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Optimization of a protein extraction method from natural rubber sheets made of Hevea brasiliensis latex

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Abstract

A method to extract proteins from natural rubber (NR) samples has been developed to characterize the protein composition of NR by SDS-PAGE electrophoresis. While the incubation of NR pieces in aqueous buffers did not provide satisfactory results (no protein bands detected by SDS-PAGE), the solubilization of NR pieces in an organic solvent, prior to centrifugation and protein extraction from the obtained pellet, was found efficient to highlight protein bands by SDS-PAGE. Pure cyclohexane was used to solubilize NR pieces and resulted in a low share of nitrogen in the pellet (29%). The addition of 10% (v/v) ethanol to cyclohexane allowed to significantly increase this share to 73% and to detect 13 protein bands by SDS-PAGE. The molecular weights of the four most intense bands suggest the presence in NR of the following proteins: rubber elongation factor, small rubber particle protein, hevamine, and β-1-3 glucanase.

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Abbreviations

ADS:

Air-dried sheet

CV:

Constant viscosity

FT-IR:

Fourier transform infrared

NR:

Natural rubber

PBS:

Phosphate-buffered saline

SDS-PAGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

SMR:

Standard Malaysian rubber

STR:

Standard Thai rubber

TEM:

Transmission electron microscope

THF:

Tetrahydrofuran

TSR:

Technically specified rubber

USS:

Unsmoked sheet

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Acknowledgements

This research is supported in part by the Graduate Program Scholarship from the Graduate School, Kasetsart University. Agropolis Foundation is thanked for the support to the creation of LipPolGreen-Asia platform in Kasetsart Agricultural and Agro-Industrial Product Improvement Institute, Bangkok. The authors thank Union Rubber Co. Ltd for providing latex in their plantation. This work was undertaken under the umbrella of Hevea Research Platform in Partnership (HRPP).

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Authors and Affiliations

  1. Department of Biotechnology, Faculty of Agro-Industry, Kasetsart University, Bangkok, 10900, Thailand

    Surattiya Srisomboon & Kittipong Rattanaporn

  2. Department of Applied Microbiology, Institute of Food Research and Product Development, Kasetsart University, Bangkok, Thailand

    Kanthida Wadeesirisak

  3. CIRAD, UMR IATE, 10900 Bangkok, Thailand

    Laurent Vaysse

  4. IATE, Univ Montpellier, CIRAD, INRAE, Institut Agro, Montpellier, France

    Laurent Vaysse, Jérôme Sainte-Beuve, Frédéric Bonfils & Céline Bottier

  5. CIRAD, UMR IATE, F-34398 Montpellier, France

    Jérôme Sainte-Beuve, Frédéric Bonfils & Céline Bottier

  6. Kasetsart Agricultural and Agro-Industrial Product Improvement Institute, Kasetsart University, Bangkok, 10900, Thailand

    Natedao Musigamart & Siriluck Liengprayoon

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  1. Surattiya Srisomboon
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Srisomboon, S., Wadeesirisak, K., Vaysse, L. et al. Optimization of a protein extraction method from natural rubber sheets made of Hevea brasiliensis latex. J Rubber Res 24, 27–39 (2021). https://doi.org/10.1007/s42464-020-00069-1

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