CN1101942A - New method for preparing F-1-P crystal protein from fresh tobacco leaves - Google Patents
New method for preparing F-1-P crystal protein from fresh tobacco leaves Download PDFInfo
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- CN1101942A CN1101942A CN 93119265 CN93119265A CN1101942A CN 1101942 A CN1101942 A CN 1101942A CN 93119265 CN93119265 CN 93119265 CN 93119265 A CN93119265 A CN 93119265A CN 1101942 A CN1101942 A CN 1101942A
- Authority
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- Prior art keywords
- fresh tobacco
- tobacco leaf
- crystallin
- protein
- crystal
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- Pending
Links
- 241000208125 Nicotiana Species 0.000 title claims abstract description 15
- 235000002637 Nicotiana tabacum Nutrition 0.000 title claims abstract description 15
- 238000000034 method Methods 0.000 title claims abstract description 13
- 101710151559 Crystal protein Proteins 0.000 title abstract description 3
- 239000012043 crude product Substances 0.000 claims abstract description 15
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims abstract description 3
- 239000011734 sodium Substances 0.000 claims abstract description 3
- 229910052708 sodium Inorganic materials 0.000 claims abstract description 3
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims abstract 2
- 239000011591 potassium Substances 0.000 claims abstract 2
- 229910052700 potassium Inorganic materials 0.000 claims abstract 2
- 239000012266 salt solution Substances 0.000 claims abstract 2
- 239000012535 impurity Substances 0.000 claims description 10
- 238000012546 transfer Methods 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 9
- 238000001556 precipitation Methods 0.000 claims description 7
- 239000000706 filtrate Substances 0.000 claims description 6
- 238000002425 crystallisation Methods 0.000 claims description 5
- 230000008025 crystallization Effects 0.000 claims description 5
- 238000010790 dilution Methods 0.000 claims description 5
- 239000012895 dilution Substances 0.000 claims description 5
- 150000003839 salts Chemical class 0.000 claims description 5
- 229910017053 inorganic salt Inorganic materials 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims description 3
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 claims description 3
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 3
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 claims description 3
- 239000011575 calcium Substances 0.000 claims description 3
- 229910052791 calcium Inorganic materials 0.000 claims description 3
- 239000011777 magnesium Substances 0.000 claims description 3
- 229910052749 magnesium Inorganic materials 0.000 claims description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 claims 1
- 230000001376 precipitating effect Effects 0.000 claims 1
- 239000013078 crystal Substances 0.000 abstract description 14
- 108090000623 proteins and genes Proteins 0.000 abstract description 8
- 102000004169 proteins and genes Human genes 0.000 abstract description 8
- 238000002360 preparation method Methods 0.000 abstract description 5
- 239000006228 supernatant Substances 0.000 abstract 2
- 239000003638 chemical reducing agent Substances 0.000 abstract 1
- 238000000151 deposition Methods 0.000 abstract 1
- 238000005265 energy consumption Methods 0.000 abstract 1
- 150000002500 ions Chemical class 0.000 abstract 1
- 238000004519 manufacturing process Methods 0.000 abstract 1
- 238000003825 pressing Methods 0.000 abstract 1
- 238000000926 separation method Methods 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 12
- 238000005119 centrifugation Methods 0.000 description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 235000018102 proteins Nutrition 0.000 description 6
- 238000003756 stirring Methods 0.000 description 5
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 239000000049 pigment Substances 0.000 description 4
- 241001464837 Viridiplantae Species 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 235000011837 pasties Nutrition 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241000921313 Phyllopodium Species 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 150000003863 ammonium salts Chemical class 0.000 description 2
- 238000000502 dialysis Methods 0.000 description 2
- 235000021245 dietary protein Nutrition 0.000 description 2
- 230000008030 elimination Effects 0.000 description 2
- 238000003379 elimination reaction Methods 0.000 description 2
- 239000001103 potassium chloride Substances 0.000 description 2
- 235000011164 potassium chloride Nutrition 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- HRZFUMHJMZEROT-UHFFFAOYSA-L sodium disulfite Chemical compound [Na+].[Na+].[O-]S(=O)S([O-])(=O)=O HRZFUMHJMZEROT-UHFFFAOYSA-L 0.000 description 2
- 229940001584 sodium metabisulfite Drugs 0.000 description 2
- 235000010262 sodium metabisulphite Nutrition 0.000 description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
- GEHJYWRUCIMESM-UHFFFAOYSA-L sodium sulfite Chemical compound [Na+].[Na+].[O-]S([O-])=O GEHJYWRUCIMESM-UHFFFAOYSA-L 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 1
- 102000030523 Catechol oxidase Human genes 0.000 description 1
