ml algal cells, chlorophyll 300. mu.g).
The hydrogen-releasing bottle was placed at 25 ℃ and 200. mu. mol phosns m-2s-1After continuous light culture for 1.5 days in a light incubator, 1.352g/L NaHSO is added by a syringe3To each hydrogen-evolving bottle; and then respectively taking different time points to detect the hydrogen quantity.
100 μ l of gas was withdrawn from above in the closed vessel with a micro-syringe and a sample of the withdrawn gas was injected into a gas phase system equipped with a thermal conductivity detector (GC, 7890A, Agilent, USA) for hydrogen detection.
The experiment was repeated three times and the results averaged.
As a result, as shown in FIG. 1, it was found that the amount of hydrogen produced was 169.58. + -. 11.57ml/L within 3 days, and that the addition of NaHSO was accompanied by a decrease in the amount of hydrogen produced3The hydrogen yield of Chlamydomonas reinhardtii 124 in the TAP culture solution is higher than that of the existing culture solution added with NaHSO3The system is improved by more than 2 times.
The embodiments described above are described to facilitate an understanding and use of the invention by those skilled in the art. It will be readily apparent to those skilled in the art that various modifications to these embodiments may be made, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above embodiments, and those skilled in the art should make improvements and modifications within the scope of the present invention based on the disclosure of the present invention.
Claims (9)
1. An iron-deficiency culture medium for improving hydrogen production of microalgae, which is characterized in that the iron-deficiency culture medium is a culture medium without FeSO in TAP culture solution4·7H2O, and adding NaHSO3。
2. The iron-deficiency culture medium for improving hydrogen production of microalgae according to claim 1, is composed of solute and solvent, wherein the solvent is water, and the solute is composed of 15 substances: NH (NH)4Cl、MgSO4·7H2O、CaCl2·2H2O、K2HPO4、KH2PO4Tris base, Na2EDTA·2H2O、ZnSO4·7H2O、H3BO3、MnCl2·4H2O、CoCl2·6H2O、CuSO4·5H2O、(NH4)6Mo7O24Glacial acetic acid and NaHSO3。
3. The iron-deficiency culture medium for increasing hydrogen production of microalgae according to claim 2,
the NH4The final concentration of Cl in the medium was 0.375 g/L;
the MgSO4·7H2The final concentration of O in the medium was 0.0925 g/L;
the CaCl is2·2H2The final concentration of O in the culture medium is 0.05 g/L;
said K2HPO4The final concentration in the medium was 0.288 g/L;
the KH2PO4The final concentration in the medium was 0.144 g/L;
the final concentration of the Tris base in the culture medium is 2.42 g/L;
the Na is2EDTA·2H2The final concentration of O in the culture medium is 0.05 g/L;
the ZnSO4·7H2The final concentration of O in the culture medium is 0.022 g/L;
said H3BO3The final concentration in the culture medium is 0.0114 g/L;
the MnCl2·4H2The final concentration of O in the medium was 0.00506 g/L;
the CoCl2·6H2The final concentration of O in the medium was 0.00161 g/L;
the CuSO4·5H2The final concentration of O in the medium was 0.00157 g/L;
said (NH)4)6Mo7O24The final concentration in the medium was 0.0011 g/L;
the final concentration of the glacial acetic acid in the culture medium is 1 ml/L;
the NaHSO3The final concentration in the medium was 1.352 g/L.
4. The iron-deficiency culture medium for increasing hydrogen production of microalgae according to claim 2, wherein the pH of the culture medium is 7.3.
5. The method for preparing the iron-deficiency culture medium for increasing the hydrogen production of the microalgae according to any one of claims 1 to 4, wherein each solute in the culture medium is dissolved in water to reach a final concentration to obtain the culture medium.
6. The use of the iron-deficient culture medium for increasing hydrogen production of microalgae according to any one of claims 1 to 4, wherein the culture medium is used for promoting hydrogen production of Chlamydomonas reinhardtii.
7. The use of the iron-deficiency culture medium for increasing hydrogen production of microalgae according to claim 6, wherein the Chlamydomonas reinhardtii is Chlamydomonas reinhardtii (Chlamydomonas reinhardtii) 124.
8. The application of the iron-deficiency culture medium for improving hydrogen production of microalgae according to claim 6, characterized in that chlamydomonas reinhardtii is cultured in the culture medium, and the culture product is collected to obtain hydrogen.
