{"title":"Construction of Recombinant E.coli Expressing Fusion Protein to Produce 1,3-Propanediol","authors":"Rosarin Rujananon, Poonsuk Prasertsan, Amornrat Phongdara, Tanate Panrat,Jibin Sun, Sugima Rappert, An-Ping Zeng","volume":52,"journal":"International Journal of Biotechnology and Bioengineering","pagesStart":228,"pagesEnd":235,"ISSN":"1307-6892","URL":"https:\/\/publications.waset.org\/pdf\/7023","abstract":"In this study, a synthetic pathway was created by\r\nassembling genes from Clostridium butyricum and Escherichia coli\r\nin different combinations. Among the genes were dhaB1 and dhaB2\r\nfrom C. butyricum VPI1718 coding for glycerol dehydratase (GDHt)\r\nand its activator (GDHtAc), respectively, involved in the conversion\r\nof glycerol to 3-hydroxypropionaldehyde (3-HPA). The yqhD gene\r\nfrom E.coli BL21 was also included which codes for an NADPHdependent\r\n1,3-propanediol oxidoreductase isoenzyme (PDORI)\r\nreducing 3-HPA to 1,3-propanediol (1,3-PD). Molecular modeling\r\nanalysis indicated that the conformation of fusion protein of YQHD\r\nand DHAB1 was favorable for direct molecular channeling of the\r\nintermediate 3-HPA. According to the simulation results, the yqhD\r\nand dhaB1 gene were assembled in the upstream of dhaB2 to express\r\na fusion protein, yielding the recombinant strain E. coliBL21\r\n(DE3)\/\/pET22b+::yqhD-dhaB1_dhaB2 (strain BP41Y3). Strain\r\nBP41Y3 gave 10-fold higher 1,3-PD concentration than E. coliBL21\r\n(DE3)\/\/pET22b+::yqhD-dhaB1_dhaB2 (strain BP31Y2) expressing\r\nthe recombinant enzymes simultaneously but in a non-fusion mode.\r\nThis is the first report using a gene fusion approach to enhance the\r\nbiological conversion of glycerol to the value added compound 1,3-\r\nPD.","references":"[1] H. Miller, \"Major fiber producers hop on corterra bandwagon,\" Int\r\nFiber J, vol. 15, pp. 14-16, 2000.\r\n[2] R. 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