CN1076488A - The preparation method of polyvinyl alcohol microbe or enzyme immobilization carrier and application thereof - Google Patents
The preparation method of polyvinyl alcohol microbe or enzyme immobilization carrier and application thereof Download PDFInfo
- Publication number
- CN1076488A CN1076488A CN92101875A CN92101875A CN1076488A CN 1076488 A CN1076488 A CN 1076488A CN 92101875 A CN92101875 A CN 92101875A CN 92101875 A CN92101875 A CN 92101875A CN 1076488 A CN1076488 A CN 1076488A
- Authority
- CN
- China
- Prior art keywords
- microorganism
- weight
- polyvinyl alcohol
- aqueous solution
- enzyme
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/10—Biological treatment of water, waste water, or sewage
Landscapes
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Abstract
(basicity is more than 70% with the PVA of microorganism or enzyme and 1-20 weight %, the polymerization degree 1000-3000) after solution mixes, place saturated boric acid solution, make it form spheroid in short period of time, contact with phosphate solution (＞5 weight %) with being about to this spheroid, it is fully hardened, and make microorganism or enzyme immobilization carrier.And the fixation support that will make, be used in and remove inorganic nitrogen and organic carbon in the waste water, and in the production sequence of biochemical products.
Description
Use immobilized microorganism or zymotechnic that all natural or synthesized polymer materials coat the microorganism intact cell effectively, since the eighties, enjoyed and gazed at.And existing much be applied to industrial success example, for example high fructose syrups, 6-APA, the production of biochemical products such as L-amino acid.Usually the representative macromolecular material that is used in fixation support is a polyacrylamide, deer horn phycocolloid (K-carrageenan), Sodium Alginate and agar etc.Polyacrylamide is inexpensive than other polymers, often is used.But, difficultly make the spheroidal particle that helps general flow reactor, and monomer whose molecule and polymerization promotor etc. all have toxicity, be unfavorable for the immobilization operation of micro-organisms living cell.Though the deer horn phycocolloid is shaped easily, toxicity is low, main drawback is that price is higher.Though Sodium Alginate is also cheap, make sphere easily, still, containing phosphoric acid salt, in the cationic reaction solutions such as sodium and potassium, colloid intensity rather unstable, even disintegration.In addition, agar colloidal physical strength is not suitable for prolonged operation a little less than disliking too again.Therefore, if desire effectively is applied to the industrialized process of biochemical commodity with the immobilized bacterium body technique, enjoy the field of waste water treatment of gazing at especially recently, the key of its success or not, be to be microorganism is not had toxicity, with low cost and have an exploitation of the solid support material of tough physical strength.
Polyvinyl alcohol (PVA), be by Vinyl Acetate Monomer through polymerization, the water soluble resin that forms of alcoholization.PVA is not had toxicity even to also general recognized as safe of human body, and system is described easily, the physical strength height, and be the industrialization macromolecule raw material of producing at a low price.Therefore, be suitable as very much the carrier of immobilized thallus.
In recent years, the patent gazette of American-European document and Japan has been announced all about utilizing PVA to carry out microbial immobilized technology.For example the PVA aqueous solution is freezed vacuum-drying or carries out gel sets (the Japanese Patent spy opens clear 57-14129, and the spy opens clear 61-139385) with the freezing method of rising again with imposing after microorganism mixes.Impose uviolizing, form the gel method (Japanese patent laid-open 1-454372) of photo-crosslinking structure.Also have in addition the PVA aqueous solution and microbial cells mixed solution, place saturated boric acid aqueous solution contact to form the gel technique (the Japanese Patent spy opens clear 61-100193) of bridge formation structure.Though aforesaid method can obtain the carrier of the colloid of high strength as immobilized thallus, still have many shortcomings to remain to be improved.In freeze-drying method, must remain under-30～-80 ℃ the low temperature material is freezing, then must drying and dehydrating to certain water ratio, the step of this kind freeze-thaw-dehydration, not only time-consuming very of a specified duration, and formality is numerous and diverse, expends the energy.The method that on the whole the photo-crosslinking rule is utilized when making film is unfavorable for the use of general thalline immobilization program.Moreover in the PVA-borate method, the mixture of microorganism-PVA then must contact dipping 12-24 hour with boric acid, can obtain the colloid of suitable intensity, otherwise the colloid fragility.
