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Purification and characterization of protocatechuate 3,4-dioxygenase from Pseudomonas pseudoalcaligenes KF707

Journal of the Korean Society for Applied Biological Chemistry volume 56, pages 401–408 (2013)Cite this article

An Erratum to this article was published on 30 October 2013

Abstract

Protocatechuate 3,4-dioxygenase was isolated and characterized from Pseudomonas pseudoalcaligenes KF707 for the purpose of developing a new anti-browning agent. The protocatechuate 3,4-dioxygenase from Pseudomonas pseudoalcaligenes KF707 was purified 296.8-fold, and showed specific activity of 121.7 U/mg. Based on the SDS-polyacrylamide and gel permeation chromatography, the molecular weight of protocatechuate 3,4-dioxygenase was 189.9 kDa and was composed of 3 αβ protomers, with molecular weights of 29.0 kDa of α subunit and 34.3 kDa of â subunit. The optimal pH and temperature were 7.5 and 38°C, respectively. Km values of catechol, protocatechuate, gallate, p-cresol, caffeic acid, catechin, L-DOPA, 4-methylcatechol and pyrogallol were 14, 17, 2, 10, 12, 20, 30, 21 and 3 μM, and the Vmax/Km (mim−1) values were 0.052, 3.06, 0.35, 0.01, 0.03, 0.02, 0.006, 0.008 and 0.11, respectively. This indicates that the enzyme is active on a wide range of phenyl compounds, in contrast to the high specificity of similar enzymes from other sources. Our data also show that the turnover number of protocatechuate 3,4-dioxygenase from Pseudomonas pseudoalcaligenes KF707 is 68 s−1, which is much higher than the known values from other sources.

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  1. Department of Biochemistry and Health Science, College of Natural Science, Changwon National University, Changwon, Gyeongnam, 641-773, Republic of Korea

    Hyun Woo Sim, MiJa Jung & Yong Kweon Cho

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  1. Hyun Woo Sim
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  2. MiJa Jung
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Correspondence to Yong Kweon Cho.

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H. W. Sim and M. J. Jung contributed equally.

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Sim, H.W., Jung, M. & Cho, Y.K. Purification and characterization of protocatechuate 3,4-dioxygenase from Pseudomonas pseudoalcaligenes KF707. J Korean Soc Appl Biol Chem 56, 401–408 (2013). https://doi.org/10.1007/s13765-013-3080-2

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