Sains Malaysiana 46(9)(2017): 1491–1498

http://dx.doi.org/10.17576/jsm-2017-4609-18

Expression and Functional Analysis of a Transgenic Cytochrome P450 Monooxygenase in Pueraria mirifica

(Analisis Ekspresi dan Fungsian Transgen Sitokrom P450 Monooksigenase dalam Pueraria mirifica)

LI XU, RUETHAITHIP KAOPONG, SUREEPORN NUALKAEW, ARTHIYA CHULLASARA

& AMORNRAT PHONGDARA*

Center for Genomics and Bioinformatics Research, Department of Molecular Biotechnology

and Bioinformatics,Faculty of Science, Prince of Songkla University, Hatyai, Songkhla 90112

Thailand

Diserahkan: 31 Ogos 2016/Diterima: 17 April 2017

ABSTRACT

Isoflavonoids are the main compound in White Kwao Krua (Pueraria mirifica), which is an effective folk medicinal plant endemic to Thailand. It has been widely used for improving human physical and treating diseases. There are substances with estrogenic activities have been isolated from P. mirifica, such as puerarin, daidzein and genistein. Isoflavone synthase (IFS) is one of the key enzymes in Leguminous plants to convert liquiritigenin, liquiritigenin C-glucoside and naringenin chalcone to isoflavonoids. The aim of this research was to enhance the production of isoflavonoids by metabolic engineering. Transgenic plants were constructed by introducing P450 gene (EgP450) which is similar to IFS from oil palm (Elaeis guineensis), into P. mirifica by a biolistic method. After the transgenic plants had proved successfully, isoflavonoids of each group plants were determined by HPLC. The contents of daidzein and genistein in transgenic plants were higher than the control plants.

Keywords: Elaeis guineensis; isoflavonoids; Pueraria mirifica; P450 gene; transgenic plants

ABSTRAK

Isoflavonoid adalah sebatian utama dalam White Kwao Krua (Pueraria mirifica) yang merupakan tumbuhan perubatan yang berkesan dan endemik di Thailand. Ia telah digunakan secara meluas untuk meningkatkan tahap fizikal manusia dan merawat penyakit. Terdapat bahan dengan aktiviti estrogen yang telah dipencil daripada P. Mirifica seperti puerarin, daidzein dan genistein. Sintas isoflavon (IFS) merupakan salah satu enzim penting dalam tumbuhan kekacang yang menukar likuiritigenin, likuiritigenin C-glukosid dan naringenin kalkon kepada isoflavonoid. Kajian ini bertujuan meningkatkan pengeluaran isoflavonoid melalui kejuruteraan metabolisma. Tumbuhan transgen telah dihasilkan dengan memperkenalkan gen P450 (EgP450) yang menyerupai IFS daripada kelapa sawit (Elaeis guineensis) kepada P. mirifica melalui kaedah biolistik. Selepas tumbuhan transgen terbukti berjaya, isoflavonoid setiap kumpulan tumbuhan telah ditentukan melalui HPLC. Kandungan daidzein dan genistein dalam tumbuhan transgen adalah lebih tinggi daripada tumbuhan kawalan.

Kata kunci: Elaeis guineensis; gen P450; isoflavonoid; Pueraria mirifica; tumbuhan transgen

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