Generally speaking, the common drawback of above-mentioned prior art has, and the first, the immobilization program is time-consuming permanent, and formality is numerous and diverse, needs large-scale production unit to produce, and causes the lifting of production cost, reduces productivity.The second, the neither environment that very is fit to microorganism existence of low temperature, vacuum and boric acid.Especially boric acid has more toxicity, and production process all may cause the reduction of microorganism active for a long time.The clear proposition once in Beiye utilizes PVA and microbial cells mixture, the method that places sulfate liquor to contact, and (the Japanese Patent spy opens clear 64-5490 to the above-mentioned shortcoming of attempt improvement, 64-5491).Though this method has shortened immobilization program required time effectively, yet employed gelating soln concentration is quite high, must carry out with 30% sodium sulfate or 70% ammoniumsulphate soln, and shaping particles is difficult for when hanging down if this concentration is crossed, and intensity is also not good.Therefore, the material requested cost is also higher; Form that high salt density also very has detrimentally affect to microbial biochemical activity in the glue process.
Boric acid proposed by the invention-phosphoric acid salt two-stage becomes the method for glue, at first places for to be shaped the 3% boric acid solution short period of time to saturation concentration, strengthens in the 3-20% phosphate solution again.It is quite easy to be shaped, and phosphoric acid salt is cheap, also is microbial energy metabolism source, does not have a bio-toxicity.In addition, the salt working concentration is low, reduces cost.And has better colloid toughness than sulphate process.
It is simple that the present invention's purpose is to propose production process, low-cost, and can obtain enhanced water resistance, tough physical strength at short notice, and the gel immobilized microorganism of PVA or the enzyme of the excellent biochemical activity of tool, and it is widely used on field of waste water treatment and the biochemical industry.
The invention provides the improvement preparation method of a kind of polyvinyl alcohol microbe or enzyme immobilization carrier, comprise that to be 3 weight % carry out to the saturated boric acid aqueous solution that gelation handles produces a gel spheres in a concentration with polyvinyl alcohol and microorganism or mixture that enzyme formed, its improvement part is that this gelation treatment time is 10 minutes to 2 hours, and this gel spheres then makes its sclerosis more than 30 minutes and obtains polyvinyl alcohol microbe or enzyme immobilization carrier with 3-20 weight % phosphoric acid or aqueous phosphatic dipping.
The principle of the inventive method utilizes that hydroxyl and the intermolecular ion bridging action of boric acid make it shaping in the PVA molecule with being characterised in that; Then, again that this is very not strong gel impregnated in phosphoric acid or the phosphate solution, utilizes the esterification between PVA and phosphoric acid or microcosmic salt to make the colloid sclerosis, the acquisition water tolerance is strong, and physical strength is high and damage the immobilized thallus of microorganism or enzyme biochemical activity hardly.The making flow process of microorganism of the present invention or enzyme immobilization carrier can be represented with accompanying drawing 1.
PVA used in the present invention, basicity are more than 70%, and the polymerization degree is 1000-3000, and it is comparatively suitable to reach polymerization degree 1500-2000 person more than 95% with basicity.The polymerization degree is crossed low then colloid instability, and too high then viscosity raises and is difficult for handling.This PVA system is mixed with this microorganism or enzyme with an aqueous solution form, and suitable PVA concentration is then at 10-20 weight %.The boric acid curring time was preferable with the about 15-30 of saturated boric acid aqueous solution minute.
