. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 144 H. MICHIBATA ET AL 254nm 030 200 300 Wavelength ( nm ). 15 20 25 30 Fraction Number F'igure 6. Elution profile of the blood cell homogenate of Ascidia aliixlon on Sephadex G-15 column chromatography. Pelleted blood cells ( mg) were homogenized in 10 mA/ glycine-HCl buffer solu- tion at pH The homogenate ( ml) was loaded onto a column ( cm 0 x 48 cm) and eluted with the same buffer solution in 3 ml fractions. Vt and Vo of the column were 85 ml and 39 ml, respectively. Ve of vanadobin was 63 ml. The inset sh
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 144 H. MICHIBATA ET AL 254nm 030 200 300 Wavelength ( nm ). 15 20 25 30 Fraction Number F'igure 6. Elution profile of the blood cell homogenate of Ascidia aliixlon on Sephadex G-15 column chromatography. Pelleted blood cells ( mg) were homogenized in 10 mA/ glycine-HCl buffer solu- tion at pH The homogenate ( ml) was loaded onto a column ( cm 0 x 48 cm) and eluted with the same buffer solution in 3 ml fractions. Vt and Vo of the column were 85 ml and 39 ml, respectively. Ve of vanadobin was 63 ml. The inset shows the UV spectrum of the peak fraction (fraction 21). Absorbance at 254 nm was observed. umn, the elution profile obtained was somewhat more complicated (Fig. 6). A scan of UV wavelengths for frac- tion 21 (Fig. 6, inset) indicates that 254 nm is the peak of absorption. A total of /^g of vanadium was eluted in fractions 17 through 26, and its peak was in frac- tion 21. As shown in Figure 7, when these vanadium-contain- ing fractions were loaded onto a column of SE-52, non- absorbed substances were first washed off the column by eluting with the glycine-HCl buffer solution, and a big peak with no vanadium was eluted with a linear gradient of KC1 dissolved in buffer solution. The vanadobin was obtained thereafter in fractions 54 through 58 with a peak in fraction 56. The vanadium content recovered was ^g. The absorption peak, observed in fraction 56, was 232 nm (Fig. 7, inset). Vanadobin was extractable from the blood cells of three other ascidian species, A. :ara OKA, Corel/a japon- ica HERDMAN, and dona intcstimilis (LINNE); the procedures and results were similar to those described above (data not shown). The process of vanadobin ex- traction by chromatography is summarized in Table I. The highest vanadium content of 8750 ng/mg wet weight (14350 ng/1640 mg wet weight) in the blood cells was observed in A. gemmata. The next highest value was 2040 ng/mg wet weight de
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