The Characterization of the Purine Nucleoside Phosphorylase from Agaricus bisporus and Its Potential Application in Reducing Purine Content in Beer

Jun Liu; Jian Lu

doi:10.3390/jof11040268

The Characterization of the Purine Nucleoside Phosphorylase from Agaricus bisporus and Its Potential Application in Reducing Purine Content in Beer

J Fungi (Basel). 2025 Mar 31;11(4):268. doi: 10.3390/jof11040268.

Authors

Jun Liu^{1

2

3

4

5}, Jian Lu^{1

2

3}

Affiliations

¹ Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
² National Engineering Research Center of Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi 214122, China.
³ Jiangsu Provincial Research Center for Bioactive Product Processing Technology, Jiangnan University, Wuxi 214122, China.
⁴ Sericultural and Agri-Food Research Institute, Guangdong Academy of Agricultural Sciences, Key Laboratory of Functional Foods, Ministry of Agriculture and Rural Affairs, Guangdong Key Laboratory of Agricultural Products Processing, Guangzhou 510610, China.
⁵ Key Laboratory for Prevention and Control of Avian Influenza and Other Major Poultry Diseases, Ministry of Agriculture and Rural Affairs, Key Laboratory of Livestock Disease Prevention of Guangdong Province, Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China.

Abstract

Beer, the most popular alcoholic beverage, poses health risks for individuals with gout and hyperuricemia due to its high purine content. Herein, we identified a novel purine nucleoside phosphorylase (AbPNP) from the edible mushroom Agaricus bisporus and heterologously expressed it in Pichia pastoris. The recombinant AbPNP exhibited optimal activity at 60 °C and pH 7.0, retaining >80% activity at pH 6.0-9.0 and >85% activity after 3 h at ≤60 °C. Kinetic analysis revealed high catalytic efficiency (kcat/Km = 2.02 × 10⁶ s^-1⋅M^-1) toward inosine, with strong resistance to metal ions except for Co²⁺ and Cu²⁺. The application of AbPNP (1.0-5.0 U/mL) during wort saccharification reduced purine nucleosides by 33.54% (from 151.53 to 100.65 mg/L) while increasing yeast utilization of free purine bases. The resulting beer showed improved fermentation performance (alcohol content increased by 3.6%) without compromising flavor profiles. This study provides the food-grade enzymatic strategy for low-purine beer production, leveraging the GRAS status of both A. bisporus and P. pastoris.

Keywords: Agaricus bisporus; beer; gout; purine nucleoside phosphorylase; purines.

Abstract

Grants and funding