The preferred concentration of phosphoric acid or aqueous phosphatic is 5-15 weight %, and dipping time needs 1-2 hour approximately, can obtain desirable finished product.This aqueous phosphatic can use sodium phosphate, sodium hydrogen phosphate, SODIUM PHOSPHATE, MONOBASIC, potassiumphosphate, potassium hydrogen phosphate, potassium primary phosphate, ammonium phosphate, ammonium hydrogen phosphate or primary ammonium phosphate.
Another better embodiment of the inventive method is that this boric acid is mixed with phosphoric acid or aqueous phosphatic, and carries out this gelation and sclerosis simultaneously, and the time is about 30 minutes to 3 hours, is preferable with 1-2 hour.
The invention is characterized at first and set up the colloid base frame in boric acid solution, because duration of contact is not long, make boric acid to microorganism or the possible injury of enzyme minimize (borate method that this prize of bridge is carried, must the contact 15-24 hour, the Japanese Patent spy opens clear 61-100193).Moreover the colloid process of setting is used the phosphate solution with shock absorption, and phosphoric also be the essential element of microbial energy metabolism, so not only harmless and may have the function that activates microorganism for microorganism.
The PVA colloid of gained produced according to the present invention is fit to be used in the being coated and fixed technology of enzyme, industrial microorganism, wastewater treatment microbial flora and animal and plant cells etc.With regard to enzyme, such as amylase, cellulase, proteolytic enzyme etc.With regard to microorganism, microorganism species such as alcohol fermentation yeast, nitrobacteria, denitrifying bacteria and active sludge, anaerobic digestion mud, methanation mud, denitration pollution.The above is all the feasible applications object of PVA gel method of the present invention.
Embodiment 1
The PVA(basicity of 15 weight % is more than 99%, the polymerization degree 2000) aqueous solution 20g, fully mix (the denitration sludge microbe is taken from the denitration groove in the biological denitrificaion program of laboratory) with concentrated solution (sludge concentration 50g/l) 20g of denitration mud.This PVA-mud mixture is splashed in the saturated boric acid aqueous solution of slow stirring, form the ball colloid of the about 3mm of diameter, and place this solution after 20 minutes, and take out in the sodium dihydrogen phosphate that is transferred to 8 weight % again and flooded 40 minutes, at last colloidal solid is taken out washing.The fixed microorganism carrier 20g that makes and contains the 80ml artificial wastewater that saltpetre and methyl alcohol are main composition (the concentration of nitrate nitrogen is 100ppm, methyl alcohol 350ppm) and mixes, and places the serum bottle of 125ml, criticizes the formula denitration under oxygen free condition.The concentration of nitrate nitrogen of artificial wastewater is reduced to 32ppm after cultivating in 2 hours.
Identical fixed microorganism carrier heavily covers batch formula denitrification test, upgrade the artificial wastewater of identical composition every day, and record the denitration speed of immobilized microorganism, after the 7th day, reach 0.65mgN/g gel/h, after this, until the 30th day operate continuously all is maintained at this speed, biochemical activity quite stable.Moreover utilize this prize of bridge (the Japanese Patent spy opens clear 61-100193) institute extracting method to prepare the immobilized thallus of identical sludge content, under the same conditions, carry out the same experiment of embodiment therewith.Criticize the formula denitration after cultivating in 2 hours, the concentration of nitrate nitrogen of artificial wastewater is 82ppm, and heavily covers the denitration speed that batch test must reach 0.55mg N/g gel/h at 15 days rears, and ongoing operation backward is also very unstable.
Embodiment 2
The PVA(basicity of 20 weight % is more than 99%, the polymerization degree 2000) aqueous solution, with the concentrated solution (sludge concentration 30g/l) of nitrifying sludge, with 1: 1 part by weight uniform mixing (the nitrifying sludge microorganism is taken from the nitrifying groove in the biological denitrificaion program of laboratory).The PVA-mud mixture carries out and the same microbial immobilized program of embodiment (1).
