CN109628502B - Method for preparing bioethanol by using bloom-forming cyanobacteria as raw material - Google Patents
Method for preparing bioethanol by using bloom-forming cyanobacteria as raw material Download PDFInfo
- Publication number
- CN109628502B CN109628502B CN201811617124.9A CN201811617124A CN109628502B CN 109628502 B CN109628502 B CN 109628502B CN 201811617124 A CN201811617124 A CN 201811617124A CN 109628502 B CN109628502 B CN 109628502B
- Authority
- CN
- China
- Prior art keywords
- bloom
- algae
- cyanobacteria
- bioethanol
- forming cyanobacteria
- 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.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/06—Ethanol, i.e. non-beverage
- C12P7/08—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method for preparing bioethanol by using bloom-forming cyanobacteria as a raw material, which comprises the following steps: 1) Salvaging and collecting the water bloom blue algae; 2) Inoculating the collected bloom-forming cyanobacteria into a photobioreactor, regulating the nitrogen-phosphorus ratio, adding a photosynthesis promoter and a metabolic interference agent, introducing air to supplement an inorganic carbon source, and performing re-culture; 3) After the re-culture is finished, filtering and concentrating by using a tubular ultrafiltration membrane to form high-concentration algae liquid, and drying to obtain algae powder; 4) Hydrolyzing algae powder under an acidic condition to obtain hydrolysate containing reducing sugar; 5) Adjusting the hydrolysate to weak acidity, and adding yeast for fermentation to prepare bioethanol. The invention adopts a re-culture method, so that the sugar content in the bloom-forming cyanobacteria is obvious, the bioavailability is increased, and the yield of the bioethanol is greatly improved. The method is simple and easy to implement, has low cost, not only solves the problem that the algae needs to be cultured in a large scale in the existing process of preparing the bioethanol, but also provides an effective resource approach for the bloom-forming cyanobacteria.
Description
Technical Field
The invention belongs to the fields of biological energy technology and environmental protection, and particularly relates to a method for preparing bioethanol by utilizing water-blooming cyanobacteria after re-culture.
Background
In recent decades, the problem of lake eutrophication has become serious increasingly, which causes the blue algae to breed in large quantities and the occurrence of disastrous water blooms frequently, thus causing serious threats to the functions and the ecological systems of lakes. Salvage is an emergency removal technology for eliminating the threat of blue algae bloom in a short time, is also a main method for treating the blue algae bloom in China at present, and is used for eutrophication lakes in China, such as Taihu lake, dian lake and nested lakeEtc., are used in a normalized manner. Taking the Taihu lake as an example, 8.5 multiplied by 10 algae water is salvaged in cities around the Taihu lake in 2007 to 2015 6 m 3 (algae content of 0.5%) corresponding to the removal of blue algae dry matter of 4.25 × 10 4 t. After the algae water salvaged ashore is subjected to two procedures of settlement and air flotation, the volume is greatly reduced, and the algae water still needs to be subjected to timely and effective harmless treatment to prevent serious secondary environmental pollution caused by algae corruption.
The most effective processing means for salvaging the blue-green algae is resource or energy utilization, so that the waste is changed into valuable, the social, resource and economic values are exerted, and the continuous operation of the work of salvaging the blue-green algae is facilitated. At present, the comprehensive utilization of blue algae resource mainly comprises the following modes: producing organic fertilizer as farmland organic fertilizer; blue algae additive is developed and added into various feeds and baits; economic components such as phycocyanin, pigment, exopolysaccharide and the like are extracted, but the resource method is difficult to popularize in practical application due to the existence of algal toxin. In the aspect of algae energy regeneration, research on preparation of biofuels such as diesel oil, ethanol, hydrogen and the like by using microalgae is widely carried out at home and abroad at present. However, most of the researches adopt artificially cultured microalgae, the culture process includes a biomass increasing stage and a lipid/carbohydrate accumulation stage, during which a large amount of manpower, material resources and energy are consumed, which results in an excessive cost, and the variety of microalgae which can be used for producing biofuel is mostly concentrated in green algae, golden algae, etc., while the content of cyanobacteria lipid (< 10%) and carbohydrate (10% -20%) is low, which is generally considered to be unsuitable as a raw material for producing biofuel. CN102242065A discloses a blue algae engineering bacterium with high cellulose yield and a preparation method and application thereof, which constructs blue algae with cesA gene deletion, wherein the cellulose content of the blue algae is increased to 13 percent from 2 percent of dry weight of cell walls, but the cellulose only accounts for a small proportion of total sugar of the blue algae. The application of transgenic algae still faces problems and challenges, such as few varieties of microalgae capable of performing specific functional gene modification, lack of application examples in large-scale culture of genetically engineered algae, lack of evaluation of biological safety, incomplete supervision system and the like.
