Regulatory phosphorylation of poly-γ-glutamic acid with phosphate salts in the culture of Bacillus subtilis (natto).

Poly-γ-glutamic acid (PGA) was easily phosphorylated by direct addition of phosphorylating agents into the culture medium of Bacillus subtilis (natto). Tetrapolyphosphate salt was the most incorporated into PGA molecules of all used reagents. Phosphorylation occurred at the α-carboxyl side chains of PGA molecule. The amounts of bound phosphate to PGA were dependent on the amounts of added phosphorylating agent. In low molecular weight regions of less than 100 kDa, a cross-linked peak was observed in the phosphorylated PGAs, whereas their peaks at approximately 1000 kDa shifted to a higher molecular weight due to the bound phosphate. The PGA derivatives had a wide range in viscosity up to 15/1000 to 15 times when compared to the native PGA, depending on the degree of phosphorylation (DP) in the PGA derivatives. The PGA with low DP had a high viscosity due to the unfolding conformation whereas highly phosphorylated PGA had aggregation with low viscosity. Heat treatment at 80 °C after the addition of phosphate salt elicited a novel collagen-like helix structure. These observations show that phosphorylation is an effective way to diversify the physicochemical properties of PGA.

Feasible protein aggregation of phosphorylated poly-γ-glutamic acid derivative from Bacillus subtilis (natto).

Poly-γ-glutamic acid (PGA) was modified with phosphorylating agents such as sodium metaphosphate and potassium metaphosphate in the culture medium of Bacillus subtilis (natto). The highly phosphorylated PGA derivatives were prepared and investigated for their chemical and physicochemical properties. The PGA derivatives had approximately 7% (W/W) inorganic phosphorus and characteristic absorbance PO2- bands at 1082cm-1 and 1260cm-1 by Fourier Transform Infrared Spectroscopy. The derivative modified by sodium metaphosphate (J-5) was easily hydrated in water and had extremely low viscosity. The shear rate-induced transition leading to the decrease of viscosity was not observed in J-5 whereas the derivative modified by potassium metaphosphate (J-6) as well as unmodified PGA (J-1) showed the typical decrease of viscosity. In circular dichroism (CD) measurement of J-5, there was a significant loss of the negative chirality CD signal, implying that protein aggregation occured at decreasing pH from 6.2 to 4.4. The thioflavin T fluorescence intensity of the aqueous solution in the J-5 was extremely high despite the absence of heat-treatment. The results indicate that the J-5 is the likeliest type of aggregation by ß-sheet cross-linking which is relevant to protein diseases like Alzheimer's disease.

Improvement on the freeze-thaw stability of corn starch gel by the polysaccharide from leaves of Corchorus olitorius L.

Effect of the polysaccharide from leaves of Corchorus olitorius L. (PLC) on the freeze-thaw (FT) stability of corn starch gel was studied. PLC was incorporated into the starch gel at 0.7% and total solid was adjusted to 6.0%. The syneresis was measured by the centrifugal-filtration method and, as a result, addition of PLC reduced effectively the syneresis of the starch gel even after 5 FT cycles, which was less than one third that of the normal starch gel. The rheological changes of the starch/PLC gel during the FT treatments were evaluated while the gel remained on the rheometer plate. The starch/PLC gel had less significant changes in the rheological parameters during the FT cycles than starch/guar gum or xanthan gum gel systems. SEM images showed that PLC stabilized the gel matrix surrounding pores, which would contribute to both a lower syneresis production and a higher stability in the rheological behavior at FT.