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Journal of Environmental BiologypISSN: 0254-8704 ; eISSN: 2394-0379 ; CODEN: JEBIDP |
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Abstract - Issue Sep 2010, 31 (5) BackPreparation and assay of C-glucosyltransferase from roots of Pueraria lobata Gang Chen1,2, Xuan
Wu1, Wen-Ling Zhou1 and 1Guangdong
Key Lab of Biotechnology for Plant Development, College of Life Sciences, 2Department
of Biology, (Received: Abstract: C-glucosyltransferase
(EC 2.4.1.X) is one of the key enzymes for the biosynthesis of puerarin. This paper describes the methodology in
purification and assay of the enzyme for the first time in Puerarin
lobata (Willd.) Ohwi. C-glucosyltransferase from roots of P. lobata
was extracted and partially purified by (NH4)2SO4
saturation. The effects of pH, temperature, and substrate concentration on the
activity of the enzyme were investigated. The properties of the puerarin produced by C-glucosyltransferase
were studied by thin layer chromatography (TLC) and high performance liquid
chromatography (HPLC). The peak activity of C-glucosyltransferase
was detected in fraction of by 80% saturation of (NH4)2SO4
and the optimal conditions for enzymatic reaction were 35.5 mmol l-1 of isoliquiritigenin and 560 mmol l-1 of UDP-G at pH 8.1, 28oC for 1 h. Mn2+ at 1 mmol l-1
and Al3+ at 1 mmol l-1
increased the enzyme activity, while Mg2+ inhibited its activity.
The enzyme activity in Nicotiana tabacum
and P. lobata were detected under the above assay
conditions. Higher activity was found in roots than in leaves and stems of P. lobata, while no enzyme activity was detected in leaves of
N. tabacum. It was the first time that activity of C-glucosyltransferase, which transforms isoliquiritigenin
to puerarin, was detected in P. lobata. Key
words: Pueraria
lobata (Willd.) Ohwi, Biosynthesis of puerarin,
C-glucosyltransferase, C-glucosylation PDF of full length paper is available online Copyright ? 2010 Triveni Enterprises. All rights reserved. No part of the Journal can be reproduced in any form
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