Effects of rubber elongation factor and small rubber particle protein from rubber-producing plants on lipid metabolism in Saccharomyces cerevisiae

Abstract

The common proteins rubber elongation factor (REF) and small rubber particle protein (SRPP) are associated with Hevea brasiliensis (Hb) rubber particles. They are involved in the stability of rubber particles and natural rubber biosynthesis. Recently, we cloned cDNAs encoding REF and SRPP from laticifers of Ficus carica (Fc). In the present study, we overexpressed REF/SRPPs (HbREF, FcREF, and FcSRPP) in Saccharomyces cerevisiae in anticipation of future rubber biosynthesis in recombinant yeast. The proteins were localized in the endoplasmic reticulum and lipid droplets (LDs), and affected LD morphology. Furthermore, their overexpression resulted in an accumulation of neutral lipids and a decrease in yeast cell size. This suggests that REF/SRPPs affect lipid metabolism and lead to a decline in the phospholipid content of yeast. We also found that expression of these proteins induced accumulation of steryl esters and triacylglycerols in yeast. This suggests that the coexpression of REF/SRPPs with key enzymes for the biosynthesis of target lipids in yeast is a promising way of increasing production of important lipids like triacylglycerols and terpenes, and that a protein complex consisting of cis-prenyltransferase (CPT), CPT-like protein, and REF/SRPPs for rubber biosynthesis could be reconstituted on yeast lipid droplets.