Abstract

Oleuropein (OE), the main polyphenol in olive leaf extract, is likely to decompose into hydroxytyrosol (HT) and elenolic acid under the action of light, acid, base, high temperature. In the enzymatic process, the content of OE in olive leaf extract and enzyme are key factors that affect the yield of HT. A selective enzyme was screened from among 10 enzymes with a high OE degradation rate. A single factor (pH, temperature, time, enzyme quantity) optimization process and a Box-Behnken design were studied for the enzymatic hydrolysis of 81.04% OE olive leaf extract. Additionally, enzymatic hydrolysis results with different substrates (38.6% and 81.04% OE) were compared and the DPPH antioxidant properties were also evaluated. The result showed that the performance of hydrolysis treatments was best using hemicellulase as a bio-catalyst, and the high purity of OE in olive extract was beneficial to biotransform OE into HT. The optimal enzymatic conditions for achieving a maximal yield of HT content obtained by the regression were as follows: pH 5, temperature 55 °C and enzyme quantity 55 mg. The experimental result was 11.31% ± 0.15%, and the degradation rate of OE was 98.54%. From the present investigation of the antioxidant activity determined by the DPPH method, the phenol content and radical scavenging effect were both decreased after enzymatic hydrolysis by hemicellulase. However, a high antioxidant activity of the ethyl acetate extract enzymatic hydrolysate (IC50 = 41.82 μg/mL) was demonstated. The results presented in this work suggested that hemicellulase has promising and attractive properties for industrial production of HT, and indicated that HT might be a valuable biological component for use in pharmaceutical products and functional foods.

Highlights

  • Olive, mainly originated from Mediterranean region, is a famous woody oil tree used to produce virgin olive oil

  • 38.6% OE olive extract sample were absorbed for 24 h by eight macroporous resins (DA201, DM130, AB-8, DM301, S-8, X-5, XAD-4, H103), and the OE content in the adsorption liquids were measured by High Performance Liquid Chromatography (HPLC)

  • This study suggested the best hydrolysis treatment was using hemicellulase as a biocatalysts

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Summary

Introduction

Olive (olea europaea L.), mainly originated from Mediterranean region, is a famous woody oil tree used to produce virgin olive oil. Olive oil is the cardinal characteristic of the Mediterranean diet, serving as the principal source of dietary fat Because it is rich in bioactive compounds (vitamins, flavonoids and polyphenols), it has been associated with lower rates of coronary heart disease (CHD), as well as reduced breast and colon cancer risk. Olive leaves contain a wide variety of phenolic compounds, for example, oleuropein (OE), hydroxytyrosol (HT), tyrosol (T), cumaric acid, ferulic acid, caffeic acid, etc These phenolic compounds have very good biological activity, i.e., antioxidative activity [1,2], in addition to antimicrobial activity against Helicobacter pylori, Campylobacter jejuni, Staphylococcus aureus [3], and Bacillus cereus, Escherichia coli and Salmonella enteritidis [4]. They used olive leaf extracts to produced large amounts of highly purified HT with a partially hyperthermophilic β-glucosidase immobilized on chitosan support (pH = 7.0, 60 °C) [20]. We investigated the phenol content and DPPH radical scavenging effect before and after the enzymatic hydrolysis

Preparation for High Purity OE
Enzyme Selection for HT Biotransformation
Substrate for HT Biotransformation
Effect of pH
Effect of Temperature
Effect of Bioconversion Time
Effect of Enzyme Quantity
Optimization of Enzymatic Hydrolysis by RSM
Antioxidant Activity
Materials
Preparation of the High Purity OE
Saturated Static Adsorption
Static Adsorption Kinetics
Static Desorption for Different Concentrations of the Eluent
Dynamic Adsorptions
Enzymatic Hydrolysis
Single Factor Experimental Design
Box-Behnken Design
Total Phenol Content Determination
Conclusions
Conflicts of Interest
Full Text
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