In-situ and real-time monitoring of two-stage enzymatic preparation of ACE inhibitory peptides from Cordyceps militaris medium residues by ultrasonic-assisted pretreatment.

A protocol for the preparation of angiotensin-Ⅰ-converting enzyme (ACE) inhibitory peptides from Cordyceps militaris medium residues (CMMR) was established by two-stage enzymatic hydrolysis (amylase and protease). In a combination (mono-, dual-, and tri-frequency) of five different frequencies (20, 28, 35, 40 and 50 kHz), ultrasound-assisted pretreatment increased ACE inhibition rate in hydrolysate by 63.30 % under the mode of 20/28 kHz. Afterwards, near-infrared (NIR) spectrometer combined with spectral preprocessing methods and multivariate analysis like partial least square (PLS), synergy interval-PLS (Si-PLS), random frog-PLS (RF-PLS) and competitive adaptive reweighted sampling (CARS-PLS) was used to monitor the ACE inhibitory activity. The performance of models was evaluated by the correlation coefficient (Rp) and root mean square error (RMSEP). CARS-PLS models achieved optimal results for both amylase and protease hydrolysis with Rp = 0.9693, RMSEP = 0.01 for the former and Rp = 0.9454, RMSEP = 0.03 for the latter. NIR spectrometer combined with CARS-PLS models may be employed for in-situ and real-time monitoring of the preparation of ACE inhibitory peptides under ultrasonic-assisted pretreatment.