FI116531B - Pullulanas, mikroorganismer, som producerar detta, förfaranden för framställning av pullulanaset och dess användning - Google Patents
Pullulanas, mikroorganismer, som producerar detta, förfaranden för framställning av pullulanaset och dess användning Download PDFInfo
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- FI116531B FI116531B FI935900A FI935900A FI116531B FI 116531 B FI116531 B FI 116531B FI 935900 A FI935900 A FI 935900A FI 935900 A FI935900 A FI 935900A FI 116531 B FI116531 B FI 116531B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2451—Glucanases acting on alpha-1,6-glucosidic bonds
- C12N9/2457—Pullulanase (3.2.1.41)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/52—Genes encoding for enzymes or proenzymes
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/74—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
- C12N15/75—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora for Bacillus
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2451—Glucanases acting on alpha-1,6-glucosidic bonds
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- 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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/16—Preparation of compounds containing saccharide radicals produced by the action of an alpha-1, 6-glucosidase, e.g. amylose, debranched amylopectin
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01041—Pullulanase (3.2.1.41)
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Virology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Mechanical Means For Catching Fish (AREA)
- Selective Calling Equipment (AREA)
- Saccharide Compounds (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Claims (28)
1. Isolerat pullulanas, kännetecknat av att det kan erhällas fran Bacillus deramificans-stammen LMG P-5 13056, varvid pullulanaset katalyserar hydrolys av a-1,6- glukosidbindningar, och har följande N-terminala sekvens (sekvens id. nr 1) i riktning amino-karboxyl, frän vänster till höger:
2. Pullulanas enligt patentkrav 1, känneteck nat av att det innehaller aminosyrasekvensen av aminosy-rorna 1 - 928 som visas i figur 4 (sekvens id. nr 11).
3. Pullulanas enligt patentkrav 2, kännetecknat av attdet syntetiseras i form av en prekursor, som 20 innehaller en ytterligare sekvens pä 29 aminosyror (sekvens id. nr 12).
4. Pullulanas enligt patentkrav 1, kännetecknat av attdet utvecklar optimal enzymaktivitet, mätt vid ;·. en temperatur pä ca 60 °C, vid ett pH pä ca 4,3.
5. Pullulanas enligt nägot av de föregäende pa- * · .,V tentkraven, kännetecknat av att det har en molekyl- vikt pä ca 100 kDa.
* » · ’·* ‘ 6. Pullulanas enligt nägot av de föregäende pa- tentkraven, kännetecknat av att det har en isoelekt-j 30 risk punkt mellan 4,1 och 4,5. ‘:i
'< 7. Pullulanas enligt nägot av de föregäende pa- tentkraven, kännetecknat av att det utvecklar en en-zymaktivitet pä mer än 80 % av den maximala enzymaktivite-ten inom ett pH-omräde pä ca 3,8 - 4,9 vid en temperatur : 35 pä 60 °C, dä den maximala enzymaktiviteten mäts vid en ‘ : temperatur pä ca 60 °C och vid pH 4,3. 116531 77
8. Pullulanas enligt nägot av de föregäende pa-tentkraven, kännetecknat av att det uppvisar en rela-tiv enzymaktivitet pä över 90 %, mätt vi en temperatur pä ca 60 °C i fränvaro av substrat, inom ett pH-omräde pä ca 5 3,8 - 5.
9. Pullulanas enligt nägot av de föregäende pa-tentkraven, kännetecknat av att det hydrolyserar a-1,6-gukosidbindningar som finns i amylopektin.
10. Pullulanas enligt nägot av de föregäende pa-10 tentkraven, kännetecknat av att det nedbryter pullu- lan tili maltotrios.
10 Asp Gly Asn Thr Thr Thr Ile Ile Val His 15 10 Tyr Phe Cys Pro Ala Gly Asp Tyr Gin Pro 15 20
11. Pullulanas enligt nägot av de föregäende pa-tentkraven, kännetecknat av att det hydrolyserar ami-nopektin för att bilda oligosackarider.
12. Pullulanas enligt nägot av de föregäende pa- tentkraven, kännetecknat av att det katalyserar mal-toskondensationsreaktionen för att bilda tetraholosider.
