title: Method developments to extract proteins from oil palm chromoplast for proteomic analysis.
abstract: Proteins from the plant chromoplast are essential for many physiological processes such as fatty acid biosynthesis. Different protein extraction methods were tested to find the most robust method to obtain oil palm chromoplast proteins for mass spectrometry analysis. Initially, two different solvents were employed to reduce the fruit lipids. Then, two plant cell wall digestive enzymes were used to acquire the protoplasts to increase the protein extraction effectiveness. A two-stage centrifugation-based fractionation approach enhanced the number of identified proteins, particularly the fatty acid biosynthetic enzymes. The effectiveness of each extraction method was assessed using protein yields and 2DE gel profiles. The ideal method was successfully used to establish the 2DE chromoplast proteome maps of low and high oleic acid mesocarps of oil palm. Further nanoLC-MS/MS analysis of the extracted chromoplast proteins led to the identification of 162 proteins, including some of the main enzymes involved in the fatty acid biosynthesis. The established procedures would provide a solid foundation for further functional studies, including fatty acid biosynthetic expression profiling and evaluation of regulatory function.
authors.affiliation: Advanced Biotechnology and Breeding Centre, Malaysian Palm Oil Board, No. 6, Persiaran Institusi, Bandar Baru Bangi, 43000 Kajang, Selangor Malaysia ; AgResearch Lincoln Research Centre, Christchurch, New Zealand ; Department of Wine, Food and Molecular Biosciences, Lincoln University, Christchurch, New Zealand.
authors.forename: Benjamin Yii Chung
authors.identifier: 0000-0001-7211-8284
authors.initials: BY
authors.lastname: Lau
authors.affiliation: AgResearch Lincoln Research Centre, Christchurch, New Zealand.
authors.forename: Santanu
authors.initials: S
authors.lastname: Deb-Choudhury
authors.affiliation: Department of Wine, Food and Molecular Biosciences, Lincoln University, Christchurch, New Zealand.
authors.forename: James D
authors.initials: JD
authors.lastname: Morton
authors.affiliation: AgResearch Lincoln Research Centre, Christchurch, New Zealand.
authors.forename: Stefan
authors.initials: S
authors.lastname: Clerens
authors.affiliation: AgResearch Lincoln Research Centre, Christchurch, New Zealand ; Department of Wine, Food and Molecular Biosciences, Lincoln University, Christchurch, New Zealand.
authors.forename: Jolon M
authors.initials: JM
authors.lastname: Dyer
authors.affiliation: Advanced Biotechnology and Breeding Centre, Malaysian Palm Oil Board, No. 6, Persiaran Institusi, Bandar Baru Bangi, 43000 Kajang, Selangor Malaysia.
authors.forename: Umi Salamah
authors.initials: US
authors.lastname: Ramli
keywords: Chromoplast; Elaeis guineensis; Fatty acid biosynthesis; Gel electrophoresis; Protein extraction; Proteomics