In China, the water bloom generated in eutrophic lakes and reservoirs is mainly blue algae water bloom. The biomass is very huge when the blue algae bloom is outbreak, and the biomass increasing stage in the microalgae culture process can be saved by directly utilizing the salvaged water-blooming blue algae to produce the biomass ethanol, so that the culture cost is greatly reduced. However, the related technology in this field is still in the blank, because the lipid and carbohydrate content of wild blue algae is lower than that of artificially cultured blue algae, for example, the lipid content of blue algae salvaged from Taihu lake is lower than 1%, and the carbohydrate content is lower than 10%, so that it is not feasible to directly use it as raw material for producing biofuel.
Disclosure of Invention
The invention aims to provide a method for preparing bioethanol by adopting re-cultured bloom-forming cyanobacteria and collecting high-concentration algae liquid by using a tubular ultrafiltration membrane for hydrolysis and fermentation.
The purpose of the invention can be realized by the following technical scheme:
a method for preparing bioethanol by using bloom-forming cyanobacteria as a raw material comprises the following steps:
step 1: salvaging and collecting the bloom-forming cyanobacteria, and removing macroscopic impurities;
and 2, step: collecting the bloom-forming cyanobacteria in the step 1, inoculating the bloom-forming cyanobacteria into a photobioreactor, adjusting lake water and supplementing inorganic nitrogen and inorganic phosphorus, adjusting the nitrogen-phosphorus ratio to be 20-1; introducing air to supplement an inorganic carbon source, and culturing again;
and step 3: collecting the bloom-forming cyanobacteria cultured in the step 2, filtering and concentrating by using a tubular ultrafiltration membrane to form a concentrated algae solution with the water content of 90-98%, and drying at low temperature for later use;
and 4, step 4: adding 1-3mol/L hydrochloric acid into the algae powder in the step 3, wherein the concentration of the algae powder is 20-200g/L, and hydrolyzing at 110-122 ℃ for 20-30 min to obtain hydrolysate containing reducing sugar;
and 5: and (4) adjusting the pH of the hydrolysate in the step (4) to 6-7, adding 120-150 ml of saccharomyces cerevisiae according to the inoculation amount of every 1g of algae powder, uniformly mixing, and fermenting at 35-38 ℃ for 48-72h at the speed of 100-200 r/min to obtain the bioethanol.
The inorganic nitrogen is NaNO 3 、KNO 3 Or NH 4 Cl; inorganic phosphorus is K 2 HPO 3 。
The introduced air is used for supplementing the inorganic carbon source to form a photoperiod, and an air pump is used for continuously supplying air to supplement CO 2 The flow rate is 2-20L/min.
The conditions for re-culturing are as follows: the illumination is 3500-8000 lx, the light dark period is 12h-20h, and the temperature is 28-35 ℃.
Step 3, collecting the bloom-forming cyanobacteria cultured in the step 2 until the sugar content accounts for at least 50% of the dry weight of the bloom-forming cyanobacteria, wherein the bloom-forming cyanobacteria can be collected; the low-temperature drying is carried out at the temperature of 20-50 ℃ for 24-48 h.
The photosynthesis promoter is choline chloride, ferric chloride or lanthanum chloride; the metabolism interference agent is salicylic acid, sodium chloride, ethylene or zinc chloride.
And NaOH solid is adopted for regulating the pH value of the hydrolysate.
In particular to saccharomyces cerevisiae (A), (B), (C)Saccharomyces cerevisiae) Purchased from China center for culture Collection of Industrial microorganisms, and has the number CICC33068. A ring of yeast is picked by an inoculating loop, inoculated into a 500ml conical flask filled with 250ml YPG liquid culture medium sterilized at the high temperature of 121 ℃ for 20 min, placed in a constant temperature shaking table, adjusted to the temperature of 30 ℃ and shake-cultured for 24-48 h under the condition of 150 r/min. Before inoculating to the fermentation medium, centrifuging the seed liquid with corresponding volume (45 ml) at 4000 r/min for 3min, pouring out the supernatant, adding pure water, shaking up, centrifuging, pouring out the supernatant, and repeating the above steps for 2 times. Inoculating the bacterial sludge into a fermentation bottle. Yeast seed medium YPG: tryptone 20g/L, glucose 20g/L and yeast powder 10g/L.
In particular, the fermentation is carried out in a closed vessel, but without the need for headspace and deoxygenation.
