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Characterization of photosystem I from spinach: effect of solution pH

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Abstract

Our previous work has demonstrated the isolation of photosystem I (PSI) from spinach using ultrafiltration with a final purity of 84 %. In order to get a higher purity of PSI and more importantly to develop a practical bioseparation process, key physiochemical properties of PSI and their dependence on operational parameters must be assessed. In this study, the effect of solution pH, one of the most important operating parameters for membrane process, on the property of PSI was examined. Following the isolation of crude PSI from spinach using n-dodecyl-beta-d-maltoside as detergent, the isoelectric point, aggregation size, zeta potential, low-temperature fluorescence, atomic force microscopy imaging, secondary structure, and thermal stability were determined. Solution pH was found to have a significant effect on the activity, aggregation size and thermal stability of PSI. The results also suggested that the activity of PSI was related to its aggregation size.

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Abbreviations

PSI:

Photosystem I

DDM:

n-Dodecyl-beta-d-maltoside

PMSF:

Phenylmethane-sulfonyl fluoride

DCMU:

3-(3,4-Dichlorophenyl)-1,1-dimethylurea

DCIP:

2,6-Dichloroindophenol

MV:

Methylviologen

EDTA:

Ethylenediaminetetraacetate

PES:

Polyethersulfone

IEF:

Isoelectric focusing

IEP:

Isoelectric point

AFM:

Atomic force microscopy

CD:

Circular dichroism

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

DLS:

Dynamic light scattering

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Acknowledgments

The authors are grateful for the financial support of National Natural Science Founds of China (no. 20906104), the Doctoral Foundation of Shandong Province (no. BS2009SW029), and the Fundamental Research for the Central Universities. Special thanks to Prof. Shuguang Zhang (Center for Biomedical Engineering, Massachusetts Institute of Technology) for his constructive suggestions.

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Authors and Affiliations

  1. Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao, 266555, People’s Republic of China

    Jianguo Liu, Xuefang Zhang, Meng Wang, Jing Liu & Meiwen Cao

  2. School of Physics and Astronomy, Manchester University, Manchester, M13 9PL, UK

    Jianren Lu

  3. Department of Engineering Science, University of Oxford, Oxford, OX1 3PJ, UK

    Zhanfeng Cui

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  1. Jianguo Liu
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  2. Xuefang Zhang
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  3. Meng Wang
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  4. Jing Liu
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  5. Meiwen Cao
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  7. Zhanfeng Cui
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Correspondence to Jianguo Liu.

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Liu, J., Zhang, X., Wang, M. et al. Characterization of photosystem I from spinach: effect of solution pH. Photosynth Res 112, 63–70 (2012). https://doi.org/10.1007/s11120-012-9737-6

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