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At least 289 records · Page 16

Branched-chain amino acid specialization drove diversification within Calditenuaceae ( Caldarchaeia ) and enables their cultivation

Many thermophiles that are abundant in high-temperature geothermal systems have never been cultivated and are poorly understood, including deeply branching members of the archaeal phylum Thermoproteota. Here, we describe the genome-guided cultivation of one such organism, Calditenuis ramacidaminiphagus, and show that it has evolved a heterotrophic metabolism focused on branched-chain amino acids (BCAAs). Initially, fluorescence in situ hybridization and nanoscale secondary ion mass spectrometry (FISH-nanoSIMS) showed that Cal. ramacidaminiphagus assimilated amino acids rapidly in casamino acid-amended enrichment cultures. Metagenome and metaproteome analyses showed a high abundance and expression of BCAA transporter genes, suggesting a BCAA-focused metabolism. This inference was supported by the subsequent enrichment of Cal. ramacidaminiphagus in BCAA-fed cultures, reaching 2.66×10 6 cells/mL and 48.7% of the community, whereas it was outcompeted when polar amino acids were included. Metabolic reconstruction and metaproteomics suggest that BCAAs are channeled into the mevalonate pathway for lipid biosynthesis and fuel ATP production through the TCA cycle coupled with aerobic respiration and through production of branched-chain organic acids by overflow metabolism. Ancestral state reconstructions and phylogenetic analyses of 62 Caldarchaeales genomes revealed multiple horizontal transfers of BCAA transporters to the ancestor of the genus Calditenuis. Our study highlights the crucial role of BCAAs in the early evolution and niche of this genus, and suggests a high degree of resource partitioning even within low-diversity thermophilic communities.

Biological and medical sciences↗

Amino Acids in the Antarctic Martian Meteorite MIL03346

The report by McKay et al. that the Martian meteorite ALH84001 contains evidence for life on Mars remains controversial. Of central importance is whether ALH84001 and other Antarctic Martian meteorites contain endogenous organic compounds. In any investigation of organic compounds possibly derived from Mars it is important to focus on compounds that play an essential role in biochemistry as we know it and that have properties such as chirality which can be used to distinguish between biotic versus abiotic origins. Amino acids are one of the few compounds that fulfill these requirements. Previous analyses of the Antarctic Martian meteorites ALH84001 and EETA79001 have shown that these meteorites contain low levels of terrestrial amino acid contamination derived from Antarctic ice meltwater. Here we report preliminary amino acid investigations of a third Antarctic Martian meteorite MIL03346 which was discovered in Antarctica during the 2003-04 ANSMET season. Additional information is included in the original extended abstract

Glavin, D. P.↗

Extraterrestrial hydroxy amino acids in CM and CR carbonaceous chondrites

The abundances, distributions, and enantiomeric ratios of a family of three- and four-carbon hydroxy amino acids (HAAs) were investigated in extracts of five CM and four CR carbonaceous chondrites by gas chromatography-mass spectrometry analyses. HAAs were detected in both the acid hydrolysates of the hot water extracts and the 6 M HCl extracts of all the CM and CR chondrites analyzed here with total hot water and HCl extractable HAA concentrations ranging from 6.94 to 315 nmol/g. The HAA analyses performed in this study revealed: (1) the combined (hot water + HCl) extracts of CR2 chondrites contained greater abundances of α-HAAs than that of CM2 chondrites and (2) the combined extracts of CM and CR chondrites contained roughly similar abundances of β- and γ-HAAs. Application of the new GC-MS method developed here resulted in the first successful chromatographic resolution of the enantiomers of an α-dialkyl HAA, d,l-α-methylserine, in carbonaceous chondrite extracts. Meteoritic α-methylserine was found to be mostly racemic within error and did not show l-enantiomeric excesses correlating with the degree of aqueous alteration, a phenomenon observed in meteoritic isovaline, another α-dialkyl amino acid. The HAAs identified in CM and CR chondrite extracts could have been produced during parent body alteration from the Strecker cyanohydrin reaction (for α-HAAs) and an ammonia-involved formose-like reaction (for β-, and γ-HAAs).

Amino acid(s)↗

Highly efficient peptide formation from N-acetylaminoacyl-AMP anhydride and free amino acid

The kinetics of formation of the N-blocked dipeptide, N-acetylglycylglycine, from N-acetylglycyl adenylate anhydride and glycine in aqueous solution at 25 C, and at various PH's are reported. The reaction is of interest in that over a physiologically relevant pH range (6-8), peptide synthesis proceeds more rapidly than hydrolysis, even at those pH's at which this compound becomes increasingly susceptible to base-catalyzed hydrolysis. Under similar conditions, the corresponding unblocked aminoacyl adenylate anhydrides are considerably more unstable, and undergo appreciable hydrlysis in the presence of free amino acid. Because N-blocked aminoacyl adenylate anhydrides serve as model compounds of peptidyl adenylate anhydrides, these results suggest that primitive amino acid polymerization systems may have operated by cyclic reactivation of the peptidyl carboxyl group, rather than that of the incoming amino acid.

Mullins, D. W., Jr.↗

Amino acids in a Fischer Tropsch type synthesis

One postulation is described for the presence of organic compounds in meteorites which states that they were formed during the condensation of the solar nebula. A viable laboratory simulation of these conditions can be modeled after the industrial Fischer Tropsch reaction, which is known to produce organic compounds called hydrocarbons. In this simulation, a mixture of carbon monoxide, hydrogen and ammonia is heated in the presence of iron meteorite. The reaction products for amino acids, a class of organic compounds important to life, were examined. A large number of these compounds is found in meteorites and other chemical evolution experiments, but only small quantities of a few amino acids were found in the present simulation work. These results are at odds with the existing literature in which many amino acids were reported.

Brown, D. L.↗

NCAP: Noncanonical Amino Acid Parameterization Software for CHARMM Potentials

Noncanonical Amino Acids (NCAAs) provide numerous avenues for introduction of novel functionality to peptides and proteins. NCAAs can be incorporated through solid phase synthesis or genetic code expansion in conjugation with heterologous expression of the encoded protein modification. Due to the difficulty of synthesis, wide chemical space and lack of empirically resolved structures modeling the effects of NCAA mutation is critical for rational protein design. To evaluate the structural and functional perturbations NCAAs introduce we utilize molecular potentials that describe the forces in protein structure. Most potentials such as CHARMM are designed to model canonical residues but can be parameterized in include novel NCAAs. Here, in this work, we introduce NCAP a software package to generate CHARMM compatible parameters from quantum chemical calculation. Unlike currently available tools NCAP is designed to recognize NCAA structure and automatically bridge the gap between DFT calculations and potential parameters. For our software we discuss workflow, validation against canonical parameter sets and comparison to published NCAA-protein structures.

59 BASIC BIOLOGICAL SCIENCES↗

Sequences Of Amino Acids For Human Serum Albumin

Sequences of amino acids defined for use in making polypeptides one-third to one-sixth as large as parent human serum albumin molecule. Smaller, chemically stable peptides have diverse applications including service as artificial human serum and as active components of biosensors and chromatographic matrices. In applications involving production of artificial sera from new sequences, little or no concern about viral contaminants. Smaller genetically engineered polypeptides more easily expressed and produced in large quantities, making commercial isolation and production more feasible and profitable.

Carter, Daniel C.↗