scholarly journals Transformation of Isopropylamine to l-Alaninol by Pseudomonas sp. Strain KIE171 Involves N-Glutamylated Intermediates

2002 ◽  
Vol 68 (5) ◽  
pp. 2368-2375 ◽  
Author(s):  
Susana I. de Azevedo Wäsch ◽  
Jan R. van der Ploeg ◽  
Tere Maire ◽  
Alice Lebreton ◽  
Andreas Kiener ◽  
...  

ABSTRACT Pseudomonas sp. strain KIE171 was able to grow with isopropylamine or l-alaninol [S-(+)-2-amino-1-propanol] as the sole carbon source, but not with d-alaninol. To investigate the hypothesis that l-alaninol is an intermediate in the degradation of isopropylamine, two mini-Tn5 mutants unable to utilize both isopropylamine and l-alaninol were isolated. Whereas mutant KIE171-BI transformed isopropylamine to l-alaninol, mutant KIE171-BII failed to do so. The two genes containing a transposon insertion were cloned, and the DNA regions flanking the insertions were sequenced. Two clusters, one comprising eight ipu (isopropylamine utilization) genes (ipuABCDEFGH) and the other encompassing two genes (ipuI and orf259), were identified. Comparisons of sequences of the deduced Ipu proteins and those in the database suggested that isopropylamine is transported into the cytoplasm by a putative permease, IpuG. The next step, the formation of γ-glutamyl-isopropylamide from isopropylamine, ATP, and l-glutamate, was shown to be catalyzed by IpuC, a γ-glutamylamide synthetase. γ-Glutamyl-isopropylamide is then subjected to stereospecific monooxygenation by the hypothetical four-component system IpuABDE, thereby yielding γ-glutamyl-l-alaninol [γ(l-glutamyl)-l-hydroxy-isopropylamide]. Enzymatic hydrolysis by a hydrolase, IpuF, was shown to finally liberate l-alaninol and to regenerate l-glutamate. No gene(s) encoding an enzyme for the next step in the degradation of isopropylamine was found in the ipu clusters. Presumably, l-alaninol is oxidized by an alcohol dehydrogenase to yield l-2-aminopropionaldehyde or it is deaminated by an ammonia lyase to propionaldehyde. Genetic evidence indicated that the aldehyde formed is then further oxidized by the hypothetical aldehyde dehydrogenases IpuI and IpuH to either l-alanine or propionic acid, compounds which can be processed by reactions of the intermediary metabolism.

Author(s):  
Vivek Kumar Ranjan ◽  
Shriparna Mukherjee ◽  
Subarna Thakur ◽  
Krutika Gupta ◽  
Ranadhir Chakraborty

1961 ◽  
Vol 155 (959) ◽  
pp. 195-201 ◽  

The lag preceding growth of Bact. lactis aerogenes (Aerobacter aerogenes) after a first transfer to a medium containing D-arabinose as sole carbon source increases with the age and decreases with the size of the inoculum. During the long lag phase the β -galactosidase activity declines steeply. In contrast with this (and with a control ageing in a glucose medium) the D-ribulose isomerase activity is maintained, although no detectable consumption of D-arabinose occurs. If the long lag of unadapted cells in D-arabinose is divided into parts by intermediate passages in glucose or lactose media, the sum of the partial lags is nearly constant and equal to that observed when there is no interruption. But the periodic passages in the other media increase the rate at which growth eventually occurs in the D-arabinose. It is concluded that during the lag a decay of the enzymes in general occurs concomitantly with the development of the specific mechanisms concerned in the utilization of the new substrate. The balance of these processes (together with varying loss or retention of diffusible metabolites) is largely responsible for the observed variations in lag and mean generation time.