- 108010031396 Catechol oxidase Proteins 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 244000208060 Lawsonia inermis Species 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 1
- 108010003581 Ribulose-bisphosphate carboxylase Proteins 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 description 1
- 238000012870 ammonium sulfate precipitation Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 238000001879 gelation Methods 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000003907 kidney function Effects 0.000 description 1
- 238000005360 mashing Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 230000022558 protein metabolic process Effects 0.000 description 1
- 230000006920 protein precipitation Effects 0.000 description 1
- 238000001742 protein purification Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 229960004249 sodium acetate Drugs 0.000 description 1
- 235000017550 sodium carbonate Nutrition 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 235000010344 sodium nitrate Nutrition 0.000 description 1
- 239000004317 sodium nitrate Substances 0.000 description 1
- 229940001516 sodium nitrate Drugs 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- 235000010265 sodium sulphite Nutrition 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000009967 tasteless effect Effects 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
Landscapes
- Peptides Or Proteins (AREA)
Abstract
A process for preparing F-1-P crystal protein from fresh tobacco leaf includes homogenizing fresh tobacco leaf in the presence of reducing agent, filter pressing, centrifugal separation to obtain supernatant, and directly depositing the coarse F-1-P from said supernatant. Then the ion strength or pH value of soluble salt solution of sodium or potassium is changed to re-dissolve the crude product and then crystallize, thus obtaining the crystal F-1-P protein with specific geometrical shape. The invention opens up a new way for the industrial preparation of the high-purity crystal F-1-P protein, and has the advantages of simple process, stable operation, easy control and no energy consumption.
Description
The present invention relates to the processing method of extracting effective components in green plants, exactly is a kind of novel method of producing the F-1-P crystallin in fresh tobacco leaf.
Contain water-soluble and water-insoluble substance in the dry (accounting for 10～20%) that the bright leaf of green plants is left after dewatering, the latter mostly is the fibrous texture material, the former is divided into two classes again, one class is that (molecular weight is no more than 10 to low molecule, 000) compound is as sugar, amino acid etc., another kind of is protein (molecular weight is more than 30,000).Protein divides two kinds again, a kind of be molecular weight 30,000～100, the mixed protein between 000, general designation F-2-P albumen is (hereinafter to be referred as P
2), another kind is single albumen, molecular weight 550,000 claims F-1-P albumen (hereinafter to be referred as P
1), it is a kind of enzyme relevant with photosynthesis, claims 1.5-bisphosphate ribose ketone carboxylase, Rubisco etc. again.Soluble proteins accounts for 20% of dry, P in the fresh tobacco leaf
1And P
2Respectively account for about 10%.
Highly purified P
1Colourless, tasteless, performances such as its amino acid composition, protein efficiency ratio, PER, water-soluble, gelation, emulsifying property, suction oil suction all are better than other food protein additives, especially be fit to renal function disease patient and protein metabolism imbalance patient take, so its application prospect are very wide.The seventies is found by obtaining crystalline P in the fresh tobacco leaf
1, and take from the generally non-crystallizable of other green plantss.
In fresh tobacco leaf, extract P
1At first all need bright leaf and phyllopodium homogenate (promptly mashing, grind to form pasty state) to discharge effective ingredient wherein, are separated homogenate then,, obtain being dissolved with P with thicker solid matter of filtering (fiber-like) and pigment material
1And P
2Brown juice.To this juice, 1, use filter membrane and dialysis tubing to carry out ultrafiltration, dialysis, form crystal after holding back macromole.2, Sephadex G-25 or G-50 column chromatography.3, ammonium sulfate precipitation, polyethylene glycol system crystallization.4, isoelectric precipitation, percrystallization again after the redissolution.Above method industrialization is had any problem.United States Patent (USP) 4268632(applying date 79.9.4) a kind of thermal induction crystalline extraction process is disclosed, bright leaf heats (48～52 ℃) to homogenate after homogenate, separate after being cooled to room temperature, elimination fiber-like and pigment material get brown juice, then juice are cooled to 8 ℃, leave standstill after 24 hours and just obtain P
1The octagon crystal.Heating can promote the cohesion of pigment material, separate more thorough, the P that crystallization is separated out
1Purer.But consumes energy is wanted in heating, and Heating temperature wants strict control, otherwise will cause protein denaturation, forms the insoluble protein precipitation.
