DOI: 10.1111/j.1432-1033.1982.tb07021.x. Purification and Properties of the Bifunctional Proline Dehydrogenase/1‐Pyrroline‐5‐Carboxylate Dehydrogenase from<i>Pseudomonas aeruginosa</i>

Purification and Properties of the Bifunctional Proline Dehydrogenase/1‐Pyrroline‐5‐Carboxylate Dehydrogenase fromPseudomonas aeruginosa

10.1111/j.1432-1033.1982.tb07021.x

Crossref journal-article

Wiley

European Journal of Biochemistry (311)

Abstract

Proline dehydrogenase/1‐pyrroline‐5‐carboxylate dehydrogenase (Pro/P5C dehydrogenase), a bifunctional enzyme catalyzing the two consecutive reactions of the oxidation of proline to glutamic acid, was purified fromPseudomonas aeruginosastrain PAO1. Pro/P5C dehydrogenase oxidized L‐proline in an FAD‐dependent reaction to L‐Δ1‐pyrroline‐5‐carboxylic acid and converted this intermediate with NAD or NADP as cosubstrates to L‐glutamic acid. The purification procedure involved DEAE‐cellulose chromatography, affinity chromatography on Matrex gel red A and gel filtration on Sephadex G‐200. It resulted, after 40‐fold purification with 11% yield, in a homogeneous preparation (< 98% pure).The molecular weight of the single subunit was determined as 119000. Gel filtration of purified Pro/P5C dehydrogenase yielded a molecular weight of 242000 while polyacrylamide gel electrophoresis under native conditions led to the appearance of two catalytically active forms of the enzyme with molecular weights of 241000 and 470000. Manual Edman degradation revealed proline, alanine and aspartic acid as theN‐terminal amino acid sequence.Pro/P5C dehydrogenase was highly specific for the L‐forms of proline and Δ1‐pyrroline‐5‐carboxylic acid. Its apparentKmvalues were 45 mM for L‐proline, 0.03 mM for NAD and 0.17 mM for NADP. The saturation function for Δ1‐pyrroline‐5‐carboxylic acid was non‐hyperbolic.

Bibliography

MEILE, L., & LEISINGER, T. (1982). Purification and Properties of the Bifunctional Proline Dehydrogenase/1âPyrrolineâ5âCarboxylate Dehydrogenase fromPseudomonas aeruginosa. European Journal of Biochemistry, 129(1), 67â75. Portico.

Authors 2

Leo MEILE (first)
Thomas LEISINGER (additional)

References 32 Referenced 21

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Dates

Type	When
Created	20 years, 6 months ago (March 3, 2005, 5:20 p.m.)
Deposited	8 months, 1 week ago (Dec. 25, 2024, 12:29 p.m.)
Indexed	8 months, 1 week ago (Dec. 25, 2024, 1:10 p.m.)
Issued	42 years, 9 months ago (Dec. 1, 1982)
Published	42 years, 9 months ago (Dec. 1, 1982)
Published Online	20 years, 6 months ago (March 3, 2005)
Published Print	42 years, 9 months ago (Dec. 1, 1982)

Funders 0

None

BibTeX

@article{MEILE_1982, title={Purification and Properties of the Bifunctional Proline Dehydrogenase/1‐Pyrroline‐5‐Carboxylate Dehydrogenase fromPseudomonas aeruginosa}, volume={129}, ISSN={1432-1033}, url={http://dx.doi.org/10.1111/j.1432-1033.1982.tb07021.x}, DOI={10.1111/j.1432-1033.1982.tb07021.x}, number={1}, journal={European Journal of Biochemistry}, publisher={Wiley}, author={MEILE, Leo and LEISINGER, Thomas}, year={1982}, month=dec, pages={67–75} }

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