Aromatic-Alcohol Dehydrogenases from Pseudomonas putida N.C.I.B. 9869

1976 ◽  
Vol 4 (3) ◽  
pp. 494-495 ◽  
Author(s):  
MALCOLM J. KEAT ◽  
DAVID J. HOPPER
Keyword(s):  
Pseudomonas Putida ◽  
Alcohol Dehydrogenases ◽  
Aromatic Alcohol

scholarly journals The aromatic alcohol dehydrogenases in Pseudomonas putida N.C.I.B. 9869 grown on 3,5-xylenol and p-cresol

1978 ◽  
Vol 175 (2) ◽  
pp. 659-667 ◽  
Author(s):  
M J Keat ◽  
D J Hopper
Keyword(s):  
Alcohol Dehydrogenase ◽  
Pseudomonas Putida ◽  
The Other ◽  
Alcohol Dehydrogenases ◽  
Whole Cells ◽  
Aromatic Alcohol ◽  
Aromatic Alcohols ◽  
Hydroxybenzyl Alcohol

Whole cells of Pseudomonas putida N.C.I.B 9869, when grown on either 3,5-xylenol or p-cresol, oxidized both m- and p-hydroxybenzyl alcohols. Two distinct NAD+-dependent m-hydroxybenzyl alcohol dehydrogenases were purified from cells grown on 3,5-xylenol. Each is active with a range of aromatic alcohols, including both m- and p-hydroxybenzyl alcohol, but differ in their relative rates with the various substrates. An NAD+-dependent alcohol dehydrogenase was also partially purified from p-cresol grown cells. This too was active with m- and p-hydroxybenzyl alcohol and other aromatic alcohols, but was not identical with either of the other two dehydrogenases. All three enzymes were unstable, but were stabilized by dithiothreitol and all were inhibited with p-chloromercuribenzoate. All were specific for NAD+ and each was shown to catalyse conversion of alcohol into aldehyde.

scholarly journals Three distinct quinoprotein alcohol dehydrogenases are expressed when Pseudomonas putida is grown on different alcohols.

1995 ◽  
Vol 177 (9) ◽  
pp. 2442-2450 ◽  
Author(s):  
H Toyama ◽  
A Fujii ◽  
K Matsushita ◽  
E Shinagawa ◽  
M Ameyama ◽  
...  
Keyword(s):  
Pseudomonas Putida ◽  
Alcohol Dehydrogenases

scholarly journals Novel Dehalogenase Mechanism for 2,3-Dichloro-1-Propanol Utilization in Pseudomonas putida Strain MC4

2012 ◽  
Vol 78 (17) ◽  
pp. 6128-6136 ◽  
Author(s):  
Muhammad Irfan Arif ◽  
Ghufrana Samin ◽  
Jan G. E. van Leeuwen ◽  
Jantien Oppentocht ◽  
Dick B. Janssen
Keyword(s):  
Energy Source ◽  
Nuclear Magnetic Resonance ◽  
Pseudomonas Putida ◽  
Soil Degradation ◽  
Potassium Ferricyanide ◽  
Cosmid Library ◽  
High Similarity ◽  
Alcohol Dehydrogenases ◽  
Content Type ◽  
Cell Lysate

ABSTRACTAPseudomonas putidastrain (MC4) that can utilize 2,3-dichloro-1-propanol (DCP) and several aliphatic haloacids and haloalcohols as sole carbon and energy source for growth was isolated from contaminated soil. Degradation of DCP was found to start with oxidation and concomitant dehalogenation catalyzed by a 72-kDa monomeric protein (DppA) that was isolated from cell lysate. ThedppAgene was cloned from a cosmid library and appeared to encode a protein equipped with a signal peptide and that possessed high similarity to quinohemoprotein alcohol dehydrogenases (ADHs), particularly ADH IIB and ADH IIG fromPseudomonas putidaHK. This novel dehalogenating dehydrogenase has a broad substrate range, encompassing a number of nonhalogenated alcohols and haloalcohols. With DCP, DppA exhibited akcatof 17 s−1.1H nuclear magnetic resonance experiments indicated that DCP oxidation by DppA in the presence of 2,6-dichlorophenolindophenol (DCPIP) and potassium ferricyanide [K3Fe(CN)6] yielded 2-chloroacrolein, which was oxidized to 2-chloroacrylic acid.

scholarly journals Functional role of lanthanides in enzymatic activity and transcriptional regulation of PQQ-dependent alcohol dehydrogenases in Pseudomonas putida KT2440