The purpose of this invention is to provide and a kind ofly simple and effectively just can obtain high-purity crystals P at normal temperatures
1The preparation method, this method can realize industrialization, and easy to operate, be easy to control.
The present invention is achieved in that and at first directly precipitates the preparation crude product in the homogenate clear liquid, recrystallize after making it to redissolve by the ionic strength that changes solution then, thereby the crystal P that obtains having geometry in particular
1Detailed process is as follows:
1, gets fresh tobacco leaf and phyllopodium, under the condition that reductive agent exists, carry out homogenate.Reductive agent can be to dredge basic ethanol, xitix, Sodium Metabisulfite etc., suppresses polyphenol oxidase product and proteinic covalent attachment.Homogenate in the pasty state, the disperse phase (B) of the solid of including fiber class (A), pigment, P
1And P
2And other impurity such as reductive agent and Polyphenols.
2, separate homogenate.A is removed in press filtration, and centrifugal or fractionation by adsorption is removed B, obtains brown or henna homogenate clear liquid, includes P
1, P
2With residual B and other impurity.
3, the homogenate clear liquid left standstill 1～5 hour, and isoelectric precipitation also separates P
1, this precipitation is amorphous, is crude product.
4, crude product is purified.The variation of the ingenious land productivity salts solution of contriver ionic strength in the present invention comes protein purification.(〉=0.05M) sodium chloride solution stirs the crude product input, transfers PH to 7.5 simultaneously, P to get finite concentration
1Dissolving, and other impurity do not dissolve, and separate and remove impurity.With clear liquid dilution (more than 1 times, promptly reducing the ionic strength of solution) or transfer pH value to 5.0～6.0 o'clock, the P of redissolution
1Recrystallize is separated out, and separates to obtain pure product crystal P
1, crystal is regular hexagon dodecahedron or square two pyramids of octagon.
Experiment showed, that except that sodium-chlor the soluble inorganic salt of sodium sulfate, SODIUMNITRATE, yellow soda ash, sodium sulphite, sodium-acetate etc. and sylvite, ammonium salt and calcium and magnesium all can be used to purifying P
1PH is 7.5 during redissolution, when the ionic strength that reduces solution or when transferring pH value 5.0～6.0, and P
1Recrystallize is separated out.
5, isolate crystal P
1After clear liquid when regulating PH to 4, P
2Precipitation is separated out, separated and collected P
2
The present invention is preparation of industrialization high-purity crystals P
1Opened up new approach, technology is simple, stable operation, control, not consumes energy easily.
Embodiment:
One, crude product P
1Preparation.
Get the 1000g fresh tobacco leaf, add 500ml1% Sodium Metabisulfite solution grinding homogenate and get the pasty state homogenate.Homogenate is removed A with the squeezer press filtration, and filtrate was removed most of B in 10 minutes in the 4000rpm centrifugation, gets the homogenate clear liquid.The homogenate clear liquid left standstill four hours, and isoelectric precipitation is separated out P
1,, get crude product P in 4000rpm centrifugation 5 minutes
1
Two, crude product is purified.
1, gets 0.5MNaCl solution 200ml, with crude product P
1Drop into and stir, transfer PH7.5, make P with 0.1NNaOH
1Dissolving, in 4000rpm centrifugation 5 minutes, elimination impurity, filtrate is transferred PH5.0～6.0, leaves standstill P 2～4 hours
1Recrystallize is separated out.Centrifugation gets 4～6g crystal P
1, crystalline form is square two pyramids of octagon.
2, get 0.3MNaCl solution 200ml, with crude product P
1Drop into and stir, transfer PH7.5, make P with 0.1NNaOH
1Dissolving, later operation transfer PH5.0～5.5 to get 4～6g crystal P with embodiment 1
1
3, get 0.1MNaCl solution 200ml, with crude product P
1Drop into and stir, transfer PH7.5, make P with 0.1NNaOH
1Dissolving, impurity is fallen in centrifugation, and P was left standstill in 2 times of filtrate water dilutions 10 hours
1Recrystallize is separated out.Centrifugation gets 4～6g crystal P
1, crystalline form is the regular hexagon dodecahedron.
