Three-dimensional
(3D)-printing techniques such as stereolithography
(SLA) are currently gaining momentum for the production of miniaturized
analytical devices and molds for soft lithography. However, most commercially
available SLA resins inhibit polydimethylsiloxane (PDMS) curing, impeding
reliable replication of the 3D-printed structures in this elastomeric
material. Here, we report a systematic study, using 16 commercial
resins, to identify a fast and straightforward treatment of 3D-printed
structures and to support accurate PDMS replication using UV and/or
thermal post-curing. In-depth analysis using Raman spectroscopy, nuclear
magnetic resonance, and high-resolution mass spectrometry revealed
that phosphine oxide-based photo-initiators, leaching out of the 3D-printed
structures, are poisoning the Pt-based PDMS catalyst. Yet, upon UV
and/or thermal treatments, photo-initiators were both eliminated and
recombined into high molecular weight species that were sequestered
in the molds.
Starch bound proteins mainly include enzymes from the starch biosynthesis pathway. Recently, new functions in starch molecular assembly or active protein targeting were also proposed for starch associated proteins. The potato genome sequence reveals 77 loci encoding starch metabolizing enzymes with the identification of previously unknown putative isoforms. Here we show by bottom-up proteomics that most of the starch biosynthetic enzymes in potato remain associated with starch even after washing with SDS or protease treatment of the granule surface. Moreover, our study confirmed the presence of PTST1 (Protein Targeting to Starch), ESV1 (Early StarVation1) and LESV (Like ESV), that have recently been identified in Arabidopsis. In addition, we report on the presence of a new isoform of starch synthase, SS6, containing both K-X-G-G-L catalytic motifs. Furthermore, multiple protease inhibitors were also identified that are cleared away from starch by SDS and thermolysin treatments. Our results indicate that SS6 may play a yet uncharacterized function in starch biosynthesis and open new perspectives both in understanding storage starch metabolism as well as breeding improved potato lines.
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