Polyphosphate-dependent enzymes in some coryneform bacteria isolated from sewage sludge

Bark, Kerstin; Kämpfer, Peter; Sponner, Alexander; Dott, Wolfgang

doi:10.1111/j.1574-6968.1993.tb06019.x

Cited by 24 publications

(6 citation statements)

References 21 publications

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“…Other researchers also found that actinobacteria comprised a large proportion of bacteria in EBPR sludge, as determined by FISH (29,51), by respiratory quinone profiles (27), and by clone library analysis (17). Additionally, a range of actinobacterial isolates has been investigated for phosphate accumulation (6,39,40,48). It will be worthwhile to follow up on the role of actinobacteria in EBPR.…”

Section: Discussionmentioning

confidence: 97%

Identification of Some of the Major Groups of Bacteria in Efficient and Nonefficient Biological Phosphorus Removal Activated Sludge Systems

Bond

Erhart

Wagner

et al. 1999

Appl Environ Microbiol

207

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To investigate the bacteria that are important to phosphorus (P) removal in activated sludge, microbial populations were analyzed during the operation of a laboratory-scale reactor with various P removal performances. The bacterial population structure, analyzed by fluorescence in situ hybridization (FISH) with oligonucleotides probes complementary to regions of the 16S and 23S rRNAs, was associated with the P removal performance of the reactor. At one stage of the reactor operation, chemical characterization revealed that extremely poor P removal was occurring. However, like in typical P-removing sludges, complete anaerobic uptake of the carbon substrate occurred. Bacteria inhibiting P removal overwhelmed the reactor, and according to FISH, bacteria of the β subclass of the class Proteobacteria other than β-1 or β-2 were dominant in the sludge (58% of the population). Changes made to the operation of the reactor led to the development of a biomass population with an extremely good P removal capacity. The biochemical transformations observed in this sludge were characteristic of typical P-removing activated sludge. The microbial population analysis of the P-removing sludge indicated that bacteria of the β-2 subclass of the class Proteobacteria and actinobacteria were dominant (55 and 35%, respectively), therefore implicating bacteria from these groups in high-performance P removal. The changes in operation that led to the improved performance of the reactor included allowing the pH to rise during the anaerobic period, which promoted anaerobic phosphate release and possibly caused selection against non-phosphate-removing bacteria.

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Section: Discussionmentioning

confidence: 97%

Identification of Some of the Major Groups of Bacteria in Efficient and Nonefficient Biological Phosphorus Removal Activated Sludge Systems

Bond

Erhart

Wagner

et al. 1999

Appl Environ Microbiol

207

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“…This indicates that at least two kinds of NAD kinase, poly( P )/ATP–NAD kinase and ATP–NAD kinase, exist in living organisms. Other NAD‐phosphorylating enzymes which utilize p ‐nitrophenylphosphate or glucose 6‐phosphate have been detected in cell extracts of Proteus mirabilis [11] and coryneform bacteria isolated from sewage sludge [17], respectively. E. coli ATP–NAD kinase, as well as poly( P )/ATP–NAD kinase [7], did not show any activity toward these phosphoryl donors.…”

