Next Article in Journal
Experimental Research of an Active Solution for Modeling In Situ Activating Selective Catalytic Reduction Catalyst
Next Article in Special Issue
Regioselective Synthesis of Lactulose Esters by Candida antarctica and Thermomyces lanuginosus Lipases
Previous Article in Journal
Controllable and Large-Scale Synthesis of Carbon Nanostructures: A Review on Bamboo-Like Nanotubes
Previous Article in Special Issue
Exploiting the Versatility of Aminated Supports Activated with Glutaraldehyde to Immobilize β-galactosidase from Aspergillus oryzae
Article Menu
Issue 9 (September) cover image

Export Article

Open AccessArticle

Characterization of a Novel Nicotine Hydroxylase from Pseudomonas sp. ZZ-5 That Catalyzes the Conversion of 6-Hydroxy-3-Succinoylpyridine into 2,5-Dihydroxypyridine

School of Food and Biological Engineering, Zhengzhou University of Light Industry, Zhengzhou 450002, China
State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China
Author to whom correspondence should be addressed.
Catalysts 2017, 7(9), 257;
Received: 11 August 2017 / Revised: 26 August 2017 / Accepted: 26 August 2017 / Published: 31 August 2017
(This article belongs to the Special Issue Biocatalysis and Biotransformations)
PDF [1452 KB, uploaded 31 August 2017]


A novel nicotine hydroxylase was isolated from Pseudomonas sp. ZZ-5 (HSPHZZ). The sequence encoding the enzyme was 1206 nucleotides long, and encoded a protein of 401 amino acids. Recombinant HSPHZZ was functionally overexpressed in Escherichia coli BL21-Codon Plus (DE3)-RIL cells and purified to homogeneity after Ni-NTA affinity chromatography. Liquid chromatography-mass spectrometry (LC-MS) analyses indicated that the enzyme could efficiently catalyze the conversion of 6-hydroxy-3-succinoylpyridine (HSP) into 2,5-dihydroxypyridine (2,5-DHP) and succinic acid in the presence of nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD). The kinetic constants (Km, kcat, and kcat/Km) of HSPHZZ toward HSP were 0.18 mM, 2.1 s−1, and 11.7 s−1 mM−1, respectively. The optimum temperature, pH, and optimum concentrations of substrate and enzyme for 2,5-DHP production were 30 °C, 8.5, 1.0 mM, and 1.0 μM, respectively. Under optimum conditions, 85.3 mg/L 2,5-DHP was produced in 40 min with a conversion of 74.9%. These results demonstrated that HSPHZZ could be used for the enzymatic production of 2,5-DHP in biotechnology applications. View Full-Text
Keywords: HSP; 2,5-DHP; Pseudomonas; HSP hydroxylase; gene cloning HSP; 2,5-DHP; Pseudomonas; HSP hydroxylase; gene cloning

Graphical abstract

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

Supplementary material


Share & Cite This Article

MDPI and ACS Style

Wei, T.; Zang, J.; Zheng, Y.; Tang, H.; Huang, S.; Mao, D. Characterization of a Novel Nicotine Hydroxylase from Pseudomonas sp. ZZ-5 That Catalyzes the Conversion of 6-Hydroxy-3-Succinoylpyridine into 2,5-Dihydroxypyridine. Catalysts 2017, 7, 257.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics



[Return to top]
Catalysts EISSN 2073-4344 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top