Lysozyme refolding at high concentration by dilution and size-exclusion chromatography |
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Authors: | Gao?Yong-gui Guan?Yi-xin Email author" target="_blank">Yao?Shan-jing?Email author Cho?Man-gi |
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Institution: | (1) Department of Chemical and Biochemical Engineering, Zhejiang University, 310027 Hangzhou, China;(2) College of Life Science, Zhejiang University, 310027 Hangzhou, China;(3) Engineering Research Center, Dong-Seo University, 69-1 Jurea-dong Sasanggu, 617-716 Busan, Korea |
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Abstract: | This study of renaturation by dilution and size exclusion chromatography (SEC) addition of urea to improve yield as well as
the initial and final protein concentrations showed that although urea decreased the rate of lysozyme refolding, it could
suppress protein aggregation to sustain the pathway of correct refolding at high protein concentration; and that there existed
an optimum urea concentration in renaturation buffer. Under the above conditions, lysozyme was successfully refolded from
initial concentration of up to 40 mg/mL by dilution and 100 mg/mL by SEC, with the yield of the former being more than 40%
and that of the latter being 34.8%. Especially, under the condition of 30 min interval time, i.e. τ>2(t
R2−tR1), the efficiency was increased by 25% and the renaturation buffer could be recycled for SEC refolding in continuous operation
of downstream process.
Project supported by Zhejiang Provincial Natural Science Foundation of China (No. 201099) and the Local-Lab Project by Korean
Institute of Science and Technology Evaluation and Planning, Korea |
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Keywords: | Lysozyme refolding Downstream process Dilution Size exclusion chromatography Urea |
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