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1. 发明专利

JPH10234364A BETA-GALACTOSIDE-ALPHA-2,6-SIALATE TRANSFERASE 失效
公开(公告)号：JPH10234364A
公开(公告)日：1998-09-08
申请号：JP4509897
申请日：1997-02-28
申请人： JAPAN TOBACCO INC
发明人： YAMAMOTO TAKESHI , NAKASHIZU MOTOKO , TERADA ICHIRO
IPC分类号： C12N15/09 , C07H21/04 , C12N1/21 , C12N9/10 , C12N15/00 , C12N15/31 , C12R1/01 , C12R1/19
摘要： PROBLEM TO BE SOLVED: To obtain a recombinant β-galactoside-α-2,6-sialate transferase having high stability and capable of adding sialic acid to oligosaccharides, complex carbohydrates, and sugar chains in high yields without side reactions, by deleting a part of the memberane-binding region. SOLUTION: This enzyme is composed of a protein having an amino acid sequence which may lack in at least a part of the membrane-binding region, or an amino acid sequence which may lack in either part consisting of amino acid sequence 499-675 or part consisting of amino acid sequence 674-675, among the amino acid position 16-675. This enzyme consists of an amino acid sequence in which at least one amino acid is deleted, substituted, or inserted in the 16-498 position in the formula or a DNA which codes the amino acid sequence corresponding to the 16-498 positions, and is obtained by culturing host cells transformed with an expression vector including DNA coding a protein having a β-galactoside-α-2,6-sialate transferase activity.

2. 发明专利

JPS63190840A PURIFICATION OF SOLANESOL 失效
公开(公告)号：JPS63190840A
公开(公告)日：1988-08-08
申请号：JP2247987
申请日：1987-02-04
申请人： JAPAN TOBACCO INC
发明人： TAWARA TAKAHIRO , MATSUSHIMA SANJI , NINOMIYA NAOYOSHI , HASHIMOTO TATSUJI , TERADA ICHIRO , SHIMIZU YOSHIAKI
IPC分类号： C07C29/76 , C07C27/00 , C07C33/02 , C07C67/00
摘要： PURPOSE:To obtain high-purity solanesol useful as a synthetic precursor for coenzyme Q used as a heart drug by a few processes, by treating an unsaponified substance of tobacco extract by the use of specific chromatography and purifying. CONSTITUTION:An unsaponified substance of tobacco extract is sent to carrier- free centrifugal liquid partition chromatography having n-hexane as a stationary phase and a hydrous alcohol such as methanol, ethanol, etc., having 5-10% water content as a mobile phase and partitioned. Then feed direction of solution is reversed and fractionation is carried out by the use of h-hexane to give high-purity solanesol. Further solanesol is preferably treated industrially by high performance liquid chromatography.

3. 发明专利

JPH08173182A PRODUCTION OF SACCHARIDES CONTAINING SIALIC ACID 失效
公开(公告)号：JPH08173182A
公开(公告)日：1996-07-09
申请号：JP32715294
申请日：1994-12-28
申请人： JAPAN TOBACCO INC
发明人： YAMAMOTO TAKESHI , NAKASHIZU MOTOKO , TERADA ICHIRO , KODAMA HISASHI
IPC分类号： C12N9/10 , C12P19/26 , C12R1/01
摘要： PURPOSE: To simply obtain saccharides containing sialic acid and facilitate the examination of physiological functions or biological meanings of saccharides containing the sialic acid by reacting cytidine monophosphate-sialic acid with an enzyme capable of transferring the sialic acid to a galactose residue, etc., in a conjugated carbohydrate saccharide chain or a free saccharide chain. CONSTITUTION: Cytidine monophosphate-sialic acid is reacted with a β- galactoside α2,6-sialyltransferase derived from Photobacterium damsela JT0160, ATCC33539, ATCC35083, etc., to transfer the sialic acid to the 6-position of the galactose residue in a conjugated carbohydrate saccharide chain or a free saccharide chain or the 6-position of a monosaccharide capable of constituting the conjugated carbohydrate derived from an animal and having hydroxyl group at the 6-position. The transferase has 64000±5000 molecular weight (measured by the gel filtration), 5-6 optimum pH and 30 deg.C optimum temperature.