The beneficial effects of the invention are:
1) During the water bloom, mechanical fishing is an effective means for removing the blue-green algae in time, a large amount of the collected blue-green algae is urgently needed to be treated so as to avoid secondary pollution to the environment, and the resource utilization of the blue-green algae is the most economic way. The invention adopts the bloom-forming cyanobacteria as the raw material, thereby achieving two purposes, solving the problems of ecology and environment and solving the problem of energy.
2) The invention saves the microalgae amplification culture stage, only keeps the sugar accumulation stage, greatly shortens the production period of the algae-derived sugar, and effectively reduces the culture cost.
3) In the re-culture stage, photosynthesis and sugar metabolism are regulated by adding promoter and metabolic interference agent, so as to raise sugar content and raise the yield of final product bioethanol.
Drawings
FIG. 1 is a flow chart of the present invention.
Detailed Description
The process of the present invention is further illustrated below with reference to several specific examples.
Example 1
The blue algae in the lake Taiwan region during water bloom outbreak is salvaged and collected, the absolute dominant species is microcystis, and macroscopic impurities are removed. Inoculating the collected blue algae to a photobioreactor, adding Taihu lake water to make chlorophyll concentration be 2-4 mg/L, adding NaNO 3 And K 2 HPO 3 The cells were cultured at 6000 lx at 28 ℃. Periodically measuring the dry weight and the yield of soluble total sugar, and calculating the sugar content; the Chemical Oxygen Demand (COD) and Biochemical Oxygen Demand (BOD) of the algae cell contents are measured, and the BOD/COD is calculated for representing the biodegradability of the algae cell contents.
Under the condition that the nitrogen-phosphorus ratio is 10, 0.01-0.05 per thousand of photosynthesis promoter and 1-3 per thousand of metabolism interference agent are added, the dry weight is increased from 0.527 g/L to 1.758 g/L, and the total soluble sugar is increased from 33.7 mg/L to 995.6 mg/L. Compared with the uncultured water-blooming cyanobacteria, the soluble total sugar content of the uncultured water-blooming cyanobacteria is increased from 6.4% to 56.6%, and the BOD/COD is increased from 0.39 to 0.86, so that the sugar content and the bioavailability of the water-blooming cyanobacteria are remarkably improved.
Example 2
1.2 g of the re-cultured algae powder was directly dissolved in 40 ml of 2mol/L hydrochloric acid solution to prepare 30 g/L algae solution. Hydrolyzing at 121 deg.C for 30 min, and filtering the hydrolysate with 0.45 μm filter membrane. The change in soluble total sugars and reducing sugars before and after hydrolysis was determined. The soluble total sugar and reducing sugar before the hydrolysis of the re-cultured algae powder are respectively 17.0 g/L and 1.9 g/L, and respectively account for 56.6 percent and 6.4 percent of the dry weight; after hydrolysis, the concentration was 17.9 g/L and 17.3 g/L, respectively, and accounted for 59.8% and 57.5% of the dry weight, respectively. After hydrolysis, the ratio of reducing sugar to total sugar increased from 10.9% to 96.2%.
Meanwhile, the algal powder which is not re-cultured is treated under the same condition, and the soluble total sugar and reducing sugar respectively account for 1.9 g/L and 1.2 g/L before hydrolysis, respectively account for 6.2 percent and 4 percent of the dry weight, respectively account for 2.1 g/L and 2.0 g/L after hydrolysis, respectively account for 6.9 percent and 6.7 percent of the dry weight, and are far lower than the re-cultured algal powder.
Example 3
The hydrolysate NaOH solids were adjusted to pH =6.5. When the OD value of the cultured yeast is about 2, centrifuging at 4000 r/min for 3min, pouring out the supernatant, adding pure water, shaking up, centrifuging, pouring out the supernatant, and repeating for 2 times. The inoculation amount of 150 ml of Saccharomyces cerevisiae (OD value is about 2) is added into every 1g of algae powder, namely, the inoculation amount of 50ml of 30 g/L algae liquid is 225 ml of yeast. Adding the hydrolysate into a 100 ml closed sample bottle, and fermenting at 35 deg.C for 48 h at a speed of 150 r/min. Filtering the fermentation liquid with 0.22 μm organic filter membrane, sealing 5ml filtrate in 20ml headspace sample bottle, and measuring ethanol content by sampling headspace gas chromatography.
