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Nicotiana benthamiana
No full text<i>2.2. Characterization of TlTPS820 expressed in N. benthamiana</i> <p> Leaves from <i>N. benthamiana</i> expressing <i>Tl</i> TPS820 were extracted with hexane and analyzed by GC-MS. The analysis yielded a sesquiterpenoid profile identical to <i>Tg</i> TPS2 (Fig. 1). The main product of <i>Tg</i> TPS2 is epikunzeaol (<b>1</b>) (Fig. 2), thus <i>Tl</i> TPS820 is also an epikunzeaol synthase and an orthologue of <i>Tg</i> TPS2. As described previously, the high injection port temperature at 250 ̊C lead to cope-rearrangement of <b>1</b> (Andersen et al., 2017). Thus, <b>1</b> was partly degraded to two unknown alcohols.</p>Published as part of <i>Andersen, Trine Bundgaard, Rasmussen, Silas Anselm, Christensen, Søren Brøgger & Simonsen, Henrik Toft, 2019, Biosynthesis of tovarol and other sesquiterpenoids in Thapsia laciniata Rouy, pp. 168-174 in Phytochemistry 157</i> on page 169, DOI: 10.1016/j.phytochem.2018.10.027, <a href="http://zenodo.org/record/10481193">http://zenodo.org/record/10481193</a>
Nicotiana benthamiana
No full text<i>2.4. Characterization of TlTPS18983 expressed in N. benthamiana</i> <p> Volatiles from <i>N. benthamiana</i> leaves expressing <i>Tl</i> TPS18983 were captured by HS-SPME and analyzed by GC-MS. The analysis of these plants showed that expression of <i>Tl</i> TPS18983 generate at least 15 products. The identification of one of the major products, farnesene (<b>5</b>), was based on comparison of the mass spectrum of the compound with NIST and Wiley GC-MS spectrum libraries (Fig. 4, MS spectrum is provided in supplementary data Fig. 3). It was not possible to identify the remaining products. Using an injection port temperature of 160 ̊C instead of 250 ̊C did not alter the composition of the chromatogram, which supports that these compounds are not a result of the GC-MS analysis method. Extraction of the products was also attempted using organic solvents such as hexane and pentane mimicking procedures that were successful for other sqTPS products. However, for <i>Tl</i> TPS18983 we did not detect any compounds by GC-MS using organic solvent extraction.</p>Published as part of <i>Andersen, Trine Bundgaard, Rasmussen, Silas Anselm, Christensen, Søren Brøgger & Simonsen, Henrik Toft, 2019, Biosynthesis of tovarol and other sesquiterpenoids in Thapsia laciniata Rouy, pp. 168-174 in Phytochemistry 157</i> on page 170, DOI: 10.1016/j.phytochem.2018.10.027, <a href="http://zenodo.org/record/10481193">http://zenodo.org/record/10481193</a>
Expression of presumptive Microbial terpene synthase like genes (MTPSLs) from Lophocolea bidentata (liverwort) in moss Physcomitrella patens
No full textTerpenoids are structurally and functionally diverse natural compounds that are voluminous and more divergent in plants and less abundant in microbes. Terpene synthases are the enzymes that proprietarily produce broad range of terpene metabolites in nature by catalyzing the conversion of prenyl diphosphate precursor molecules. Putative microbial terpene synthase like genes (MTPSLs) were identified predominantly from non-seed plants but not in seed plants. This study is aimed at heterologous expression and biochemical characterization of presumptive microbial terpene synthase like genes (LbMTPSL1,3,4and 5) from untapped liverwort species Lophocolea bidentata in model plant Physcomitrella patens by exploiting its indispensable natural in vivo homologous recombination machinery. A new promising protocol using a blend of enzymes Cellulase R-10 and Macerozyme R-10 was utilized for protoplast preparation from P. patens. Homology based structural models for LbMTPSL1,4 have closest structural similarity to Epi-isozizaene synthase from Streptomyces coelicolor and LbMTPSL3,5 have identical stereoview as that of Selinadiene synthase from Streptomyces pristinaespiralis both belonging to C15 subclass, sesquiterpene synthases. Structure based function analysis predicted the active site binding affinity to Farnesyl diphosphate (FPP) for LbMTPSL1,3,5 and its analogue FsPP (Farnesyl thiopyrophosphate) for LbMTPSL4. The three basic residues along with phenol containing amino acid involved in substrate recognition motif and three aromatic residues with cation-π interactions for stabilizing carbocation intermediates were identified from the active site contour of all enzymes. It is believed that this knowledge on structural and chemical biology of LbMTPSLs built on the basis of computational data analysis could be sourceful for future mechanistic characterization and subsequent manipulations of FPP cyclization trajectory by rational design approach to alter the product profile significantly
Nicotiana benthamiana
No full text<i>2.3. Characterization of TlTPS509 expressed in N. benthamiana</i> <p> HS-SPME-GC-MS analysis of <i>N. benthamiana</i> leaves expressing <i>Tl</i> TPS509 resulted in the detection of two major sesquiterpene alcohols, guaiol (<b>3</b>) (RI: 1585, 34% of the area) and bulnesol (<b>4</b>) (RI: 1654, 20% of the area) (Fig. 3). Compound <b>3</b> and <b>4</b> exhibited similar EI MS spectrum to those of bulnesol and guaiol in NIST and Wiley GC-MS spectrum libraries. The product profile showed several additional compounds, and we could identify two minor products; guaiene (RI 1418, Rt: 8.6)) and bulnesene (RI 1487, Rt: 9.18) (Fig. 3). Several peaks with a <i>m/z</i> of 220 were also observed but not identified. A hexane extract of <i>N. benthamiana</i> leaves, expressing <i>Tl</i> TPS509 was analyzed by GC-MS using a Programmed Temperature Vaporising injector (PTV) port. By using a low initial temperature of 60 ̊C followed by a gradual increase to 250 ̊C it was expected that thermal rearrangements were avoided. Again, <b>3</b> and <b>4</b> were detected as major compounds, but not guaiene and bulnesene, which were probably below the detection limit. Furthermore, neither of the unknown compounds with <i>m/z</i> 220 were detected in the PTV analysis.</p>Published as part of <i>Andersen, Trine Bundgaard, Rasmussen, Silas Anselm, Christensen, Søren Brøgger & Simonsen, Henrik Toft, 2019, Biosynthesis of tovarol and other sesquiterpenoids in Thapsia laciniata Rouy, pp. 168-174 in Phytochemistry 157</i> on page 169, DOI: 10.1016/j.phytochem.2018.10.027, <a href="http://zenodo.org/record/10481193">http://zenodo.org/record/10481193</a>
Meet the Moss - User Testing Our First Fragrance
Get PDFVideo highlights of our first public outing with the Fragrant Moss. We demoed the product to passers by at Westfield Mall and Indie.Bio in San Francisc
Four novel geminally dialkylated, non-aromatic acetophenone derivatives from <em>Melicope coodeana</em>
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