The moss Physcomitrella patens, has been genetically engineered to produce patchoulol and β-santalene, two valuable sesquiterpenoid ingredients in the fragrance industry. The highest yield of patchoulol achieved was 1.34 mg/g dry weight. This was achieved by non-targeted transformation of the patchoulol synthase and either a yeast or P. patens HMGR gene under the control of a 35S promoter. Santalene synthase targeted to the plastids yielded 0.039 mg/g dry weight of α/β santalene; cytosolic santalene synthase and 35S controlled HMGR afforded 0.022 mg/g dry weight. It has been observed that the final yield of the fragrance molecules is dependent on the expression of the synthase. This is the first report of heterologous production of sesquiterpenes in moss and it opens up a promising source for light-driven production of valuable fragrance ingredients.
Mercury ions (Hg(2+)) can specifically interact with the thymine-rich Hg(2+) aptamer and malachite green (MG) to form the Hg(2+) aptamer-MG-Hg(2+) complex, inducing the increase of resonance scattering (RS) intensity at 611 nm, which enables the label-free detection of Hg(2+) in aqueous solution with high selectivity and a detection limit of 1.7 nM.
Heterologous and stable expression of genes encoding terpenoid biosynthetic enzymes in planta is an important tool for functional characterization and is an attractive alternative to expression in microbial hosts for biotechnological production. Despite improvements to the procedure, such as streamlining of large scale Agrobacterium infiltration and upregulation of the upstream pathways, transient in planta heterologous expression quickly reaches limitations when used for production of terpenoids. Stable integration of transgenes into the nuclear genome of the moss Physcomitrella patens has already been widely recognized as a viable alternative for industrial-scale production of biopharmaceuticals. For expression of terpenoid biosynthetic genes, and reconstruction of heterologous pathways, Physcomitrella has unique attributes that makes it a very promising biotechnological host. These features include a high native tolerance to terpenoids, a simple endogenous terpenoid profile, convenient genome editing using homologous recombination, and cultivation techniques that allow up-scaling from single cells in microtiter plates to industrial photo-bioreactors. Beyond its use for functional characterization of terpenoid biosynthetic genes, engineered Physcomitrella can be a green biotechnological platform for production of terpenoids. Here, we describe two complementary and simple procedures for stable nuclear transformation of Physcomitrella with terpenoid biosynthetic genes, selection and cultivation of transgenic lines, and metabolite analysis of terpenoids produced in transgenic moss lines. We also provide tools for metabolic engineering through genome editing using homologous recombination.
Four silicate glasses were prepared by the fusion of about 1 kg powder each of a basalt, syenite, soil and andesite to provide reference materials of natural composition for microanalytical work. These glasses are referred to as ‘Chinese Geological Standard Glasses’ (CGSG) ‐1, ‐2, ‐4 and ‐5. Micro and bulk analyses indicated that the glasses are well homogenised with respect to major and trace elements. Some siderophile/chalcophile elements (e.g., Sn, Pt, Pb) may be heterogeneously distributed in CGSG‐5. This paper provides the first analytical data for the CGSG reference glasses using a variety of analytical techniques (wet chemistry, XRF, EPMA, ICP‐AES, ICP‐MS, LA‐ICP‐MS) performed in nine laboratories. Most data agree within uncertainty limits of the analytical techniques used. Discrepancies in the data for some siderophile/chalcophile elements exist, mainly because of possible heterogeneities of these elements in the glasses and/or analytical problems. From the analytical data, preliminary reference and information values for fifty‐five elements were calculated. The analytical uncertainties [2 relative standard error (RSE)] were estimated to be between about 1% and 20%.
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