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1. 发明专利

JPH10327869A CHAPERONINE ALPHA AND BETA SUBUNIT GENE DERIVED FROM SUPERTHERMOPHILIC MICROORGANISM BELONGING TO GENUS THERMOCOCCUS 失效
公开(公告)号：JPH10327869A
公开(公告)日：1998-12-15
申请号：JP14170697
申请日：1997-05-30
申请人： KAIYO BIO TECH LAB
发明人： IIDA TOSHIMOCHI , MARUYAMA TADASHI , YOSHIDA TAKAO , YOUDA MASABUMI , ENDO ISAO , KAGAWA YASUO
IPC分类号： C12N15/09 , C07K14/47 , C12N1/21 , C12N15/00 , C12P21/02 , C12R1/01 , C12R1/91
摘要： PROBLEM TO BE SOLVED: To obtain the subject new protein, comprising a chaperonine α subunit, etc., having a specific amino acid sequence, having high stability and useful as a stabilizer for other proteins and for an improvement, etc., in the recovery ratio of a soluble fraction of an expressed protein. SOLUTION: This new gene is capable of coding a protein, comprising an amino acid sequence described in the formula or an amino acid sequence in which one or several amino acids are deleted, substituted or added in the amino acid sequence described in the formula and having functions simular to those of a chaperonine α subunit. The protein prepared from the gene is useful as a feed source for a reagent having high stability and a stabilizer for other coexisting proteins or for an improvement, etc., in the recovery ratio of a soluble fraction of proteins produced by a genetic engineering technique. The gene is obtained by separating a genomic DNA from a microorganism of the genus Thermococcus, preparing a DNA library and screening the resultant library with a chaperonine gene fragment prepared by a polymerase chain reactional(PCR) method.

2. 发明专利

JPH09238684A HEAT RESISTANT ATPASE ALPHA AND BETA SUBUNIT GENE 失效
公开(公告)号：JPH09238684A
公开(公告)日：1997-09-16
申请号：JP4926196
申请日：1996-03-06
申请人： KAIYO BIO TECH LAB
发明人： IIDA TOSHIMOCHI , SUZUKI KIYOKO , MARUYAMA TADASHI
IPC分类号： C12N15/09 , C07H21/04 , C12N9/14 , C12N15/00 , C12R1/01
摘要： PROBLEM TO BE SOLVED: To obtain new subject genes each having a specific base arrangement, coding for a heat resistant ATPase subunit and useful as an ATP supplying enzyme in a heat resistant reagent and a bioreactor. SOLUTION: The new genes possess DNA base sequences expressed by formulas I and II, or DNA base sequences essentially equal to those expressed by formulas I and II and code for a heat resistant ATPase α-subunit and β-subunit respectively and useful for producing ATPase useful as an ATP supplying enzyme in a heat resistant reagent and a bioreactor. The genes are obtained by extracting genome DNA from Thermococcus KI strain, a super thermophilic sulfur dependent bacteria, in a usual method and cloning by PCR by using a primer comprising a part of heat resistant ATPase α-subunit and β-subunit and the genome DNA as a template.

3. 发明专利

JPH08131167A HEAT-RESISTANT ATPASE AND ITS PRODUCTION 失效
公开(公告)号：JPH08131167A
公开(公告)日：1996-05-28
申请号：JP27945194
申请日：1994-11-14
申请人： KAIYO BIO TECH LAB
发明人： IIDA TOSHIMOCHI , HOAKI TOSHIHIRO , MARUYAMA TADASHI
IPC分类号： C12N9/14 , C12R1/01
摘要： PURPOSE: To obtain the subject highly thermally stable new enzyme having high optimal temperature, useful for reagents or bioreactors, by culturing microbes belonging to the genus Thermococcus and having heat-resistant ATPase productivity. CONSTITUTION: Microbes belonging to the genus Thermococcus and having ATPase productivity [e.g. Thermococcus K1 strain (FERM P-14622) ] is subjected to stationary culture anaerobically in a medium at 90 deg.C for 8hr followed by centrifugation to collect the resultant microbes, which are then crushed in a buffer solution followed by cetrifugation to separate a membrane fraction. The membrane fraction is put to extraction with a surfactant-contg. buffer solution, and the resultant extract is purified by column chromatography, thus obtaining the objective new heat-resistant ATPase having the following characteristics: hydrolyzing nucleotide triphosphate into nucleotide diphosphate and inorganic phosphoric acid; requiring Mg , Mn , Co , Ni , Zn , Ca as cofactor; rising activity by HSO 3 , S2 O 4 , HCO 3 ; optimal pH: 6-6.5; optimal temperature; 90-95 deg.C; molecular weight: 600kDa.

