5 research outputs found
Synthesis of a Chloroalkene Dipeptide Isostere-Containing Peptidomimetic and Its Biological Application
The first rapid and
efficient chemical synthesis of a cyclic Arg-Gly-Asp
(RGD) peptide containing a chloroalkene dipeptide isostere (CADI)
is reported. By a developed synthetic method, an <i>N</i>-<i>tert</i>-butylsulfonyl protected CADI was obtained
utilizing diastereoselective allylic alkylation as a key reaction.
This CADI was also transformed into an <i>N</i>-Fmoc protected
CADI in a few steps. The CADI was used in Fmoc-based solid-phase peptide
synthesis. The first synthesis of a CADI-containing cyclic RGD peptide
was successful, and the synthesized CADI-containing peptidomimetic
was found to be a more potent inhibitor against integrin-mediated
cell attachment than the parent cyclic peptide
Combined sensory, volatilome and transcriptome analyses identify a limonene terpene synthase as a major contributor to the characteristic aroma of a Coffea arabica L. specialty coffee
International audienceBackground The fruity aromatic bouquet of coffee has attracted recent interest to differentiate high value market produce as specialty coffee. Although the volatile compounds present in green and roasted coffee beans have been extensively described, no study has yet linked varietal molecular differences to the greater abundance of specific substances and support the aroma specificity of specialty coffees. Results This study compared four Arabica genotypes including one, Geisha Especial, suggested to generate specialty coffee. Formal sensory evaluations of coffee beverages stressed the importance of coffee genotype in aroma perception and that Geisha Especial-made coffee stood out by having fine fruity, and floral, aromas and a more balanced acidity. Comparative SPME-GC-MS analyses of green and roasted bean volatile compounds indicated that those of Geisha Especial differed by having greater amounts of limonene and 3-methylbutanoic acid in agreement with the coffee cup aroma perception. A search for gene ontology differences of ripening beans transcriptomes of the four varieties revealed that they differed by metabolic processes linked to terpene biosynthesis due to the greater gene expression of prenyl-pyrophosphate biosynthetic genes and terpene synthases. Only one terpene synthase (CaTPS10-like) had an expression pattern that paralleled limonene loss during the final stage of berry ripening and limonene content in the studied four varieties beans. Its functional expression in tobacco leaves confirmed its functioning as a limonene synthase. Conclusions Taken together, these data indicate that coffee variety genotypic specificities may influence ripe berry chemotype and final coffee aroma unicity. For the specialty coffee variety Geisha Especial, greater expression of terpene biosynthetic genes including CaTPS10-like, a limonene synthase, resulted in the greater abundance of limonene in green beans, roasted beans and a unique citrus note of the coffee drink
Structural Insights into the CotB2-Catalyzed Cyclization of Geranylgeranyl Diphosphate to the Diterpene Cyclooctat-9-en-7-ol
The diterpene cyclase
CotB2 catalyzes the cyclization of geranylgeranyl
diphosphate (GGPP) to the tricyclic cyclooctat-9-en-7-ol, which is
characterized by a 5-8-5-fused ring skeleton. We have previously proposed
a cyclization cascade involving a unique carbon–carbon bond
rearrangement combined with multiple hydride shifts, all occurring
at a single active site. Here, we report the first high-resolution
X-ray crystal structure of CotB2 with bound substrate analog geranylgeranyl
thiodiphosphate (GGSPP). In the GGSPP-bound form, GGSPP folds into
a unique S-shaped conformation that probably reflects the substrate-bound
state prior to ionization of the substrate GGPP. The folded framework
of GGSPP is surrounded by hydrophobic residues and several aromatic
and asparagine residues that are well-positioned to stabilize a series
of reactive carbocation intermediates through a combination of cation−π
and dipole charge interactions. The combined crystal structures and
mutagenesis-based biochemical assays provide a structural basis for
exquisite control of ring formation and stereochemistry during CotB2
catalysis
Structural Insights into the CotB2-Catalyzed Cyclization of Geranylgeranyl Diphosphate to the Diterpene Cyclooctat-9-en-7-ol
The diterpene cyclase
CotB2 catalyzes the cyclization of geranylgeranyl
diphosphate (GGPP) to the tricyclic cyclooctat-9-en-7-ol, which is
characterized by a 5-8-5-fused ring skeleton. We have previously proposed
a cyclization cascade involving a unique carbon–carbon bond
rearrangement combined with multiple hydride shifts, all occurring
at a single active site. Here, we report the first high-resolution
X-ray crystal structure of CotB2 with bound substrate analog geranylgeranyl
thiodiphosphate (GGSPP). In the GGSPP-bound form, GGSPP folds into
a unique S-shaped conformation that probably reflects the substrate-bound
state prior to ionization of the substrate GGPP. The folded framework
of GGSPP is surrounded by hydrophobic residues and several aromatic
and asparagine residues that are well-positioned to stabilize a series
of reactive carbocation intermediates through a combination of cation−π
and dipole charge interactions. The combined crystal structures and
mutagenesis-based biochemical assays provide a structural basis for
exquisite control of ring formation and stereochemistry during CotB2
catalysis