scholarly journals Key Enzymes Enabling the Growth of Arthrobacter sp. Strain JBH1 with Nitroglycerin as the Sole Source of Carbon and Nitrogen

2012 ◽  
Vol 78 (10) ◽  
pp. 3649-3655 ◽  
Author(s):  
Johana Husserl ◽  
Joseph B. Hughes ◽  
Jim C. Spain
Keyword(s):  
Heterologous Expression ◽  
Nitrogen Source ◽  
Sole Source ◽  
Glycerol Kinase ◽  
Enzyme Assays ◽  
Content Type ◽  
Key Enzymes ◽  
Carbon And Nitrogen ◽  
Subsequent Transformation

ABSTRACTFlavoprotein reductases that catalyze the transformation of nitroglycerin (NG) to dinitro- or mononitroglycerols enable bacteria containing such enzymes to use NG as the nitrogen source. The inability to use the resulting mononitroglycerols limits most strains to incomplete denitration of NG. Recently,Arthrobacterstrain JBH1 was isolated for the ability to grow on NG as the sole source of carbon and nitrogen, but the enzymes and mechanisms involved were not established. Here, the enzymes that enable theArthrobacterstrain to incorporate NG into a productive pathway were identified. Enzyme assays indicated that the transformation of nitroglycerin to mononitroglycerol is NADPH dependent and that the subsequent transformation of mononitroglycerol is ATP dependent. Cloning and heterologous expression revealed that a flavoprotein catalyzes selective denitration of NG to 1-mononitroglycerol (1-MNG) and that 1-MNG is transformed to 1-nitro-3-phosphoglycerol by a glycerol kinase homolog. Phosphorylation of the nitroester intermediate enables the subsequent denitration of 1-MNG in a productive pathway that supports the growth of the isolate and mineralization of NG.

scholarly journals Genes of theN-Methylglutamate Pathway Are Essential for Growth of Methylobacterium extorquens DM4 with Monomethylamine

2014 ◽  
Vol 80 (11) ◽  
pp. 3541-3550 ◽  
Author(s):  
Christelle Gruffaz ◽  
Emilie E. L. Muller ◽  
Yousra Louhichi-Jelail ◽  
Yella R. Nelli ◽  
Gilles Guichard ◽  
...  
Keyword(s):  
Carbon Source ◽  
Nitrogen Source ◽  
Gene Clusters ◽  
Sole Source ◽  
Nadh:Ubiquinone Oxidoreductase ◽  
Methylotrophic Bacteria ◽  
Methylobacterium Extorquens ◽  
Content Type ◽  
Carbon And Nitrogen ◽  
In The Wild

ABSTRACTMonomethylamine (MMA, CH3NH2) can be used as a carbon and nitrogen source by many methylotrophic bacteria.Methylobacterium extorquensDM4 lacks the MMA dehydrogenase encoded bymaugenes, which inM. extorquensAM1 is essential for growth on MMA. Identification and characterization of minitransposon mutants with an MMA-dependent phenotype showed that strain DM4 grows with MMA as the sole source of carbon, energy, and nitrogen by theN-methylglutamate (NMG) pathway. Independent mutations were found in a chromosomal region containing the genesgmaS,mgsABC, andmgdABCDfor the three enzymes of the pathway, γ-glutamylmethylamide (GMA) synthetase, NMG synthase, and NMG dehydrogenase, respectively. Reverse transcription-PCR confirmed the operonic structure of the two divergent gene clustersmgsABC-gmaSandmgdABCDand their induction during growth with MMA. The genesmgdABCDandmgsABCwere found to be essential for utilization of MMA as a carbon and nitrogen source. The genegmaSwas essential for MMA utilization as a carbon source, but residual growth of mutant DM4gmaSgrowing with succinate and MMA as a nitrogen source was observed. Plasmid copies ofgmaSand thegmaShomolog METDI4690, which encodes a protein 39% identical to GMA synthetase, fully restored the ability of mutants DM4gmaSand DM4gmaSΔmetdi4690 to use MMA as a carbon and nitrogen source. Similarly, chemically synthesized GMA, the product of GMA synthetase, could be used as a nitrogen source for growth in the wild-type strain, as well as in DM4gmaSand DM4gmaSΔmetdi4690 mutants. The NADH:ubiquinone oxidoreductase respiratory complex component NuoG was also found to be essential for growth with MMA as a carbon source.