4. 发明专利

JPH08154673A NEW BETA-GALACTOSIDE-ALPHA2,6-SIALIC ACID TRANSFERASE AND ITS PRODUCTION 失效
公开(公告)号：JPH08154673A
公开(公告)日：1996-06-18
申请号：JP30409094
申请日：1994-12-07
申请人： JAPAN TOBACCO INC
发明人： YAMAMOTO TAKESHI , NAKASHIZU MOTOKO , TERADA ICHIRO , KODAMA HISASHI
IPC分类号： C12N9/10 , C12N1/20 , C12R1/01
摘要： PURPOSE: To produce in large quantities at low cost a new β-galactoside-α2,6- sialic acid transferase derived from microorganisms. CONSTITUTION: This new β-galactoside-α2,6-sialic acid transferase is derived from a Photobacterium sp. microorganism, having the following physicochemical properties: (1) action and specificity; transferring of sialic acid from cytidine monophosphate sialic acid to the 6-site in galactose residue in complex saccharides or free sugar chain, or to the 6-site in a monosaccharide capable of constituting a complex saccharide derived from an animal and having hydroxyl group at the 6-site carbon, (2) the optimum PH; 5-6, (3) the optimum temperature; 30 deg.C and (4) molecular weight; 64000±5000(measured by the gel filtration method).

5. 发明专利

JPH07322886A PRODUCTION OF OLIGOSACCHARIDE AND MONOSACCHARIDE 失效
公开(公告)号：JPH07322886A
公开(公告)日：1995-12-12
申请号：JP12042094
申请日：1994-06-01
申请人： JAPAN TOBACCO INC
发明人： SUGANO YASUSHI , TERADA ICHIRO , YAMAZAKI YOSHINARI , KODAMA HISASHI , NOMA MASAKATA
IPC分类号： C12P19/14
摘要： PURPOSE:To obtain the subject saccharides having preventing effects on retrogradation of starch by reacting a substrate containing agarose with a '-agarase acting on both a polysaccharide and an oligosaccharide derived from the agarose and an alpha-agarase acting on the oligosaccharide derived from the agarose. CONSTITUTION:This method for producing an oligosaccharide and a monosaccharide is to react a substrate containing agarose with an enzyme having the following physico- chemical properties: capable of catalyzing reaction for hydrolyzing agar and the agarose into an endo type and reducing the molecular weight without acting on carrageenan and alginic acid; having Moptimum pH of 7-8.6 (30 deg.C) and 30 deg.C optimum temperature; stable at pH6-9; retaining about 85% maximum activities by heating at 50 deg.C for 15min, having about 6.3 isoelectric point, about 95000 molecular weight [measured by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS- PAGE)] and an amino acid sequence expressed by the formula at the amino terminal as a '-agarase acting on both of a polysaccharide and an oligosaccharide derived from the agarose and an alpha-agarase specifically acting on the oligosaccharide derived from the agarose. Thereby, the objective oligosaccharide or monosaccharide or both having preventing effects on retrogradation of starch and a product containing the starch are effectively obtained.

6. 发明专利

JPH03151880A MANIFESTATION VECTOR CONTAINING GENE CODING AQUALYCIN I AND HIGHLY THERMOPHILIC GRAM-NEGATIVE BACTERIUM 失效
公开(公告)号：JPH03151880A
公开(公告)日：1991-06-28
申请号：JP28881389
申请日：1989-11-08
申请人： JAPAN TOBACCO INC
发明人： TERADA ICHIRO , OTA TAKAHISA , MATSUZAWA HIROSHI , TAGUCHI HAYAO , HIGASHIHARA NAMI
IPC分类号： C12N15/09 , C12N1/21 , C12N9/52 , C12N15/57 , C12N15/74 , C12R1/01
摘要： PURPOSE:To prevent the contamination of various bacteria by integrating the precursor gene of aqualycin I together with a promoter region and a tail region into a highly thermophilic bacterium plasmid vector. CONSTITUTION:The DNA of Thermus.aquaticus YT-1 (ATC 25104) is perfectly cleaved with restriction enzymes XbaI and SacI into a fragment containing a promoter region, a SD region, a signal region and the N terminal side of a region coding the aqualycin I and a fragment containing the C terminal side of the region coding the aqualycin I and a tail region. The fragments are cloned in vector plasmid pUC 12 to prepare the precursor gene of the aqualycin I. The gene is inserted into a vector pYK 101 for a highly thermophilic gram-negative bacterium to prepare a manifestation vector containing a promoter region active in the highly thermophilic gram-negative bacterium, a SD region existing at the downstream of the promoter region, a signal region, a pro region, a region coding the aqualycin I and a tail region.