13. Pullulanas enligt nägot av de föregäende pa-tentkraven, kännetecknat av att det har producerats 20 av Bacillus deramificans-stammen LMG P-13056 eller ett de-rivat eller en mutant därav.
14. Pullulanas enligt nägot av patentkraven 1 - 12, kännetecknat av att det producerats heterologt av ;»"t en mikroorganism av släktet Bacillus, vilken innehäller en * · . 25 gen som kodar för ett alkaliskt proteas dä den befinner * · ;*,* sig i vilt tillständ, varvid genen som kodar för alkaliskt » · ;* proteas har avlägsnats frän mikro-organismen av släktet I ( | ’·' * Bacillus.
15. Förfarande för framställning av ett pullula- · 30 nas enligt nägot av patentkraven 1 - 13, kännetecknat I i * : av att det omfattar att man odlar en stam av Bacillus de- ,···. ramificans eller ett derivat därav, som förmär producera * · · pullulanaset i ett lämpligt näringsmedium innehällande G* koi- och kvävekällor och mineralsalter under aeroba be- 35 tingelser, och tar tillvara pullulanaset som sälunda er- ; \i hälls. 116531 78
16. Förfarande för framställning av ett pullula-nas enligt nägot av patentkraven 1 - 13, kännetecknat av att det omfattar att man isolerar ett DNA-fragment soin kodar för pullulanas, infogar DNA-fragmentet i en lämplig 5 vektor, introducerar vektorn i en lämplig värd eller in-troducerar DNA-fragmentet i kromosomen hos en lämplig värd, odlar värden, uttrycker pullulanaset och tar tillva-ra pullulanaset.
17. Användning av ett pullulanas enligt nägot av 10 patentkraven 1-14 för försockring av stärkelse.
18. DNA-molekyl, kännetecknad av att den in-nehäller nukleotidsekvensen (sekvens id. nr 10) som kodar för pullulanas enligt nägot av patentkraven 1-14.
19. DNA-molekyl enligt patentkrav 18, känne-15 tecknad av att den innehäller hela pullulanas-genen (sekvens id. nr 8) av Bacillus deramificans LMG P-13056.
20. Expressionsvektor, kännetecknad av att den innehäller DNA-molekylen enligt patentkrav 18.
21. Expressionsvektor enligt patentkrav 20, 20 kännetecknad av att den är pUBDEBRAl, som visas i fi- gur 1.
22. Kromosomal integrationsvektor, känneteck-nad av att den innehäller DNA-molekylen enligt patentkrav .:i:j 18.
23. Kromosomal integrationsvektor enligt patent- krav 22, kännetecknad av att den är pUBCDEBRAllDNSI, som visas i figur 3.
’ 24. Transformerad stam av Bacillus licheniformis, kännetecknad av att den innehäller DNA-molekylen en- i.i · 30 ligt patentkrav 18. > · > :.(,ί
25. Transformerad stam enligt patentkrav 24 av Bacillus licheniformis, kännetecknad av att den inne- » » * häller expressionsvektorn enligt patentkrav 20 eller den I · kromosomala integrationsvektorn enligt patentkrav 22. * · it» 116531 79
26. Transformerad stam enligt patentkrav 25 av Bacillus licheniformis, kännetecknad av att den inne-häller expressionsvektorn pUBDEBRAl, som visas i figur 1 eller den kromosomala integrationsvektorn pUBCDEBRAllDNSI, 5 som visas i figur 3.