scholarly journals A Mannose Family Phosphotransferase System Permease and Associated Enzymes Are Required for Utilization of Fructoselysine and Glucoselysine in Salmonella enterica Serovar Typhimurium

2015 ◽  
Vol 197 (17) ◽  
pp. 2831-2839 ◽  
Author(s):  
Katherine A. Miller ◽  
Robert S. Phillips ◽  
Paul B. Kilgore ◽  
Grady L. Smith ◽  
Timothy R. Hoover
Keyword(s):  
Nitrogen Source ◽  
Nitrogen Sources ◽  
Maillard Reaction Products ◽  
Reaction Products ◽  
Phosphotransferase System ◽  
Carbon And Nitrogen Sources ◽  
Content Type ◽  
Carbon And Nitrogen ◽  
Serovar Typhimurium ◽  
Food Borne Illness

ABSTRACTSalmonella entericserovar Typhimurium, a major cause of food-borne illness, is capable of using a variety of carbon and nitrogen sources. Fructoselysine and glucoselysine are Maillard reaction products formed by the reaction of glucose or fructose, respectively, with the ε-amine group of lysine. We report here thatS. Typhimurium utilizes fructoselysine and glucoselysine as carbon and nitrogen sources via a mannose family phosphotransferase (PTS) encoded bygfrABCD(glucoselysine/fructoselysine PTS components EIIA, EIIB, EIIC, and EIID; locus numbers STM14_5449 to STM14_5454 inS. Typhimurium 14028s). Genes coding for two predicted deglycases within thegfroperon,gfrEandgfrF, were required for growth with glucoselysine and fructoselysine, respectively. GfrF demonstrated fructoselysine-6-phosphate deglycase activity in a coupled enzyme assay. The biochemical and genetic analyses were consistent with a pathway in which fructoselysine and glucoselysine are phosphorylated at the C-6 position of the sugar by the GfrABCD PTS as they are transported across the membrane. The resulting fructoselysine-6-phosphate and glucoselysine-6-phosphate subsequently are cleaved by GfrF and GfrE to form lysine and glucose-6-phosphate or fructose-6-phosphate. Interestingly, althoughS. Typhimurium can use lysine derived from fructoselysine or glucoselysine as a sole nitrogen source, it cannot use exogenous lysine as a nitrogen source to support growth. Expression ofgfrABCDEFwas dependent on the alternative sigma factor RpoN (σ54) and an RpoN-dependent LevR-like activator, which we designated GfrR.IMPORTANCESalmonellaphysiology has been studied intensively, but there is much we do not know regarding the repertoire of nutrients these bacteria are able to use for growth. This study shows that a previously uncharacterized PTS and associated enzymes function together to transport and catabolize fructoselysine and glucoselysine. Knowledge of the range of nutrients thatSalmonellautilizes is important, as it could lead to the development of new strategies for reducing the load ofSalmonellain food animals, thereby mitigating its entry into the human food supply.

scholarly journals Aerobic Biodegradation of 2,4-Dinitroanisole by Nocardioides sp. Strain JS1661

2014 ◽  
Vol 80 (24) ◽  
pp. 7725-7731 ◽  
Author(s):  
Tekle Tafese Fida ◽  
Shannu Palamuru ◽  
Gunjan Pandey ◽  
Jim C. Spain
Keyword(s):  
Microbial Transformation ◽  
Growth Yield ◽  
Sole Source ◽  
Aerobic Biodegradation ◽  
Enzyme Assays ◽  
Content Type ◽  
Dead End ◽  
End Products ◽  
Subsequent Degradation ◽  
Genes Encoding

ABSTRACT2,4-Dinitroanisole (DNAN) is an insensitive munition ingredient used in explosive formulations as a replacement for 2,4,6-trinitrotoluene (TNT). Little is known about the environmental behavior of DNAN. There are reports of microbial transformation to dead-end products, but no bacteria with complete biodegradation capability have been reported.Nocardioidessp. strain JS1661 was isolated from activated sludge based on its ability to grow on DNAN as the sole source of carbon and energy. Enzyme assays indicated that the first reaction involves hydrolytic release of methanol to form 2,4-dinitrophenol (2,4-DNP). Growth yield and enzyme assays indicated that 2,4-DNP underwent subsequent degradation by a previously established pathway involving formation of a hydride-Meisenheimer complex and release of nitrite. Identification of the genes encoding the key enzymes suggested recent evolution of the pathway by recruitment of a novel hydrolase to extend the well-characterized 2,4-DNP pathway.

