biostudies-literatureKohler TTechnische Universität DresdenBundesministerium für Bildung und ForschungDeutsche ForschungsgemeinschaftEuropean Research Council40588-40596https://www.ebi.ac.uk/biostudies/studies/S-EPMC9057574biostudies-literatureUnknown10(66)In the present work, microgels were utilized as a cell-free reaction environment to produce a functional malonyl-CoA synthetase (deGFP-MatB) under geometry-controlled transcription and translation. Our approach combines the straight-forward optimization of overall protein yield of an <i>E. coli</i>-based cell-free protein synthesis (CFPS) system based on concentration screening of magnesium and potassium glutamate, DNA as well as polyethylene glycol (PEG), and its innovative usage in microgel-based production of a key enzyme of the polyketide synthesis pathway. After partial modification of the carboxyl groups of hyaluronic acid (HA) with 5'-methylfuran groups <i>via</i> 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM)-activation, these were further functionalizeRSC advancesCell-free protein synthesis and <i>in situ</i> immobilization of deGFP-MatB in polymer microgels for malonate-to-malonyl CoA conversion.PMC9057574GRK 1865031A360C852065TH 2037/1-1Weigel NThiele JHoefgen SKohler THeida TValiante VfalseCell-free protein synthesis and <i>in situ</i> immobilization of deGFP-MatB in polymer microgels for malonate-to-malonyl CoA conversion.In the present work, microgels were utilized as a cell-free reaction environment to produce a functional malonyl-CoA synthetase (deGFP-MatB) under geometry-controlled transcription and translation. Our approach combines the straight-forward optimization of overall protein yield of an <i>E. coli</i>-based cell-free protein synthesis (CFPS) system based on concentration screening of magnesium and potassium glutamate, DNA as well as polyethylene glycol (PEG), and its innovative usage in microgel-based production of a key enzyme of the polyketide synthesis pathway. After partial modification of the carboxyl groups of hyaluronic acid (HA) with 5'-methylfuran groups <i>via</i> 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM)-activation, these were further functionalize2020-01-01T00:00:00Z2020 Nov2025-04-19T09:20:15.651Z2025-04-19T09:20:15.651ZS-EPMC90575743552086810.1039/d0ra06702d