9. The use of the iron-deficient culture medium for increasing hydrogen production of microalgae according to claim 8, wherein the culture temperature is 25 ℃, the culture time is 3 days, and the illumination intensity of the culture is 200 μmol photons m-2s-1。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111438479.3A CN114317275A (en) | 2021-11-30 | 2021-11-30 | Iron-deficiency culture medium for improving hydrogen yield of microalgae and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111438479.3A CN114317275A (en) | 2021-11-30 | 2021-11-30 | Iron-deficiency culture medium for improving hydrogen yield of microalgae and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114317275A true CN114317275A (en) | 2022-04-12 |
Family
ID=81047669
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111438479.3A Pending CN114317275A (en) | 2021-11-30 | 2021-11-30 | Iron-deficiency culture medium for improving hydrogen yield of microalgae and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114317275A (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101775407A (en) * | 2010-02-26 | 2010-07-14 | 上海师范大学 | Method for improving chlamydomonas reinhardtii hydrogen production amount of leghemoglobin ferrous chelate enzyme gene |
| CN102181369A (en) * | 2011-02-24 | 2011-09-14 | 中国科学院植物研究所 | Genetic engineering chlamydomonas having increased hydrogen producing capacity and application thereof |
| CN102925490A (en) * | 2011-08-12 | 2013-02-13 | 中国科学院植物研究所 | Culture medium for improving hydrogen production amount of Chlamydomonas reinhardtii, and its preparation method |
| CN103789365A (en) * | 2014-01-20 | 2014-05-14 | 湖南大学 | Method for recycling microbial oil from recycled light oil |
| CN106520897A (en) * | 2017-01-10 | 2017-03-22 | 上海师范大学 | Method for improving hydrogen production of chlamydomonas |
| CN110144368A (en) * | 2019-06-20 | 2019-08-20 | 哈尔滨工业大学 | A method of it maintains persistently to produce hydrogen after chlorella cells are dead |
-
2021
- 2021-11-30 CN CN202111438479.3A patent/CN114317275A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101775407A (en) * | 2010-02-26 | 2010-07-14 | 上海师范大学 | Method for improving chlamydomonas reinhardtii hydrogen production amount of leghemoglobin ferrous chelate enzyme gene |
| CN102181369A (en) * | 2011-02-24 | 2011-09-14 | 中国科学院植物研究所 | Genetic engineering chlamydomonas having increased hydrogen producing capacity and application thereof |
| CN102925490A (en) * | 2011-08-12 | 2013-02-13 | 中国科学院植物研究所 | Culture medium for improving hydrogen production amount of Chlamydomonas reinhardtii, and its preparation method |
| CN103789365A (en) * | 2014-01-20 | 2014-05-14 | 湖南大学 | Method for recycling microbial oil from recycled light oil |
| CN106520897A (en) * | 2017-01-10 | 2017-03-22 | 上海师范大学 | Method for improving hydrogen production of chlamydomonas |
| CN110144368A (en) * | 2019-06-20 | 2019-08-20 | 哈尔滨工业大学 | A method of it maintains persistently to produce hydrogen after chlorella cells are dead |
Non-Patent Citations (2)
| Title |
|---|
| 张亚琴 等: "绿藻光合产氢的研究进展", 浙江大学学报（理学版）, vol. 50, no. 1, pages 69 - 82 * |
| 高敏强: "缺铁提高NaHSO3诱导的微藻光合产氢的功能机理研究", 中国优秀硕士学位论文全文数据库基础科学辑, no. 07, pages 30 - 31 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chang et al. | Cultivation of Spirulina platensis for biomass production and nutrient removal from synthetic human urine | |
| Lam et al. | Biohydrogen production from algae | |
| Liu et al. | Enhanced bio-hydrogen production by the combination of dark-and photo-fermentation in batch culture | |
| Yang et al. | Nutrients removal and lipids production by Chlorella pyrenoidosa cultivation using anaerobic digested starch wastewater and alcohol wastewater | |
| Silva et al. | Sulphate removal from industrial wastewater using a packed-bed anaerobic reactor | |
| Lo et al. | Sequential dark–photo fermentation and autotrophic microalgal growth for high-yield and CO2-free biohydrogen production | |
| Chen et al. | Phototrophic hydrogen production in photobioreactors coupled with solar-energy-excited optical fibers | |
| Lee et al. | Nitrogen removal from wastewaters by microalgae without consuming organic carbon sources | |
| Nayak et al. | Biohydrogen production from algal biomass (Anabaena sp. PCC 7120) cultivated in airlift photobioreactor | |
| Hwang et al. | Photoheterotrophic microalgal hydrogen production using acetate-and butyrate-rich wastewater effluent | |
| WO2001002595A1 (en) | Microbial process for producing hydrogen | |
| Ouyang et al. | Effect of light intensity on the capability of different microalgae species for simultaneous biogas upgrading and biogas slurry nutrient reduction | |
| Xie et al. | The kinetic characterization of photofermentative bacterium Rhodopseudomonas faecalis RLD-53 and its application for enhancing continuous hydrogen production | |
| CN107663529B (en) | Method for improving hydrogen production of photosynthetic microalgae through algal bacteria co-culture | |
| Palamae et al. | Production of renewable biohydrogen by Rhodobacter sphaeroides S10: a comparison of photobioreactors | |
| Yang et al. | Enhanced hydrogen production performance of Rubrivivax gelatinosus M002 using mixed carbon sources | |
| Xie et al. | Feasibility studies on continuous hydrogen production using photo-fermentative sequencing batch reactor | |
| Cardeña et al. | Optimization of volatile fatty acids concentration for photofermentative hydrogen production by a consortium | |
| Wimonsong et al. | Comparison of different catalyst for fermentative hydrogen production | |
| He et al. | Production of coenzyme Q10 by purple non-sulfur bacteria: current development and future prospect | |
| Kaushik et al. | Integrating photobiological hydrogen production with dye–metal bioremoval from simulated textile wastewater | |
| Xiaolong et al. | Effect of sodium ion concentration on hydrogen production from sucrose by anaerobic hydrogen-producing granular sludge | |
| Chen et al. | Characterization of cell growth and photobiological H2 production of Chlamydomonas reinhardtii in ASSF industry wastewater | |
| Whitton et al. | Influence of light regime on the performance of an immobilised microalgae reactor for wastewater nutrient removal | |
| Mona et al. | Prolonged hydrogen production by Nostoc in photobioreactor and multi-stage use of the biological waste for column biosorption of some dyes and metals |
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 |