485-80-3Relevant academic research and scientific papers
Directed Evolution of a Halide Methyltransferase Enables Biocatalytic Synthesis of Diverse SAM Analogs
Tang, Qingyun,Grathwol, Christoph W.,Aslan-üzel, A?k?n S.,Wu, Shuke,Link, Andreas,Pavlidis, Ioannis V.,Badenhorst, Christoffel P. S.,Bornscheuer, Uwe T.
supporting information, p. 1524 - 1527 (2020/11/30)
Biocatalytic alkylations are important reactions to obtain chemo-, regio- and stereoselectively alkylated compounds. This can be achieved using S-adenosyl-l-methionine (SAM)-dependent methyltransferases and SAM analogs. It was recently shown that a halide methyltransferase (HMT) from Chloracidobacterium thermophilum can synthesize SAM from SAH and methyl iodide. We developed an iodide-based assay for the directed evolution of an HMT from Arabidopsis thaliana and used it to identify a V140T variant that can also accept ethyl-, propyl-, and allyl iodide to produce the corresponding SAM analogs (90, 50, and 70 % conversion of 15 mg SAH). The V140T AtHMT was used in one-pot cascades with O-methyltransferases (IeOMT or COMT) to achieve the regioselective ethylation of luteolin and allylation of 3,4-dihydroxybenzaldehyde. While a cascade for the propylation of 3,4-dihydroxybenzaldehyde gave low conversion, the propyl-SAH intermediate could be confirmed by NMR spectroscopy.
An enzymatic Finkelstein reaction: Fluorinase catalyses direct halogen exchange
Lowe, Phillip T.,Cobb, Steven L.,O'Hagan, David
supporting information, p. 7493 - 7496 (2019/08/20)
The fluorinase enzyme from Streptomyces cattleya is shown to catalyse a direct displacement of bromide and iodide by fluoride ion from 5′-bromodeoxyadenosine (5′-BrDA) and 5′-iododeoxyadenosine (5′-IDA) respectively to form 5′-fluorodeoxyadenosine (5′-FDA) in the absence of l-methionine (l-Met) or S-adenosyl-l-methionine (SAM). 5′-BrDA is the most efficient substrate for this enzyme catalysed Finkelstein reaction.
Preparation, Assay, and Application of Chlorinase SalL for the Chemoenzymatic Synthesis of S-Adenosyl-L-Methionine and Analogs
Davis, Tony D.,Kunakom, Sylvia,Burkart, Michael D.,Eustaquio, Alessandra S.
, p. 367 - 388 (2018/06/16)
S-adenosyl-L-methionine (SAM) is universal in biology, serving as the second most common cofactor in a variety of enzymatic reactions. One of the main roles of SAM is the methylation of nucleic acids, proteins, and metabolites. Methylation often imparts regulatory control to DNA and proteins, and leads to an increase in the activity of specialized metabolites such as those developed as pharmaceuticals. There has been increased interest in using SAM analogs in methyltransferase-catalyzed modification of biomolecules. However, SAM and its analogs are expensive and unstable, degrading rapidly under physiological conditions. Thus, the availability of methods to prepare SAM in situ is desirable. In addition, synthetic methods to generate SAM analogs suffer from low yields and poor diastereoselectivity. The chlorinase SalL from the marine bacterium Salinispora tropica catalyzes the reversible, nucleophilic attack of chloride at the C5′ ribosyl carbon of SAM leading to the formation of 5′-chloro-5′-deoxyadenosine (ClDA) with concomitant displacement of L-methionine. It has been demonstrated that the in vitro equilibrium of the SalL-catalyzed reaction favors the synthesis of SAM. In this chapter, we describe methods for the preparation of SalL, and the chemoenzymatic synthesis of SAM and SAM analogs from ClDA and L-methionine congeners using SalL. In addition, we describe procedures for the in situ chemoenzymatic synthesis of SAM coupled to DNA, peptide, and metabolite methylation, and to the incorporation of isotopes into alkylated products.
A Tandem Enzymatic sp2-C-Methylation Process: Coupling in Situ S-Adenosyl-l-Methionine Formation with Methyl Transfer
Sadler, Joanna C.,Humphreys, Luke D.,Snajdrova, Radka,Burley, Glenn A.
, p. 992 - 995 (2017/06/13)
A one-pot, two-step biocatalytic platform for the regiospecfic C-methylation and C-ethylation of aromatic substrates is described. The tandem process utilises SalL (Salinospora tropica) for in situ synthesis of S-adenosyl-l-methionine (SAM), followed by alkylation of aromatic substrates by the C-methyltransferase NovO (Streptomyces spheroides). The application of this methodology is demonstrated for the regiospecific labelling of aromatic substrates by the transfer of methyl, ethyl and isotopically labelled 13CH3, 13CD3 and CD3 groups from their corresponding SAM analogues formed in situ.
A general NMR-based strategy for the in Situ characterization of sugar-nucleotide-dependent biosynthetic pathways
Singh, Shanteri,Peltier-Pain, Pauline,Tonelli, Marco,Thorson, Jon S.
supporting information, p. 3220 - 3223 (2014/07/08)
A simple method for the study of sugar-nucleotide-dependent multienzyme cascades is highlighted where the use of selectively 13C-labeled sugar nucleotides and inverse 13C detection NMR offers fast, direct detection and quantification of reactants and products and circumvents the need for chromatographic separation. The utility of the method has been demonstrated by characterizing four previously uncharacterized sugar nucleotide biosynthetic enzymes involved in calicheamicin biosynthesis.
Biocatalytic Friedel-Crafts alkylation using non-natural cofactors
Stecher, Harald,Tengg, Martin,Ueberbacher, Bernhard J.,Remler, Peter,Schwab, Helmut,Griengl, Herfried,Gruber-Khadjawi, Mandona
supporting information; experimental part, p. 9546 - 9548 (2010/03/25)
A novel biocatalytic protocol for C -C bond formation is described and is an equivalent to Friedel-Crafts alkylation. S-Adenosyl-L-methionine (SAM), the major Chemical Equation Presentation methyl donor for biological methylation catalyzed by methyltransferases (Mtases), can perform alkylations (see scheme). These enzymes can accept non-natural cofactors and transfer functionalities other than methyl onto aromatic substrates
The fluorinase from Streptomyces cattleya is also a chlorinase
Deng, Hai,Cobb, Steven L.,McEwan, Andrew R.,McGlinchey, Ryan P.,Naismith, James H.,O'Hagan, David,Robinson, David A.,Spencer, Jonathan B.
, p. 759 - 762 (2007/10/03)
(Chemical Equation Presented) Choices choices: The fluorinase enzyme from Streptomyces cattleya (catalyzes the formation of a C-F bond from fluoride ions) also has the capacity to utilize a chloride ion although it has a clear preference for the fluoride ion. The enzyme mediates a nucleophilic chlorination reaction, which is an unusual mechanism for enzymatic chlorination.
