The citrus pectin by-product (CPB), generated from pectin industry, is a rich-source of flavanones, but not explored until now. As most of these compounds are inside vacuoles or bound to cell wall matrix, enzymatic hydrolysis was applied on their recovery, followed by hydroalcoholic and ultrasound extraction. Different parameters were studied: enzymes (β-glucosidase, tannase, and cellulase), their concentration (5, 10, and 20 U g−1 CPB), and reaction time (6, 12, and 24 h). Extracts were characterized in total phenolic content (TPC), antioxidant capacity (ORAC and DPPH assays), and polyphenolic profile (HPLC–DAD). All enzymatic treatments significantly improved CPB antioxidant capacity and TPC, compared with hydroalcoholic and ultrasound extraction. β-glucosidase (5 U) for 24 h was the most effective in polyphenol extraction and bioconversion, followed by β-glucosidase (5 U) for 12 h and tannase (5 U) for 24 h. Thus, the concentration of these enzymes was increased (10 and 20 U) to improve flavanones extraction. β-glucosidase at 20 U offered the highest amount of naringenin (77.63 mg 100 g−1 of CPB) and hesperetin (766.44 mg 100 g−1) obtained so far by biological processes. According to Person’s correlation analysis, TPC and antioxidant activity were highly correlated with CPB contents of hesperetin and naringenin. The aglycone flavanones are rarely found in natural sources and have higher biological potential than their glycosylated forms. Our results indicated enzyme-assisted extraction as a good choice for recovering aglycone flavanones from CPB, and increased knowledge on the biological activity of this agroindustrial waste, amplifying their application in food and pharmaceutical field.
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Andrić P, Meyer AS, Jensen PA, Dam-Johansen K (2010) Reactor design for minimizing product inhibition during enzymatic lignocellulose hydrolysis: I. Significance and mechanism of cellobiose and glucose inhibition on cellulolytic enzymes. Biotechnol Adv 28:308–324. https://doi.org/10.1016/J.BIOTECHADV.2010.01.003
Barbosa PPM, Ruviaro AR, Macedo GA (2018) Comparison of different Brazilian citrus by-products as source of natural antioxidants. Food Sci Biotechnol 27:1301–1309. https://doi.org/10.1007/s10068-018-0383-4
Barreca D, Gattuso G, Bellocco E et al (2017) Flavanones: citrus phytochemical with health-promoting properties. BioFactors 43:495–506. https://doi.org/10.1002/biof.1363
Battestin V, Macedo GA (2007) Tannase production by Paecilomyces variotii. Bioresour Technol 98:1832–1837. https://doi.org/10.1016/J.BIORTECH.2006.06.031
Bondet V, Brand-Williams W, Berset C (1997) Kinetics and mechanisms of antioxidant activity using the DPPH.Free radical method. LWT - Food Sci Technol 30:609–615. https://doi.org/10.1006/FSTL.1997.0240
Caridi D, Trenerry VC, Rochfort S et al (2007) Profiling and quantifying quercetin glucosides in onion (Allium cepa L.) varieties using capillary zone electrophoresis and high performance liquid chromatography. Food Chem 105:691–699. https://doi.org/10.1016/J.FOODCHEM.2006.12.063
Cerda A, Martínez ME, Soto C et al (2013) The enhancement of antioxidant compounds extracted from Thymus vulgaris using enzymes and the effect of extracting solvent. Food Chem 139:138–143. https://doi.org/10.1016/j.foodchem.2012.12.044
