Efficient purification protocol for bioengineering allophycocyanin trimer with N-terminus Histag
Joint Authors
Gao, Na
Tang, Zhihong
Song, Lufei
Pu, Yang
Li, Wenjun
Qin, Song
Source
Saudi Journal of Biological Sciences
Issue
Vol. 24, Issue 3 (31 Mar. 2017), pp.451-458, 8 p.
Publisher
Publication Date
2017-03-31
Country of Publication
Saudi Arabia
No. of Pages
8
Main Subjects
Topics
Abstract EN
Allophycocyanin plays a key role for the photon energy transfer from the phycobilisome to reaction center chlorophylls with high efficiency in cyanobacteria.
Previously, the high soluble self-assembled bioengineering allophycocyanin trimer with N-terminus polyhistidine from Synechocystis sp.
PCC 6803 had been successfully recombined and expressed in Escherichia coli strain.
The standard protocol with immobilized metal-ion affinity chromatography with chelating transition metal ion (Ni2+) was used to purify the recombinant protein.
Extensive optimization works were performed to obtain the desired protocol for high efficiency, low disassociation, simplicity and fitting for large-scale purification.
In this study, a 33 full factorial response surface methodology was employed to optimize the varied factors such as pH of potassium phosphate (X1), NaCl concentration (X2), and imidazole concentration (X3).
A maximum trimerization ratio (Y1) of approximate A650 nm/A620 nm at 1.024 was obtained at these optimum parameters.
Further examinations, with absorbance spectra, fluorescence spectra and SDS-PAGE, confirmed the presence of bioengineering allophycocyanin trimer with highly trimeric form.
All these results demonstrate that optimized protocol is efficient in purification of bioengineering allophycocyanin trimer with Histag.
American Psychological Association (APA)
Li, Wenjun& Pu, Yang& Gao, Na& Tang, Zhihong& Song, Lufei& Qin, Song. 2017. Efficient purification protocol for bioengineering allophycocyanin trimer with N-terminus Histag. Saudi Journal of Biological Sciences،Vol. 24, no. 3, pp.451-458.
https://search.emarefa.net/detail/BIM-761724
Modern Language Association (MLA)
Li, Wenjun…[et al.]. Efficient purification protocol for bioengineering allophycocyanin trimer with N-terminus Histag. Saudi Journal of Biological Sciences Vol. 24, no. 3 (Mar. 2017), pp.451-458.
https://search.emarefa.net/detail/BIM-761724
American Medical Association (AMA)
Li, Wenjun& Pu, Yang& Gao, Na& Tang, Zhihong& Song, Lufei& Qin, Song. Efficient purification protocol for bioengineering allophycocyanin trimer with N-terminus Histag. Saudi Journal of Biological Sciences. 2017. Vol. 24, no. 3, pp.451-458.
https://search.emarefa.net/detail/BIM-761724
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references : p. 457-458
Record ID
BIM-761724