The Experts below are selected from a list of 216 Experts worldwide ranked by ideXlab platform
John M. Ward - One of the best experts on this subject based on the ideXlab platform.
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The substrate specificity, enantioselectivity and structure of the (R)-selective amine : pyruvate Transaminase from Nectria haematococca
FEBS Journal, 2014Co-Authors: C. Sayer, Ruben J. Martinez-torres, Nina Richter, Michail N. Isupov, Helen C. Hailes, Jennifer A. Littlechild, John M. WardAbstract:During the last decade the use of Transaminases for the production of pharmaceutical and fine chemical intermediates has attracted a great deal of attention. Transaminases are versatile biocatalysts for the efficient production of amine intermediates and many have (S)-enantiospecificity. Transaminases with (R)-specificity are needed to expand the applications of these enzymes in biocatalysis. In this work we have identified a fungal putative (R)-specific Transaminase from the Eurotiomycetes Nectria haematococca, cloned a synthetic version of this gene, demonstrated (R)-selective deamination of several substrates including (R)-α-methylbenzylamine, as well as production of (R)-amines, and determined its crystal structure. The crystal structures of the holoenzyme and the complex with an inhibitor gabaculine offer the first detailed insight into the structural basis for substrate specificity and enantioselectivity of the industrially important class of (R)-selective amine : pyruvate Transaminases. Database The atomic coordinates and structure factors for the Nectria TAm in holoenzyme and gabaculine-bound forms have been deposited in the PDB as entries 4cmd and 4cmf respectively. Structured digital abstract • TAm and TAm bind by x-ray crystallography (View interaction)
J-s Shin - One of the best experts on this subject based on the ideXlab platform.
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Purification, characterization, and molecular cloning of a novel amine:pyruvate Transaminase from Vibrio fluvialis JS17.
Applied microbiology and biotechnology, 2003Co-Authors: J-s Shin, H Yun, J-w Jang, I Park, B-g KimAbstract:A Transaminase from Vibrio fluvialis JS17 showing activity toward chiral amines was purified to homogeneity and its enzymatic properties were characterized. The Transaminase showed an apparent molecular mass of 100 kDa as determined by gel filtration chromatography and a subunit mass of 50 kDa by MALDI-TOF mass spectrometry, suggesting a dimeric structure. The enzyme had an isoelectric point of 5.4 and its absorption spectrum exhibited maxima at 320 and 405 nm. The optimal pH and temperature for enzyme activity were 9.2 and 37 degrees C, respectively. Pyruvate and pyridoxal 5'-phosphate increased enzyme stability whereas (S)-alpha-methylbenzylamine reversibly inactivated the enzyme. The Transaminase gene was cloned from a V. fluvialis JS17 genomic library. The deduced amino acid sequence (453 residues) showed significant homology with omega-amino acid:pyruvate Transaminases (omega-APT) from various bacterial strains (80 identical residues with four omega-APTs). However, of 159 conserved residues in the four omega-APTs, 79 were not conserved in the Transaminase from V. fluvialis JS17. Taken together with the sequence homology results, and the lack of activity toward beta-alanine (a typical amino donor for the omega-APT), the results suggest that the Transaminase is a novel amine:pyruvate Transaminase that has not been reported to date.
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Purification, characterization, and molecular cloning of a novel amine:pyruvate Transaminase from Vibrio fluvialis JS17
Applied Microbiology and Biotechnology, 2003Co-Authors: J-s Shin, H Yun, Jungmin Jang, In-suk Park, Byoung-joon KimAbstract:A Transaminase from Vibrio fluvialis JS17 showing activity toward chiral amines was purified to homogeneity and its enzymatic properties were characterized. The Transaminase showed an apparent molecular mass of 100 kDa as determined by gel filtration chromatography and a subunit mass of 50 kDa by MALDI-TOF mass spectrometry, suggesting a dimeric structure. The enzyme had an isoelectric point of 5.4 and its absorption spectrum exhibited maxima at 320 and 405 nm. The optimal pH and temperature for enzyme activity were 9.2 and 37°C, respectively. Pyruvate and pyridoxal 5′-phosphate increased enzyme stability whereas (S)-α-methylbenzylamine reversibly inactivated the enzyme. The Transaminase gene was cloned from a V. fluvialis JS17 genomic library. The deduced amino acid sequence (453 residues) showed significant homology with ω-amino acid:pyruvate Transaminases (ω-APT) from various bacterial strains (80 identical residues with four ω-APTs). However, of 159 conserved residues in the four ω-APTs, 79 were not conserved in the Transaminase from V. fluvialis JS17. Taken together with the sequence homology results, and the lack of activity toward β-alanine (a typical amino donor for the ω-APT), the results suggest that the Transaminase is a novel amine:pyruvate Transaminase that has not been reported to date.
Byoung-joon Kim - One of the best experts on this subject based on the ideXlab platform.