27. Kultur, kännetecknad av att den är en kultur av Bacillus dermificans LMG P-13056.
28. Enzymkomposition, kännetecknad av att den innehäller ett pullulanas enligt nägot av patentkraven 10 1 - 14. • · I · * · I I I » 1 1 « 1 » I * 1 ♦ • · I t · i I 1 ♦ · ’ 1 · • 1 · I · * t 1 ( » 1
Applications Claiming Priority (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
BE9201156 | 1992-12-28 | ||
BE9201156A BE1006483A3 (fr) | 1992-12-28 | 1992-12-28 | Pullulanase, microorganismes la produisant, procedes de preparation de cette pullulanase, utilisations et compositions comprenant celle-ci. |
BE9300744 | 1993-07-15 | ||
BE9300744A BE1007313A3 (fr) | 1993-07-15 | 1993-07-15 | Pullulanase, microorganismes la produisant, procedes de preparation de cette pullulanase et utilisations de celle-ci. |
BE9301278 | 1993-11-19 | ||
BE9301278A BE1007723A6 (fr) | 1993-11-19 | 1993-11-19 | Pullulanase, microorganismes la produisant, procedes de preparation de cette pullulanase et utilisations de celle-ci. |
Publications (3)
Publication Number | Publication Date |
---|---|
FI935900A0 FI935900A0 (fi) | 1993-12-28 |
FI935900A FI935900A (sv) | 1994-06-29 |
FI116531B true FI116531B (sv) | 2005-12-15 |
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Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
FI935900A FI116531B (sv) | 1992-12-28 | 1993-12-28 | Pullulanas, mikroorganismer, som producerar detta, förfaranden för framställning av pullulanaset och dess användning |
Country Status (13)
Country | Link |
---|---|
US (6) | US5817498A (sv) |
EP (1) | EP0605040B1 (sv) |
JP (1) | JPH06217770A (sv) |
KR (1) | KR100319442B1 (sv) |
CN (1) | CN1061089C (sv) |
AT (1) | ATE183236T1 (sv) |
AU (2) | AU686574B2 (sv) |
CA (1) | CA2112028C (sv) |
DE (1) | DE69325981T2 (sv) |
DK (1) | DK0605040T3 (sv) |
ES (1) | ES2137222T3 (sv) |
FI (1) | FI116531B (sv) |
MX (1) | MX9400123A (sv) |
Families Citing this family (63)
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CN113490740A (zh) | 2019-03-18 | 2021-10-08 | 诺维信公司 | 适合用于液化中的具有普鲁兰酶活性的多肽 |
EP4055157A1 (en) | 2019-11-08 | 2022-09-14 | Novozymes A/S | Stabilized liquid enzyme compositions for brewing |
CN111560077A (zh) * | 2020-05-21 | 2020-08-21 | 中国海洋大学 | 一种酶及其在合成普鲁兰多糖中的作用 |
WO2023225459A2 (en) | 2022-05-14 | 2023-11-23 | Novozymes A/S | Compositions and methods for preventing, treating, supressing and/or eliminating phytopathogenic infestations and infections |
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US4560651A (en) * | 1981-04-20 | 1985-12-24 | Novo Industri A/S | Debranching enzyme product, preparation and use thereof |
JPS57174089A (en) * | 1981-04-20 | 1982-10-26 | Novo Industri As | Chain dividing enzyme product |
US4628031A (en) * | 1984-09-18 | 1986-12-09 | Michigan Biotechnology Institute | Thermostable starch converting enzymes |
US4828994A (en) * | 1984-09-21 | 1989-05-09 | Genex Corporation | Bacillus strains with reduced extracellular protease levels |
US4612287A (en) * | 1985-05-23 | 1986-09-16 | Cpc International Inc. | Plasmids containing a gene coding for a thermostable pullulanase and pullulanase-producing strains of Escherichia coli and Bacillus subtilis containing the plasmids |
US4628028A (en) * | 1985-05-23 | 1986-12-09 | Cpc International Inc. | Novel thermostable pullulanase enzyme and method for its production |
CA1285896C (en) * | 1986-09-26 | 1991-07-09 | Kuniharu Nakai | .alpha.-1, 6-GLUCOSIDASE AND PROCESS FOR PRODUCING THE SAME |
JPH0223872A (ja) * | 1988-07-12 | 1990-01-26 | Ezaki Glico Co Ltd | 組換えプラスミド,それにより形質転換された枯草菌及びそれによる耐熱性プルラナーゼの製造法 |