Chamorro S, Viveros A, Alvarez I et al (2012) Changes in polyphenol and polysaccharide content of grape seed extract and grape pomace after enzymatic treatment. Food Chem 133:308–314. https://doi.org/10.1016/j.foodchem.2012.01.031
Delgado CHO, Fleuri LF (2016) Orange and mango by-products: agro-industrial waste as source of bioactive compounds and botanical versus commercial description: a review. Food Rev Int 32:1–14. https://doi.org/10.1080/87559129.2015.1041183
Di Majo D, Giammanco M, La Guardia M et al (2005) Flavanones in citrus fruit: structure–antioxidant activity relationships. Food Res Int 38:1161–1166. https://doi.org/10.1016/j.foodres.2005.05.001
FAO (2018) Food and Agriculture Organization of the United Nations - FAOSTAT. In: Crop Prod. Data Base. http://www.fao.org/faostat/en/#data/QC. Accessed 20 Mar 2019
Ferreira LR, Macedo JA, Ribeiro ML, Macedo GA (2013) Improving the chemopreventive potential of orange juice by enzymatic biotransformation. Food Res Int 51:526–535. https://doi.org/10.1016/j.foodres.2013.01.018
Khan MK, Zill-E-Huma Dangles O (2014) A comprehensive review on flavanones, the major citrus polyphenols. J Food Compos Anal 33:85–104. https://doi.org/10.1016/j.jfca.2013.11.004
Khan MK, Abert-Vian M, Fabiano-Tixier AS et al (2010) Ultrasound-assisted extraction of polyphenols (flavanone glycosides) from orange (Citrus sinensis L.) peel. Food Chem 119:851–858. https://doi.org/10.1016/j.foodchem.2009.08.046
Lachos-Perez D, Baseggio AM, Mayanga-Torres PC et al (2018) Subcritical water extraction of flavanones from defatted orange peel. J Supercrit Fluids 138:7–16. https://doi.org/10.1016/j.supflu.2018.03.015
Li BB, Smith B, Hossain MM (2006) Extraction of phenolics from citrus peels: II. Enzyme-assisted extraction method. Sep Purif Technol 48:189–196. https://doi.org/10.1016/J.SEPPUR.2005.07.019
M’hiri N, Ioannou I, Ghoul M, Boudhrioua NM (2014) Extraction methods of citrus peel phenolic compounds. Food Rev Int 30:265–290. https://doi.org/10.1080/87559129.2014.924139
Madeira JV, Macedo GA (2015) Simultaneous extraction and biotransformation process to obtain high bioactivity phenolic compounds from Brazilian citrus residues. Biotechnol Prog 31:1273–1279. https://doi.org/10.1002/btpr.2126
Madeira JV, Teixeira CB, Macedo GA (2013) Biotransformation and bioconversion of phenolic compounds obtainment: an overview. Crit Rev Biotechnol 35:75–81. https://doi.org/10.3109/07388551.2013.803020
Madeira JV, Nakajima VM, Macedo JA, Macedo GA (2014) Rich bioactive phenolic extract production by microbial biotransformation of Brazilian Citrus residues. Chem Eng Res Des 92:1802–1810. https://doi.org/10.1016/J.CHERD.2014.07.014
Mamma D, Christakopoulos P (2014) Biotransformation of citrus by-products into value added products. Waste Biomass Valorization 5:529–549. https://doi.org/10.1007/s12649-013-9250-y
Martins IM, Roberto BS, Blumberg JB et al (2016) Enzymatic biotransformation of polyphenolics increases antioxidant activity of red and white grape pomace. Food Res Int 89:533–539. https://doi.org/10.1016/J.FOODRES.2016.09.009
Matsuura M, Sasaki J, Murao S (1995) Studies on β-glucosidases from soybeans that hydrolyze daidzin and genistin: isolation and characterization of an isozyme. Biosci Biotechnol Biochem 59:1623–1627. https://doi.org/10.1271/bbb.59.1623
Miller GL, Blum R, Glennon WE, Burton AL (1960) Measurement of carboxymethylcellulase activity. Anal Biochem 1:127–132. https://doi.org/10.1016/0003-2697(60)90004-X
