Expression, purification, and characterization of a novel acid phosphatase that displays protein tyrosine phosphatases activity from Metarhizium anisopliae strain CQMa102

The research was conducted at Chongqing key laboratory of soil multi-scale interfacial process, College of Resources and Environment, Southwest University.

Abstract

The protein tyrosine phosphatase (PTPase) plays an important role in insect immune system. Our group has purified a type of acid phosphatase that could specifically dephosphorylate trans-Golgi p230 in vitro. In order to study this phosphatase further, we have identified and cloned the phosphatase gene from a locust specific Metarhizium anisopliae Strain CQMa102. The CQMa102 phosphatase was expressed in Pichia pastoris to verify its protease activity. The molecular weight (M_W) and the isoelectric point (pI) of the phosphatase were about 85 kDa and 6.15, respectively. Substrate specificity evaluation showed that the purified enzyme exhibited high activity on O-phospho-L-tyrosine. At its optimal pH of 6.5 and optimum temperature of 70 °C, the protein showed the highest activity respectively. It can be activated by Ca²⁺, Mg²⁺, Mn²⁺, Ba²⁺, Co²⁺ and phosphate analogs, but inhibited by Zn²⁺, Cu²⁺, fluoride, dithiothreitol, β-mercaptoethanol and N-ethylmaleimide.

Graphical abstract

The expression, purification and characterization of a novel acid phosphatase that displays protein tyrosine phosphatases activity.

Key words:

• Metarhizium anisopliae
• protein tyrosine phosphatase
• characterization

Notes

The research was conducted at Chongqing key laboratory of soil multi-scale interfacial process, College of Resources and Environment, Southwest University.