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Research Article

Academia Journal of Biotechnology 6(8): 236-243, August 2018
DOI: 10.15413/ajb.2018.0127
ISSN: 2315-7747
2018 Academia Publishing

Abstract


Isolation and purification of Dragline Silk protein from Crossopriza lyoni Web
 

Accepted 8th August, 2018
 
N.G Shing Yeng* and Johan Ariff Bin Mohtar

Universiti Malaysia Perlis, Kampus Uniciti Alam Sg. Chuchuh, 02100 Padang Besar, Perlis, Malaysia.

Spider silk is a robust biomaterial due to its desirable tensile strength, biocompatibility and biodegradability. It is composed of spidroins which are recently used in wide technical applications. At present, dragline spidroins from Nephila clavipes are heavily studied, thus, neglecting the presence of the protein in other species. The aim of this study is to isolate and purify dragline silk protein from Crossopriza lyoni web. Spider webs were pre-treated with 0.02 M of sodium carbonate and followed by spidroin extraction using 20% (w/v) SDS-ethanol extraction solution. Two-level full factorial design was applied to analyze the significant effect of parameters namely; temperature (C), agitation speed (rpm) and incubation time (h) on maximum spidroin recovery. Optimization study was performed by Response Surface Methodology (RSM) through Box-Behnken Design (BBD). The crude sample was purified by gel filtration chromatography and characterized by one-dimensional SDS-PAGE for dragline spidroin presence. Agitation speed, incubation time and temperature posed significant effect on spidroin extraction with p < 0.0500 in decreasing order, respectively. From the optimization study, the highest spidroin concentration (1210.78 0.974 g/ml) was obtained at 90C, 102.5 rpm and 4.5 h with a fitted model at p value of 0.0001. Validation experiment was conducted and the result showed that the model was good to fit to the experimental data with percentage error less than 1%. Purification of the crude sample resulted in the dragline silk protein putative peaks on the chromatogram at 280 nm with 0.1 ml of column volume. SDS-PAGE analysis further confirmed the presence of the dragline silk protein with a band of molecular weight of more than 250 kDa. Isolation and purification of dragline spidroin in the present study is anticipated to bridge the gap of knowledge in the dragline spidroins from non-araneoid species. This would be an initiative to expand the spider silk technology at national level in the near future.

Key words: Spider silk, Spidroin, Crossopriza lyoni, screening, optimization, purification.
 

This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Cite this article as:
Yeng NGS, Mohtar JAB (2018). Isolation and purification of Dragline Silk protein from Crossopriza lyoni Web. Acad. J. Biotechnol. 6(8): 236-243.

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