NDLI: Alanine dehydrogenase from the psychrophilic bacterium strain PA-43: overexpression, molecular characterization, and sequence analysis

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Extremophiles : Volume 7, Issue 1, February 2003

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Extremophiles : Volume 4

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Extremophiles : Volume 2

Extremophiles : Volume 1

Alanine dehydrogenase from the psychrophilic bacterium strain PA-43: overexpression, molecular characterization, and sequence analysis

Content Provider	Springer Nature Link
Author	Irwin, Jane A. Lynch, Susan V. Coughlan, Suzie Baker, Patrick J. Gudmundsson, Haflidi M. Alfredsson, Gudni A. Rice, David W. Engel, Paul C.
Copyright Year	2003
Abstract	The gene encoding alanine dehydrogenase (AlaDH; EC 1.4.1.1) from the marine psychrophilic bacterium strain PA-43 was cloned, sequenced, and overexpressed in Escherichia coli. The primary structure was deduced on the basis of the nucleotide sequence. The enzyme subunit contains 371 amino acid residues, and the sequence is 90% and 77% identical, respectively, to AlaDHs from Shewanella Ac10 and Vibrio proteolyticus. The half-life of PA-43 AlaDH at 52 °C is 9 min, and it is thus more thermolabile than the AlaDH from Shewanella Ac10 or V. proteolyticus. The enzyme showed strong specificity for NAD+ and l-alanine as substrates. The apparent K m for NAD+ was temperature dependent (0.04 mM–0.23 mM from 15 °C to 55 °C). A comparison of the PA-43 deduced amino acid sequence to the solved three-dimensional structure of Phormidium lapideum AlaDH showed that there were likely to be fewer salt bridges in the PA-43 enzyme, which would increase enzyme flexibility and decrease thermostability. The hydrophobic surface character of the PA-43 enzyme was greater than that of P. lapideum AlaDH, by six residues. However, no particular modification or suite of modifications emerged as being clearly responsible for the psychrophilic character of PA-43 AlaDH.
Starting Page	135
Ending Page	143
Page Count	9
File Format	PDF
ISSN	14310651
Journal	Extremophiles
Volume Number	7
Issue Number	2
Language	English
Publisher	Springer-Verlag
Publisher Date	2003-02-04
Publisher Place	Tokyo
Access Restriction	One Nation One Subscription (ONOS)
Content Type	Text
Resource Type	Article
Subject	Medicine Molecular Medicine Microbiology

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