Kiyoshi Yasukawa

Summary

Affiliation: Kyoto University
Country: Japan

Publications

  1. doi request reprint Characterization of Moloney murine leukaemia virus/avian myeloblastosis virus chimeric reverse transcriptases
    Kiyoshi Yasukawa
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    J Biochem 145:315-24. 2009
  2. ncbi request reprint Insight into the mechanism of the stabilization of moloney murine leukaemia virus reverse transcriptase by eliminating RNase H activity
    Masaki Mizuno
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Japan
    Biosci Biotechnol Biochem 74:440-2. 2010
  3. ncbi request reprint Effects of organic solvents on the reverse transcription reaction catalyzed by reverse transcriptases from avian myeloblastosis virus and Moloney murine leukemia virus
    Kiyoshi Yasukawa
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan
    Biosci Biotechnol Biochem 74:1925-30. 2010
  4. doi request reprint Kinetic analysis of reverse transcriptase activity of bacterial family A DNA polymerases
    Kiyoshi Yasukawa
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto 606 8502, Japan
    Biochem Biophys Res Commun 427:654-8. 2012
  5. ncbi request reprint Comparison of the thermal stabilities of reverse transcriptases from avian myeloblastosis virus and Moloney murine leukaemia virus
    Kiyoshi Yasukawa
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto, Japan
    J Biochem 143:261-8. 2008
  6. doi request reprint Increase in thermal stability of Moloney murine leukaemia virus reverse transcriptase by site-directed mutagenesis
    Kiyoshi Yasukawa
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kitashirakawa, Sakyo ku, Kyoto 606 8502, Japan
    J Biotechnol 150:299-306. 2010
  7. doi request reprint Effects of the mutational combinations on the activity and stability of thermolysin
    Masayuki Kusano
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    J Biotechnol 147:7-16. 2010
  8. doi request reprint Effects of introducing negative charges into the molecular surface of thermolysin by site-directed mutagenesis on its activity and stability
    Teisuke Takita
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    Biochim Biophys Acta 1784:481-8. 2008
  9. ncbi request reprint Effects of mutations of thermolysin, as N116 to asp and asp150 to glu, on salt-induced activation and stabilization
    Evans Menach
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan
    Biosci Biotechnol Biochem 77:741-6. 2013
  10. pmc Effects of site-directed mutagenesis in the N-terminal domain of thermolysin on its stabilization
    Yuichi Kawasaki
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto 606 8502, Japan
    J Biochem 153:85-92. 2013

Collaborators

Detail Information

Publications51

  1. doi request reprint Characterization of Moloney murine leukaemia virus/avian myeloblastosis virus chimeric reverse transcriptases
    Kiyoshi Yasukawa
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    J Biochem 145:315-24. 2009
    ....
  2. ncbi request reprint Insight into the mechanism of the stabilization of moloney murine leukaemia virus reverse transcriptase by eliminating RNase H activity
    Masaki Mizuno
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Japan
    Biosci Biotechnol Biochem 74:440-2. 2010
    ..These results suggest that elimination of RNase H activity enhanced the intrinsic thermal stability of MMLV RT rather than its affinity toward T/P...
  3. ncbi request reprint Effects of organic solvents on the reverse transcription reaction catalyzed by reverse transcriptases from avian myeloblastosis virus and Moloney murine leukemia virus
    Kiyoshi Yasukawa
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan
    Biosci Biotechnol Biochem 74:1925-30. 2010
    ..The effects of DMSO and formamide appeared to be accompanied by decreases in the melting temperatures of the primers, and the effect of glycerol was due to increases in the thermal stabilities of AMV RT and MMLV RT...
  4. doi request reprint Kinetic analysis of reverse transcriptase activity of bacterial family A DNA polymerases
    Kiyoshi Yasukawa
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto 606 8502, Japan
    Biochem Biophys Res Commun 427:654-8. 2012
    ..This suggests that high concentrations of DNA-primed RNA template decrease the efficiency of cDNA synthesis with bacterial family A DNA polymerases...
