Chromatographic separation of cytidine triphosphate from fermentation broth of yeast using anion-exchange cryogel.
A novel separation method was developed to isolate directly cytidine triphosphate (CTP) from fermentation broth of yeast using anion-exchange supermacroporous cryogel. The anion-exchange cryogel with tertiary amine groups was prepared by graft polymerization. The breakthrough characteristics and elution performance of pure CTP in the cryogel bed were investigated experimentally and the CTP binding capacity was determined. Then the separation experiments of CTP from crude fermentation broth of yeast using the cryogel column were carried out using deionized water and 0.01 M HCl as washing buffer, respectively. The chromatographic behavior was monitored and analyzed. The purity and concentration of the obtained CTP in these processes were determined quantitatively by HPLC. The maximal purity of CTP obtained at the condition of 0.01 M HCl as washing buffer and 0.5 M NaCl in 0.01 M HCl as elution buffer reached 93%.
Wang
Lianghua
L
Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou, China.
Shen
Shaochuan
S
Yun
Junxian
J
Yao
Kejian
K
Yao
Shan-Jing
SJ
D001798:Blood Proteins; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D059985:Cryogels; D017077:Culture Media, Conditioned / Q000737:chemistry*; D003570:Cytidine Triphosphate / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D005285:Fermentation; D005353:Fibronectins; D020100:Hydrogels; D012441:Saccharomyces cerevisiae / Q000378:metabolism*
D001798:Blood Proteins; D059985:Cryogels; D017077:Culture Media, Conditioned; D005353:Fibronectins; D020100:Hydrogels; D003570:Cytidine Triphosphate