INVESTIGATION OF THE COMPOSITION AND GROWTH OF IN VITRO NATURAL RUBBER USING HIGH RESOLUTION SIZE EXCLUSION CHROMATOGRAPHY

Monitoring the growth of in vitro natural rubber (NR) was accomplished by high-resolution Size Exclusion Chromatography SEC. Washed rubber particles (WRP)s isolated from H. brasiliensis latex, containing the rubber transferase enzyme, were used to catalyze the polymerization of synthetic isopentenyl pyrophosphate (IPP) monomer in the presence of farnesyl pyrophosphate (FPP) initiator.  The high resolution SEC was able to detect the formation of new rubber. Changes in the low molecular weight fraction were also detected. Gravimetric analysis revealed ~30% mass gain after the in vitro synthesis. The overall gel content was found to be reduced, which further supported the formation of new rubber. This is the first report that utilizes high resolution SEC to monitor in vitro NR growth without the use of radio-labeling.