Hesperidinase encapsulation towards hesperitin production targeting improved bioavailability

Abstract

Hesperidin (hesperitin-7-O-rutinoside) and hesperitin (hesperitin-7-O-glucoside) show anti-inflammatory, antimicrobial, antioxidant, and anticarcinogenic effects and prevent bone loss. However, hesperidin has a low bioavailability compared to hesperitin due to the rutinoside moiety attached to the flavonoid. The removal of the rhamnose group to yield the corresponding flavonoid glucoside (hesperetin-7-glucoside) improved the bioavailability of the aglycone, hesperetin, in humans. In line with these assumptions, the aim of this work was the enzymatic production of hesperitin from hesperidin with hesperidinase. Despite the low hesperidin solubility in the reaction medium, the enzymatic bioconversion was carried with hesperidin soluble at lower concentrations (≤0.05 mg ml(-1)) and insoluble for high concentrations (>0.1-50 mg ml(-1)). A twofold increase in maximum reaction rates overtook the expected values, pointing to the enzyme ability to degrade insoluble hesperidin. To improve the bioprocess, hesperidinase was tested soluble and immobilized in calcium alginate (2%), k-carrageenan (2%), and chitosan (2%) beads. The immobilization was carried out by adsorption and encapsulation. Chitosan was cross-linked with glutaraldehyde (1% and 2%) and sodium sulfate (13.5% and 15%) in acetate buffer (0.02 M, pH 4.0). The relation between bioprocessing conditions and hesperidinase stability was studied. A residual activity of 193% was obtained with immobilized hesperidinase compared to the soluble form. A half-life of 770 min was attained with hesperidinase encapsulated in calcium alginate beads. The results presented in this work highlight the potential of hesperidinase encapsulation towards hesperitin production with insoluble substrate. To our knowledge, this work presents for the first time the potential of hesperidinase encapsulation on hydrogels for hesperitin production. This is an important achievement for pharmaceutical and nutraceutical applications of hesperitin because this compound presents a higher bioavailability compared to hesperidin.

Chemical Characterization, Antioxidant and Cytotoxic Activities of the Edible Fruits of Brosimun gaudichaudii Trécul, a Native Plant of the Cerrado Biome

“…The elimination of an additional hexose (162 Da) or the direct loss of the rutinose residue results in the aglycone ion (hesperetin) at m/z 303 ( Scheme 1). [31,32] Hesperetin‐7‐ O ‐rutinoside is reported as an active compound that reduces blood pressure in healthy volunteers [33] and prevents bone loss [34] …”

“…[31,32] Hesperetin-7-O-rutinoside is reported as an active compound that reduces blood pressure in healthy volunteers [33] and prevents bone loss. [34] In the same manner, peak 5 [M + H] + = 465.1027 (C 21 H 20 O 12 ) was proposed as quercetin-O-glucoside. In the MS spectrum, the loss of hexose (162 Da) moieties gave a fragment ion at m/z 303, which corresponds to the quercetin aglycone structure (Figure 2A).…”

Protective Effect of Hesperetin and Naringenin against Apoptosis in Ischemia/Reperfusion-Induced Retinal Injury in Rats

“…Antioxidant activity of the flavonoids favors by the presence of a 3-hydroxyl group in the heterocyclic ring and a catechol group in ring B, which are also the structural features of hesperetin and naringenin [16, 27]. Although this study implemented a treatment method of intraperitoneal injection of flavonoids, their intestinal absorbable forms include a 2-hydroxyl group (hesperidin and naringin), which appears in citrus fruits naturally and are converted to biologically active forms (hesperetin and naringenin) in vivo [16, 28]. As those have 3-hydroxyl group, hesperetin and naringenin have been suggested to be more effective antioxidants than ascorbic acid and α -tocopherol, particularly for urgent antioxidative treatment needs [29].…”

Citrus Flavonoids as Promising Phytochemicals Targeting Diabetes and Related Complications: A Systematic Review of In Vitro and In Vivo Studies

“…Hesperetin ((S) -2,3-dihydro-5,7-dihydroxy-2- (3-hydroxy-4-methoxyphenyl) -4-benzopyran), an important citrus flavonoid and aglycone form of hesperidin, is a bitter compound mainly found in bitter oranges and lemons [ 90 ]. It is interesting to note that hesperetin has a higher bioavailability compared to hesperidin due to the rutinoside moiety attached to the flavonoid, and this seems to contribute to its superior anti-inflammatory and antioxidant properties [ 91 ]. Hesperetin is widely studied in several pathological conditions and exhibits neuroprotective effects [ 92 ], anticancer properties [ 93 ], anti-neuroinflammatory potential [ 94 ], antioxidant effects [ 95 ], and anti-inflammatory [ 96 ] activities, among others.…”