2017 ◽  
Author(s):  
Matthias Wehrmann ◽  
Patrick Billard ◽  
Audrey Martin Meriadec ◽  
Asfaw Zegeye ◽  
Janosch Klebensberger
Keyword(s):  
Transcriptional Regulation ◽  
Pseudomonas Putida ◽  
Regulatory Network ◽  
Model Organism ◽  
Adaptive Strategy ◽  
Lanthanide Ions ◽  
Methylotrophic Bacterium ◽  
Methylotrophic Bacteria ◽  
Pseudomonas Putida Kt2440 ◽  
Alcohol Dehydrogenases

ABSTRACTThe oxidation of alcohols and aldehydes is crucial for detoxification and efficient catabolism of various volatile organic compounds (VOCs). Thus, many Gram-negative bacteria have evolved periplasmic oxidation systems, based on pyrroloquinoline quinone-dependent alcohol dehydrogenases (PQQ-ADHs), which are often functionally redundant. Using purified enzymes from the soil-dwelling model organism Pseudomonas putida KT2440, the present study reports the first description and characterization of a lanthanide-dependent PQQ-ADH (PedH) in a non-methylotrophic bacterium. PedH exhibits enzyme activity on a similar substrate range as its Ca2+-dependent counterpart PedE, including linear and aromatic primary and secondary alcohols as well as aldehydes, however, only in the presence of lanthanide ions including La3+, Ce3+, Pr3+, Sm3+ or Nd3+. Reporter assays revealed that PedH not only has a catalytic function, but is also involved in the transcriptional regulation of pedE and pedH, most likely acting as a sensory module. Notably, the underlying regulatory network is responsive to as little as 1 – 10 nM of lanthanum, a concentration assumed to be of ecological relevance. The present study further demonstrates that the PQQ-dependent oxidation system is crucial for efficient growth with a variety of volatile alcohols. From these results we conclude that functional redundancy and inverse regulation of PedE and PedH represents an adaptive strategy of P. putida KT2440 to optimize growth with volatile alcohols in response to different lanthanide availability.IMPORTANCEDue to their low bioavailability, lanthanides have long been considered as biologically inert. In recent years however, the identification of lanthanides as a cofactor in methylotrophic bacteria has attracted tremendous interest among various biological fields. The present study reveals that one of the two PQQ-ADHs produced by the model organism P. putida KT2440 also utilizes lanthanides as a cofactor, thus expanding the scope of lanthanide employing bacteria beyond the methylotrophs. Similar to methyloptrophic bacteria, a complex regulatory network is involved in the lanthanide-responsive switch between the two PQQ-ADHs encoded by P. putida KT2440. We further show that functional production of at least one of the enzymes is crucial for efficient growth with several volatile alcohols. Overall, our study provides a novel understanding for the redundancy of PQQ-ADHs observed in many organisms and further highlights the importance of lanthanides for bacterial metabolism, particularly in soil environments.

scholarly journals Functional Role of Lanthanides in Enzymatic Activity and Transcriptional Regulation of Pyrroloquinoline Quinone-Dependent Alcohol Dehydrogenases in Pseudomonas putida KT2440

mBio ◽  
2017 ◽  
Vol 8 (3) ◽  
Author(s):  
Matthias Wehrmann ◽  
Patrick Billard ◽  
Audrey Martin-Meriadec ◽  
Asfaw Zegeye ◽  
Janosch Klebensberger
Keyword(s):  
Transcriptional Regulation ◽  
Pseudomonas Putida ◽  
Regulatory Network ◽  
Model Organism ◽  
Pyrroloquinoline Quinone ◽  
Lanthanide Ions ◽  
Bacterial Metabolism ◽  
Methylotrophic Bacteria ◽  
Alcohol Dehydrogenases ◽  
Complex Regulatory Network

