A beta-fructofuranosidase (EC 3.2.1.26) was purified to homogeneity from Aspergillus japonicus TIT-KJ1. The enzyme had an optimum pH for activity of 5.4 and pH stability at 7.0-8.4. The optimum temperature at pH 5.4 was 60 degrees C. The enzyme had a molecular weight of 236,000 with two subunits and an isoelectric point of pH 4.0. The enzyme was inactivated by 5 mM Hg2+ and Ag+. The enzyme had a high transfructosylating activity. Treatment of 50% (w/v) sucrose with the enzyme under optimum conditions afforded more than 55% fructooligosaccharides.
C–H–O interactions of a self-assembled triple helix based on the 1-acetamido-3(2-pyrazinyl)-imidazolium cation has been probed by high pressure. The infrared spectroscopic profiles and ab initio calculations allow us to make a vibrational assignment of this compound. The C–H bonds forming C–H–O interactions shorten as the pressure was elevated, while free C–H vibration modes show low sensitivity to high pressure. The pressure-dependent results can be attributed to the strengthening of C–H–O electrostatic–dispersion interactions upon increasing pressure. The appearance of the free-NH infrared absorption indicates that the conventional N–H–O hydrogen bond does not dominate the inter-strand packing in the compound. It is proposed that the charge-enhanced C–H–O interactions, forming a helical hydrogen-bonding network, disturb the formation of inter-strand N–H–O hydrogen-bonding in order to form a maximum number of hydrogen bonds. Applying high-pressure seems not to change the C=O bond length in contrast to the trend of blue-shift in frequency of C–H vibrations. London dispersion energy is suggested to be required for understanding the pressure-dependent results, although more additional terms, such as the effect in the presence of charge, are needed for the correct explanation. This work demonstrates that high-pressure studies may have the potential to provide insight into the C–H–O structural properties of biological related systems.
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