Identification and Characterization of Terpene Synthases Potentially Involved in the Formation of Volatile Terpenes in Carrot (Daucus carotaL.) Roots

2015 ◽  
Vol 63 (19) ◽  
pp. 4870-4878 ◽  
Author(s):  
Mosaab Yahyaa ◽  
Dorothea Tholl ◽  
Guy Cormier ◽  
Roderick Jensen ◽  
Philipp W. Simon ◽  
...  
Keyword(s):  
Terpene Synthases ◽  
Volatile Terpenes ◽  
Identification And Characterization

Identification and characterization of a kunzeaol synthase from Thapsia garganica: implications for the biosynthesis of the pharmaceutical thapsigargin

2012 ◽  
Vol 448 (2) ◽  
pp. 261-271 ◽  
Author(s):  
Benjamin Pickel ◽  
Damian P. Drew ◽  
Tom Manczak ◽  
Corinna Weitzel ◽  
Henrik T. Simonsen ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Antagonistic Effect ◽  
Farnesyl Diphosphate ◽  
Fragmentation Pattern ◽  
Terpene Synthases ◽  
Engineered Yeast ◽  
Thapsia Garganica ◽  
Identification And Characterization

Thapsigargin is a major terpenoid constituent of Thapsia garganica root. Owing to its potent antagonistic effect on the sarcoplasmic/endoplasmic reticulum Ca2+-ATPase, thapsigargin has been widely used to study Ca2+ signalling and is also a potential drug for prostate cancer. Despite its importance, thapsigargin biosynthesis in T. garganica remains unknown. In order to decipher thapsigargin biosynthesis, deep transcript sequencing (454 and Illumina) of the T. garganica root was performed, and two terpene synthases (TgTPS1/2) were identified. Functional characterization of their encoded enzymes in a metabolically engineered yeast revealed that TgTPS1 synthesized δ-cadinene, whereas TgTPS2 produced ten distinct terpenoids. However, cultivation of the TgTPS2-expressing yeast in pH-maintained conditions (pH 6–7) yielded one major oxygenated sesquiterpenoid, suggesting that formation of multiple terpenoids was caused by acidity. The major terpene product from TgTPS2 was identified as 6β-hydroxygermacra-1(10),4-diene (kunzeaol) by mass-fragmentation pattern, retention index, the nature of its acid-induced degradation and NMR. Also, recombinant TgTPS2 efficiently catalysed the synthesis of kunzeaol in vitro from farnesyl diphosphate with a Km of 2.6 μM and a kcat of 0.03 s−1. The present paper is the first report of a kunzeaol synthase, and a mechanism for the transformation of kunzeaol into the thapsigargin backbone is proposed.

Identification and characterization of aldose reductase in cultured rat mesangial cells

Diabetes ◽  
1992 ◽  
Vol 41 (9) ◽  
pp. 1165-1171 ◽  
Author(s):  
R. Kikkawa ◽  
K. Umemura ◽  
M. Haneda ◽  
N. Kajiwara ◽  
S. Maeda ◽  
...  
Keyword(s):  
Aldose Reductase ◽  
Mesangial Cells ◽  
Identification And Characterization
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