Phenylalanine Ammonia Lyase Catalyzed Synthesis of Amino Acids by an MIO‐Cofactor Independent Pathway

Phenylalanine ammonia lyases (PALs) belong to a family of 4‐methylideneimidazole‐5‐one (MIO) cofactor dependent enzymes which are responsible for the conversion of L ‐phenylalanine into trans‐cinnamic acid in eukaryotic and prokaryotic organisms. Under conditions of high ammonia concentration, this deamination reaction is reversible and hence there is considerable interest in the development of PALs as biocatalysts for the enantioselective synthesis of non‐natural amino acids. Herein the discovery of a previously unobserved competing MIO‐independent reaction pathway, which proceeds in a non‐stereoselective manner and results in the generation of both L ‐ and D ‐phenylalanine derivatives, is described. The mechanism of the MIO‐independent pathway is explored through isotopic‐labeling studies and mutagenesis of key active‐site residues. The results obtained are consistent with amino acid deamination occurring by a stepwise E₁cB elimination mechanism.     

Enzymatic Fabrication of High‐Density RNA Arrays

A powerful new strategy for the fabrication of high‐density RNA arrays is described. A high‐density DNA array is fabricated by standard photolithographic methods, the surface‐bound DNA molecules are enzymatically copied into their RNA complements from a surface‐bound RNA primer, and the DNA templates are enzymatically destroyed, leaving behind the desired RNA array. The strategy is compatible with 2′‐fluoro‐modified (2′F) ribonucleoside triphosphates (rNTPs), which may be included in the polymerase extension reaction to impart nuclease resistance and other desirable characteristics to the synthesized RNAs. The use and fidelity of the arrays are explored with DNA hybridization, DNAzyme cleavage, and nuclease digestion experiments.     

Amino Acid Modified RNA Bases as Building Blocks of an Early Earth RNA-Peptide World

Fossils of extinct species allow us to reconstruct the process of Darwinian evolution that led to the species diversity we see on Earth today. The origin of the first functional molecules able to undergo molecular evolution and thus eventually able to create life, are largely unknown. The most prominent idea in the field posits that biology was preceded by an era of molecular evolution, in which RNA molecules encoded information and catalysed their own replication. This RNA world concept stands against other hypotheses, that argue for example that life may have begun with catalytic peptides and primitive metabolic cycles. The question whether RNA or peptides were first is addressed by the RNA-peptide world concept, which postulates a parallel existence of both molecular species. A plausible experimental model of how such an RNA-peptide world may have looked like, however, is absent. Here we report the synthesis and physicochemical evaluation of amino acid containing adenosine bases, which are closely related to molecules that are found today in the anticodon stem-loop of tRNAs from all three kingdoms of life. We show that these adenosines lose their base pairing properties, which allow them to equip RNA with amino acids independent of the sequence context. As such we may consider them to be living molecular fossils of an extinct molecular RNA-peptide world.     

Optically Triggered Control of the Charge Carrier Density in Chemically Functionalized Graphene Field Effect Transistors

Field effect transistors (FETs) based on 2D materials are of great interest for applications in ultrathin electronic and sensing devices. Here we demonstrate the possibility to add optical switchability to graphene FETs (GFET) by functionalizing the graphene channel with optically switchable azobenzene molecules. The azobenzene molecules were incorporated to the GFET channel by building a van der Waals heterostructure with a carbon nanomembrane (CNM), which is used as a molecular interposer to attach the azobenzene molecules. Under exposure with 365 nm and 455 nm light, azobenzene molecules transition between cis and trans molecular conformations, respectively, resulting in a switching of the molecular dipole moment. Thus, the effective electric field acting on the GFET channel is tuned by optical stimulation and the carrier density is modulated.     

Efficient Preparation of γ-Hydroxynitriles via Nitrile Enolate-Epoxide Reactions: Scope and Diastereoselectivity

The nucleophilic opening of epoxides by nitrile enolates using an efficient, convenient protocol is described. The diastereoselectivity of this reaction was explored and found to give syn:anti ratios ranging from 1.1:1.0 to 4.8:1.0.

     

On the photoelectron spectrum of sulphur trioxide

He(I), He(II) and variable temperature Ne(I) photoelectron spectra of sulphur trioxide are reported. All six of the expected ionic states have been detected and assigned; the assignment differs from a previous one. Ab initio calculations in a medium basis set have been carried out and spectral intensities have been calculated; agreement between the calculations and the spectra is good. The calculations show a substantial S(3d) character in the S-O bond, and the spectra are consistent with this.     

Reversed-phase poly(styrene-divinylbenzene) materials optimised for large scale preparative and process purification of synthetic peptides and recombinant proteins

Rigid macroporous copolymers of styrene and divinylbenzene have been designed for large-scale preparative and process-scale purification of synthetic peptides and recombinant proteins. The polymeric particles are mechanically stable and hence able to operate at the required high linear velocities. The pore size and pore morphology has been optimised to enable unhindered solute diffusion whilst providing maximum available surface area to enhance loading capacity. A 100 A pore size has been developed for synthetic peptides and a 300 A pore size for recombinant proteins. Precise control of particle size, within the range 10 to 20 microm, is possible which together with the very narrow particle size distribution enables maximum resolution/loading to be obtained within the pressure limits of the instrumentation being used. The chemical stability of the polymer enables cleaning in place with 1 M sodium hydroxide without particle dissolution or a deterioration in selectivity. These materials can be packed into compression hardware and are manufactured as single lots up to 100 kg (300 l) batch size.