  5. ncbi request reprint Comparison of the thermal stabilities of reverse transcriptases from avian myeloblastosis virus and Moloney murine leukaemia virus
    Kiyoshi Yasukawa
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto, Japan
    J Biochem 143:261-8. 2008
    ..Thermodynamic analysis indicates that thermal inactivation of AMV RT and MMLV RT is due to the large entropy change of activation for thermal inactivation...
  6. doi request reprint Increase in thermal stability of Moloney murine leukaemia virus reverse transcriptase by site-directed mutagenesis
    Kiyoshi Yasukawa
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kitashirakawa, Sakyo ku, Kyoto 606 8502, Japan
    J Biotechnol 150:299-306. 2010
    ..The highest temperature at which the triple and quadruple variants exhibited cDNA synthesis activity was 60°C, which was again higher than for WT (54°C). Thus, highly stable MMLV RT variants were generated by this mutation strategy...
  7. doi request reprint Effects of the mutational combinations on the activity and stability of thermolysin
    Masayuki Kusano
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    J Biotechnol 147:7-16. 2010
    ..Its k(cat)/K(m) values in the hydrolysis of FAGLA and ZDFM were 8.6 and 10.2 times higher than those of wild-type thermolysin (WT), respectively, and its rate constant for thermal inactivation at 80 degrees C was 60% of that of WT...
  8. doi request reprint Effects of introducing negative charges into the molecular surface of thermolysin by site-directed mutagenesis on its activity and stability
    Teisuke Takita
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    Biochim Biophys Acta 1784:481-8. 2008
    ..The stability of WT was enhanced by CaCl(2) and reached the level of S53D and S65D at 100 mM, suggesting that S53D and S65D might be stabilized by reinforcement of the Ca(2+)-binding structures...
  9. ncbi request reprint Effects of mutations of thermolysin, as N116 to asp and asp150 to glu, on salt-induced activation and stabilization
    Evans Menach
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan
    Biosci Biotechnol Biochem 77:741-6. 2013
    ..These results indicate that unlike Asn116→Asp, Asp150→Glu reduced NaCl-induced activation and stabilization, suggesting that the binding of ions with certain residues of thermolysin is involved in the activation and stabilization...
  10. pmc Effects of site-directed mutagenesis in the N-terminal domain of thermolysin on its stabilization
    Yuichi Kawasaki
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto 606 8502, Japan
    J Biochem 153:85-92. 2013
    ..9, 17 and 7.2 mM, respectively, suggesting that Gly8→Cys/Asn60→Cys and Ser65→Pro stabilize thermolysin by improving its affinity to calcium ions, most probably the one at the Ca²⁺-binding site III in the N-terminal domain...
  11. doi request reprint Effects of amines and aminoalcohols on bovine intestine alkaline phosphatase activity
    Satoshi Sekiguchi
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto 606 8502, Japan
    Enzyme Microb Technol 49:171-6. 2011
    ..2M (0.18±0.02 mM), while that at 1.0M triethanolamine (0.31±0.01 mM) was similar as that at 0.2M (0.25±0.01 mM), suggesting that the mechanisms of BIALP activation are different between the aminoalcohols...
  12. ncbi request reprint A new method for the extracellular production of recombinant thermolysin by co-expressing the mature sequence and pro-sequence in Escherichia coli
    Kiyoshi Yasukawa
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    Protein Eng Des Sel 20:375-83. 2007
    ..The results also demonstrate clearly that the covalent linking between the pro- and mature sequences is not necessary for the proper folding of the mature sequence by the propeptide in thermolysin...
  13. ncbi request reprint Effects of site-directed mutagenesis of the surface residues Gln128 and Gln225 of thermolysin on its catalytic activity
    Chika Tatsumi
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    J Biochem 141:835-42. 2007
    ..Thus, site-directed mutagenesis of surface residues of thermolysin, including apparently thermodynamically unfavorable introduction of hydrophobic residues, should be explored to improve its activity and stability...
