Preparation of Inositol
Tetrakisphosphate and Its Application in Modification of Porcine Hemoglobin
HONG Min, LI Zhen, LI Shi-Yun, YUAN Zhong-Yi*
(Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai
200031, China)
Abstract Modification
of hemoglobin using inositol tetrakisphosphate (IP4) can improve the oxygen
affinity of hemoglubin. Phytase was extracted from wheat bran and purified by
ammonium sulphate fractionation, followed by Sephadex G-50 gel filtration and
Mono Q chromatography. The purified phytase was used for hydrolysis of phytic
acid under controled conditions. IP4, as a major composition of the
hydrolysate, was further purified on resin 714 column. The purified IP4 was oxidized by periodate to
obtain dialdehyde-IP4. By the reaction between the aldehyde group of IP4 derivative and the amino
group of porcine hemoglobin (pHb), pHb-IP4 conjugate was formed and was
found to have better ability of O2 binding and releasing than the native
hemoglobin.
Key words phytase； inositol
tetrakisphosphate；
blood substitutes
Received: June 18,
1999 Accepted: August 12, 1999
* Corresponding author: Tel, 86-21-64374430; Fax, 86-21-64338357; e-mail, [email protected]