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Patterns of protein expression upon adding sugar and elicitor to the cell culture of Eschscholtzia californica

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Abstract

The optimal timing of elicitation was determined for the production of benzophenanthridine alkaloids (BPAs) by Eschscholtzia californica cell culture. Upon elicitation, 7-day old cells produced more alkaloids than 14-day old cells (5.1 times for sanguinarine and 2.7 times for dihydrosanguinarine). We presumed that these alkaloids are growth-rate-associated secondary metabolites in E. californica cell culture. Although the specific productivity of alkaloids were higher in 7-day old culture, the total cell mass of 7-day old culture was about half that of 14-day old culture. In order to increase the overall productivity, sucrose was added to the 14-day old culture before the addition of elicitor. By this way, cells in the stationary phase (14-day old culture) could be switched to the cells in the logarithmic growth phase (similar to 7-day old culture). Total production of alkaloids was increased by adding sucrose; especially the production of sanguinarine was increased as high as 5.7 times of the control. To find out the protein level changed by the elicitation, proteins extracted from whole cell were separated by using two-dimensional gel electrophoresis. The patterns of the gels were different and little correlation among the proteins could be observed. And Western blotting was employed to check the expression level of selected five enzymes, these enzymes believed to be involved in BPAs production, resulting in up-regulated with elicitor addition.

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Abbreviations

4′OMT:

3-hydroxy-N-methylcoclaurine 4-O-methyltransferase

6OMT:

norcoclaurine 6-O-methyltransferase

BPA:

benzophenanthridine alkaloid

BBE:

berbeine bridge enzyme

CYP80B1:

N-methylcoclaurine 3′-hydroxylase

DHBO:

dihydrobenzophenanthridine oxidase

DBPA:

Dihydroform benzophenanthridine alkaloid

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Acknowledgement

This work was financially supported by the KOSEF through the AEBRC at POSTECH.

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Authors and Affiliations

  1. Department of Chemical Engineering, Pohang University of Science and Technology, San31, Pohang, Kyungbuk, 790-784, South Korea

    Jeong-Jin Park, Seok Young Son, Hong Soon Rhee & Jong Moon Park

  2. Institute of Environmental and Energy Technology, Pohang University of Science and Technology, Pohang, Kyungbuk, 790-784, South Korea

    Sung-Yong H. Yoon

  3. School of Environmental Engineering, Pohang University of Science and Technology, Pohang, Kyungbuk, 790-784, South Korea

    Hwa Young Cho

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  1. Jeong-Jin Park
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  2. Sung-Yong H. Yoon
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Correspondence to Jong Moon Park.

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Park, JJ., Yoon, SY.H., Cho, H.Y. et al. Patterns of protein expression upon adding sugar and elicitor to the cell culture of Eschscholtzia californica . Plant Cell Tiss Organ Cult 86, 257–269 (2006). https://doi.org/10.1007/s11240-006-9115-1

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  • DOI: https://doi.org/10.1007/s11240-006-9115-1

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