Abstract
BackgroundSago palm (Metroxylon sagu) is a versatile crop and has been hailed as the next viable commodity in Sarawak, Malaysia. Sago palm can thrive in the harsh swampy peat, from low flooded areas to uplands and in acidic to neutral soils. Sago palm has been neglected and unfortunately, very little is known about the sago palm proteome. This study aimed to determine the best protein extraction method of Metroxylon sagu for the two-dimensional gel electrophoresis (2-DE) and its comparative analysis.ResultsTo perform good proteome research, the most critical step is to establish a method that gives the best quality of extracted total proteins. Five different protein extraction protocols: polyethylene glycol (PEG) fractionation method, phenol extraction method, TCA–acetone method, the combination of phenol and TCA–acetone extraction method and imidazole method were compared to develop an optimized protein extraction method for two-dimensional gel electrophoresis analysis of Metroxylon sagu. The PEG fractionation method was found to give the most reproducible gels with the highest number of spots and highest protein concentration followed by phenol extraction method. The lowest number of spots was observed in the imidazole method. The PEG fractionation method provides improved resolution and reproducibility of 2-DE and reduces the time required to analyze samples. Partitioning Rubisco by polyethylene glycol (PEG) fractionation provides clearer detection of low abundance protein. Hence, the results from this study propose PEG fractionation as the effective protein extraction method for 2-DE proteomic studies of Metroxylon sagu.ConclusionIn this study, the PEG fractionation method is considered as the best extraction method for 2-DE proteomic studies of Metroxylon sagu in terms of yield, gel quality, spot numbers, and quantities of proteins.
Highlights
Sago palm (Metroxylon sagu) is a versatile crop and has been hailed as the viable commodity in Sarawak, Malaysia
The higher protein concentration of 8.9 μg/μl and 4.6 μg/μl were obtained from polyethylene glycol (PEG) fractionation and phenol extraction method, respectively (Table 1), followed by combined phenol and Trichloroacetic acid (TCA)–acetone method 3.9 μg/μl and TCA– acetone method 2.5 μg/μl
The TCA–acetone extraction method resulted in the lowest concentration and fewer spots detected in Two-dimensional gel electrophoresis (2-DE) separation
Summary
Sago palm (Metroxylon sagu) is a versatile crop and has been hailed as the viable commodity in Sarawak, Malaysia. The sago palm (Metroxylon sagu Rottb.) is a versatile plant that can tolerate many biotic and abiotic stresses, during its growth stages. It belongs to family Arecaceae, and among fourteen starch producing genera, the genus Metroxylon is the most productive among all of them. Sago palm is a starch producing plant and can accumulate high carbohydrate content in the trunk. It is socioeconomically important for sustainable agriculture and considered as one approach for rural development in
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