ABSTRACT The oxidation of alcohols and aldehydes is crucial for detoxification and efficient catabolism of various volatile organic compounds (VOCs). Thus, many Gram-negative bacteria have evolved periplasmic oxidation systems based on pyrroloquinoline quinone-dependent alcohol dehydrogenases (PQQ-ADHs) that are often functionally redundant. Here we report the first description and characterization of a lanthanide-dependent PQQ-ADH (PedH) in a nonmethylotrophic bacterium based on the use of purified enzymes from the soil-dwelling model organism Pseudomonas putida KT2440. PedH (PP_2679) exhibits enzyme activity on a range of substrates similar to that of its Ca2+-dependent counterpart PedE (PP_2674), including linear and aromatic primary and secondary alcohols, as well as aldehydes, but only in the presence of lanthanide ions, including La3+, Ce3+, Pr3+, Sm3+, or Nd3+. Reporter assays revealed that PedH not only has a catalytic function but is also involved in the transcriptional regulation of pedE and pedH, most likely acting as a sensory module. Notably, the underlying regulatory network is responsive to as little as 1 to 10 nM lanthanum, a concentration assumed to be of ecological relevance. The present study further demonstrates that the PQQ-dependent oxidation system is crucial for efficient growth with a variety of volatile alcohols. From these results, we conclude that functional redundancy and inverse regulation of PedE and PedH represent an adaptive strategy of P. putida KT2440 to optimize growth with volatile alcohols in response to the availability of different lanthanides. IMPORTANCE Because of their low bioavailability, lanthanides have long been considered biologically inert. In recent years, however, the identification of lanthanides as a cofactor in methylotrophic bacteria has attracted tremendous interest among various biological fields. The present study reveals that one of the two PQQ-ADHs produced by the model organism P. putida KT2440 also utilizes lanthanides as a cofactor, thus expanding the scope of lanthanide-employing bacteria beyond the methylotrophs. Similar to the system described in methylotrophic bacteria, a complex regulatory network is involved in lanthanide-responsive switching between the two PQQ-ADHs encoded by P. putida KT2440. We further show that the functional production of at least one of the enzymes is crucial for efficient growth with several volatile alcohols. Overall, our study provides a novel understanding of the redundancy of PQQ-ADHs observed in many organisms and further highlights the importance of lanthanides for bacterial metabolism, particularly in soil environments. IMPORTANCE Because of their low bioavailability, lanthanides have long been considered biologically inert. In recent years, however, the identification of lanthanides as a cofactor in methylotrophic bacteria has attracted tremendous interest among various biological fields. The present study reveals that one of the two PQQ-ADHs produced by the model organism P. putida KT2440 also utilizes lanthanides as a cofactor, thus expanding the scope of lanthanide-employing bacteria beyond the methylotrophs. Similar to the system described in methylotrophic bacteria, a complex regulatory network is involved in lanthanide-responsive switching between the two PQQ-ADHs encoded by P. putida KT2440. We further show that the functional production of at least one of the enzymes is crucial for efficient growth with several volatile alcohols. Overall, our study provides a novel understanding of the redundancy of PQQ-ADHs observed in many organisms and further highlights the importance of lanthanides for bacterial metabolism, particularly in soil environments.

Enzimaktivitások és a fluoreszkáló pszeudomonasz csíraszámok változása a fehér lóhere (Trifolium repens L.) rizoszférájában sókezelés (NaCl) hatására

2004 ◽  
Vol 53 (3-4) ◽  
pp. 367-376 ◽  
Author(s):  
A. A. Khalif ◽  
H. Abdorhim ◽  
Hosam E. H. T. Bayoumi ◽  
Anna Füzy ◽  
Mihály Kecskés
Keyword(s):  
Pseudomonas Putida ◽  
Trifolium Repens ◽  
Trifolium Repens L

Üvegházi körülmények között savanyú barna erdotalajban nevelt fehér here (Trifolium repens L.) növények rizoszférájának sókezelés hatására bekövetkezo változását ellenoriztük. Megvizsgáltuk a különbözo sókoncentrációknak (0, 0,2, 0,4, 0,6 és 0,8 tömeg %) a baktériumnépesség összetételére és a különbözo talajenzimek aktivitására gyakorolt hatását.  Megállapítottuk, hogy a talaj sótartalma közvetlenül befolyásolta a rizoszférában található fluoreszkáló pszeudomonaszok csíraszámát. A legsurubb populáció a 0,2% NaCl-ot tartalmazó talajban volt mérheto, ahol a fluoreszkáló pszeudomonaszok között a Pseudomonas putida és a P. fluorescens fordultak elo a legnagyobb számban. A pszeudomonaszok ily módon jól tolerálják a talaj magas NaCl-tartalmát, és gyökérkolonizáló tevékenységet képesek kifejteni a magas NaCl-tartalmú talajban is. A sókoncentráció növelésével kezdetben (a 0,2-0,4%-os tartományban) jelentosen növekedett a dehidrogenáz, kataláz, és ureáz enzimek aktivitása. A proteáz enzimek aktivitásmaximuma a 0,1-0,2% NaCl-koncentráció tartományba esett. A 0,4%-nál magasabb koncentrációkban a kontrollhoz hasonló mértékure csökkent mind a négy enzim aktivitása, és a baktériumok száma is. A foszfatáz- és a b-glükozidáz-tevékenység viszont a NaCl-dózis növelése következtében a koncentrációval arányosan, jelentosen csökkent a kontrollhoz viszonyítva.  Feltételezésünk szerint az enzimaktivitások változását is a sókezelés hatására bekövetkezo mikrobióta összetételének megváltozása okozta.

Exemplar Abstract for Pseudomonas putida (Trevisan 1889) Migula 1895 (Approved Lists 1980).

2003 ◽  
Author(s):  
Charles Thomas Parker ◽  
Dorothea Taylor ◽  
George M Garrity
Keyword(s):  
Pseudomonas Putida
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