  14. ncbi request reprint Improving the activity and stability of thermolysin by site-directed mutagenesis
    Kiyoshi Yasukawa
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    Biochim Biophys Acta 1774:1281-8. 2007
    ..Thermodynamic analysis suggests that the single mutation of Leu144-->Ser and the triple mutation of Gly8-->Cys, Asn60-->Cys, and Ser65-->Pro are independent...
  15. doi request reprint Effects of site-directed mutagenesis of Asn116 in the β-hairpin of the N-terminal domain of thermolysin on its activity and stability
    Evans Menach
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    J Biochem 152:231-9. 2012
    ..These results suggest that Asn116 plays an important role in the activity and stability of thermolysin presumably by stabilizing this β-hairpin structure...
  16. ncbi request reprint Stabilization of bovine intestine alkaline phosphatase by sugars
    Satoshi Sekiguchi
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto, Japan
    Biosci Biotechnol Biochem 76:95-100. 2012
    ..These results suggest that not only trehalose, which is considered the most effective stabilizer of enzymes, but also sucrose, glucose, and fructose can be used as stabilizers of BIALP...
  17. ncbi request reprint Engineering, expression, purification, and production of recombinant thermolysin
    Kuniyo Inouye
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    Biotechnol Annu Rev 13:43-64. 2007
    ..In this review, we aim to outline recombinant thermolysins associated with their engineering, expression, purification, and production...
  18. ncbi request reprint Improvement in performance of affinity gels containing Gly-D-Phe as a ligand to thermolysin due to increasing the spacer chain length
    Kuniyo Inouye
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan
    Biosci Biotechnol Biochem 71:2083-6. 2007
    ..These results indicate that the performance of affinity gels is improved by increasing the spacer chain length to 13 carbon-and-oxygen atoms...
  19. ncbi request reprint Effects of conversion of the zinc-binding motif sequence of thermolysin, HEXXH, to that of dipeptidyl peptidase III, HEXXXH, on the activity and stability of thermolysin
    Evans Menach
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University
    Biosci Biotechnol Biochem 77:1901-6. 2013
    ..Our results are in contrast to ones reported previously, that DPP III variants bearing H(450)ELGH(455) exhibit activity. ..
  20. ncbi request reprint Effects of site-directed mutagenesis of the loop residue of the N-terminal domain Gly117 of thermolysin on its catalytic activity
    Evans Menach
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan
    Biosci Biotechnol Biochem 74:2457-62. 2010
    ..This suggests that Gly117 plays an important role in the activity and stability of thermolysin, presumably by affecting the geometries of the Asn112-Trp115 and Ser118-Tyr122 strands...
  21. ncbi request reprint Engineering of the pH-dependence of thermolysin activity as examined by site-directed mutagenesis of Asn112 located at the active site of thermolysin
    Masayuki Kusano
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502
    J Biochem 139:1017-23. 2006
    ....
  22. ncbi request reprint Comparison of the thermal stabilities of the αβ heterodimer and the α subunit of avian myeloblastosis virus reverse transcriptase
    Atsushi Konishi
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto, Japan
    Biosci Biotechnol Biochem 75:1618-20. 2011
    ..Unlike the αβ heterodimer, the α subunit was not stabilized by template-primer. These results suggest that interaction between the α and β subunits is important for AMV RT stability...
  23. doi request reprint Insights into the catalytic roles of the polypeptide regions in the active site of thermolysin and generation of the thermolysin variants with high activity and stability
    Masayuki Kusano
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    J Biochem 145:103-13. 2009
    ..Thermal inactivation of thermolysin at 80 degrees C was greatly suppressed with the D150H, D150W, I168A, I168H, N227A, N227H and S234A. The evidence might provide the insights into the activation and stabilization of thermolysin...
  24. ncbi request reprint The cleavage site preference of the porcine pepsin on the N-terminal α1 chain of bovine type I collagen: a focal analysis with mass spectrometry
    Jun Qian
    a Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan
    Biosci Biotechnol Biochem . 2016
    ..These results suggest that pepsin preferentially cleaves Tyr6-Asp7 and less preferentially Gly12-Ile13. They also suggest that the former cleavage requires native collagen structure, while the latter cleavage does not...