30 The concentration of bioethanol produced by the algae powder which is subjected to the re-culture in g/L is 6.52 g/L, and the ethanol yield is 21.7%, while the concentration of bioethanol produced by the algae powder which is not subjected to the re-culture in 30 g/L is only 0.85 g/L, and the ethanol yield is 2.8%. The ethanol yield of the re-cultured algae powder is obviously improved.
Example 4
Different areas (poplar, sand tea, etc.) of Taihu lake water bloom blue algae are collected, 4 sampling points are counted, and the re-culture, hydrolysis and fermentation are carried out according to the methods of the embodiments 1, 2 and 3. The soluble total sugar of the re-cultured algae powder accounts for 48.5%, 58.9%, 54.2% and 55.5% of the dry weight respectively; after hydrolysis, soluble total sugars were 50.8%, 61.0%, 56.5% and 59.8% and reducing monosaccharides were 98.7%, 99.2%, 96.2% and 96.0% of the soluble total sugars, respectively, on a dry weight basis. 30 The concentration of ethanol after the fermentation of the g/L algae powder is respectively 6.01 g/L, 6.73 g/L, 5.79 g/L and 7.07 g/L, and the yield of the ethanol is respectively 20%, 22.3%, 19.3% and 23.5%.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention, and the present invention should not be limited by the disclosure of the preferred embodiments. Therefore, it is intended that all equivalents and modifications which are within the spirit of the present disclosure be protected.
Claims (1)
1. A method for preparing bioethanol by using bloom-forming cyanobacteria as a raw material is characterized by comprising the following steps:
step 1: salvaging and collecting the bloom-forming cyanobacteria, and removing macroscopic impurities;
and 2, step: collecting the bloom-forming cyanobacteria in the step 1, inoculating the bloom-forming cyanobacteria into a photobioreactor, adjusting lake water to ensure that the chlorophyll concentration is 2-4 mg/L, supplementing inorganic nitrogen and inorganic phosphorus, adjusting the nitrogen-phosphorus ratio to be 20-1, and adding 0.01-0.05 thousandth of a photosynthesis promoter and 1-3 thousandth of a metabolic interference agent; introducing air to supplement an inorganic carbon source, and re-culturing;
and 3, step 3: collecting the bloom-forming cyanobacteria cultured in the step 2, filtering and concentrating by using a tubular ultrafiltration membrane to form a concentrated algae solution with the water content of 90-98%, and drying at low temperature for later use;
and 4, step 4: adding 1-3mol/L hydrochloric acid into the algae powder in the step 3, wherein the concentration of the algae powder is 20-200g/L, and hydrolyzing at 110-122 ℃ for 20-30 min to obtain hydrolysate containing reducing sugar;
and 5: adjusting the pH of the hydrolysate obtained in the step 4 to be 6-7, adding 120-150 ml of saccharomyces cerevisiae into every 1g of algae powder, uniformly mixing, and fermenting at the temperature of 35-38 ℃ for 48-72h at the speed of 100-200 r/min to obtain bioethanol; wherein:
the inorganic nitrogen is NaNO 3 、KNO 3 Or NH 4 Cl; inorganic phosphorus is K 2 HPO 3 ；
The introduced air is used for supplementing the inorganic carbon source to form a photoperiod, and an air pump is used for continuously supplying air to supplement CO 2 The flow rate is 2-20L/min;
the conditions for re-culturing are as follows: the illumination is 3500-8000 lx, the light dark period is 12h-20h, and the temperature is 28-35 ℃;
step 3, collecting the water-blooming cyanobacteria cultured in the step 2 until the sugar content accounts for at least 50% of the dry weight of the water-blooming cyanobacteria, wherein the water-blooming cyanobacteria can be collected; the low-temperature drying is carried out at the temperature of 20-50 ℃ for 24-48 h;
the photosynthesis promoter is choline chloride, ferric chloride or lanthanum chloride; the metabolism interference agent is salicylic acid, sodium chloride, ethylene or zinc chloride.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811617124.9A CN109628502B (en) | 2018-12-28 | 2018-12-28 | Method for preparing bioethanol by using bloom-forming cyanobacteria as raw material |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811617124.9A CN109628502B (en) | 2018-12-28 | 2018-12-28 | Method for preparing bioethanol by using bloom-forming cyanobacteria as raw material |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109628502A CN109628502A (en) | 2019-04-16 |
| CN109628502B true CN109628502B (en) | 2022-10-04 |
Family
ID=66078607
