Creatinine decomposing enzymes in Pseudomonas putida.

Tsuru, Daisuke; Oka, Imao; Yoshimoto, Tadashi

doi:10.1271/bbb1961.40.1011

Cited by 24 publications

(12 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This reaction is the first step of the creatinine-metabolizing pathway found in Pseudomonas putida. 1 Creatine is then degraded by creatinase (EC 3.5.3.3), 2 sarcosine dehydrogenase (EC 1.5.3.1), 3 and formaldehyde dehydrogenase (EC 1.1. 1.159) 4 in this microorganism.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Substitution of Glu122 by Glutamine Revealed the Function of the Second Water Molecule as a Proton Donor in the Binuclear Metal Enzyme Creatininase

Yamashita

Nakajima

Matsushita

et al. 2010

Journal of Molecular Biology

Self Cite

View full text Add to dashboard Cite

Creatininase is a binuclear zinc enzyme and catalyzes the reversible conversion of creatinine to creatine. It exhibits an open-closed conformational change upon substrate binding, and the differences in the conformations of Tyr121, Trp154, and the loop region containing Trp174 were evident in the enzyme-creatine complex when compared to those in the ligand-free enzyme. We have determined the crystal structure of the enzyme complexed with a 1-methylguanidine. All subunits in the complex existed as the closed form, and the binding mode of creatinine was estimated. Site-directed mutagenesis revealed that the hydrophobic residues that show conformational change upon substrate binding are important for the enzyme activity. We propose a catalytic mechanism of creatininase in which two water molecules have significant roles. The first molecule is a hydroxide ion (Wat1) that is bound as a bridge between the two metal ions and attacks the carbonyl carbon of the substrate. The second molecule is a water molecule (Wat2) that is bound to the carboxyl group of Glu122 and functions as a proton donor in catalysis. The activity of the E122Q mutant was very low and it was only partially restored by the addition of ZnCl(2) or MnCl(2). In the E122Q mutant, k(cat) is drastically decreased, indicating that Glu122 is important for catalysis. X-ray crystallographic study and the atomic absorption spectrometry analysis of the E122Q mutant-substrate complex revealed that the drastic decrease of the activity of the E122Q was caused by not only the loss of one Zn ion at the Metal1 site but also a critical function of Glu122, which most likely exists for a proton transfer step through Wat2.

show abstract

Section: Introductionmentioning

confidence: 99%

“…1 Later, it was found that the enzyme activity was increased approximately 4-fold by the addition of manganese. 5 The gene encoding creatininase was cloned from P. putida and over-expressed using an Escherichia coli system.…”

Section: Introductionmentioning

confidence: 99%

Substitution of Glu122 by Glutamine Revealed the Function of the Second Water Molecule as a Proton Donor in the Binuclear Metal Enzyme Creatininase

Yamashita

Nakajima

Matsushita

et al. 2010

Journal of Molecular Biology

Self Cite

View full text Add to dashboard Cite

show abstract

“…Creatinine amidohydrolase is widely distributed binuclear Zn 2+ enzyme responsible for the hydrolysis of creatinine to form creatine 15,16 . Interestingly, within this family are the two subfamilies FAPy deformylase and the mycofactocin peptidase, both of which are associated with cofactor or putative cofactor biosynthesis.…”

mentioning

confidence: 99%

Mycofactocin Biosynthesis Proceeds through 3-Amino-5-[(p-hydroxyphenyl)methyl]-4,4-dimethyl-2-pyrrolidinone (AHDP); Direct Observation of MftE Specificity toward MftA*

et al. 2018

View full text Add to dashboard Cite

The structure of the ribosomally synthesized and post-translationally modified peptide product, mycofactocin is unknown. Recently, the first step in mycofactocin biosynthesis was shown to be catalyzed by MftC in two S-adenosylmethionine dependent steps. In the first step, MftC catalyzes the oxidative decarboxylation of the MftA peptide to produce the styrene containing intermediate MftA**, followed by a subsequent C-C bond formation to yield the lactam containing MftA*. Here, we demonstrate the subsequent biosynthetic step catalyzed by MftE is specific for MftA*. The hydrolysis of MftA* leads to the formation of MftA(1–28) and 3-amino-5-[(p-hydroxyphenyl) methyl]-4,4-dimethyl-2-pyrrolidinone (AHDP). The hydrolysis reaction is Fe2+ dependent and addition of the metal to the reaction leads to a kobs ~ 0.2 min−1. Lastly, we validate the structure of AHDP by a 1H, 13C, and COSY NMR techniques as well as mass spectrometry.

show abstract

“…Although a peptidase is not encoded by the Mft cluster, we were intrigued by the presence of a gene that encodes a protein with similarity to the creatininase subfamily of amidohydrolases. The prototypical creatininase hydrolyzes an amide bond in the cyclic metabolite creatinine to produce creatine (23)(24)(25). We reasoned that it might be possible that the creatininase homolog encoded in this cluster potentially processes the decarboxylated MftA to a smaller metabolite en route to the mature natural product.…”

mentioning

confidence: 99%

The Creatininase Homolog MftE from Mycobacterium smegmatis Catalyzes a Peptide Cleavage Reaction in the Biosynthesis of a Novel Ribosomally Synthesized Post-translationally Modified Peptide (RiPP)

Bruender¹,

Bandarian

2017

Journal of Biological Chemistry

View full text Add to dashboard Cite

Most ribosomally synthesized and post-translationally modified peptide (RiPP) natural products are processed by tailoring enzymes to create complex natural products that are still recognizably peptide-based. However, some tailoring enzymes dismantle the peptide en route to synthesis of small molecules. A small molecule natural product of as yet unknown structure, mycofactocin, is thought to be synthesized in this way via the gene cluster found in many strains of mycobacteria. This cluster harbors at least six genes, which appear to be conserved across species. We have previously shown that one enzyme from this cluster, MftC, catalyzes the oxidative decarboxylation of the C-terminal Tyr of the substrate peptide MftA in a reaction that requires the MftB protein. Herein we show that encodes a creatininase homolog that catalyzes cleavage of the oxidatively decarboxylated MftA peptide to liberate its final two residues, including the C-terminal decarboxylated Tyr (VY*). Unlike MftC, which requires MftB for function, MftE catalyzes the cleavage reaction in the absence of MftB. The identification of this novel metabolite, VY*, supports the notion that the cluster is involved in generating a small molecule from the MftA peptide. The ability to produce VY* from MftA by reconstitution of the activities of MftB, MftC, and MftE sets the stage for identification of the novel metabolite that results from the proteins encoded by the cluster.

show abstract

Creatinine decomposing enzymes in Pseudomonas putida.

Cited by 24 publications

References 0 publications

Substitution of Glu122 by Glutamine Revealed the Function of the Second Water Molecule as a Proton Donor in the Binuclear Metal Enzyme Creatininase

Substitution of Glu122 by Glutamine Revealed the Function of the Second Water Molecule as a Proton Donor in the Binuclear Metal Enzyme Creatininase

Mycofactocin Biosynthesis Proceeds through 3-Amino-5-[(p-hydroxyphenyl)methyl]-4,4-dimethyl-2-pyrrolidinone (AHDP); Direct Observation of MftE Specificity toward MftA*

The Creatininase Homolog MftE from Mycobacterium smegmatis Catalyzes a Peptide Cleavage Reaction in the Biosynthesis of a Novel Ribosomally Synthesized Post-translationally Modified Peptide (RiPP)

Contact Info

Product

Resources

About