subsequently an aliquot of 10 u03bcl was transferred from each mixture into a well of a microwell plate 5 u03bcl of a solution of a test peptide r r l i e d a e py a a r g seq id no 1 1 mm in water and 10 u03bcl of a buffer containing 0 1 mm cacl 2 50 mm tris ph 7 0 were added to each well and incubated for 30 min at 37 u00b0 c free phosphate resulting from the enzymatic hydrolysis was then detected via addition of 100 u03bcl of a 1 m hcl solution additionally containing 0 034 w v malachite green 10 mm sodium molybdate and 3 4 v v ethanol the detection was performed for 10 min at rt while constantly agitating the microwell plate relative concentrations of free phosphate were detected following the determination of the extinction 620 nm of each reaction well subsequently an aliquot of 10 u03bcl was transferred from each mixture to a microwell plate 5 u03bcl of a solution of a test peptide r r l i e d a e py a a r g seq id no 1 1 mm in water and 10 u03bcl of a buffer containing 0 1 mm cacl 2 50 mm tris ph 7 0 were added to each well and incubated for 30 min at 37 u00b0 c free phosphate resulting from the enzymatic hydrolysis was then detected via addition of 100 u03bcl of a 1m hcl solution additionally containing 0 034 w v malachite green 10 mm sodium molybdate and 3 4 v.