For example, the term “comprising a nitrile oxido-reductase” includes single or plural nitrile oxido-reductase molecules and is considered equivalent to the phrase “comprising at least one nitrile oxido-reductase.” The term “or” refers to a single element of stated alternative elements or a combination of two or more elements, unless the context clearly indicates otherwise. Users are In some examples, the reaction includes salt, salt such as NaCl or KCl. Synthesis and Characterisation of α-Glycosyloxyamides Derive... Boron-Catalyzed Silylative Reduction of Nitriles in Accessing Primary Amines and Imines, Method for synthesising bis[3-(N,N-dialkylamino)propyl]ethers, Photochemical reactions of ?,?-unsaturated ?,?-epoxy ketones and nitriles in the presence of amine. SEQ ID NO: 29 is an exemplary nitrile oxido-reductase amino acid sequence (GenBank Accession no. A Domino Reaction of α,β-Acetylenic γ-Hydroxy Nitriles with Arenecarboxylic Acids: An Unexpected Facile Shortcut to 4-Cyano-3(2H)-furanones. Methods of reducing a nitrile containing compound to an amine are provided. BLASTN is used to compare nucleic acid sequences, while BLASTP is used to compare amino acid sequences. Biol. Laura Leemans, Marc D. Walter, Frank Hollmann, Anett Schallmey, Luuk M. van Langen. Synthetic methods : Part (ii) Oxidation and reduction methods. in the Biosciences 8, 155-65, 1992; and Pearson et al., Meth. 5) Nitriles to aldehydes or primary amines: The nitriles are either reduced to aldehydes or primary amines depending on the amount of DIBAL-H consumed. Methods of producing variant sequences are known in the art. YP—217894). SEQ ID NO: 9 is an exemplary nitrile oxido-reductase amino acid sequence. without permission from the American Chemical Society. Thus, E. coli, B. subtilis, S. cerevisiae, K. lactis, Candida blankii, and Pichia pastoris are microorganisms and can be used as described herein. Primary amines contain the -NH 2 group. YP—208721). When less than the entire sequence is being compared for sequence identity, homologs will typically possess at least 75% sequence identity over short windows of 10-20 amino acids, and can possess sequence identities of at least 85%, at least 90%, at least 95% or at least 98% depending on their identity to the reference sequence. The demands of an increasingly environmentally conscientious society and industries need to maximize its bottom line have driven a rapid increase in the use of enzymes and microorganisms for mediating industrially important chemical transformations (Schmid et al., Nature 409: 258-68, 2001; Ritter, Chem. 11 is a schematic drawing showing exemplary nitrile containing compounds. FolE is part of a structural superfamily of functionally distant pterin/purine binding proteins which utilize a common oligomerization of the characteristic T-fold (Tunneling-fold), comprised of an antiparallel β-sheet and two helices, to form a β2nαn barrel (Colloc'h et al. NADPH regeneration systems are known in the art, for example Woodyear et al. For clarity and space, only sequences from select organisms are shown from among 60 sequences in the original alignment, and the N-terminii have been truncated. For example, a nitrile oxido-reductase nucleic acid sequence can contain at least 24 contiguous nucleotides of a nitrile oxido-reductase nucleic acid sequence. Microbiol. SEQ ID NO: 40 is an exemplary nitrile oxido-reductase amino acid sequence (GenBank Accession no. To learn more, see our tips on writing great answers. 326:503-16, 2003). AAQ66412). In the accompanying sequence listing: SEQ ID NO: 1 is a nucleic acid sequence of a B. subtilis QueF (GeneID No. NP—421453). SEQ ID NO: 70 is an exemplary nitrile oxido-reductase amino acid sequence from Pseudomonas syringae. In other particular examples, the variant nitrile oxido-reductase includes an E98A, E98L, E98I, E98M, E98V, E98Q, E98N, E98K, or E98R substitution (wherein the substitution refers to the sequence shown in SEQ ID NO: 2, but one skilled in the art can identify the corresponding Glu residue, for example by using FIGS. These reactions are typically non-selective, requiring the use of protecting groups when other reducible functional groups are present, and can result in the formation of unwanted byproducts. Crystals were screened for diffraction quality at beam line 11-1 of the Stanford Synchrotron Research Laboratory (SSRL), using a robot for mounting of crystals (using the SSRL crystal freezing cassette kit which holds 96 crystals at a time). NP—10841). 338:224-38, 2004; Schmidt et al. It has a standard Ramachandran plot with 90.2% of residues falling in its favored regions, and 8.3%, 1.5% in the allowed and generously allowed regions, respectively. One-step amino-functionalization of milled carbon fibre for enhancement of thermo-physical properties of epoxy composites. SEQ ID NO: 30 is an exemplary nitrile oxido-reductase amino acid sequence (GenBank Accession no. ) and its derivatives. Environ. Inverting lower triangular matrix in time n^2. ZP—00272373). Analysis of velocity data with variable NADPH and constant, saturating preQ0 provided a KM for NADPH of 36 μM, consistent with the KM values for other bacterial NADPH-dependent oxidoreductases (Smith et al. Examples of particular substitutions which can be made to a nitrile oxido-reductase, while still retaining nitrile oxido-reductase activity, include, but are not limited to: L9T, Y22P, K58M, K58I, I103T, D132V, K139A, E149W and N154Y of SEQ ID NO: 2, and A11E, G32P, H52Q, Y82A, K126E, T156E, or E174L of SEQ ID NO: 4, as well as combinations of these substitutions (similar substitutions can be made to SEQ ID NOS: 6-95, using the alignment provided in FIGS. NP—623163). When aligning short peptides (fewer than around 30 amino acids), the alignment should be performed using the Blast 2 sequences function, employing the PAM30 matrix set to default parameters (open gap 9, extension gap 1 penalties). In addition, the disclosure provides isolated nitrile oxido-reductase nucleic acid sequences which contain a variation of in the sequence. Xinzhi Chen, Shaodong Zhou, Yuehan Chen, Zehan Dong, Yeyu Gao, Chao Qian, Chaohong He.
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