Abstract
The objective of this study was to establish a fast approach (< 1 h) for the evaluation of technical enzyme preparations (TEPs). An automated photometric analyzer (GalleryTM Plus) was equipped with 32 synthetic and natural substrates to measure aminopeptidase, carboxypeptidase, dipeptidyl peptidase and endopeptidase activities distinguishably and the proteolytic activity towards lupine protein of TEPs. The established so-called “activity fingerprints” (AFPs) delivered detailed information about the substrate spectra and peptidase side activities, noticing furthermore batch variations of Flavourzyme1000L. Based on their AFPs, particular TEPs were selected for lupine protein hydrolysis and the hydrolysates were analyzed regarding the degree of hydrolysis and the free amino acids. It was demonstrated that the information of the AFPs were applicable to predict important properties of the resulting hydrolysates. Consequently, the hydrolysis efficiency was improved (increase of 47%). The system introduced enables the targeted selection of TEPs for enzymatic protein hydrolysis, resulting in specific food protein hydrolysates.
Highlights
The worldwide market offers various technical enzyme preparations (TEPs) and, due to their wide range of application,1 3 Vol.:(0123456789)European Food Research and Technology (2019) 245:1695–1708 new preparations are added continuously [1, 2]
The values represent the enzyme activity in nkat m L−1 TEP, measured at 37 °C, pH 7.0, substrate concentration 1 mM for the synthetic substrates and 2.5 g L−1 for lupine protein. b Batch hydrolysis of 10% (w/v) lupine protein with 6% (w/v) P278, 6% (w/v) DZM and a combination of 3% (w/v) P278 and 3% (w/v) DZM at 37 °C, pH 7.0. c Free amino acid profiles of lupine protein hydrolysates using
The values represent the enzyme activity in nkat mL−1 TEP, measured at 37 °C, pH 7.0, substrate concentration 1 mM for the synthetic substrates and 2.5 g L−1 for lupine protein. b Batch hydrolysis of 10% (w/v) lupine protein with 6% (w/v) FP51, 6% (w/v) PepR and a combination of 3% (w/v) FP51 and 3% (w/v) PepR at 37 °C, pH 7.0. c Free amino acid profiles of lupine protein hydrolysates using FP51, PepR and a combination of FP51 and PepR, measured by ultra-performance liquid chromatography (UPLC)
Summary
The worldwide market offers various technical enzyme preparations (TEPs) and, due to their wide range of application (detergent, pharmaceutical, food and beverage industry),1 3 Vol.:(0123456789)European Food Research and Technology (2019) 245:1695–1708 new preparations are added continuously [1, 2]. The worldwide market offers various technical enzyme preparations (TEPs) and, due to their wide range of application (detergent, pharmaceutical, food and beverage industry),. TEPs are standardized either on one specific or an unspecific peptidase activity. TEPs have been described as containing several practice-relevant activities [19, 20], a reliable approach for the fast determination of important specific activities is missing. Due to this lack, TEPs are commonly chosen without detailed information about their process performance or are termed imprecisely as exopeptidases or endopeptidases [21,22,23,24]
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