ORIGINAL PAPER Proteomic analysis of cell suspension cultures of Boesenbergia rotunda induced by phenylalanine: identification of proteins involved in flavonoid and phenylpropanoid biosynthesis pathways Eng Chong Tan • Saiful Anuar Karsani • Gen Teck Foo • Sher Ming Wong • Noorsaadah Abdul Rahman • Noorzulaani Khalid • Shatrah Othman • Rohana Yusof Received: 14 February 2012 / Accepted: 2 June 2012 Ó Springer Science+Business Media B.V. 2012 Abstract Boesenbergia rotunda belongs to the Zingi- beraceae family. It is widely found throughout Southeast Asia and is commonly used as a food ingredient and in folk medicine. Extracts from this plant contain a number of important bioactive compounds such as boesenbergin, cardamonin, pinostrobin, pinocembrin, panduratin A and 4-hydroxypanduratin A. These compounds have been shown to exhibit anti-HIV protease, anti-dengue NS2B/ NS3 protease, antibacterial, antifungal, anti-inflammatory, anticancer, and antioxidant activity. Here we report the use of proteomic approaches to identify proteins that may be involved in the biosynthesis of these compounds. Protein expressions of B. rotunda suspension cultures for phenyl- alanine-treated and normal callus were compared by two- dimensional gel electrophoresis. Following image analysis, protein spots whose expressions were found to be regulated were identified using Matrix Assisted Laser Desorption- Ionization tandem mass spectrometry. In all, thirty four proteins were identified. These proteins were categorized into nine functional categories—defence mechanism, pro- tein biosynthesis, metabolism, terpenoid biosynthesis, cell division, cell organization, energy-related, signaling pro- cesses and proteins of unknown function. Eleven of the proteins involved in the phenylpropanoid biosynthetic pathway are related to the biosynthesis of cyclohexenyl chalcone derivatives. Keywords Boesenbergia rotunda Á Flavonoids Á Panduratin A Á Proteomics Á 4-Hydroxypanduratin A Introduction Boesenbergia rotunda is a small perennial plant belonging to the Zingiberaceae family. It is widely found in South East Asian countries with local names such as Chinese keys, finger root, and temu kunci. It is commonly used as a food ingredient and folk medicine to treat diseases such as aphthous ulcer, stomach discomfort, leucorrhea, dysentery, rheumatism and muscular pain. It has been shown that the primary bioactive compounds of this ginger are boesen- bergin, cardamonin, pinostrobin, pinocembrin, panduratin Electronic supplementary material The online version of this article (doi:10.1007/s11240-012-0188-8) contains supplementary material, which is available to authorized users. E. C. Tan (&) Á S. Othman Á R. Yusof (&) Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia e-mail: techong@siswa.um.edu.my R. Yusof e-mail: rohana@um.edu.my E. C. Tan Á S. A. Karsani (&) Á N. Abdul Rahman Á S. Othman Á R. Yusof Drug Design and Development Research Group, University of Malaya, 50603 Kuala Lumpur, Malaysia e-mail: saiful72@um.edu.my S. A. Karsani Á G. T. Foo Á S. M. Wong Á N. Khalid Faculty of Science, Institute of Biological Sciences, University of Malaya, 50603 Kuala Lumpur, Malaysia S. A. Karsani University of Malaya Centre for Proteomics Research (UMCPR), Kuala Lumpur, Malaysia G. T. Foo Á S. M. Wong Á N. Khalid Biotechnology and Bioproduct Research Cluster (UMBIO), University of Malaya, 50603 Kuala Lumpur, Malaysia N. Abdul Rahman Department of Chemistry, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia 123 Plant Cell Tiss Organ Cult DOI 10.1007/s11240-012-0188-8