A phycocyanin·phellandrene synthase fusion enhances recombinant protein expression and β-phellandrene (monoterpene) hydrocarbons production in Synechocystis (cyanobacteria)

摘要: Cyanobacteria can be exploited as photosynthetic platforms for heterologous generation of terpene hydrocarbons with industrial applications. Transformation Synechocystis and expression the β-phellandrene synthase (PHLS) gene alone is necessary sufficient to confer ability divert intermediate terpenoid metabolites generate monoterpene during photosynthesis. However, synthases, including PHLS, have a slow Kcat (low Vmax) necessitating high levels enzyme concentration enable meaningful rates yield product formation. Here, novel approach was applied increase PHLS protein alleviating limitations in rate generation. Different fusion constructs were generated endogenous cpcB sequence, encoding abundant cyanobacteria phycocyanin β-subunit, expressed under native cpc operon promoter. In one these constructs, CpcB·PHLS accumulated approaching 20% total cellular protein, i.e., substantially higher than expressing same The retained activity catalyzed synthesis, yielding an average 3.2 mg g(-1) dry cell weight (dcw) versus 0.03 g(-1)dcw measured low-expressing 100-fold improvement. conclusion, fusion-protein promising, as, this case, it increased amount cyanobacteria, commensurately improved production microorganisms.