EP0405283A3 (en) * | 1989-06-26 | 1991-03-06 | Enzyme Bio-Systems Ltd. | Novel thermoduric and aciduric pullulanase enzyme and method for its production |
US5055403A (en) * | 1989-06-26 | 1991-10-08 | Enzyme Bio-Systems, Ltd. | Thermoduric and aciduric pullulanase enzyme and method for its production |
DK47291D0 (da) * | 1991-03-15 | 1991-03-15 | Novo Nordisk As | Pullulanase |
JP3026857B2 (ja) * | 1991-07-05 | 2000-03-27 | ナガセ生化学工業株式会社 | 新規プルラナーゼおよびその製造法 |
JP3173849B2 (ja) * | 1992-02-26 | 2001-06-04 | 天野エンザイム株式会社 | 新規な枝切り酵素及びその製造法 |
JPH05236959A (ja) * | 1992-02-28 | 1993-09-17 | Yoshiyuki Takasaki | プルラナーゼ、その製造法及び該酵素を用いる澱粉の 糖化法 |
DE4244107C2 (de) * | 1992-12-24 | 1996-02-08 | Hirschmann Richard Gmbh Co | Hochfrequenz-Übertrager |
DE69325981T2 (de) * | 1992-12-28 | 2000-02-03 | Genencor Int | Pullulanase, Mikroorganismen die sie produzieren, Verfahren zur Herstellung und ihre Anwendung |
GB2279955B (en) * | 1993-07-15 | 1998-02-18 | Solvay | Xylanase derived from a Bacillus species, expression vectors for such xylanase and other proteins, host organisms therefor and use thereof |
-
1993
- 1993-12-20 DE DE69325981T patent/DE69325981T2/de not_active Expired - Lifetime
- 1993-12-20 EP EP93203593A patent/EP0605040B1/fr not_active Expired - Lifetime
- 1993-12-20 ES ES93203593T patent/ES2137222T3/es not_active Expired - Lifetime
- 1993-12-20 DK DK93203593T patent/DK0605040T3/da active
- 1993-12-20 AT AT93203593T patent/ATE183236T1/de active
- 1993-12-28 FI FI935900A patent/FI116531B/sv not_active IP Right Cessation
- 1993-12-28 CN CN93121736A patent/CN1061089C/zh not_active Expired - Lifetime
- 1993-12-28 JP JP5337202A patent/JPH06217770A/ja active Pending
- 1993-12-28 KR KR1019930030410A patent/KR100319442B1/ko not_active IP Right Cessation
- 1993-12-29 CA CA002112028A patent/CA2112028C/fr not_active Expired - Lifetime
- 1993-12-30 AU AU52759/93A patent/AU686574B2/en not_active Ceased
-
1994
- 1994-01-03 MX MX9400123A patent/MX9400123A/es not_active IP Right Cessation
-
1995
- 1995-06-07 US US08/478,341 patent/US5817498A/en not_active Expired - Lifetime
- 1995-06-07 US US08/472,293 patent/US5731174A/en not_active Expired - Lifetime
- 1995-06-07 US US08/474,140 patent/US5721127A/en not_active Expired - Lifetime
- 1995-06-07 US US08/474,545 patent/US5736375A/en not_active Expired - Lifetime
- 1995-06-07 US US08/477,630 patent/US5721128A/en not_active Expired - Lifetime
-
1997
- 1997-12-23 US US08/996,733 patent/US6074854A/en not_active Expired - Lifetime
-
1998
- 1998-05-11 AU AU64831/98A patent/AU6483198A/en not_active Abandoned
Also Published As
Publication number | Publication date |
---|---|
CN1061089C (zh) | 2001-01-24 |
AU5275993A (en) | 1994-07-07 |
AU6483198A (en) | 1998-07-30 |
FI935900A0 (fi) | 1993-12-28 |
KR100319442B1 (ko) | 2002-11-18 |
JPH06217770A (ja) | 1994-08-09 |
US5721127A (en) | 1998-02-24 |
CA2112028A1 (fr) | 1994-06-29 |
DE69325981D1 (de) | 1999-09-16 |
US5731174A (en) | 1998-03-24 |
US5721128A (en) | 1998-02-24 |
US6074854A (en) | 2000-06-13 |
ATE183236T1 (de) | 1999-08-15 |
FI935900A (sv) | 1994-06-29 |
US5817498A (en) | 1998-10-06 |
KR940014776A (ko) | 1994-07-19 |
ES2137222T3 (es) | 1999-12-16 |
CN1090325A (zh) | 1994-08-03 |
CA2112028C (fr) | 2003-03-11 |
DK0605040T3 (da) | 2000-03-13 |
DE69325981T2 (de) | 2000-02-03 |
EP0605040B1 (fr) | 1999-08-11 |
AU686574B2 (en) | 1998-02-12 |
MX9400123A (es) | 1994-07-29 |
EP0605040A1 (fr) | 1994-07-06 |
US5736375A (en) | 1998-04-07 |
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