Folding thermodynamics of PET-hydrolyzing enzyme Cut190 depending on Ca2+ concentration

The enzyme, cutinase from Saccharomonospora viridis AHK190 (Cut190), can hydrolyze the inner block of polyethylene terephthalate (PET). Cut190 has a unique feature that both its activity and thermal stability are increased upon Ca²⁺ binding. In consideration of the glass transition temperature of PET, which is between 60 and 65 °C, the increased activity and thermal stability are of great interest to apply for PET bio-recycling. Our previous mutational analysis showed that the S226P/R228S mutant (Cut190*) has a higher activity and thermal stability than those of the wild type. In this study, we analyzed the folding thermodynamics of the inactive mutant of Cut190*, Cut190*S176A, using circular dichroism and differential scanning calorimetry. The results show that the denaturation temperature increases from 54 to 71 °C due to the addition of 250 mM Ca²⁺, in a Ca²⁺ concentration-dependent manner. The increased thermal stability is mainly due to the increased enthalpy change, partially compensated by the increased entropy change. Based on the crystal structure of Cut190*S176A bound to Ca²⁺, molecular dynamics simulations were carried out to analyze the effects of Ca²⁺ on the structural dynamics, showing that the Ca²⁺-bound structure fluctuated less than the Ca²⁺-free structure. Structural analysis indicates that Ca²⁺ binding increases the intramolecular interactions of the enzyme, while decreasing its fluctuation, which are in good correlation with the experimental results of the folding thermodynamics.

     

Thermodynamic interactions in rubbery polymer/gas systems

The Flory–Huggins interaction parameters χ for 23 gases (He, Ne, Ar, Kr, Xe, H₂, N₂, O₂, N₂O, CO₂, CH₄, C₂H₄, C₂H₆, C₃H₆, C₃H₈, 1,3-C₄H₆, four C₄H₈'s, n-C₄H₁₀, iso-C₄H₁₀, and n-C₅H₁₂) in five rubbery polymers (1,2-polybutadiene (PB), poly(ethylene-co-vinyl acetate)) (EVAc), polyethylene (PE), polypropylene (PP), and poly(dimethyl siloxane) (PDMS) were determined from either literature data on Henry's law coefficient and partial molar volume or those on sorptive dilation for each polymer/gas system. Values of χ for the gases increased in the order of PDMS < PP ≡ PB < EVAc ≡ PE. Among the gases except He and H₂ whose χ values are not reliable, Ne and Xe have respectively the highest and the lowest values of χ for the polyolefins. The χ values of the hydrocarbons were compared together with previously reported χ values of n-alkanes C₃-C₁₀. The dependencies of χ upon concentration and temperature were discussed on the basis of the literature data. © 1997 John Wiley & Sons, Inc. J Polym Sci B: Polym Phys 35 : 1049–1053, 1997     

Solubilization of oligosaccharides into chloroform by their incorporation into polystyrene as pendant groups

Solubilization of hydrophilic saccharide chains into organic solvents has been attempted by incorporating saccharide-substituted styrene unit into polystyrene main chain. Lactose-, maltopentaose, and amylose-substituted styrene monomers were copolymerized with styrene. Resulting chloroform-soluble copolymers were characterized, and structural formation was investigated. Copolymers of lactose-substituted styrene and maltopentaose-substituted styrene with styrene were dissolved into chloroform. The chloroform-soluble polymers contained about 12 disaccharide lactose chains or 1.7 maltopentaose chains as the pendant groups in one polystyrene molecule. Chloroform-insoluble methyl orange was dissolved into chloroform with the help of chloroform-soluble polystyrene having some saccharide chains. On the other hand, when an amylose-substituted styrene unit was inserted in a polystyrene chain, the resulting polymer became insoluble into chloroform. Amylose polysaccharide of DP_n = ∼24 was not dissolved into chloroform by this method.     

A Highly Water‐Tolerant Magnesium(II) Coordination Polymer Derived from a Flexible Layered Structure

A two‐dimensional (2D) layered Mg^II coordination polymer (CP) with a high tolerance for H₂O was designed, synthesised, and crystallographically characterised. The synthesis was achieved by the introduction of a flexible 2D layered structure composed of Mg^II ions and isonicotinate N‐oxide ligands. Owing to its high H₂O tolerance, the obtained 2D layered structure has the flexibility to repeatedly adsorb a large amount of H₂O associated with interlayer expansion and enable the removal of H₂O from a H₂O/2‐propanol mixed vapour. These results indicate that the CP could be an excellent dehydrating agent.     

Williamson Ether Synthesis with Phenols at a Tertiary Stereogenic Carbon: Formal Enantioselective Phenoxylation of β‐Keto Esters

The enantioselective formation of α‐aryloxy‐β‐keto esters is described for the first time. Lewis acid catalyzed enantioselective chlorination of β‐keto esters and subsequent S_N2 reactions with phenols yielded α‐aryloxy‐β‐keto esters with up to 96 % ee. Favorskii rearrangement of α‐chloro‐β‐keto esters was also found to give 1,2‐diesters with slightly reduced enantiopurity.     

Preparation of HPLC chiral packing materials using cellulose tris(4-methylbenzoate) for the separation of chrysanthemate isomers

We investigated the separation of chrysanthemate isomers ( 1 ), particularly the (1R)-trans form, by high-performance liquid chromatography (HPLC) using polysaccharide derivatives, such as the phenylcarbamates and benzoates of cellulose and amylose, as the chiral stationary phases (CSPs). The chiral packing materials (CPMs) having a high chiral recognition for the chrysanthemic acid ethyl ester ( 1a ) were prepared by coating cellulose tris(4-methylbenzoate) ( 2a ) dissolved in solvents containing methyl benzoate or acetophenone as an additive on silica gel. The separation factor for 1a significantly depended on the preparation conditions of CPM 2a , such as the coating amount of 2a and the type and amount of additives. The chiral recognition ability created by imprinting the additives was lost when the CPM was heated at a high temperature, and was recovered by contacting it with the additive in a packed column. The structural change in 2a during these treatments was not clearly detected by spectroscopic methods. © 2006 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 44: 5087–5097, 2006     

Weak isoscalar response of 11B

Weak-interaction responses of ¹¹B were studied by measuring the ¹¹B(³He, t) and ¹¹B(p, p′) reactions. Obtained nuclear transition matrix elements B(GT), B(στ), and B(σ) were compared with the shell-model calculations. The shell-model calculations, which explained the isovector parts B(GT) and B(στ) reasonably well if the quenching factors of 0.5–0.7 were taken into account, did not describe the isoscalar part B(σ).