Metaproteomic investigation of enzyme profile in daqu used for the production of Nongxiangxing baijiu

Enzymes and microbiota in daqu are essential for the brewing of Nongxiangxing baijiu. Uncover the key enzymes and functional strains in daqu is beneficial to improve the flavor and quality of Nongxiangxing baijiu. In this study, metaproteome technology was employed to determine the enzyme profiles in Nongxiangxing daqu, and strains with high saccharification activity were screened and identified. 933 proteins were identified in daqu, of which 463 belonged to enzymes, including 140 oxidoreductases, 98 transferases, 91 hydrolases, 49 ligases, 41 lyases and 27 isomerases, and hydrolase is the enzyme with the highest abundance in baijiu brewing. Among hydrolases, a total of 36 carbohydrate metabolism-related enzymes (CMEs) were identified, and 12 of them were key enzymes related to glycoside hydrolysis. Four major glycoside hydrolysis enzymes glucoamylase (EC 3.2.1.3), glucan 1,4-alpha-glucosidase (EC 3.2.1.3), glucanase (EC 3.2.1.-) and β-glucosidase (EC 3.2.1.21) were revealed, and their sources were Byssochlamys spectabilis, Lichtheimia ramosa and Thermoascus aurantiacus, respectively. Then, strains Aspergillus A2, A3, A7, Lichtheimia L1, L4, L5, and Saccharomycopsis S2, S4, S6 with high saccharifying enzyme-producing capacity were screened through culture-dependent approach. Resents presented in this study can further reveal the enzyme profiles and identify the main functional strains in daqu, which can provide theoretical support for the brewing of Nongxiangxing baijiu.