scholarly journals Histidine Utilization Is a Critical Determinant of Acinetobacter Pathogenesis

2020 ◽  
Vol 88 (7) ◽  
Author(s):  
Zachery R. Lonergan ◽  
Lauren D. Palmer ◽  
Eric P. Skaar
Keyword(s):  
Nitrogen Source ◽  
Treatment Options ◽  
Energy Generation ◽  
Zinc Homeostasis ◽  
Critical Determinant ◽  
Content Type ◽  
Carbon And Nitrogen ◽  
Amino Acid Histidine ◽  
Tract Infections

ABSTRACT Acinetobacter baumannii is a nosocomial pathogen capable of causing a range of diseases, including respiratory and urinary tract infections and bacteremia. Treatment options are limited due to the increasing rates of antibiotic resistance, underscoring the importance of identifying new targets for antimicrobial development. During infection, A. baumannii must acquire nutrients for replication and survival. These nutrients include carbon- and nitrogen-rich molecules that are needed for bacterial growth. One possible nutrient source within the host is amino acids, which can be utilized for protein synthesis or energy generation. Of these, the amino acid histidine is among the most energetically expensive for bacteria to synthesize; therefore, scavenging histidine from the environment is likely advantageous. We previously identified the A. baumannii histidine utilization (Hut) system as being linked to nutrient zinc homeostasis, but whether the Hut system is important for histidine-dependent energy generation or vertebrate colonization is unknown. Here, we demonstrate that the Hut system is conserved among pathogenic Acinetobacter and regulated by the transcriptional repressor HutC. In addition, the Hut system is required for energy generation using histidine as a carbon and nitrogen source. Histidine was also detected extracellularly in the murine lung, demonstrating that it is bioavailable during infection. Finally, the ammonia-releasing enzyme HutH is required for acquiring nitrogen from histidine in vitro, and strains inactivated for hutH are severely attenuated in a murine model of pneumonia. These results suggest that bioavailable histidine in the lung promotes Acinetobacter pathogenesis and that histidine serves as a crucial nitrogen source during infection.

scholarly journals Carbaryl as a Carbon and Nitrogen Source: an Inducible Methylamine Metabolic Pathway at the Biochemical and Molecular Levels inPseudomonassp. Strain C5pp

2018 ◽  
Vol 84 (24) ◽  
Author(s):  
Kamini ◽  
Rakesh Sharma ◽  
Narayan S. Punekar ◽  
Prashant S. Phale
Keyword(s):  
Carbon Source ◽  
Nitrogen Source ◽  
Sole Source ◽  
Biogeochemical Cycles ◽  
Ecological Systems ◽  
Suitable Candidate ◽  
Carbon And Nitrogen ◽  
Metabolic Versatility ◽  
Soil Isolate ◽  
Water Ecosystems

ABSTRACTCarbaryl is the most widely used carbamate family pesticide, and its persistent nature causes it to pollute both soil and water ecosystems. Microbes maintain the Earth’s biogeochemical cycles by metabolizing various compounds present in the matter, including xenobiotics, as a sole source of carbon, nitrogen, and energy. Soil isolatePseudomonassp. strain C5pp metabolizes carbaryl efficiently as the carbon source. Periplasmic carbaryl hydrolase catalyzes the conversion of carbaryl to 1-naphthol and methylamine. 1-Naphthol was further used as a carbon source via gentisate, whereas the metabolic fate of methylamine is not known. Here, we demonstrate that strain C5pp showed efficient growth on carbaryl when supplied as a carbon and nitrogen source, suggesting that the methylamine generated was used as the nitrogen source. Genes involved in the methylamine metabolism were annotated and characterized at the biochemical and molecular level. Transcriptional and enzyme activity studies corroborate that the γ-glutamylmethylamide/N-methylglutamate (GMA/NMG) pathway is involved in the metabolism of carbaryl and methylamine as a nitrogen source. Compared to carbaryl, methylamine was found to be an effective inducer for the metabolic and transporter genes. Strain C5pp also harbored genes involved in sarcosine metabolism that were cotranscribed and induced by sarcosine. The presence of inducible pathways for metabolism of carbaryl as a nitrogen and carbon source helps in complete and efficient mineralization of carbaryl by strain C5pp, thereby maintaining the biogeochemical cycles.IMPORTANCEThe degradation of xenobiotics plays a significant role in the environment to maintain ecological systems as well as to prevent the imbalance of biogeochemical cycles via carbon-nitrogen cycling. Carbaryl is the most widely used pesticide from the carbamate family.Pseudomonassp. strain C5pp, capable of utilizing carbaryl as a carbon and nitrogen source for its growth, subsequently helps in complete remediation of carbaryl. Thus, it maintains the ecosystem by balancing the biogeochemical cycles. The metabolic versatility and genetic diversity of strain C5pp for the transformation of contaminants like carbaryl and 1-naphthol into less harmful products make it a suitable candidate from the perspective of bioremediation.