7. 发明专利

JPH0292288A GENE CODING AQUALYSIN I PRECURSOR, EXPRESSION VECTOR CONTAINING SAME GENE, E.COLI CONTAINING SAME EXPRESSION VECTOR AND PRODUCTION OF AQUALYSIN I USING SAME E.COLI 失效
公开(公告)号：JPH0292288A
公开(公告)日：1990-04-03
申请号：JP24398188
申请日：1988-09-30
申请人： JAPAN TOBACCO INC
发明人： TERADA ICHIRO , OTA TAKAHISA , MATSUZAWA HIROSHI , KEN SHIYAKUTAI , MIYATA HIROKO
IPC分类号： C12N1/20 , C12N1/21 , C12N9/52 , C12N15/09 , C12N15/57 , C12R1/01 , C12R1/19
摘要： PURPOSE:To produce aqualysin I in a mass at a low cost by using a gene having a signal peptide code region and a pro region at the upper stream of an aqyalysin I structural gene and a tail region at the lower stream of the structural gene and expressing the gene in E.coli. CONSTITUTION:A gene capable of producing a heat-resistant alkaline proteinase aqualysin I useful as an additive for detergent, etc., is produced from E.coli by gene recombination using a gene having a signal peptide code region at the uppermost stream of an aqualysin I structural gene, a pro region at the down stream of the code region and a tail region coding a part inhibiting the activity of aqualysin at the downstream of the aqualysin structural gene. The gene is integrated into an expression vector, an E.coli transformed with the vector is cultured and the bacterial cells are heat-treated to obtain aqualysin I.

8. 发明专利

JPH07322878A NEW ALPHA-AGARASE AND ITS PRODUCTION 失效
公开(公告)号：JPH07322878A
公开(公告)日：1995-12-12
申请号：JP12041994
申请日：1994-06-01
申请人： JAPAN TOBACCO INC
发明人： SUGANO YASUSHI , TERADA ICHIRO , YAMAZAKI YOSHINARI , KODAMA HISASHI , NOMA MASAKATA
IPC分类号： C12N9/24 , C12R1/63
摘要： PURPOSE:To obtain an alpha-agarase, having the activity in hydrolyzing alpha-1,3 bonds in an oligosaccharide derived from agarose and capable of producing an oligosaccharide having a low polymerization degree or an odd number of carbon atoms useful for preventing starch and a product containing the starch from retrograding. CONSTITUTION:This alpha-agarase is obtained from a culture of Vibrio sp. JT0l07-L4 (FERM BP-4541), etc., and capable of hydrolyzing alpha-1,3 bonds between D-galactose of a

9. 发明专利

JPH10234374A SIGNAL PEPTIDE 失效
公开(公告)号：JPH10234374A
公开(公告)日：1998-09-08
申请号：JP4510397
申请日：1997-02-28
申请人： JAPAN TOBACCO INC
发明人： YAMAMOTO TAKESHI , NAKASHIZU MOTOKO , TERADA ICHIRO
IPC分类号： C12N15/09 , C07H21/04 , C07K14/195 , C12N1/21 , C12N9/10 , C12N15/00 , C12R1/01 , C12R1/19
摘要： PROBLEM TO BE SOLVED: To obtain a gene capable of coding the subject peptide useful for secreting a β-galactosido-α 2,6-sialic transferase according to genetic engineering and facilitating the purification. SOLUTION: This signal peptide is a gene capable of coding a signal peptide containing an amino acid sequence of amino acid residue Nos.1 to 15 in an amino acid sequence represented by the formula. The amino acid sequence represented by the formula indicates an ST0160 protein [a sialic transferase 0160 produced by Photobacterium damsela JT0160] and contains an extracellular region of amino acid residue Nos.16 to 498 other than a signal sequence. The gene capable of coding the ST0160 protein can be obtained from a gene library prepared by using a genomic DNA of the JT0160 cultured in a nutrient broth (manufactured by Oxoid Ltd.) by utilizing a plaque hybridization method, etc., and the signal sequence region can be cut off therefrom.

10. 发明专利

JPH10234373A GENE CAPABLE OF CODING BETA-GALACTOSIDO-ALPHA2,6-SIALIC TRANSFERASE 失效
公开(公告)号：JPH10234373A
公开(公告)日：1998-09-08
申请号：JP4508797
申请日：1997-02-28
申请人： JAPAN TOBACCO INC
发明人： YAMAMOTO TAKESHI , NAKASHIZU MOTOKO , TERADA ICHIRO
IPC分类号： C12N15/09 , C07H21/04 , C12N1/21 , C12N9/10 , C12N15/00 , C12N15/54 , C12R1/01 , C12R1/19
摘要： PROBLEM TO BE SOLVED: To obtain the subject gene useful for mass-producing a β-galactosido-α 2,6-sialic transferase according to genetic engineering. SOLUTION: This gene is capable of coding a ST0160 protein having an amino acid sequence represented by the formula [a sialic transferase 0160 produced by Photobacterium damusela JT0160]. The gene can be obtained from a gene library prepared by using a genomic DNA of the JT0160 cultured with a nutrient broth (manufactured by Oxoid Ltd.) by a plaque hybridization method, etc.

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