TY - JOUR
T1 - Gene cloning and characterization of aldehyde dehydrogenase from a petroleum-degrading bacterium, strain HD-1
AU - Okibe, Naoko
AU - Amada, Kei
AU - Hirano, Shin Ichi
AU - Haruki, Mitsuru
AU - Imanaka, Tadayuki
AU - Morikawa, Masaaki
AU - Kanaya, Shigenori
N1 - Funding Information:
This work was supported by a grant from the Program for Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN).
Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 1999
Y1 - 1999
N2 - The hd-ald gene encoding aldehyde dehydrogenase (hd-ALDH) from an mixotrophic petroleum-degrading bacterium, strain HD-1 was cloned and sequenced, hd-ALDH (506 amino acids) is a member of the NAD+-dependent aldehyde dehydrogenase group. The hd-ald gene was expressed in Escherichia coli, and the recombinant enzyme was purified and characterized biochemically and enzymatically. The molecular weight of the enzyme was estimated to be 55,000 by SDS-PAGE, and 224,000 by gel filtration chromatography, suggesting that it acts as a tetramer. The CD spectrum suggests that the helical content of the enzyme is 10%. hd-ALDH was active on various aliphatic aldehyde substrates. The K(m) values of the enzyme were 6.4 μM for acetaldehyde, 4.2 μM for hexanal, 2.8 μM for octanal, and 0.84 μM for decanal, whereas the k(cat) values for these substrates were nearly equal (51-64 min-1). These results indicate that hd-ALDH acts preferentially on long-chain aliphatic aldehydes.
AB - The hd-ald gene encoding aldehyde dehydrogenase (hd-ALDH) from an mixotrophic petroleum-degrading bacterium, strain HD-1 was cloned and sequenced, hd-ALDH (506 amino acids) is a member of the NAD+-dependent aldehyde dehydrogenase group. The hd-ald gene was expressed in Escherichia coli, and the recombinant enzyme was purified and characterized biochemically and enzymatically. The molecular weight of the enzyme was estimated to be 55,000 by SDS-PAGE, and 224,000 by gel filtration chromatography, suggesting that it acts as a tetramer. The CD spectrum suggests that the helical content of the enzyme is 10%. hd-ALDH was active on various aliphatic aldehyde substrates. The K(m) values of the enzyme were 6.4 μM for acetaldehyde, 4.2 μM for hexanal, 2.8 μM for octanal, and 0.84 μM for decanal, whereas the k(cat) values for these substrates were nearly equal (51-64 min-1). These results indicate that hd-ALDH acts preferentially on long-chain aliphatic aldehydes.
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U2 - 10.1016/S1389-1723(99)80167-4
DO - 10.1016/S1389-1723(99)80167-4
M3 - Article
C2 - 16232565
AN - SCOPUS:0032866859
SN - 1389-1723
VL - 88
SP - 7
EP - 11
JO - Journal of Bioscience and Bioengineering
JF - Journal of Bioscience and Bioengineering
IS - 1
ER -