Production of highly soluble native human paraoxonase 2 with potential anti-biofilm property.

Fauzia Parween, Priyamedha Yadav, Kalyani Singh, Rinkoo Devi Gupta

Preparative biochemistry & biotechnology 2023

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Paraoxonase 2 (PON2) is considered as a potential anti-biofilm agent due to the highest lactonase activity among the PON family members implicating quorum quenching in gram-negative bacteria. However, PON2 is expressed mostly in insoluble fractions in the bacterial expression host which limits its application as an anti-biofilm agent. Therefore, obtaining the native human PON2 (HuPON2) protein in soluble form, better protein yield, stability, and enzymatic activities is essential. In this study, procedures for obtaining a high yield of the native form of HuPON2 in soluble and active forms were optimized. Guanidinium hydrochloride solubilized the HuPON2 protein, however, it is lethal for several bacteria, and thus a major problem for studying the various downstream application of the protein. Therefore, another refolding process for native HuPON2 was optimized. Owing to the promiscuous nature of HuPON2, we hypothesized that it could inhibit the biofilm formation in Mycobacterium smegmatis also. Interestingly, we observed a significant inhibition of the biofilm formation by HuPON2_Rf. However, the primary target of HuPON2 and the probable mechanism behind the quorum quenching in M. smegmatis need to be further explored, which would help widen the scope of HuPON2 as a potential anti-biofilm agent beyond the gram-negative bacteria.

Citation

Fauzia Parween, Priyamedha Yadav, Kalyani Singh, Rinkoo Devi Gupta. Production of highly soluble native human paraoxonase 2 with potential anti-biofilm property. Preparative biochemistry & biotechnology. 2023;53(5):465-474