Biochemical Properties of a Partially Purified Protease from Bacillus sp. CL18 and Its Use to Obtain Bioactive Soy Prote
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Biochemical Properties of a Partially Purified Protease from Bacillus sp. CL18 and Its Use to Obtain Bioactive Soy Protein Hydrolysates Andréia Monique Lermen 1 & Naiara Jacinta Clerici 1 & Daniel Joner Daroit 1 Received: 28 February 2020 / Accepted: 22 May 2020/ # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract
Microbial proteases are relevant biocatalysts with diverse applications. Production of protein hydrolysates is recently focused, since they might display biological activities. Therefore, the extracellular protease from Bacillus sp. CL18 was partially purified through ammonium sulfate precipitation (25–50% saturation) and gel filtration chromatography, with a 60.7-fold purification (40,593 U/mg protein) and 21.3% recovery. The partially purified protease (PPP) was characterized as a serine protease, with optimal activity at 51–59 °C and pH 7.4–8.8 and low thermal stability. Thermal inactivation followed first-order kinetics. PPP depended on Ca2+ for higher thermal stability, depicted by increases in half-lives (t1/2), activation energy (Ea), and free energy (ΔG#) for kinetic inactivation. PPP preferentially hydrolyzed casein > soy protein isolate (SPI) >>> keratinous materials. SPI hydrolysis by PPP was further investigated, and the obtained hydrolysates exhibited increased in vitro bioactivities. Hydrolysates displayed antioxidant capacities through the scavenging of synthetic organic radicals and Fe3+-reducing ability. In addition, hydrolysates inhibited the activities of dipeptidyl peptidase IV (DPP IV) and angiotensin-converting enzyme (ACE), suggesting antidiabetic and antihypertensive potentials, respectively. From its biochemical properties, PPP might be used to produce protein hydrolysates with multifunctional bioactivities. Both PPP and SPI hydrolysates can find applications in food biotechnology. Keywords Enzyme . Characterization . Thermal inactivation . Biocatalysis . Protein hydrolysate . Bioactivity Abbreviations Residual activity during thermal inactivation of partially purified protease A/A0 Abs Absorbance Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12010-02003355-1) contains supplementary material, which is available to authorized users.
* Daniel Joner Daroit [email protected] Extended author information available on the last page of the article
Applied Biochemistry and Biotechnology
ΔG# ΔH# ΔS# DMSO Ea EDTA h k Kb SPI HSPI HSPI2 HSPI4 P PMSF PPP R SDS SDS-PAGE t T t1/2 TCA U
Free energy change for the thermal inactivation of partially purified protease Enthalpy change for the thermal inactivation of partially purified protease Entropy change for the thermal inactivation of the partially purified protease Dimethyl sulfoxide Activation energy for thermal inactivation of partially purified protease Ethylenediaminetetraacetic acid Planck constant (6.6262 × 10−34 J/s) Inactivation rate of the partially purified protease at a given temperature Boltzmann constant (1.3806 × 10−23 J/K) Soy protein isolate
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