It is the bio-reactor of 10l that the microbial fixed carrier of making (particle diameter is 3mm) places the operation volume, the artificial wastewater (influx 30l/ days) who becomes a mandarin continuously and contain the 200ppm ammonia nitrogen, and the carrier packing fraction is 25%, air flow 20l/ minute.After operate continuously in 10 days, the ammonia nitrogen concentration that goes out flowing water is 9ppm, and has 92% ammonia nitrogen to be converted into nitrate nitrogen.
Embodiment 3
With the cause waste water of raising pigs (COD concentration 1500-2000ppm, total nitrogen concentration 200-300ppm) domestication reaches 1 month active sludge, through centrifugal and thickened sludge solution, mix 18 weight %PVA(basicities more than 99% with 1: 1 part by weight, the polymerization degree 2000) the aqueous solution, this uniform mixing drop goes into to contain in the mixing solutions of 5 weight % boric acid and 10 weight % potassium primary phosphates, floods the ball colloidal solid that formed the 3mm particle diameter in 1 hour.The fixed microorganism carrier of making places and the same bio-reactor of embodiment (2), the processing of the cause of raising pigs waste water, and reactor operating condition is identical with embodiment (2).After operate continuously in 20 days, the COD concentration that goes out flowing water is reduced to 200-300ppm, and total nitrogen concentration is also reduced to 120-180ppm.
Embodiment 4
The PVA(basicity of 20 weight % is more than 99%, the polymerization degree 2000) aqueous solution 10g, 10g fully mixes with the centrifugal concentrated solution of cereuisiae fermentum (Saccharomyces cerevisia) (cell concn 30g/l), and PVA-yeast thalline mixture carries out and the same microbial immobilized program of embodiment (1).The fixed cell carrier of making (particle diameter is 2mm) 15g mixes with the substratum 150ml that contains 3 weight % glucose, places triangular flask, and in 30 ℃ of following shaking culture, after 8 hours, it is 10.2g/l that the alcohol of nutrient solution produces concentration.And, under same culture condition, carry out the cultivation of free cell with identical biomass, after 8 hours, it is 10.6g/l that the alcohol in the nutrient solution produces concentration.
Embodiment 5
The PVA(basicity of 20 weight % is more than 99%, the polymerization degree 2000) aqueous solution 10g, 10g fully mixes with Arthrobacter simplex (Arthrobacter simplex) concentrated solution (cell concn 20g/l), and this mixture carries out microbial immobilized program similarly to Example 3.The fixed microorganism carrier of making (particle diameter is about 2mm) 15g mixes with the substratum 150ml that contains the 0.2g/l hydrocortisone, place the triangular flask shaking culture of volume 500ml, carry out the △ 1-dehydrogenation biochemical reaction of steroid, after cultivating in 5 hours, there is 90% matrix to be transformed and generates prednisolone (prednisolone).
Embodiment 6
15 weight %PVA(basicities are more than 99%, the polymerization degree 2000) aqueous solution 10g, with the isoamylase fermented liquid of 3g and the beta-amylase fermented liquid thorough mixing of 2g, and carry out enzyme immobilization with the same gel program of embodiment (1).The fixation support of making (the about 2mm of particle diameter) 15g mixes with the matrix solution 150ml that contains the 50g/l liquefying starch, place the triangular flask vibration of volume 500ml to stir, carry out the starch hydrolysis reaction, maltose concentration can reach 41g/l in the solution after reacting 3 hours, and substrate conversion is about 82%.
Claims (16)
1, the improvement preparation method of a kind of polyvinyl alcohol microbe or enzyme immobilization carrier, comprise that to be 3 weight % carry out to the saturated boric acid aqueous solution that gelation handles produces a gel spheres in a concentration with polyvinyl alcohol and microorganism or the formed mixture of enzyme, it is characterized in that this gelation treatment time is 10 minutes to 2 hours, this gel spheres then makes its sclerosis more than 30 minutes and obtains the polyvinyl alcohol microbe fixation support with 3-20 weight % phosphoric acid or aqueous phosphatic dipping.
