Characterization of Spaghetti Prepared Under Different Drying Conditions.

Spaghetti dried at low (max. 50 °C), high (max. 70 °C), and very high (max. 85 °C) temperatures were characterized by their color, surface structure, rupture strength, texture analysis, and sauce retention capacity. The texture and sauce retention capacity were estimated for cooked spaghetti. The color of the spaghetti's methanol extracts, as evaluated through absorbance at 440 and 466 nm, did not depend on the drying temperature. A trend was observed in the surface texture of spaghetti, as estimated by atomic force microscopy and mercury intrusion porosimetry, where the surface was rougher when dried at higher temperatures than at low temperatures. Furthermore, the rupture strength was also higher for the spaghetti dried at higher temperatures. This result can be ascribed to the formation of stronger gluten networks, promoted by denaturation of gluten at the higher temperatures. However, the hardness of cooked spaghetti was not affected by the drying temperature, a result attributable to the action of water sorption to offset any differences in hardness among the spaghettis dried at the 3 different temperatures. The sauce retention capacity of cooked spaghetti was evaluated using a dextran solution as a simulated sauce, and by this method, the capacity of the spaghetti dried at a low temperature was shown to be significantly lower than that of the spaghetti dried at higher temperatures. This can be ascribed to the smoother surface of cooked spaghetti dried at the lower temperature and also to the leakage of amylose onto the surface during cooking.

Heterosubunit composition and crystal structures of a novel bacterial M16B metallopeptidase.

Three subfamilies of metallopeptidase family M16 enzymes--M16A, M16B, and M16C--are widely distributed among eukaryotes and prokaryotes. SPH2681, a periplasmic M16B protein found in Sphingomonas sp. strain A1, contains an HXXEH motif essential for Zn(2+) binding and catalytic activity. SPH2682 is another member of M16B, which lacks the metal-binding motif but conserves an active-site R/Y pair commonly found in the C-terminal half of M16 enzymes. Two genes coding for SPH2681 and SPH2682 assemble into a single operon in the bacterial genome. This study determined SPH2681 to be constitutively expressed in strain A1 cells grown on different carbon sources, suggesting a more general cellular function. SPH2681 and SPH2681/SPH2682 were overexpressed in Escherichia coli, purified, and characterized. SPH2681 was found to associate with SPH2682, forming a heterosubunit enzyme with peptidase activity, while SPH2681 alone exhibited no enzymatic activity. X-ray crystallography of the SPH2681/SPH2682 complex revealed two conformations (open and closed heterodimeric forms) within the same crystal. Compared with the closed form, the open form contains two subunits rotated away from each other by approximately 8°, increasing the distance between the zinc ion and active-site residues by up to 8 Å. In addition, many hydrogen bonds are formed or broken on change between the conformations of the heterodimers, suggesting that subunit dynamics is a prerequisite for catalysis. To our knowledge, this is the first report on both conformational forms of the same M16 peptidase, providing a unique insight into the general proteolytic mechanism of M16 proteases.