l-Penicillamine is a mechanism-based inhibitor of serine palmitoyltransferase by forming a pyridoxal-5′-phosphate-thiazolidine adduct

Lowther, Jonathan; Beattie, Ashley E.; Langridge‐Smith, Patrick R. R.; Clarke, David J.; Campopiano, Dominic J.

doi:10.1039/c2md20020a

Cited by 16 publications

(15 citation statements)

References 29 publications

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“…11d) determined by UV absorbance at 340 nm (Supplementary Fig. 11a), with concomitant loss of the ketoenamine peak at 390 nm, as observed previously by Lowther et al 44. for serine palmitoyltransferase.…”

Section: Resultssupporting

confidence: 78%

An L-threonine transaldolase is required for L-threo-β-hydroxy-α-amino acid assembly during obafluorin biosynthesis

et al. 2017

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β-Lactone natural products occur infrequently in nature but possess a variety of potent and valuable biological activities. They are commonly derived from β-hydroxy-α-amino acids, which are themselves valuable chiral building blocks for chemical synthesis and precursors to numerous important medicines. However, despite a number of excellent synthetic methods for their asymmetric synthesis, few effective enzymatic tools exist for their preparation. Here we report cloning of the biosynthetic gene cluster for the β-lactone antibiotic obafluorin and delineate its biosynthetic pathway. We identify a nonribosomal peptide synthetase with an unusual domain architecture and an L-threonine:4-nitrophenylacetaldehyde transaldolase responsible for (2S,3R)-2-amino-3-hydroxy-4-(4-nitrophenyl)butanoate biosynthesis. Phylogenetic analysis sheds light on the evolutionary origin of this rare enzyme family and identifies further gene clusters encoding L-threonine transaldolases. We also present preliminary data suggesting that L-threonine transaldolases might be useful for the preparation of L-threo-β-hydroxy-α-amino acids.

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Section: Resultssupporting

confidence: 78%

An L-threonine transaldolase is required for L-threo-β-hydroxy-α-amino acid assembly during obafluorin biosynthesis

et al. 2017

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“…From the configuration of K233 and Y123 in the Ls -MalY structural model and the results of the present mutational analysis, the Ls -MalY catalyzed racemase reaction appears to proceed through a two-base mechanism similar to that of the well-known alanine racemase ( Watanabe et al, 2002 ). Some PLP-dependent enzymes are known to be inhibited by L -Cys and/or D- Cys through the formation of thiazolidine derivatives ( Schonbeck et al, 1975 ; Dunlop and Neidle, 2005 ; Lowther et al, 2012 ). The spectral features of Ls -MalY when reacted with D- Cys are in agreement with such reports, suggesting that Ls -MalY forms a dea D- end product, namely, a thiazolidine adduct, and that Ls -MalY can catalyze β-lyase reactions against L -Cys but not racemase reactions between L -Cys and D- Cys.…”

Section: Discussionmentioning

confidence: 99%

A Novel Bifunctional Amino Acid Racemase With Multiple Substrate Specificity, MalY From Lactobacillus sakei LT-13: Genome-Based Identification and Enzymological Characterization

Kato

Oikawa

2018

Front. Microbiol.

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The Lactobacillus sakei strain LK-145 isolated from Moto, a starter of sake, produces potentially large amounts of three D-amino acids, D-Ala, D-Glu, and D-Asp, in a medium containing amylase-digested rice as a carbon source. The comparison of metabolic pathways deduced from the complete genome sequence of strain LK-145 to the type culture strain of Lactobacillus sakei strain LT-13 showed that the L- and D-amino acid metabolic pathways are similar between the two strains. However, a marked difference was observed in the putative cysteine/methionine metabolic pathways of strain LK-145 and LT-13. The cystathionine β-lyase homolog gene malY was annotated only in the genome of strain LT-13. Cystathionine β-lyase is an important enzyme in the cysteine/methionine metabolic pathway that catalyzes the conversion of L-cystathionine into L-homocysteine. In addition to malY, most genome-sequenced strains of L. sakei including LT-13 lacked the homologous genes encoding other putative enzymes in this pathway. Accordingly, the cysteine/methionine metabolic pathway likely does not function well in almost all strains of L. sakei. We succeeded in cloning and expressing the malY gene from strain LT-13 (Ls-malY) in the cells of Escherichia coli BL21 (DE3) and characterized the enzymological properties of Ls-MalY. Spectral analysis of purified Ls-MalY showed that Ls-MalY contained a pyridoxal 5′-phosphate (PLP) as a cofactor, and this observation agreed well with the prediction based on its primary structure. Ls-MalY showed amino acid racemase activity and cystathionine β-lyase activity. Ls-MalY showed amino acid racemase activities in various amino acids, such as Ala, Arg, Asn, Glu, Gln, His, Leu, Lys, Met, Ser, Thr, Trp, and Val. Mutational analysis revealed that the 𝜀-amino group of Lys233 in the primary structure of Ls-MalY likely bound to PLP, and Lys233 was an essential residue for Ls-MalY to catalyze both the amino acid racemase and β-lyase reactions. In addition, Tyr123 was a catalytic residue in the amino acid racemase reaction but strongly affected β-lyase activity. These results showed that Ls-MalY is a novel bifunctional amino acid racemase with multiple substrate specificity; both the amino acid racemase and β-lyase reactions of Ls-MalY were catalyzed at the same active site.

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“…l -Penicillamine has also been shown to inhibit other PLP enzymes such as alanine aminotransferase and glutamate decarboxylase. 150 d -Penicillamine can also bind to the PLP cofactor, but is a weaker inhibitor due to its inverse stereochemistry. 151 Similarly, SPT can be inhibited by d -serine, which forms an external aldimine which cannot subsequently be deprotonated.…”

Section: Serine Palmitoyltransferase (Spt)mentioning

confidence: 99%

Sphingolipid biosynthesis in man and microbes

2018

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l-Penicillamine is a mechanism-based inhibitor of serine palmitoyltransferase by forming a pyridoxal-5′-phosphate-thiazolidine adduct

Abstract: Post-print of a peer-reviewed article published by the Royal Society of Chemistry.

Cited by 16 publications

References 29 publications

An L-threonine transaldolase is required for L-threo-β-hydroxy-α-amino acid assembly during obafluorin biosynthesis

An L-threonine transaldolase is required for L-threo-β-hydroxy-α-amino acid assembly during obafluorin biosynthesis

A Novel Bifunctional Amino Acid Racemase With Multiple Substrate Specificity, MalY From Lactobacillus sakei LT-13: Genome-Based Identification and Enzymological Characterization

Sphingolipid biosynthesis in man and microbes

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