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Purification, characterization, and molecular cloning of a novel amine:pyruvate Transaminase from Vibrio fluvialis JS17
Applied Microbiology and Biotechnology, 2003Co-Authors: J-s Shin, H Yun, Jungmin Jang, In-suk Park, Byoung-joon KimAbstract:A Transaminase from Vibrio fluvialis JS17 showing activity toward chiral amines was purified to homogeneity and its enzymatic properties were characterized. The Transaminase showed an apparent molecular mass of 100 kDa as determined by gel filtration chromatography and a subunit mass of 50 kDa by MALDI-TOF mass spectrometry, suggesting a dimeric structure. The enzyme had an isoelectric point of 5.4 and its absorption spectrum exhibited maxima at 320 and 405 nm. The optimal pH and temperature for enzyme activity were 9.2 and 37°C, respectively. Pyruvate and pyridoxal 5′-phosphate increased enzyme stability whereas (S)-α-methylbenzylamine reversibly inactivated the enzyme. The Transaminase gene was cloned from a V. fluvialis JS17 genomic library. The deduced amino acid sequence (453 residues) showed significant homology with ω-amino acid:pyruvate Transaminases (ω-APT) from various bacterial strains (80 identical residues with four ω-APTs). However, of 159 conserved residues in the four ω-APTs, 79 were not conserved in the Transaminase from V. fluvialis JS17. Taken together with the sequence homology results, and the lack of activity toward β-alanine (a typical amino donor for the ω-APT), the results suggest that the Transaminase is a novel amine:pyruvate Transaminase that has not been reported to date.
B-g Kim - One of the best experts on this subject based on the ideXlab platform.
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Purification, characterization, and molecular cloning of a novel amine:pyruvate Transaminase from Vibrio fluvialis JS17.
Applied microbiology and biotechnology, 2003Co-Authors: J-s Shin, H Yun, J-w Jang, I Park, B-g KimAbstract:A Transaminase from Vibrio fluvialis JS17 showing activity toward chiral amines was purified to homogeneity and its enzymatic properties were characterized. The Transaminase showed an apparent molecular mass of 100 kDa as determined by gel filtration chromatography and a subunit mass of 50 kDa by MALDI-TOF mass spectrometry, suggesting a dimeric structure. The enzyme had an isoelectric point of 5.4 and its absorption spectrum exhibited maxima at 320 and 405 nm. The optimal pH and temperature for enzyme activity were 9.2 and 37 degrees C, respectively. Pyruvate and pyridoxal 5'-phosphate increased enzyme stability whereas (S)-alpha-methylbenzylamine reversibly inactivated the enzyme. The Transaminase gene was cloned from a V. fluvialis JS17 genomic library. The deduced amino acid sequence (453 residues) showed significant homology with omega-amino acid:pyruvate Transaminases (omega-APT) from various bacterial strains (80 identical residues with four omega-APTs). However, of 159 conserved residues in the four omega-APTs, 79 were not conserved in the Transaminase from V. fluvialis JS17. Taken together with the sequence homology results, and the lack of activity toward beta-alanine (a typical amino donor for the omega-APT), the results suggest that the Transaminase is a novel amine:pyruvate Transaminase that has not been reported to date.
H Yun - One of the best experts on this subject based on the ideXlab platform.
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Purification, characterization, and molecular cloning of a novel amine:pyruvate Transaminase from Vibrio fluvialis JS17.
Applied microbiology and biotechnology, 2003Co-Authors: J-s Shin, H Yun, J-w Jang, I Park, B-g KimAbstract:A Transaminase from Vibrio fluvialis JS17 showing activity toward chiral amines was purified to homogeneity and its enzymatic properties were characterized. The Transaminase showed an apparent molecular mass of 100 kDa as determined by gel filtration chromatography and a subunit mass of 50 kDa by MALDI-TOF mass spectrometry, suggesting a dimeric structure. The enzyme had an isoelectric point of 5.4 and its absorption spectrum exhibited maxima at 320 and 405 nm. The optimal pH and temperature for enzyme activity were 9.2 and 37 degrees C, respectively. Pyruvate and pyridoxal 5'-phosphate increased enzyme stability whereas (S)-alpha-methylbenzylamine reversibly inactivated the enzyme. The Transaminase gene was cloned from a V. fluvialis JS17 genomic library. The deduced amino acid sequence (453 residues) showed significant homology with omega-amino acid:pyruvate Transaminases (omega-APT) from various bacterial strains (80 identical residues with four omega-APTs). However, of 159 conserved residues in the four omega-APTs, 79 were not conserved in the Transaminase from V. fluvialis JS17. Taken together with the sequence homology results, and the lack of activity toward beta-alanine (a typical amino donor for the omega-APT), the results suggest that the Transaminase is a novel amine:pyruvate Transaminase that has not been reported to date.
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Purification, characterization, and molecular cloning of a novel amine:pyruvate Transaminase from Vibrio fluvialis JS17
Applied Microbiology and Biotechnology, 2003Co-Authors: J-s Shin, H Yun, Jungmin Jang, In-suk Park, Byoung-joon KimAbstract:A Transaminase from Vibrio fluvialis JS17 showing activity toward chiral amines was purified to homogeneity and its enzymatic properties were characterized. The Transaminase showed an apparent molecular mass of 100 kDa as determined by gel filtration chromatography and a subunit mass of 50 kDa by MALDI-TOF mass spectrometry, suggesting a dimeric structure. The enzyme had an isoelectric point of 5.4 and its absorption spectrum exhibited maxima at 320 and 405 nm. The optimal pH and temperature for enzyme activity were 9.2 and 37°C, respectively. Pyruvate and pyridoxal 5′-phosphate increased enzyme stability whereas (S)-α-methylbenzylamine reversibly inactivated the enzyme. The Transaminase gene was cloned from a V. fluvialis JS17 genomic library. The deduced amino acid sequence (453 residues) showed significant homology with ω-amino acid:pyruvate Transaminases (ω-APT) from various bacterial strains (80 identical residues with four ω-APTs). However, of 159 conserved residues in the four ω-APTs, 79 were not conserved in the Transaminase from V. fluvialis JS17. Taken together with the sequence homology results, and the lack of activity toward β-alanine (a typical amino donor for the ω-APT), the results suggest that the Transaminase is a novel amine:pyruvate Transaminase that has not been reported to date.
