Heterogenous expression of Synechocystis sp. PCC 6803 deg proteases and their possible roles on phycobiliprotein degradation in vitro

Abstract

The Synechocystis sp. PCC 6803 genome harbours a Deg gene family consisting of three members, htrA (degP, slr1204), hhoA (degQ, sll1679) and hhoB (degS, sll1427). This work provided biochemical characterization of HhoA, HtrA and HhoB from Synechocystis sp. PCC 6803. Firstly mature HhoA, HhoB and HtrA from Synechocystis sp. PCC 6803 were cloned and expressed as soluble recombinant his-tagged fusion protein in Escherichia coli. Then the proteolytic activity of HhoA, HhoB and HtrA was tested using casein, bovine serum albumin, five recombinant chromoproteins and cyanobacterial phycocyanin as substrates in vitro. The experimental results showed that HhoA and HtrA had proteolytic activity on casein, five recombinant chromoproteins and cyanobacterial phycocyanin. No proteolytic activity of HhoB was found using all substrates in vitro, indicating functional difference among Deg proteases from Synechocystis sp. PCC 6803. Therefore, the results indicated the biochemical properties of HhoA and HtrA on hydrolysis of proteins and phycobiliproteins in vitro, which implicated that they were proteases possibly involved in phycobiliprotein degradation in vivo.

Keywords

Deg protease / proteolytic activity / phycobiliprotein / recombinant chromoprotein / Phycobilisome(PBS) degradation

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