scholarly journals Genome Analysis ofFimbriiglobus ruberSP5T, a Planctomycete with Confirmed Chitinolytic Capability

2018 ◽  
Vol 84 (7) ◽  
Author(s):  
Nikolai V. Ravin ◽  
Andrey L. Rakitin ◽  
Anastasia A. Ivanova ◽  
Alexey V. Beletsky ◽  
Irina S. Kulichevskaya ◽  
...  
Keyword(s):  
Sole Source ◽  
Major Constituent ◽  
Special Importance ◽  
Gene Encoding ◽  
Fungal Cell ◽  
Content Type ◽  
Carbon And Nitrogen ◽  
Type Iv ◽  
The Family ◽  
Weak Growth

ABSTRACTMembers of the bacterial orderPlanctomycetaleshave often been observed in associations with Crustacea. The ability to degrade chitin, however, has never been reported for any of the cultured planctomycetes although utilization ofN-acetylglucosamine (GlcNAc) as a sole carbon and nitrogen source is well recognized for these bacteria. Here, we demonstrate the chitinolytic capability of a member of the familyGemmataceae,Fimbriiglobus ruberSP5T, which was isolated from a peat bog. As revealed by metatranscriptomic analysis of chitin-amended peat, the pool of 16S rRNA reads fromF. ruberincreased in response to chitin availability. Strain SP5Tdisplayed only weak growth on amorphous chitin as a sole source of carbon but grew well with chitin as a source of nitrogen. The genome ofF. ruberSP5Tis 12.364 Mb in size and is the largest among all currently determined planctomycete genomes. It encodes several enzymes putatively involved in chitin degradation, including two chitinases affiliated with the glycoside hydrolase (GH) family GH18, GH20 family β-N-acetylglucosaminidase, and the complete set of enzymes required for utilization of GlcNAc. The gene encoding one of the predicted chitinases was expressed inEscherichia coli, and the endochitinase activity of the recombinant enzyme was confirmed. The genome also contains genes required for the assembly of type IV pili, which may be used to adhere to chitin and possibly other biopolymers. The ability to use chitin as a source of nitrogen is of special importance for planctomycetes that inhabit N-depleted ombrotrophic wetlands.IMPORTANCEPlanctomycetes represent an important part of the microbial community inSphagnum-dominated peatlands, but their potential functions in these ecosystems remain poorly understood. This study reports the presence of chitinolytic potential in one of the recently described peat-inhabiting members of the familyGemmataceae,Fimbriiglobus ruberSP5T. This planctomycete uses chitin, a major constituent of fungal cell walls and exoskeletons of peat-inhabiting arthropods, as a source of nitrogen in N-depleted ombrotrophicSphagnum-dominated peatlands. This study reports the chitin-degrading capability of representatives of the orderPlanctomycetales.

