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Characterization of the Asiatic Acid Glucosyltransferase, UGT73AH1, Involved in Asiaticoside Biosynthesis in Centella asiatica (L.) Urban.

ABSTRACT: Centella asiatica (L.) Urban contains two ursane-type triterpene saponins, asiaticoside and madecassoside, as major secondary metabolites. In order to select candidate genes encoding UDP-glucosyltransferases (UGTs) involved in asiaticoside biosynthesis, we performed transcriptomic analysis of leaves elicited by methyl jasmonate (MeJA). Among the unigenes, 120 isotigs and 13 singletons of unique sequences were annotated as UGTs, including 37 putative full-length cDNAs, and 15 of the putative UGT genes were named according to the UGT committee nomenclature protocols. One of them, UGT73AH1, was characterized by heterologous expression in Escherichia coli BL21 (DE3) cells. After induction with IPTG, a total protein extract was assayed with UDP-glucose and asiatic acid. UPLC-QTOF/MS analysis showed that UGT73AH1 catalyzes the glycosylation of asiatic acid to its monoglucoside. It remains unclear whether glycosylation occurs on the triterpene C-2?, C-3?, C-23, or C-28 position. However, it is very likely that UGT73AH1 glucosylates the C-28 position, because only C-28 bears a glucose moiety in the final pathway product of asiatic acid, while C-2?, C-3?, and C-23 remain un-conjugated.

SUBMITTER: Kim OT

PROVIDER: S-EPMC5751233 | biostudies-literature | 2017 Dec

REPOSITORIES: biostudies-literature

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Characterization of the Asiatic Acid Glucosyltransferase, UGT73AH1, Involved in Asiaticoside Biosynthesis in Centella asiatica (L.) Urban.

Kim Ok Tae OT Jin Mei Lan ML Lee Dae Young DY Lee Dae Young DY Jetter Reinhard R

International journal of molecular sciences 20171206 12

<i>Centella asiatica</i> (L.) Urban contains two ursane-type triterpene saponins, asiaticoside and madecassoside, as major secondary metabolites. In order to select candidate genes encoding UDP-glucosyltransferases (UGTs) involved in asiaticoside biosynthesis, we performed transcriptomic analysis of leaves elicited by methyl jasmonate (MeJA). Among the unigenes, 120 isotigs and 13 singletons of unique sequences were annotated as UGTs, including 37 putative full-length cDNAs, and 15 of the putati ...[more]

PMID: 29210992

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Project description:BackgroundEndophytes are a rich source of novel, distinct, and applicable compounds of interest in agricultural, medical, cosmetic, and pharmaceutical industries. In this respect, they have been attracting growing interest in the past few years. Endophytes are defined as microorganisms such as bacteria and fungi which have a mutualistic relationship with their host plants without causing any harm to their host. In this study, we isolated and identified bacterial endophytes from Centella asiatica collected in Western Cape, South Africa.ResultsTwenty bacterial endophytes were isolated from Centella asiatica and characterized by using morphological and molecular techniques. Based on molecular traits, the isolates were identified as Pseudomonas sp. strain SGM1, Pseudomonas sp. strain SGM2, Pseudomonas sp. strain SGM3, Pseudomonas sp. strain SGM4, Pseudomonas sp. strain SGM5, Pseudomonas sp. strain SGM6, Pseudomonas sp. strain SGM7, Novosphingobium sp. strain SGM8, Pseudomonas sp. strain SGM9, Pseudomonas sp. strain SGM10, Chryseobacterium sp. strain SGM11, Enterobacter sp. strain SGM12, Enterobacter sp. strain SGM13, Pseudomonas sp. strain SGM14, Enterobacter sp. strain SGM15, Enterobacter sp. strain SGM16, Agrobacterium sp. strain SGM17, Pantoea sp. strain SGM18, Paraburkholderia sp. strain SGM19, and Pseudomonas sp. strain SGM20. Pseudomonas genus was dominant with eleven isolates. Morphological trait results showed that all isolates were gram-negative rod-shaped bacteria.ConclusionAccording to our understanding, this study revealed the first twenty endophytic bacteria isolated from Centella asiatica growing in the Western Cape Province, South Africa. Data obtained in the current study will increase the knowledge of the already existing microbial diversity associated with Centella asiatica. Further work is needed to evaluate the antioxidant and antibacterial activities in vitro and assess the growth and medicinal compounds of the identified endophytic bacteria in a laboratory scale bioreactors.

Dataset Information

Characterization of the Asiatic Acid Glucosyltransferase, UGT73AH1, Involved in Asiaticoside Biosynthesis in Centella asiatica (L.) Urban.

Publications

Characterization of the Asiatic Acid Glucosyltransferase, UGT73AH1, Involved in Asiaticoside Biosynthesis in Centella asiatica (L.) Urban.

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