2, the improvement preparation method of a kind of polyvinyl alcohol microbe or enzyme immobilization carrier, comprise that to be 3 weight % carry out to the saturated boric acid aqueous solution that gelation handles produces a gel spheres in a concentration with polyvinyl alcohol and microorganism or the formed mixture of enzyme, it is characterized in that containing in this boric acid aqueous solution 3-20 weight % phosphoric acid or phosphoric acid salt and this gel spheres of hardening, and the time that this gelation and hardening treatment are carried out simultaneously is 30 minutes to 3 hours.
3, method according to claim 1, wherein this boric acid aqueous solution is saturated boric acid aqueous solution, and this gelation treatment time is 15 to 30 minutes.
4, method according to claim 1, wherein the concentration of this phosphoric acid or aqueous phosphatic is 5-15 weight %, and the time of flooding this gel spheres is 1-2 hour.
5, method according to claim 2, wherein this boric acid aqueous solution contains phosphoric acid or the phosphoric acid salt of saturated boric acid and 5-15 weight %.
6, method according to claim 2, wherein this gelation and the time of carrying out simultaneously of hardening are 1-2 hour.
7, method according to claim 1 and 2, wherein this phosphoric acid or aqueous phosphatic are selected from sodium phosphate, sodium hydrogen phosphate, SODIUM PHOSPHATE, MONOBASIC, potassiumphosphate, potassium hydrogen phosphate, potassium primary phosphate, ammonium phosphate, ammonium hydrogen phosphate, primary ammonium phosphate and phosphoric acid.
8, method according to claim 1 and 2, wherein this polyethenol series with-10-20 weight % aqueous solution form is mixed with this microorganism, and this polyvinyl alcohol has polymerization degree 1000-3000 and basicity 70-98+%.
9, method according to claim 8, wherein the polymerization degree of this polyvinyl alcohol is that 1500-2000 and basicity are 95-98+%.
10, method according to claim 1 and 2, wherein the mixed weight ratio of this polyvinyl alcohol water solution and microorganism or enzyme aqueous solution is 1: 2 to 2: 1.
11, method according to claim 1 and 2, wherein this microorganism is bacterium, mushroom, algae, protozoon or their mixture.
12, method according to claim 1 and 2, wherein this microorganism is an active sludge microorganism.
13, method according to claim 12, wherein this active sludge microorganism is the active sludge microorganism of agricultural or trade effluent domestication.
14, method according to claim 1 and 2, wherein microorganism is cereuisiae fermentum (Saccharomyces cerevisia).
15, method according to claim 1 and 2, wherein this microorganism is Arthrobacter simplex (Arthrobacter simplex).
16, method according to claim 1 and 2, wherein this enzyme is amylase, cellulase, proteolytic enzyme or glucose isomerase.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN92101875A CN1035194C (en) | 1992-03-17 | 1992-03-17 | Method for preparation of polyvinyl alcohol microbe or enzyme immobilization carrier and its use |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN92101875A CN1035194C (en) | 1992-03-17 | 1992-03-17 | Method for preparation of polyvinyl alcohol microbe or enzyme immobilization carrier and its use |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1076488A true CN1076488A (en) | 1993-09-22 |
| CN1035194C CN1035194C (en) | 1997-06-18 |
Family
ID=4939343
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN92101875A Expired - Fee Related CN1035194C (en) | 1992-03-17 | 1992-03-17 | Method for preparation of polyvinyl alcohol microbe or enzyme immobilization carrier and its use |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1035194C (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1046549C (en) * | 1996-12-25 | 1999-11-17 | 中国科学院长春应用化学研究所 | Method for biological sensor produced by latex embedding enzyme |
| CN103013885A (en) * | 2012-12-26 | 2013-04-03 | 重庆绿色智能技术研究院 | Benzene composite degrading microbe and immobilized benzene composite microbial inoculum, and preparation method thereof |
