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HCHL expression in hairy roots of Beta vulgaris yields a high accumulation of p-hydroxybenzoic acid (pHBA) glucose ester,and linkage of pHBA into cell walls
Authors:Laiq ur Rahman  Hitomi Kouno  Yuya Hashiguchi  Hirobumi Yamamoto  Arjan Narbad  Adrian Parr  Nicholas Walton  Toshihiko Ikenaga  Yoshie Kitamura
Affiliation:1. School of Pharmaceutical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan;2. Faculty of Environmental Studies, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan;3. Institute of Food Research, Norwich Research Park, Norwich NR4 7UA, UK
Abstract:As part of a study to explore the potential for new or modified bio-product formation, Beta vulgaris (sugar beet) has been genetically modified to express in root-organ culture a bacterial gene of phenylpropanoid catabolism. The HCHL gene, encoding p-hydroxycinnamoyl-CoA hydratase/lyase, was introduced into B. vulgaris under the control of a CaMV 35S promoter, using Agrobacterium rhizogenes LBA 9402. Hairy root clones expressing the HCHL gene, together with non-expressing clones, were analysed and revealed that one expression-positive clone accumulated the glucose ester of p-hydroxybenzoic acid (pHBA) at about 14% on a dry weight basis. This is the best yield achieved in plant systems so far. Determination of cell-wall components liberated by alkaline hydrolysis confirmed that the ratio of pHBA to ferulic acid was considerably higher in the HCHL-expressing clones, whereas only ferulic acid was detected in a non-expressing clone. The change in cell-wall components also resulted in a decrease in tensile strength in the HCHL-expressing clones.
Keywords:Beta vulgaris   Metabolic engineering   p-Hydroxycinnamoyl-CoA hydratase/lyase   p-Hydroxybenzoic acid   Cell wall
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