Section: Discussionmentioning

confidence: 99%

Molecular characterization of Escherichia coli NAD kinase

Kawai

Mori

Mukai

et al. 2001

European Journal of Biochemistry

121

123

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NAD kinase was purified to homogeneity from Escherichia coli MG1655. The enzyme was a hexamer consisting of 30 kDa subunits and utilized ATP or other nucleoside triphosphates as phosphoryl donors for the phosphorylation of NAD, most efficiently at pH 7.5 and 60 8C. The enzyme could not use inorganic polyphosphates as phosphoryl donors and was designated as ATP±NAD kinase. The N-terminal amino-acid sequence of the purified enzyme was encoded by yfjB, which had been deposited as a gene of unknown function in the E. coli whole genomic DNA sequence database. yfjB was cloned and expressed in E. coli BL21(DE3)pLysS. The purified product (YfjB) showed NAD kinase activity, and was identical to ATP±NAD kinase purified from E. coli MG1655 in molecular structure and other enzymatic properties. ) have been purified to homogeneity. Among them, the enzymes of M. flavus and M. tuberculosis have been shown to utilize inorganic polyphosphate [poly(P)] in addition to ATP, and were designated as poly(P)/ATP±NAD kinase' [7]. However, the gene for NAD kinase, to the best of our knowledge, had never been cloned from any organism before we isolated the gene for the enzyme from M. tuberculosis [7].The lack of information about the NAD kinase gene has hindered definative understanding of NADP metabolism in organisms. To overcome this, and to obtain new insights regarding the regulation of the cellular redox system, we have attempted to isolate the NAD kinase gene of Escherichia coli. NAD kinase has been partially purified from E. coli and some properties of the enzyme have been preliminarily reported [8]. In this study, we identified the gene for NAD kinase of E. coli through purification and analysis of the primary structure of the enzyme, and revealed that the E. coli NAD kinase can utilize ATP and other nucleoside triphosphates, but not poly(P), and designated the enzyme as ATP±NAD kinase to distinguish it from poly(P)/ATP±NAD kinase of M. flavus and M. tuberculosis. M A T E R I A L S A N D M E T H O D SBacterial strains E. coli MG1655 was cultured at 37 8C in YGD medium comprising 0.1% (NH4) 2 SO4, 0.05% MgSO 4 7H 2 O, 0.1% KH 2 PO4, 0.4% Na 2 HPO 4 , 0.5% yeast extract, and 0.5% glucose (pH 7.2). As a host for plasmid amplification, E. coli DH5a (Toyobo, Osaka, Japan) was routinely cultured at 37 8C in Luria±Bertani medium [9] supplemented with ampicillin (100 mg´mL 21 ). The growth conditions for the derivative strains of E. coli BL21(DE3) pLysS (Novagen, Darmstadt, Germany) are described below.Assays NAD kinase was assayed by means of a two-step method as described previously [10] in a reaction mixture (1.0 mL) consisting of 5.0 mm NAD, 5.0 mm MgCl 2 , 100 mm Tris/ HCl pH 7.0, and 5.0 mm ATP. NADP was determined enzymatically with isocitrate dehydrogenase [11]. One unit of enzyme activity was defined as 1.0 nmol of NADP produced in 1 min at 37 8C, and specific activity was expressed in U´mg protein 21 . K m , V max and Hill coefficients (h) were determined by means of Lineweaver±Burk and Hill plots, respectively. The rate constan...

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“…Although some of the isolates were shown to store polyphosphate to a certain extent, a polyphosphate-dependent enzyme system, as reported for Acinetobacter [7,8,33] could not be detected (unpublished results).…”

Section: The Role Of Acinetobacter and Aeromonas In Epr Treatment Plamentioning

confidence: 90%

“…Different Grampositive organisms have already been described in the literature [44,46] showing high amounts of P accumulation. Recently, additional polyphosphate-dependent enzymes were found in coryneform bacteria isolated from sewage sludge [8], which is additional evidence for the importance of Gram-positive bacteria with a high G+C content as P-removing organisms.…”

Section: The Role Of Acinetobacter and Aeromonas In Epr Treatment Plamentioning

confidence: 99%

Characterization of bacterial communities from activated sludge: Culture-dependent numerical identification versus in situ identification using group- and genus-specific rRNA-targeted oligonucleotide probes

et al. 1996

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The structures of bacterial communities were studied in activated sludge samples obtained from the aerobic and anaerobic zones of a wastewater treatment plant showing enhanced phosphorous removal. Samples were analyzed by in situ hybridization with oligonucleotide probes complementary to selected regions of the 16S and 23S ribosomal RNA (rRNA) characteristic for defined phylogenetic entities (genera and larger groups). The microbial community structures revealed by molecular techniques were compared with the compositions of culturable bacterial communities, obtained from the characterization of 255 isolates from tryptone-soy (TS) agar and R2A agar. These isolates were characterized by 89 physiological tests and their cellular fatty acid patterns, and identified. Culture-dependent techniques indicated the following distribution: different Aeromonas spp. (2.7-8.3% on R2A agar; 45.0-63.7% on TS agar), Acinetobacter spp. (5.4-9.0% on R2A agar; 5.0-9.1% on TS agar), Pseudomonas spp. (up to 10% on R2A agar) and Shewanella putrefaciens (up to 3.0% on R2A agar), all members of the gamma subclass of Proteobacteria, were isolated most frequently. The relatively rare isolates of the beta subclass were identified as Acidovorax spp., Alcaligenes spp., and Comamonas spp.. The Gram-positive bacteria (high DNA G+C) were assigned mainly to Arthrobacter spp., Microbacterium spp., and Mycobacterium phlei. In order to assess the in situ abundance of the most frequently isolated genus, Aeromonas, two rRNA-targeted oligonucleotide probes were developed. The two

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Polyphosphate-dependent enzymes in some coryneform bacteria isolated from sewage sludge

Cited by 24 publications

References 21 publications

Identification of Some of the Major Groups of Bacteria in Efficient and Nonefficient Biological Phosphorus Removal Activated Sludge Systems

Identification of Some of the Major Groups of Bacteria in Efficient and Nonefficient Biological Phosphorus Removal Activated Sludge Systems

Molecular characterization of Escherichia coli NAD kinase

Characterization of bacterial communities from activated sludge: Culture-dependent numerical identification versus in situ identification using group- and genus-specific rRNA-targeted oligonucleotide probes

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