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811617124.9A Active CN109628502B (en) | 2018-12-28 | 2018-12-28 | Method for preparing bioethanol by using bloom-forming cyanobacteria as raw material |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109628502B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010095895A2 (en) * | 2009-02-23 | 2010-08-26 | University - Industry Cooperation Foundation Kangwon National University | Method for producing ethanol using freshwater blue-green algae |
| CN104073524A (en) * | 2014-06-19 | 2014-10-01 | 中国科学院广州能源研究所 | Method for preparing fuel ethanol by saccharifying and fermenting carbon-rich microalgae solid acid |
| CN105600942A (en) * | 2015-09-24 | 2016-05-25 | 中国水产科学研究院渔业机械仪器研究所 | Method for formation of bioflocs by cyanobacterial bloom |
-
2018
- 2018-12-28 CN CN201811617124.9A patent/CN109628502B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010095895A2 (en) * | 2009-02-23 | 2010-08-26 | University - Industry Cooperation Foundation Kangwon National University | Method for producing ethanol using freshwater blue-green algae |
| CN104073524A (en) * | 2014-06-19 | 2014-10-01 | 中国科学院广州能源研究所 | Method for preparing fuel ethanol by saccharifying and fermenting carbon-rich microalgae solid acid |
| CN105600942A (en) * | 2015-09-24 | 2016-05-25 | 中国水产科学研究院渔业机械仪器研究所 | Method for formation of bioflocs by cyanobacterial bloom |
Non-Patent Citations (3)
| Title |
|---|
| 氯化胆碱在钝顶螺旋藻 （ Sp irulina p latensis）养殖中的应用初探;王大志等;《厦门大学学报（自然科学版）》;19990531;第38卷(第3期);第480-484 * |
| 水杨酸对蛋白核小球藻(Chlorella pyrenoidosa)生长及抗逆相关基因的影响;丁聪聪等;《海洋与湖沼》;20151130;第46卷(第6期);第1451-1460 * |
| 盐胁迫对地木耳和葛仙米生理生化特性的影响;李运广,高坤山;《水生生物学报》;20030531;第27卷(第3期);第227-231 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109628502A (en) | 2019-04-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102676589B (en) | Method for producing, separating and purifying butanol by coupling fermenting with gas stripping | |
| CN102863115A (en) | Method for treating fermentation industry waste water and producing algae powder by using microalgae | |
| CN109576314B (en) | Method for preparing microalgae grease through mixed culture | |
| CN102864188A (en) | Method for producing biodiesel from lignocellulose | |
| Zhang et al. | Ammonia-nitrogen and orthophosphate removal by immobilized Chlorella sp. isolated from municipal wastewater for potential use in tertiary treatment | |
| CN102746991B (en) | Method for carrying out heterotrophic culture on chlorella with acid-producing wastewater | |
| CN110591918A (en) | Method for producing algae-derived sugar by purifying secondary tail water or sludge supernatant in coupling mode | |
| CN103421850A (en) | Method used for producing bioethanol with Scenedesmusabundans | |
| CN109880856B (en) | Open type microalgae grease production method | |
| CN109628315A (en) | A method of culture chlorella simultaneously promotes its biomass and lipid-producing | |
| Hariz et al. | Growth and biomass production of native microalgae Chlorella sp., chlamydomonas sp. and Scenedesmus sp. cultivated in Palm Oil Mill Effluent (POME) at different Cultivation conditions | |
| CN104232559B (en) | The method of cultivating microalgae and the method for producing grease | |
| CN109628502B (en) | Method for preparing bioethanol by using bloom-forming cyanobacteria as raw material | |
| Varela-Bojórquez et al. | Production of bioethanol from biomass of microalgae Dunaliella tertiolecta | |
| CN105713951B (en) | Method for preparing microalgae grease | |
| CN116200286B (en) | Clostridium thermocellum capable of efficiently saccharifying cellulose and application thereof | |
| CN105400697A (en) | Method for purifying undiluted anaerobic fermentation tail liquid by growing microalgae in carbon dioxide environment | |
| CN105836890A (en) | Application of self-flocculating Chlorella Minutissima HB-1 in purification of piggery sewage anaerobic effluent | |
| CN106701587A (en) | Method used for recycling microalgae residue and producing spirulina rich in polysaccharides | |
| CN102311923B (en) | Microalgae cultivation method | |
| CN112028251A (en) | Method for improving ammonia nitrogen removal effect of microalgae on livestock and poultry breeding wastewater | |
| CN201381258Y (en) | High-concentration organic wastewater treatment system | |
| CN104261630A (en) | Comprehensive utilization method of wastewater generated through ramie biological degumming and boiling (high temperature sterilization) | |
| CN110894512A (en) | Method for directly producing ethanol by using microalgae | |
| CN103773694B (en) | A kind of method of high-efficient culture oil-containing microorganism |
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 |