Nadar SS, Rao P, Rathod VK (2018) Enzyme assisted extraction of biomolecules as an approach to novel extraction technology: a review. Food Res Int 108:309–330. https://doi.org/10.1016/J.FOODRES.2018.03.006
Nakajima VM, Madeira JV, Macedo GA, Macedo JA (2016) Biotransformation effects on anti lipogenic activity of citrus extracts. Food Chem 197:1046–1053. https://doi.org/10.1016/j.foodchem.2015.11.109
Panja P (2017) Green extraction methods of food polyphenols from vegetable materials. Curr Opin Food Sci. https://doi.org/10.1016/J.COFS.2017.11.012
Prior RL, Hoang H, Gu L et al (2003) Assays for hydrophilic and lipophilic antioxidant capacity (oxygen radical absorbance capacity (ORACFL)) of plasma and other biological and food samples. J Agric Food Chem 51:3273–3279. https://doi.org/10.1021/jf0262256
Rice-Evans CA, Miller NJ, Paganga G (1996) Structure-antioxidant activity relationships of flavonoids and phenolic acids. Free Radic Biol Med 20:933–956. https://doi.org/10.1016/0891-5849(95)02227-9
Río Segade S, Pace C, Torchio F et al (2015) Impact of maceration enzymes on skin softening and relationship with anthocyanin extraction in wine grapes with different anthocyanin profiles. Food Res Int 71:50–57. https://doi.org/10.1016/j.foodres.2015.02.012
Ruviaro AR, de Barbosa PPM, Macedo GA (2018) Enzyme-assisted biotransformation increases hesperetin content in citrus juice by-products. Food Res Int. https://doi.org/10.1016/j.foodres.2018.05.004
Shahidi F, Yeo J (2016) Insoluble-bound phenolics in food. Molecules 21:1216. https://doi.org/10.3390/molecules21091216
Sharma S, Bhat TK, Dawra RK (2000) A spectrophotometric method for assay of tannase using rhodanine. Anal Biochem 279:85–89. https://doi.org/10.1006/ABIO.1999.4405
Shin K-C, Nam H-K, Oh D-K (2013) Hydrolysis of flavanone glycosides by β-glucosidase from pyrococcus furiosus and its application to the production of flavanone aglycones from citrus extracts. J Agric Food Chem 61:11532–11540. https://doi.org/10.1021/jf403332e
Singleton VL, Rossi JA (1965) Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. Am J Enol Vitic 16:144 LP–158
Sun Y, Qiao L, Shen Y et al (2013) Phytochemical profile and antioxidant activity of physiological drop of citrus fruits. J Food Sci 78:C37–C42. https://doi.org/10.1111/j.1750-3841.2012.03002.x
Timell TE (1964) The acid hydrolysis of glycosides: I. General conditions and the effect of the nature of the aglycone. Can J Chem 42:1456–1472. https://doi.org/10.1139/v64-221
Wang A-Y, Zhou M-Y, Lin W-C (2011) Antioxidative and anti-inflammatory properties of Citrus sulcata extracts. Food Chem 124:958–963. https://doi.org/10.1016/j.foodchem.2010.07.035
Winotapun W, Opanasopit P, Ngawhirunpat T, Rojanarata T (2013) One-enzyme catalyzed simultaneous plant cell disruption and conversion of released glycoside to aglycone combined with in situ product separation as green one-pot production of genipin from gardenia fruit. Enzyme Microb Technol 53:92–96. https://doi.org/10.1016/J.ENZMICTEC.2013.05.001
We thank FAPESP (Grant Number 2015/04555-2) for the financial support and CNPq for the scholarships.
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Barbosa, P.d.P.M., Ruviaro, A.R. & Macedo, G.A. Conditions of enzyme-assisted extraction to increase the recovery of flavanone aglycones from pectin waste. J Food Sci Technol 58, 4303–4312 (2021). https://doi.org/10.1007/s13197-020-04906-4
- Enzyme-assisted extraction
- Citrus by-product
- Bioactive compounds
- Antioxidant activity