Utilization of single L-amino acids as sole source of carbon and nitrogen by bacteria

1972 ◽  
Vol 18 (11) ◽  
pp. 1647-1650 ◽  
Author(s):  
Harvest Halvorson
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Nitrogen Source ◽  
Sole Source ◽  
Nutrient Agar ◽  
Single Amino Acid ◽  
Field Soil ◽  
Carbon And Nitrogen ◽  
Large Numbers ◽  
Ecological Aspects

The utilization of single L-amino acids as sole source of carbon and nitrogen by bacteria was studied and was found to be very common and widely distributed over many genera. Field soil contained large numbers of such bacteria; 7.7–79.2% of the colony count obtained on nutrient agar could be achieved by using a single amino acid as sole carbon and nitrogen source. Twenty taxonomically known bacteria which could use one or more amino acids were examined for the range of amino acids over which they could grow. Organisms which grew on one amino acid usually grew on several. Some ecological aspects of these findings are discussed.

scholarly journals Draft Genome Sequence of the Mucin Degrader Clostridium tertium WC0709

2021 ◽  
Vol 10 (32) ◽  
Author(s):  
Eliana Musmeci ◽  
Francesco Candeliere ◽  
Alberto Amaretti ◽  
Maddalena Rossi ◽  
Stefano Raimondi
Keyword(s):  
Nitrogen Source ◽  
Genome Sequence ◽  
Genome Analysis ◽  
Pure Culture ◽  
Draft Genome ◽  
Draft Genome Sequence ◽  
Comparative Genome Analysis ◽  
Content Type ◽  
Carbon And Nitrogen ◽  
The Relationship

The draft genome sequence of Clostridium tertium WC0709, a gut bacterium able to use mucin in pure culture as the sole carbon and nitrogen source, is presented here. The genome sequence of C. tertium will provide valuable references for comparative genome analysis and for studying the relationship with the host.

scholarly journals Mineralization of the Cyclic Nitramine Explosive Hexahydro-1,3,5-Trinitro-1,3,5-Triazine by Gordonia and Williamsia spp

2005 ◽  
Vol 71 (12) ◽  
pp. 8265-8272 ◽  
Author(s):  
Karen T. Thompson ◽  
Fiona H. Crocker ◽  
Herbert L. Fredrickson
Keyword(s):  
Nitrogen Source ◽  
Surface Soil ◽  
Nitrogen Sources ◽  
Sole Source ◽  
Ribosomal Gene ◽  
Aerobic Bacteria ◽  
Fatty Acid Profiles ◽  
Bacterial Strains ◽  
Carbon And Nitrogen ◽  
Pure Bacterial Cultures

ABSTRACT Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) is a cyclic nitroamine explosive that is a major component in many military high-explosive formulations. In this study, two aerobic bacteria that are capable of using RDX as the sole source of carbon and nitrogen to support their growth were isolated from surface soil. These bacterial strains were identified by their fatty acid profiles and 16S ribosomal gene sequences as Williamsia sp. KTR4 and Gordonia sp. KTR9. The physiology of each strain was characterized with respect to the rates of RDX degradation and [U-14C]RDX mineralization when RDX was supplied as a sole carbon and nitrogen source in the presence and absence of competing carbon and nitrogen sources. Strains KTR4 and KTR9 degraded 180 μM RDX within 72 h when RDX served as the only added carbon and nitrogen source while growing to total protein concentrations of 18.6 and 16.5 μg/ml, respectively. Mineralization of [U-14C]RDX to 14CO2 was 30% by strain KTR4 and 27% by KTR9 when RDX was the only added source of carbon and nitrogen. The addition of (NH4)2SO4 greatly inhibited KTR9's degradation of RDX but had little effect on that of KTR4. These are the first two pure bacterial cultures isolated that are able to use RDX as a sole carbon and nitrogen source. These two genera possess different physiologies with respect to RDX mineralization, and each can serve as a useful microbiological model for the study of RDX biodegradation with regard to physiology, biochemistry, and genetics.

Access to 2-Arylquinazolines via Catabolism/Reconstruction of Amino Acids with Insertion of Dimethyl Sulfoxide

2021 ◽  
Author(s):  
Jin-Tian Ma ◽  
Li-Sheng Wang ◽  
Zhi Chai ◽  
Xin-Feng Chen ◽  
Bo-Cheng Tang ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Dimethyl Sulfoxide ◽  
Nitrogen Source ◽  
Carbon And Nitrogen ◽  
First Time

Quinazoline skeletons are synthesized by amino acids catabolism/reconstruction combined with dimethyl sulfoxide insertion/cyclization for the first time. The amino acid acts as a carbon and nitrogen source through HI-mediated catabolism...

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