  25. doi request reprint Improving the thermal stability of avian myeloblastosis virus reverse transcriptase α-subunit by site-directed mutagenesis
    Atsushi Konishi
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    Biotechnol Lett 34:1209-15. 2012
    ....
  26. doi request reprint Thermodynamic analysis of ionizable groups involved in the catalytic mechanism of human matrix metalloproteinase 7 (MMP-7)
    Hitoshi Takeharu
    Kyoto University, Kyoto, Japan
    Biochim Biophys Acta 1814:1940-6. 2011
    ..On the other hand, no arginine or lysine residues are found in the active site of MMP-7. We proposed a possibility that a protein-bound water is the ionizable group for pK(e2)...
  27. doi request reprint Two crystal structures of lysyl-tRNA synthetase from Bacillus stearothermophilus in complex with lysyladenylate-like compounds: insights into the irreversible formation of the enzyme-bound adenylate of L-lysine hydroxamate
    Haruko Sakurama
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan
    J Biochem 145:555-63. 2009
    ..In support of other structural comparisons, the result suggested that Glu411 plays a key role in the arrangement of PPi for the nucleophilic attack...
  28. ncbi request reprint Effects of neutral salts and alcohols on the activity of Streptomyces caespitosus neutral protease
    Kuniyo Inouye
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan
    J Biochem 142:317-24. 2007
    ..The activities recovered completely by the dilution of alcohols, suggesting that the ScNP inhibition by alcohols is reversible. These characteristics of ScNP are compared with those of human matrix metalloproteinase 7 and thermolysin...
  29. doi request reprint Expression of moloney murine leukemia virus reverse transcriptase in a cell-free protein expression system
    Yuta Katano
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan
    Biotechnol Lett 38:1203-11. 2016
    ..To characterize Moloney murine leukemia virus (MMLV) reverse transcriptases (RTs) expressed in a cell-free system and in Escherichia coli...
  30. doi request reprint Inhibition of the DNA polymerase and RNase H activities of HIV-1 reverse transcriptase and HIV-1 replication by Brasenia schreberi (Junsai) and Petasites japonicus (Fuki) components
    Tetsuro Hisayoshi
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto, 606 8502, Japan
    J Nat Med 69:432-40. 2015
    ..These results suggest that Brasenia schreberi and Petasites japonicus contain substances that block HIV-1 replication by inhibiting the DNA polymerase activity and/or RNase H activity of HIV-1 RT. ..
  31. doi request reprint Preparation and characterization of the RNase H domain of Moloney murine leukemia virus reverse transcriptase
    Kosaku Nishimura
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    Protein Expr Purif 113:44-50. 2015
    ..The activity levels of WT, ΔC1, and ΔC2 were estimated to be 1%, 0.01%, and 0.01% of full-length MMLV RT activity, indicating that the C helix is important, but not critical, for the activity of the isolated RNase H domain...
  32. doi request reprint Enzymatic characterization of human immunodeficiency virus type 1 reverse transcriptase for use in cDNA synthesis
    Atsushi Konishi
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    Appl Biochem Biotechnol 169:77-87. 2013
    ..The high substrate affinity and low susceptibility to formamide of HIV-1 RT might be advantageous for its use in cDNA synthesis...
  33. ncbi request reprint Effects of polyethylene glycol on bovine intestine alkaline phosphatase activity and stability
    Satoshi Sekiguchi
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan
    Biosci Biotechnol Biochem 75:2132-7. 2011
    ..These results indicate that the addition of PEG2000 and pegylation increase BIALP activity without affecting its stability, suggesting that they can be used in enzyme immunoassay with BIALP to increase sensitivity and rapidity...
