Display of Functionally Active PHB Depolymerase on Escherichia Coli Cell Surface

作者：Hiraishi Tomohiro^*; Yamashita Koichi; Sakono Masafumi; Nakanishi Jun; Tan Liu Tzea; Sudesh Kumar; Abe Hideki; Maeda Mizuo

来源：Macromolecular Bioscience, 2012, 12(2): 218-224.

DOI：10.1002/mabi.201100273

• bioengineering
• cell surface display
• degradation
• PHB depolymerase
• poly[(R)-3-hydroxybutyrate]

The display of PHB depolymerase (PhaZRpiT1) from R. pickettii T1 on the surface of E. coli JM109 cells is realized using OprI of P. aeruginosa as the anchoring motif. The fusion protein is stably expressed and its surface localization is verified by immunofluorescence microscopy. The displayed PhaZRpiT1 retains its cleaving ability for soluble substrates as well as its ability to adsorb to the PHB surface, and also remains catalycically active in the degradation of insoluble polyester materials, in spite of the possible suppression of the enzyme movement on the polymer surface. The results demonstrate that PhaZRpiT1-displaying E. coli shows potential for use as a whole-cell biocatalyst for the production of (R)-3-hydroxybutyrate monomers from insoluble PHB materials.

• 出版日期2012-2