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The article presents a study that evaluates the bacterial surface display of w-transminase as a co-factor for efficient transamination and is functional in the dimeric state. The study is undertaken through constructing CotG-w-TA fusion protein expression by double digested polymerase chain reaction (PCR) product and through the use of protease accessibility test. The results of the study show a successful display of co-factor containing, which consists phosphate, and dimer-forming enzyme.

Abstract: We report a new membrane surface display system based on molecular chaperon, prsA, of Bacillus subtilis. Clostridium thermocellum cellulase, celA, was fused to C-terminal end of PrsA. Cellulase activity of B. subtilis protoplast, which expressed PrsA–CelA was 15 times higher compared to control strain. More than 85% of total cellulase activity was observed in surface displayed format and less than 15% of total cellulase activity was found in supernatant. Flow cytometric analysis of protoplast of PrsA–CelA fusion expressing bacteria provided another proof of uniform expression of fusion protein onto cytoplasmic membrane of B. subtilis. Without lysozyme treatment, only part of cellulase activity (10%) was observed in whole cell fraction. [Copyright &y& Elsevier]