  34. ncbi request reprint Analysis of the mechanism of inhibition of human matrix metalloproteinase 7 (MMP-7) activity by green tea catechins
    Tomoko Miyake
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan
    Biosci Biotechnol Biochem 75:1564-9. 2011
    ..Our results suggest that these differences result from that in the B ring, EGCG and GCG have phenol hydroxyl groups at the 3', 4', and 5' positions, while ECG and CG have them at the 3' and 4' positions...
  35. pmc Rapid detection of Mycobacterium tuberculosis in respiratory samples by transcription-reverse transcription concerted reaction with an automated system
    Shunji Takakura
    Department of Clinical Laboratory Medicine, Kyoto University Graduate School of Medicine, 54 Shogoin Kawaharacho, Sakyo ku, Kyoto 6068507, Japan
    J Clin Microbiol 43:5435-9. 2005
    ..The high sensitivity and specificity together with shorter detection time (within 1 h) of the TRC method allow it to be proposed as a useful method for the rapid detection of MTC in respiratory samples...
  36. pmc Isothermal RNA sequence amplification method for rapid antituberculosis drug susceptibility testing of Mycobacterium tuberculosis
    Shunji Takakura
    Department of Clinical Laboratory Medicine, Graduate School of Medicine, Kyoto University, 54 Shogoin Kawaharacho, Kyoto, 606 8507, Japan
    J Clin Microbiol 43:2489-91. 2005
    ..Agreement with the proportion method on Ogawa egg medium and the BACTEC MGIT 960 system was 100 and 87% for rifampin, 93 and 100% for isoniazid, 60 and 53% for ethambutol, and 80 and 80% for streptomycin, respectively...
  37. doi request reprint Effects of thermal treatment on the coagulation of soy proteins induced by subtilisin Carlsberg
    Kuniyo Inouye
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Japan
    J Agric Food Chem 57:717-23. 2009
    ..These findings suggest that the thermal treatment at 80 degrees C most effectively changes the secondary structure of soy proteins and renders them coagulate when hydrolyzed by subtilisin Carlsberg...
  38. ncbi request reprint Characterization of Gly-D-Phe, Gly-L-Leu, and D-Phe as affinity ligands to thermolysin
    Kiyoshi Yasukawa
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto 606 8502, Japan
    Protein Expr Purif 46:332-6. 2006
    ..These results indicate that Gly-D-Phe is more suitable than Gly-L-Leu and D-Phe as an affinity ligand for purification of thermolysin...
  39. doi request reprint Enhanced detection of RNA by MMLV reverse transcriptase coupled with thermostable DNA polymerase and DNA/RNA helicase
    Hiroyuki Okano
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    Enzyme Microb Technol 96:111-120. 2017
    ..Our results suggest that DNA polymerase with RT activity and DNA/RNA helicase are useful to increase the sensitivity of cDNA synthesis...
  40. doi request reprint Effects of salts on the interaction of 8-anilinonaphthalene 1-sulphonate and thermolysin
    Vimbai Samukange
    a Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Japan
    Biosci Biotechnol Biochem 78:1522-8. 2014
    ..Our results also suggest that the activation and stabilization of thermolysin by NaCl are partially brought about by the binding of Na(+) and/or Cl(-) ions with thermolysin. ..
  41. doi request reprint Effects of heparin and cholesterol sulfate on the activity and stability of human matrix metalloproteinase 7
    Vimbai Samukange
    a Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan
    Biosci Biotechnol Biochem 78:41-8. 2014
    ..These results indicate that heparin increases the activity and stability of MMP-7, while CS decreases them...
  42. doi request reprint Interaction of 8-anilinonaphthalene 1-sulphonate (ANS) and human matrix metalloproteinase 7 (MMP-7) as examined by MMP-7 activity and ANS fluorescence
    Vimbai Samukange
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    J Biochem 151:533-40. 2012
    ..These results suggest that at pH 4.5-9.5, the sulphonic group of ANS binds with MMP-7 through electrostatic interaction, whereas at pH 3.5, the anilinonaphthalene group of ANS binds with MMP-7 through hydrophobic interaction...