1998 ◽  
Vol 64 (12) ◽  
pp. 4629-4636 ◽  
Author(s):  
K. D. Goodwin ◽  
J. K. Schaefer ◽  
R. S. Oremland

ABSTRACT Bacterial oxidation of14CH2Br2 and14CH3Br was measured in freshwater, estuarine, seawater, and hypersaline-alkaline samples. In general, bacteria from the various sites oxidized similar amounts of14CH2Br2 and comparatively less 14CH3Br. Bacterial oxidation of14CH3Br was rapid in freshwater samples compared to bacterial oxidation of 14CH3Br in more saline waters. Freshwater was also the only site in which methyl fluoride-sensitive bacteria (e.g., methanotrophs or nitrifiers) governed brominated methane oxidation. Half-life calculations indicated that bacterial oxidation of CH2Br2 was potentially significant in all of the waters tested. In contrast, only in freshwater was bacterial oxidation of CH3Br as fast as chemical removal. The values calculated for more saline sites suggested that bacterial oxidation of CH3Br was relatively slow compared to chemical and physical loss mechanisms. However, enrichment cultures demonstrated that bacteria in seawater can rapidly oxidize brominated methanes. Two distinct cultures of nonmethanotrophic methylotrophs were recovered; one of these cultures was able to utilize CH2Br2 as a sole carbon source, and the other was able to utilize CH3Br as a sole carbon source.


1976 ◽  
Vol 54 (10) ◽  
pp. 854-865 ◽  
Author(s):  
Kwok-Him Yeung ◽  
Gillian Chaloner-Larssgn ◽  
Hiroshi Yamazaki

When Bacillus megaterium cells are grown on D-galactose as the sole carbon source, the cells actively synthesize β-galactosidase (β-D-galactoside galactohydroIase, EC 3.2.1.23). However, D-galactose, when added to a glucose-grown culture, did not induce β-galactosidase, apparently because of the glucose inhibition of the transport of galactose. On the other hand, when glucose was added to a galactose-grown culture, the transport of galactose continued at a reduced but significant rate, whereas further synthesis of β-galactosidase was halted. Adenosine 3′,5′-cyclic monophosphate (cAMP) or guanosine 3′,5′-cyclic monophosphate (cGMP) did not relieve the glucose inhibition of β-galactosidase synthesis in the preinduced culture. A method which gave a reproducible assay of c[32P]AMP in Escherichia coli did not detect cAMP or cGMP in a B. megaterium culture undergoing β-galactosidase induction, but revealed the extracellular accumulation of two unknown phosphorylated compounds. Cell-free extracts prepared from galactose-grown cells did not catalyze the degradation of cAMP or cGMP.


2006 ◽  
Vol 72 (3) ◽  
pp. 1873-1877 ◽  
Author(s):  
Zhankun Wang ◽  
Qingsheng Qi ◽  
Peng George Wang

ABSTRACT The cyclodextrin glucanotransferase (CGTase) gene (cgt) from Bacillus circulans 251 was cloned into plasmid pYD1, which allowed regulated expression, secretion, and detection. The expression of CGTase with a-agglutinin at the N-terminal end on the extracellular surface of Saccharomyces cerevisiae was confirmed by immunofluorescence microscopy. This surface-anchored CGTase gave the yeast the ability to directly utilize starch as a sole carbon source and the ability to produce the anticipated products, cyclodextrins, as well as glucose and maltose. The resulting glucose and maltose, which are efficient acceptors in the CGTase coupling reaction, could be consumed by yeast fermentation and thus facilitated cyclodextrin production. On the other hand, ethanol produced by the yeast may form a complex with cyclodextrin and shift the equilibrium in favor of cyclodextrin production. The yeast with immobilized CGTase produced 24.07 mg/ml cyclodextrins when it was incubated in yeast medium supplemented with 4% starch.