4. 发明专利

JPH09313184A CYCLOPHILIN TYPE PPI-ASE GENE ORIGINATING FROM HALOPHILIC ARCHAEBACTERIUM 失效
公开(公告)号：JPH09313184A
公开(公告)日：1997-12-09
申请号：JP13335396
申请日：1996-05-28
申请人： KAIYO BIO TECH LAB
发明人： IIDA TOSHIMOCHI , FURUYA MASAHIRO , SUZUKI KIYOKO , IWABUCHI NORIO , MARUYAMA TADASHI
IPC分类号： C12N15/09 , C12N9/90
摘要： PROBLEM TO BE SOLVED: To obtain a novel cyclophilin type PPI-ase gene which has a specific base sequence, codes a cyclophilin type PPI-ase originating from halophilic archaebacteria and can produce PPI-ase useful in regeneration of denatured protein, stabilization of proteins and for immunosuppressant. SOLUTION: This gene is a novel gene of cyclophilin type PPI-ase (peptidyl prolyl cis-trans isomerase) originating from halophilic archaebacteria having the DNA base sequence represented by the formula or a substantially identical DNA base sequence to the sequence represented by the formula and is used in the production of PPI-ase which is useful for regeneration of denatured proteins, stabilization of protein reagents, production of recombinant protein, development of novel immunosuppressant and physiologically active substances. This gene is obtained by amplifying the genome DNA of Halobacterium cutirubrum DSM669 strain, one of archaebacteria, with the PCR technique using a primer comprising a part of the gene of PPI-ase followed by cloning.

5. 发明专利

JPH1175855A HEAT-RESISTANT F TYPE ATPASE AND GENE CODING FOR THE SAME 失效
公开(公告)号：JPH1175855A
公开(公告)日：1999-03-23
申请号：JP25092697
申请日：1997-09-16
申请人： KAIYO BIO TECH LAB
发明人： IIDA TOSHIMOCHI , SUZUKI KIYOKO , MARUYAMA TADASHI
IPC分类号： C12N15/09 , C07K14/195 , C12N9/14 , C12N15/00 , C12R1/01
摘要： PROBLEM TO BE SOLVED: To obtain the subject new enzyme, composed of protein having a plurality of ATPase sub-units containing a specific amino acid sequence, resistant to heat, and useful as the ATP supply source or the like for reagents and bio-reactors to be supplied with ATP. SOLUTION: This new heat-resistant enzyme of F type ATPase is composed of 3 types of protein having an amino acid sequence shown by formula I, II or III, one or more amino acid types of which may be lost, substituted or added, and functioning as the sub-units for ATPase. It has the most suitable reaction temperature of around 80 deg.C, is retarded by NBD-Cl, DCCD or an azide, and useful as the ATP-supplying enzyme or the like for heat-resistant reagents and bioreactors. It is obtained by a series of steps of (1) culturing a Thermotoga neapolitana DSM4359 strain, (2) collecting and crushing the bacteria grown by a ultrasonic crusher, (3) centrifugal separation of the effluent, (4) ultracentrifugal separation of the supernatant liquid, and (5) recovering the precipitates corresponding to the membrane fractions.

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