4, get 0.05NaCl solution 200ml, with crude product P
1Drop into and stir, transfer PH7.5, make P with 0.1NNaOH
1Dissolving, later operation is with embodiment 3, but only dilutes 1 times during dilute with water.Get 4～6g crystal P
1
Concerning the NaCl solution of same concentration, after crude product redissolves, or dilution, or transfer PH, but equal recrystallize, productive rate basically identical, stable operation.
NaCl is the easiest to be obtained, and cheap, is suitable for industrial use most.In fact the soluble salt of other salt of sodium and sylvite, ammonium salt and calcium, magnesium all can be used to the P that purifies
1, because the ionic strength of salts solution or PH can control.
Three, reclaim P
2
Isolate crystallization P
1After clear liquid transfer PH to 4.0, P
2Precipitation is separated out, and centrifugation gets P
2P
2It also is a kind of food protein additive.
Claims (5)
1, a kind of novel method of producing the F-1-P crystallin in fresh tobacco leaf is made up of homogenate, precipitation, crystallization, sepn process, it is characterized in that:
(1), obtains the F-1-P crude product by directly precipitating to separate out and separate in the homogenate clear liquid;
(2), crude product is dissolved in concentration more than or equal to 0.05M, PH is the soluble inorganic salt solution of the salt of 7.5 sodium or potassium or ammonium or calcium or magnesium, remove by filter impurity;
(3), remove filtrate water dilution behind the impurity more than 1 times or transfer pH value between 5.0～6.0, separate obtaining the F-1-P crystallin that crystallization is separated out.
2, a kind of novel method of producing the F-1-P crystallin in fresh tobacco leaf according to claim 1, it is characterized in that: described inorganic salt are sodium-chlor or Repone K.
3, a kind of novel method of producing the F-1-P crystallin in fresh tobacco leaf according to claim 1 and 2, it is characterized in that: described inorganic salt are sodium-chlor.
4, a kind of novel method of producing the F-1-P crystallin in fresh tobacco leaf according to claim 1 is characterized in that: 2 times of the filtrate water dilutions behind the removal impurity.
5, a kind of novel method of producing the F-1-P crystallin in fresh tobacco leaf according to claim 1 is characterized in that: the filtrate behind the removal impurity is transferred pH value 5.0～5.5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 93119265 CN1101942A (en) | 1993-10-19 | 1993-10-19 | New method for preparing F-1-P crystal protein from fresh tobacco leaves |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 93119265 CN1101942A (en) | 1993-10-19 | 1993-10-19 | New method for preparing F-1-P crystal protein from fresh tobacco leaves |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1101942A true CN1101942A (en) | 1995-04-26 |
Family
ID=4992841
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 93119265 Pending CN1101942A (en) | 1993-10-19 | 1993-10-19 | New method for preparing F-1-P crystal protein from fresh tobacco leaves |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1101942A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100376684C (en) * | 2006-03-28 | 2008-03-26 | 华南理工大学 | Process for preparing bacteriostatic peptide by discarded tobacco leaf protein |
| CN105188422A (en) * | 2013-03-14 | 2015-12-23 | R.J.雷诺兹烟草公司 | Protein-enriched tobacco-derived composition |
-
1993
- 1993-10-19 CN CN 93119265 patent/CN1101942A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100376684C (en) * | 2006-03-28 | 2008-03-26 | 华南理工大学 | Process for preparing bacteriostatic peptide by discarded tobacco leaf protein |
| CN105188422A (en) * | 2013-03-14 | 2015-12-23 | R.J.雷诺兹烟草公司 | Protein-enriched tobacco-derived composition |
| CN112137158A (en) * | 2013-03-14 | 2020-12-29 | R.J.雷诺兹烟草公司 | Protein-enriched tobacco-derived compositions |
| US11166485B2 (en) | 2013-03-14 | 2021-11-09 | R.J. Reynolds Tobacco Company | Protein-enriched tobacco-derived composition |
| US11375741B2 (en) | 2013-03-14 | 2022-07-05 | R.J. Reynolds Tobacco Company | Protein-enriched tobacco-derived composition |
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| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C01 | Deemed withdrawal of patent application (patent law 1993) | ||
| WD01 | Invention patent application deemed withdrawn after publication |