2014 ◽  
Vol 64 (1) ◽  
pp. 138-146
Author(s):  
Dorota Wolicka ◽  
Andrzej Borkowski

Abstract Sulphidogenous microorganism communities were isolated from soil polluted by crude oil. The study was focused on determining the influence of 1) copper (II) concentration on the activity of selected microorganism communities and 2) the applied electron donor on the course and evolution of mineral-forming processes under conditions favouring growth of sulphate-reducing bacteria (SRB). The influence of copper concentration on the activity of selected microorganism communities and the type of mineral phases formed was determined during experiments in which copper (II) chloride at concentrations of 0.1, 0.2, 0.5 and 0.7 g/L was added to SRB cultures. The experiments were performed in two variants: with ethanol (4 g/L) or lactate (4 g/L) as the sole carbon source. In order to determine the taxonomic composition of the selected microorganism communities, the 16S rRNA method was used. Results of this analysis confirmed the presence of Desulfovibrio, Desulfohalobium, Desulfotalea, Thermotoga, Solibacter, Gramella, Anaeromyxobacter and Myxococcus sp. in the stationary cultures. The post-culture sediments contained covelline (CuS) and digenite (Cu9S5 ). Based on the results, it can be stated that the type of carbon source applied during incubation plays a crucial role in determining the mineral composition of the post-culture sediments. Thus, regardless of the amount of copper ion introduced to a culture with lactate as the sole carbon source, no copper sulphide was observed in the post-culture sediments. Cultures with ethanol as the sole carbon source, on the other hand, yielded covelline or digenite in all post-culture sediments.


2004 ◽  
Vol 72 (7) ◽  
pp. 4172-4187 ◽  
Author(s):  
Richard A. Moore ◽  
Shauna Reckseidler-Zenteno ◽  
Heenam Kim ◽  
William Nierman ◽  
Yan Yu ◽  
...  

ABSTRACT Burkholderia pseudomallei is the causative agent of melioidosis. Burkholderia thailandensis is a closely related species that can readily utilize l-arabinose as a sole carbon source, whereas B. pseudomallei cannot. We used Tn5-OT182 mutagenesis to isolate an arabinose-negative mutant of B. thailandensis. Sequence analysis of regions flanking the transposon insertion revealed the presence of an arabinose assimilation operon consisting of nine genes. Analysis of the B. pseudomallei chromosome showed a deletion of the operon from this organism. This deletion was detected in all B. pseudomallei and Burkholderia mallei strains investigated. We cloned the B. thailandensis E264 arabinose assimilation operon and introduced the entire operon into the chromosome of B. pseudomallei 406e via homologous recombination. The resultant strain, B. pseudomallei SZ5028, was able to utilize l-arabinose as a sole carbon source. Strain SZ5028 had a significantly higher 50% lethal dose for Syrian hamsters compared to the parent strain 406e. Microarray analysis revealed that a number of genes in a type III secretion system were down-regulated in strain SZ5028 when cells were grown in l-arabinose, suggesting a regulatory role for l-arabinose or a metabolite of l-arabinose. These results suggest that the ability to metabolize l-arabinose reduces the virulence of B. pseudomallei and that the genes encoding arabinose assimilation may be considered antivirulence genes. The increase in virulence associated with the loss of these genes may have provided a selective advantage for B. pseudomallei as these organisms adapted to survival in animal hosts.


2009 ◽  
Vol 61 (2) ◽  
pp. 159-164 ◽  
Author(s):  
B. Jovcic ◽  
Jelena Begovic ◽  
Jelena Lozo ◽  
L. Topisirovic ◽  
Milan Kojic

Pseudomonas sp. ATCC19151 harbors a gene encoding a putative alkylsulfatase (sdsA). Here we report a growth ability of this strain in minimal media containing 0.5, 0.75, and 1% sodium dodecyl sulfate as the sole carbon source. The most prominent growth was detected for the minimal medium with 0.5% SDS, so this concentration of SDS was used to monitor Pseudomonas sp. ATCC19151 SDS biodegradation dynamics. Bacterial growth coincided with the disappearance of SDS. Antibiotic susceptibility was tested as well. Pseudomonas sp. ATCC19151 was resistant to six out of nine tested antibiotics, including ampicillin, tetracycline, chloramphenicol, tobramycin, nalidixic acid, and gentamycin.


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