Pathway of lysine degradation in Fusobacterium nucleatum

Barker, H. A.; Kahn, J; Hedrick, L. R.

doi:10.1128/jb.152.1.201-207.1982

Cited by 62 publications

(18 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, no enzyme has been reported for the reductive amination of β-keto acids, except l - erythro -3,5-diaminohexanoate dehydrogenase (3,5-DAHDH), which was identified to catalyze the oxidative deamination of (3 S ,5 S )-diaminohexanoate ((3 S ,5 S )-DAH) to 3-keto-5-aminohexanoate in the fermentation pathway of lysine . All reported 3,5-DAHDHs have strict substrate specificity toward (3 S ,5 S )-DAH and show hardly any activity toward other β-amino acids, even the other three stereoisomers of 3,5-DAH . In the present study, a 3,5-DAHDH (NCBI reference sequence: YP_001741409.1) from Candidatus Cloacamonas acidaminovorans (3,5-DAHDHcca) was expressed in Escherichia coli BL21(DE3) host cells as a soluble protein (Supporting Information Figure S1) and engineered to β-amino acid dehydrogenase for the reductive amination of β-keto acids.…”

mentioning

confidence: 99%

Development of β-Amino Acid Dehydrogenase for the Synthesis of β-Amino Acids via Reductive Amination of β-Keto Acids

et al. 2015

View full text Add to dashboard Cite

An L-erythro-3,5-diaminohexanoate dehydrogenase from Candidatus Cloacamonas acidaminovorans (3,5-DAHDHcca) was engineered to β-amino acid dehydrogenase (β-AADH) by screening a domain scanning mutagenesis library. The best β-AADH mutant displayed about 200-fold activity toward (R)-β-homomethionine compared with the wild-type enzyme. (R)-β-Homomethionine, (R)-β-phenylalanine, and (S)-β-aminobutyric acid were obtained via β-AADHcatalyzed reductive amination of the corresponding β-keto acids, which were generated from β-keto-nitriles or β-ketoesters with nitrilase or lipase. This is the first example of the direct reductive amination of β-keto acids catalyzed by β-AADH to give β-amino acids, opening a new venue for the synthesis of chiral β-amino acids.

show abstract

mentioning

confidence: 99%

Development of β-Amino Acid Dehydrogenase for the Synthesis of β-Amino Acids via Reductive Amination of β-Keto Acids

et al. 2015

View full text Add to dashboard Cite

show abstract

“…But other pathways like lysine, glutarate, and 4-aminobutyrate pathways also contribute to butyrate production 74 . These pathways are prevalent in Firmicutes and some other phyla, such as Fusobacteria and Bacteroidetes 75, 76 . It is tempting to speculate that other butyrate-producing pathways may operate to contribute to the total butyrate pool.…”

Section: Discussionmentioning

confidence: 99%

AUF-1 knock down in mice overarches butyrate driven hypo-cholesteraemia by conjuring AUF-1-Dicer-1-miR122 hierarchy

Das

Kundu

Ghosh

et al. 2022

Preprint

View full text Add to dashboard Cite

This discourse probes the mechanistic molecular details of butyrate action in maintaining host- cholesterol balance. Hepatic miR122 being the most indispensable regulator of cholesterol metabolic enzymes, we studied upstream players of miR122 biogenesis in the presence and absence of butyrate in Huh7 cells and mice model. We showed that butyrate treatment caused upregulation of RNA-binding protein, AUF-1 resulting in RNase-III nuclease, Dicer-1 instability, and significant diminution of miR122. We proved its importance of AUF-1 and sequential downstream players in AUF-1-knock-down mice. We synthesized unique self-transfecting GMO (guanidinium-morpholino- oligonucleotides) linked PMO (Phosphorodiamidate-Morpholino Oligonucleotides)-based antisense reagent and injection of which in mouse caused near absence of AUF-1 coupled with increased Dicer- 1 and miR122, and reduced serum cholesterol regardless of butyrate treatment indicating that butyrate acts though AUF-1. The roster of intracellular players was as follows: AUF-1-Dicer-1-miR122 for triggering butyrate driven hypocholesterlaemia. To our knowledge this is the first report linking AUF-1 with cholesterol biogenesis.

show abstract

“…Fnn 25586 and Fnn 23726 DMTases allow for the development of the first genetic system in Fnn 25586. Fnn 25586 is one of the classical strains that has been studied for more than four decades 67 , yet molecular studies have not been possible because of the inability to be transformed. Our goal was to use the same system we developed previously for gene knockouts in Fnn 23726 64 .…”

Section: In Vivo Methylation Of Plasmids Increases Transformation Of ...mentioning

confidence: 99%

DNA methyltransferase enhanced Fusobacterium nucleatum genetics

Umaña

Nguyen

Sanders

et al. 2022

Preprint

View full text Add to dashboard Cite

Bacterial restriction-modification (R-M) systems are a first line immune defense against foreign DNA from viruses and other bacteria. While R-M systems are critical in maintaining genome integrity, R-M nucleases unfortunately present significant barriers to targeted genetic modification. Bacteria of the genus Fusobacterium are oral, Gram-negative, anaerobic, opportunistic pathogens that are implicated in the progression and severity of multiple cancers and tissue infections, yet our understanding of their direct roles in disease have been severely hindered by their genetic recalcitrance. Here, we demonstrate a path to overcome these barriers in Fusobacterium by using native DNA methylation as a host mimicry strategy to bypass R-M system cleavage of user introduced plasmid DNA. We report the identification, characterization, and successful use of Fusobacterium nucleatum (Fn) Type II and III DNA methyltransferase (DMTase) enzymes to produce a multi-fold increase in gene knockout efficiency in the strain Fusobacterium nucleatum subsp. nucleatum 23726 (Fnn 23726), as well as the first efficient gene knockouts and complementations in Fnn 25586. We show plasmid protection can be accomplished in vitro with purified enzymes, as well as in vivo in an E. coli host that constitutively expresses Fnn DMTase enzymes. By characterizing specific DMTases that are critical for bypassing R-M systems, we have enhanced our understanding of potential enzyme combinations, with the goal of expanding these studies to genetically modify clinical isolates of Fusobacterium that have thus far been inaccessible to molecular characterization. This proof-of-concept study provides a roadmap to guide molecular microbiology efforts of the scientific community to facilitate the discovery of new Fusobacterium virulence genes, thereby leading to a new era of characterizing how an oral opportunistic pathogen contributes to an array of human infections and diseases.IMPORTANCEFusobacterium nucleatum is an oral opportunistic pathogen associated with diseases including cancer and preterm birth. Our understanding of how this bacterium modulates human disease has been hindered by a lack of genetic systems. Here we show that F. nucleatum DNA methyltransferase modified plasmid DNA overcomes the transformation barrier and allows the development of genetic systems in previously inaccessible strains. We present a strategy that can be expanded to enable the genetic modification of clinical isolates, thereby fostering investigational studies to uncover novel host-pathogen interactions in Fusobacterium.

show abstract

Pathway of lysine degradation in Fusobacterium nucleatum

Cited by 62 publications

References 21 publications

Development of β-Amino Acid Dehydrogenase for the Synthesis of β-Amino Acids via Reductive Amination of β-Keto Acids

Development of β-Amino Acid Dehydrogenase for the Synthesis of β-Amino Acids via Reductive Amination of β-Keto Acids

AUF-1 knock down in mice overarches butyrate driven hypo-cholesteraemia by conjuring AUF-1-Dicer-1-miR122 hierarchy

DNA methyltransferase enhanced Fusobacterium nucleatum genetics

Contact Info

Product

Resources

About