| CN108642032A (en) * | 2018-07-02 | 2018-10-12 | 深圳市长隆科技有限公司 | Embedded immobilization microbial carrier and preparation method thereof and sewage water treatment method |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS645490A (en) * | 1987-06-26 | 1989-01-10 | Komatsu Mfg Co Ltd | Production of microorganism-immobilized support |
| JPH1045491A (en) * | 1996-07-31 | 1998-02-17 | Ibiden Co Ltd | Heat insulation cylinder for silicon single crystal pulling device |
-
1992
- 1992-03-17 CN CN92101875A patent/CN1035194C/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1046549C (en) * | 1996-12-25 | 1999-11-17 | 中国科学院长春应用化学研究所 | Method for biological sensor produced by latex embedding enzyme |
| CN103013885A (en) * | 2012-12-26 | 2013-04-03 | 重庆绿色智能技术研究院 | Benzene composite degrading microbe and immobilized benzene composite microbial inoculum, and preparation method thereof |
| CN108642032A (en) * | 2018-07-02 | 2018-10-12 | 深圳市长隆科技有限公司 | Embedded immobilization microbial carrier and preparation method thereof and sewage water treatment method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1035194C (en) | 1997-06-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US5290693A (en) | Immobilization of microorganisms or enzymes in polyvinyl alcohol beads | |
| US6153416A (en) | Immobilization of microbial cells and enzymes in calcium alginate-polyethylene glycol-polyethylene imide beads | |
| JPS60120987A (en) | Production of immobilized microbial cell or enzyme | |
| JPS5923791B2 (en) | Method for producing immobilized microorganisms | |
| CN111961659A (en) | Immobilization material, biological denitrification material, preparation method and application | |
| CN115925131A (en) | Composite carbon source and production process thereof | |
| CN1035194C (en) | Method for preparation of polyvinyl alcohol microbe or enzyme immobilization carrier and its use | |
| CN114835267B (en) | Microbial activity promoting method for biological wastewater treatment | |
| Wittlich et al. | Entrapment in LentiKats®: Encapsulation of various biocatalysts—bacteria, fungi, yeast or enzymes into polyvinyl alcohol based hydrogel particles | |
| CN1970747A (en) | Process for preparing spherical immobilized cell and enzyme particulate | |
| JPH05130867A (en) | Biocatalyst-immobilized gel | |
| JPH072114B2 (en) | Method for producing immobilized biocatalyst using polyvinyl alcohol | |
| CN1491922A (en) | Active humic acid biological organic compound fertilizer and its preparing method | |
| CN101270378B (en) | Method for preparing miglitol midbody N-substituted-1-deoxidization nojirimycin derivative | |
| CN116216919B (en) | Treatment method of phosphorus-containing rural sewage and composite material used by same | |
| JPH05123694A (en) | Treatment of organic waste water with nitrogen fixation bacterial | |
| TW199906B (en) | Polyvinyl alcohol as fixed support of microorganism or enzyme and method of preparing thereof | |
| KR900002839B1 (en) | Composition and method for immobilizing cells and enzymes in a carrier matrix | |
| JP3465419B2 (en) | Wastewater treatment agent and wastewater treatment method | |
| CN117089543B (en) | Water treatment microbial inoculum and application thereof in aquaculture wastewater treatment | |
| JPS61100193A (en) | Preparation of immobilized enzyme, immobilized microorganism and group of immobilized microorganism | |
| KR100453388B1 (en) | The Seed culture delivery system for foul water and wastewater and their manufacturing method | |
| CN1088756C (en) | Method for preparing micro-organism fixation carrier | |
| Dong et al. | Synthesis and application of polyurethane Hydrogel carrier on nitrobacteria immobilization | |
| Zakaria et al. | Optimum conditions for the production of lipase by alginate-immobilized bacteria |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C15 | Extension of patent right duration from 15 to 20 years for appl. with date before 31.12.1992 and still valid on 11.12.2001 (patent law change 1993) | ||
| OR01 | Other related matters | ||
| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |