MS/MS fragmentation-guided search of TMG-chitooligomycins and their structure-activity relationship in specific β-N-acetylglucosaminidase inhibition

The reducing tetrasaccharide TMG-chitotriomycin (1) is an inhibitor of β-N-acetylglucosaminidase (GlcNAcase), produced by the actinomycete Streptomyces anulatus NBRC13369. The inhibitor shows a unique inhibitory spectrum, that is, selectivity toward enzymes from chitin-containing organisms such as insects and fungi. Nevertheless, its structure-selectivity relationship remains to be clarified. In this study, we conducted a structure-guided search of analogues of 1 in order to obtain diverse N,N,N-trimethylglucosaminium (TMG)-containing chitooligosaccharides. In this approach, the specific fragmentation profile of 1 on ESI-MS/MS analysis was used for the selective detection of desired compounds. As a result, two new analogues, named TMG-chitomonomycin (3) and TMG-chitobiomycin (2), were obtained from a culture filtrate of 1-producing Streptomyces. Their enzyme-inhibiting activity revealed that the potency and selectivity depended on the degree of polymerization of the reducing end GlcNAc units. Furthermore, a computational modeling study inspired the inhibitory mechanism of TMG-related compounds as a mimic of the substrate in the Michaelis complex of the GH20 enzyme. This study is an example of the successful application of a MS/MS experiment for structure-guided isolation of natural products.     

Peptide bond formation by aminolysin-A catalysis: a simple approach to enzymatic synthesis of diverse short oligopeptides and biologically active puromycins

A new S9 family aminopeptidase derived from the actinobacterial thermophile Acidothermus cellulolyticus was cloned and engineered into a transaminopeptidase by site-directed mutagenesis of catalytic Ser(491) into Cys. The engineered biocatalyst, designated aminolysin-A, can catalyze the formation of peptide bonds to give linear homo-oligopeptides, hetero-dipeptides, and cyclic dipeptides using cost-effective substrates in a one-pot reaction. Aminolysin-A can recognize several C-terminal-modified amino acids, including the l- and d-forms, as acyl donors as well as free amines, including amino acids and puromycin aminonucleoside, as acyl acceptors. The absence of amino acid esters prevents the formation of peptides; therefore, the reaction mechanism involves aminolysis and not a reverse reaction of hydrolysis. The aminolysin system will be a beneficial tool for the preparation of structurally diverse peptide mimetics by a simple approach.     

Fluorous solvent for cell culture

Incubation of mouse melanoma B16 cells in fluorous solvents with low boiling point such as perfluoromethylcyclohexane, 1,1,1,3,3,3-hexafluoro-2-propanol, ethylpentafluoropropionate resulted in cell death. However, cells lived up to 2 days in fluorous alcohols such as 2,2,3,3,4,4,5,5-octafluoro-1-pentanol and 3,3,4,4,5,5,6,6,6-nonafluoro-1-hexanol with relatively higher fluorine content. Remarkably, cells survived deprived of nutrition up to 4 days when incubated in 2,2,3,3,4,4,5,5,6,6,6-undecafluoro-1-hexanol or in 2,2,3,3,4,4,5,5,6,6,7,7-dodecafluoroheptanol that have the most number of fluorine atoms (11 and 12, respectively) among the perfluoroalkyl alcohols used, and with boiling points of 128 °C and 169 °C, respectively.     

First principles based mean field model for oxygen reduction reaction

A first principles-based mean field model was developed for the oxygen reduction reaction (ORR) taking account of the coverage- and material-dependent reversible potentials of the elementary steps. This model was applied to the simulation of single crystal surfaces of Pt, Pt alloy and Pt core-shell catalysts under Ar and O(2) atmospheres. The results are consistent with those shown by past experimental and theoretical studies on surface coverages under Ar atmosphere, the shape of the current-voltage curve for the ORR on Pt(111) and the material-dependence of the ORR activity. This model suggests that the oxygen associative pathway including HO(2)(ads) formation is the main pathway on Pt(111), and that the rate determining step (RDS) is the removal step of O(ads) on Pt(111). This RDS is accelerated on several highly active Pt alloys and core-shell surfaces, and this acceleration decreases the reaction intermediate O(ads). The increase in the partial pressure of O(2)(g) increases the surface coverage with O(ads) and OH(ads), and this coverage increase reduces the apparent reaction order with respect to the partial pressure to less than unity. This model shows details on how the reaction pathway, RDS, surface coverages, Tafel slope, reaction order and material-dependent activity are interrelated.     

