Enzymatic synthesis of chondroitin and its derivatives catalyzed by hyaluronidase

The enzymatic polymerization to provide synthetic chondroitin and its derivatives is reported here, the first example of such in vitro synthesis to date. N-Acetylchondrosine (GlcAbeta(1-->3)GalNAc) oxazoline (1a) and its derivatives (1b-1f) were designed and synthesized as novel transition state analogue substrate monomers for catalysis by hyaluronidase. Hyaluronidase is a hydrolysis enzyme of chondroitin that also catalyzes the formation of repeated glycosidic bonds in in vitro synthesis, rather than in the catabolic direction. Monomers of 2-methyl (1a), 2-ethyl (1b), and 2-vinyl (1f) oxazoline derivatives were polymerized using this enzyme, via ring-opening polyaddition with total control of regioselectivity and stereochemistry. These reactions provided the corresponding synthetic chondroitin (natural type; N-acetyl, 2a) and the derivatives (unnatural type) with N-propionyl (2b) and N-acryloyl (2f) functional groups at the C2 position of all the galactosamine units, in good yields. Monomers of 2-n-propyl (1c) and 2-isopropyl (1d) oxazoline derivatives were polymerized to produce 2c and 2d in low yield. The 2-phenyl oxazoline derivative (1e) did not afford any enzyme-catalyzed products. M(n) values of 2a and 2b reached 4800 and 4000, respectively. The M(n) value of 2a corresponds to that of the naturally occurring chondroitin. Thus, hyaluronidase catalysis allows the in vitro production of not only natural type but also the formation of unnatural type chondroitins.     

Anomalous volumetric behavior of water-hexane and water-decane mixtures in the vicinity of the critical region as studied by infrared spectroscopy

Infrared spectra of binary mixtures of water with hexane and decane were measured at temperatures and pressures in the 473-648 K and 70-350 bar ranges, respectively. Volumetric concentrations of water and the hydrocarbons in the mixtures were obtained from absorption intensities of the fundamental OH stretching band of HDO and combination transitions of the hydrocarbons. Using both the concentrations, densities of the aqueous mixtures were estimated and compared with densities before mixing, which were calculated using literature densities of the neat liquids. It is found that anomalously large volume expansion on mixing occurs in the vicinity of the critical region of the mixtures.     

Photochemistry of 4-pyrimidinones in aqueous solution. Isolation of reversible photohydrates

Both the irradiation of 4-pyrimidinones ($\text{la-c}$) and the reaction of the isolated Dewar 4-pyrimidinones ($\text{2a-c}$) in aqueous solution gave the corresponding photohydrates ($\text{3a-c}$) which reverted spontaneously to the starting $\text{la-c}$ in the dark reaction. The photohydrates were isolated in crystalline form and their physical properties were determined.     

A practical process for large-scale synthesis of (S)-5-hydroxy-6-trans-8,11,14-cis-eicosatetraenoic acid (5-HETE)

($\text{S}$)-5-Hydroxy-6-$\text{trans}$-8,11,14-$\text{cis}$-eicosatetraenoic acid (5-HETE) (1) and its enantiomer are readily available by a chemical synthesis from arachidonic acid which includes a chromatographic separation of diastereomeric urethanes (3) made from (±)-5-HETE methyl ester and the isocyanate 4 derived from dehydroabietylamine.     

Three new types of indoloditerpenes, emindole PA-PC, from Emericella purpurea. Revision of the structure of emindole PA

Three new type indoloditerpenes, emindoles PA (1), PB (2), and PC (3), were isolated from the mycelium of Emericella purpurea along with the sesterterpenes variecolol and variecolactone, and the dicyanide derivatives epurpurins A to C. The structures of 1-3 were confirmed by the spectroscopic investigation. The structure of emindoles PA (1) was revised from our preliminary report. Emindoles PA (1), PB (2), and PC (3) are the indoloditerpenes having a new type of carbon skeleton.     

Fisher information for spike-based population decoding

We evaluate the Fisher information of a population of model neurons that receive dynamical input and interact via spikes. With spatially independent threshold noise, the spike-based Fisher information that summarizes the information carried by individual spike timings has a particularly simple analytical form. We calculate the loss of information caused by abandoning spike timing and study the effect of synaptic connections on the Fisher information. For a simple spatiotemporal input, we derive the optimal recurrent connectivity that has a local excitation and global inhibition structure. The optimal synaptic connections depend on the spatial or temporal feature of the input that the system is designed to code.     

Effect of solvent on an NMR chemical shift difference between glycyl geminal alpha-protons as a probe of beta-turn formation of short peptides

Proton NMR spectra of short peptides with a glycyl (Gly) or N-methylglycyl (sarcosyl, Sar) residue were measured in various mixed solvents with a wide range of dielectric constants: 78.3-2.3. From analyses of the octet and quartet signals of the geminal alpha-protons of Gly and Sar residues, respectively, we have estimated chemical shift differences between the two alpha-protons, Delta delta(alpha/alpha'). It is found that the Delta delta(alpha/alpha') values increase with decreasing solvent polarity and the increasing rates depend significantly on amino acid sequences. By referring to infrared spectra and chemical shift of the terminal NH protons, deltaNH, of the peptides, the Delta delta(alpha/alpha') values were found to be a good probe of beta-turn formation. From solvent-dependent change of Delta delta(alpha/alpha'), we estimated the free energies for the beta-turn formation and compared the results with those estimated from deltaNH. Using the resulting free energies, we have discussed effects of solvent on the beta-turn formation.     

Spectroscopic study of mutual solubilities of water and benzene at high temperatures and pressures

Near-infrared and ultraviolet absorption of water-benzene mixtures has been measured at temperatures and pressures in the ranges of 323-673 K and 50-400 bar, respectively. Concentrations of water and benzene in both the water-rich phase and the benzene-rich phase of the mixtures were obtained from absorption intensities of near-infrared bands of water and benzene and ultraviolet bands of benzene. Mutual solubilities in molar fractions increase remarkably with increasing temperature at pressures in the two-liquid-phase coexistence region, and are consistent with previously reported values. It proves that the solubility of benzene in water is an order of magnitude smaller than that of water in benzene throughout the two-phase region. In addition, it is found that effect of pressure on the solubilities is opposite between water in benzene and benzene in water. These solubility properties are discussed on the basis of a cavity-based solvation model. It is suggested that the asymmetry in the mutual solubility and the opposite direction of the pressure effect are caused by difference in molecular size and difference in thermal compressibility, respectively, between water and benzene.     

A new, highly sensitive time-of-flight mass spectrometer consisting of a flangeon-type conical ion lens system and a proto-type Daly detector for exhaust gas analysis based on the Jet-REMPI technique.

For the analysis of trace amounts of hazardous organic compounds, we developed a new ion detection system for supersonic jet resonance-enhanced multiphoton laser ionization mass spectrometry. High sensitivity and selectivity have been achieved by combining a proto-type Daly detector, a newly designed conical ion lens, and a potential switch that can perform the function of a mass selector. This ion detection system enables us to bring the jet nozzle closer to the ionization point. The detection sensitivity has thus been improved totally by more than 100 times compared with that obtained by the parallel-plate electrodes and micro-channel plate. We succeeded in assembling a flangeon-type ion extraction electrode consisting of a conical ion lens and a potential switch.