  43. ncbi request reprint Extracellular production of recombinant thermolysin expressed in Escherichia coli, and its purification and enzymatic characterization
    Kuniyo Inouye
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto 606 8502, Japan
    Protein Expr Purif 46:248-55. 2006
    ..The results demonstrate that our expression system should be useful for structural and functional analysis of thermolysin...
  44. ncbi request reprint Purification to homogeneity of a neutral metalloproteinase from Streptomyces caespitosus
    Kuniyo Inouye
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan
    Biosci Biotechnol Biochem 71:1773-6. 2007
    ..We also purified another protease with a molecular mass of 40 kDa from the culture supernatants. The pure preparation of ScNP obtained by this procedure is suitable for spectrophotometric measurement of its catalytic activity...
  45. pmc Rapid and specific detection of tdh, trh1, and trh2 mRNA of Vibrio parahaemolyticus by transcription-reverse transcription concerted reaction with an automated system
    Yoshitsugu Nakaguchi
    Graduate School of Medicine, Center for Southeast Asian Studies, Kyoto University, 46 Shimoadachi cho, Yoshida, Sakyo ku, Kyoto 606 8501, Japan
    J Clin Microbiol 42:4284-92. 2004
    ..Interestingly, the levels of TDH produced from the strains carrying both tdh and trh genes were lower than those carrying only the tdh gene, whereas the levels of mRNA did not significantly differ between the two groups...
  46. doi request reprint Kinetic Analysis of the Digestion of Bovine Type I Collagen Telopeptides with Porcine Pepsin
    Jun Qian
    Authors Qian, Okada, Ogura, Tanaka, Hattori, Ito, Satoh, Takita, and Yasukawa are with Div of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto Univ, Sakyo ku, Kyoto 606 8502, Japan
    J Food Sci 81:C27-34. 2016
    ..1 to 4.5 and 30 to 40 °C, respectively. These results suggest that the method might be useful for the kinetic analysis of the digestion of collagen telopeptides with pepsin. ..
  47. doi request reprint Amino acid substitutions away from the RNase H catalytic site increase the thermal stability of Moloney murine leukemia virus reverse transcriptase through RNase H inactivation
    Atsushi Konishi
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    Biochem Biophys Res Commun 454:269-74. 2014
    ..These results suggest that the mutations, E286R, E302K, and L435R increase the thermostability of MMLV RT not by increasing its affinity for T/P but by abolishing its RNase H activity...
  48. doi request reprint Stabilization of Moloney murine leukemia virus reverse transcriptase by site-directed mutagenesis of surface residue Val433
    Atsushi Konishi
    a Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan
    Biosci Biotechnol Biochem 78:75-8. 2014
    ....
  49. doi request reprint A possibility of a protein-bound water molecule as the ionizable group responsible for pKe at the alkaline side in human matrix metalloproteinase 7 activity
    Aiko Morishima
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo ku, Kyoto 606 8502, Japan
    J Biochem 151:501-9. 2012
    ..3-0.5 pH unit, and those of P217S, P217V and P217G were 10.1 ± 0.1, 9.8 ± 0.1 and 9.7 ± 0.1, respectively, different from that of WT by 0.1-0.2 pH unit. These results suggest a possibility of W1 or W2 as the ionizable group for pK(e2)...
  50. ncbi request reprint Chemiluminescent enzyme immunoassay for measuring leptin
    Satoshi Sekiguchi
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Japan
    Biosci Biotechnol Biochem 75:752-6. 2011
    ..The results also suggest that a sensitive enzyme immunoassay can be constructed by using only one polyclonal antibody...
  51. ncbi request reprint Intercalation activating fluorescence DNA probe and its application to homogeneous quantification of a target sequence by isothermal sequence amplification in a closed vessel
    Takahiko Ishiguro
    Tokyo Research Laboratories, Tosoh Corporation, 2743 1 Hayakawa, Ayase shi, Kanagawa 252 1123, Japan
    Anal Biochem 314:77-86. 2003
    ..bovis BCG during cultivation with antibiotics. The present method can potentially realize rapid antimicrobial susceptibility testing of slowly growing organisms, such as tuberculosis...