Elastic scattering of 56 MeV polarized deuterons on 4He

The differential cross section and the vector and tensor analyzing powers A_y, A_xx, A_yy and A_xz were measured for the d-⁴He elastic scattering at 56 MeV. The measurement of A_xz was performed using a deuteron beam polarized in the horizontal plane. An optical-model analysis of the experimental data was carried out. The magnitude of the tensor analyzing powers could not be reproduced without the tensor potential. By including the T_R type tensor potential, the optical-model calculations give a reasonable reproduction of the experimental data at θ_c.m. < 120°. The obtained T_R tensor potential was much stronger than that predicted by the folding model. The strength of the real T_R potential was roughly in accordance with that obtained from the optical-model analysis of d-⁴He elastic scattering at 20.2 MeV.     

Changes in the Burgers vector of perfect dislocation loops without contact with the external dislocations

We report the observations of a new type of changing process in the Burgers vector of dislocations by in situ transmission electron microscopy. Small interstitial-type perfect dislocation loops in bcc iron with diameters less than approximately 50 nm are transformed from a 1/2<111> loop to another 1/2<111> one or an energetically unfavorable <100> one; furthermore, a <100> loop is transformed to a 1/2<111> one. These transformations occurred on high-energy electron irradiation or simple heating without contact with external dislocations. The origin of these phenomena is discussed.     

Structure and mechanical properties of nylon 6.12 prepared by temperature slope crystallization. I. Crystallization of oriented spherulitic textures

Oriented spherulitic textures of a rod-shaped nylon 6.12 sample were crystallized by the temperature slope method. Crystallization conditions were compared by changing temperatures and growth rates. Three types of textures (negative spherulites, positive spherulites, and spherulitic aggregates) were observed by this method. The negative textures appeared when the growth rate was less than 0.1 mm/h. Crystalline orientation and mechanical properties of the textures were investigated by x-ray diffraction and micro-hardness measurements, respectively. The hydrogen-bonded (010) planes were perpendicular to the growth direction in the negative spherulite, while they were parallel to the growth direction in the positive spherulite. In the spherulitic aggregates, the b axis was parallel to the growth direction, while the (010) planes formed roughly an angle of 45° with the growth direction. Due to the orientation of the hydrogen-boned planes, the negative texture exhibited an anisotropy, with hardness values of 106 MPa and 137 Mpa when measured perpendicular or parallel to the growth direction, respectively.     

Nuclear moments as a probe of electronic structure in material, exotic nuclear structure and fundamental symmetry

We report our studies in various fields of Physics through nuclear moments utilizing the β-NMR technique, including material sciences, nuclear structures and fundamental symmetries. Especially, we focus on the recent progress in the studies on the electronic structure in Pt through Knight shifts of various impurities, lattice locations of impurities, electric field gradients, the analysis of nuclear spin in terms of its components, anomaly in the spin expectation value for ⁹C-⁹Li mirror pair, the G-parity conservation law, and the Ramsey resonance on UCN for future neutron EDM measurements.     

Study of Tensor Correlations in 4He via the 4He(p, dp)d and 4He(p, dp)pn Reactions

Tensor correlations in ⁴He were studied via the (p, dp) reaction at the incident energy of 392 MeV with a focus on spin configurations of correlated pn pairs in ⁴He at high relative momenta ${(P_{\rm rel}^{\rm cor})}$ . The preliminary results show that the correlated pn pair at ${P_{\rm rel}^{\rm cor} = 310 {\rm MeV/c}}$ predominantly has the channel spin S = 1, which is consistent with the characteristics of tensor correlations.