No abstract

The nature of the active site cofactor and the amino acid sequence flanking this structure have been determined in a range of copper amine oxidases. For enzymes from porcine plasma, porcine kidney, and pea seedlings, proteolytic digestion was performed on phenylhydrazone or p-nitrophenylhydrazone derivatives. Thermolysin treatment leads to relatively small active site peptides, which have been characterized by Edman degradation and by resonance Raman spectroscopy. Resonance Raman spectra of peptides show identical peak positions and intensities relative to each other and to a model p-nitrophenylhydrazone derivative of topaquinone hydantoin, establishing topaquinone as the cofactor in each instance. Edman degradation of peptides provides active site sequences for comparison to previous determinations with bovine serum and yeast amine oxidases. The available data establish a consensus sequence of Asn, Topa, Asp/Glu. Trypsin leads to significantly longer peptides, which reveal a high degree of sequence identity between plasma proteins from bovine and porcine sources (89%), with significantly decreased identity between the porcine serum and intracellular amine oxidases (56%). A lower degree of identity (45%) is observed between the pea seedling and mammalian enzymes. As an alternative to the isolation of active site peptides for topaquinone identification, visible spectra of intact proteins have been investigated. It is shown that p-nitrophenylhydrazone derivatives of native enzymes, active site-derived peptides, and a topaquinone model exhibit identical behavior, absorbing at 457-463 nm at neutral pH (pH 7.2) and at 575-587 nm in basic solution (1-2 M KOH). These spectral properties, which appear unique to topaquinone, provide a rapid and simple test for the presence of this cofactor in intact enzymes.(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of calcium pretreatment, vacuum level, and apple variety on the texture of apple chips, processed using a combination of air drying and vacuum microwave dehydration (VMD), were determined. Pretreatment of apple slices by immersion in 1-5% aqueous CaCl 2 significantly increased crispness of chips as determined by instrumental and sensory analysis; however above 1%, chips were perceived as bitter. Higher vacuum applied during VMD significantly lowered density and increased crispness of chips. This effect was mediated by the vaporization of water in the interior of the chip, which caused expansion of the tissue. Chips made from Fuji apples had higher calcium contents, and were crisper than Red and Golden Delicious apple chips. Microstructure of the chips evaluated by scanning electron microscopy indicated that chips with thicker cell walls and large internal voids were crisper.

Basil (Ocimum basilicum L.) was dried using conventional hot air or the recently developed vacuum-microwave dryers. The effect of the drying method on the relative abundance of major flavor volatiles, rehydration rate, color, and structural integrity of the plant was evaluated. Dynamic headspace analysis of volatiles present in fresh or dried basil revealed that linalool and methylchavicol (estragole) were the two major headspace volatile compounds of the plant sample. Vacuum-microwave dehydrated basil yielded approximately 2.5 times the linalool and 1.5 times the methylchavicol of the air-dried samples. Furthermore, the vacuum-microwave-treated samples yielded more volatiles than fresh basil, due to chemical reactions during drying. Air-dried samples of basil had darker and fewer green hues than those prepared by vacuum microwave. Vacuum-microwave-dried samples had a higher rehydration rate, whereas the potential of the plant material to rehydrate was hindered in air-dried samples. This is likely attributed to the dramatic and pronounced structural collapse of the air-dried cells as revealed by the scanning electron microscope.