Recovery of amylolytic enzymes from triticale malt (X. Triticosecale Wittmack) using two-phase aqueous systems

Abstract

Abstract Introduction: Triticale malt has shown higher amylolytic activity than other cereal malts, a characteristic of great importance for the brewing and starch industry. The scope of this work was to obtain concentrated enzymatic aqueous extracts containing β-amylase, α-amylase, and amyloglucosidase from triticale malts of Bicentenario and Siglo-XXI varieties, using aqueous two-phase systems (ATPS) for enzymes recovery. Methodology: The malts produced had 5 days of germination and were dried at 50°C. The extracts were prepared by mixing ground malt with deionized water (1:10), stirred at 180 rpm, testing three stirring times (30, 120, and 270 min) and two temperatures (30 and 40°C) (12 treatments). The extracts were centrifugated, freeze-dried and purified, varying the concentrations of alcohol and Na3C6H5O7. Enzyme quantification was performed using: BETAMYL-3® (β-amylase); AOAC 2002.0I, AACC22-02.0I (α-amylase), and McCleary et al., (1991) (amyloglucosidase). Results: In relation to the malt, an increase in extract enzyme quantification was achieved, mainly α-amylase for Siglo-XXI malt, initially with 99.56 CU/g to 1,268.89 CU/mL (507.5 CU/g dry matter) in 30 min/30°C extract. For the same treatment, the best ATPS for enzymatic recovery was 30% alcohol/18% Na3C6H5O7, α-amylase predominantly at the inferior phase 1,514.03 CU/mL (605.6 CU/g dry matter) and β-amylase 51.43 BU/mL (10.2 BU/g dry matter) at the upper phase. Conclusion: Aqueous amylase extraction from triticale Siglo-XXI malt in 30 min/30°C conditions is a suitable option for production of β-amylase and α-amylase in combination with the use of ethanol and Na3C6H5O7 ATPS to recovery amylolytic enzymes.