Immature citrus with peel was enzymatically treated for production of a hydrolysate with enriched bioactive components and higher antioxidant activity. The effects of reaction factors, including enzyme concentration, reaction time, and temperature on hesperetin and naringenin contents, total phenolic content (TPC), and antioxidant activity were investigated using response surface methodology. The models were adequate, and the enzyme concentration, temperature, and time positively affected hesperetin and naringenin contents and TPC, but negatively affected DPPH radical scavenging capacity. The reaction conditions for maximizing hesperetin, naringenin, and total phenol production and ferric reducing antioxidant power were optimized with the combination of enzyme concentration at 4%, 51 °C and 18 h. The hydrolysate at the optimized conditions contained higher hesperetin and naringenin contents and TPC compared with those before hydrolysis, by 251.7-, 45.5-, and 2.6-fold, respectively. This hydrolysate can be utilized in the production of functional beverages with high added values.
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Ahn SC, Kim MS, Lee SY, Kang JH, Kim BH, Oh WK, Ahn JS. Increase of bioactive flavonoid aglycone extractable from Korean citrus peel by carbohydrate-hydrolysing enzymes. Microbiology and Biotechnology Letters. 33: 288-295 (2005)
Assefa AD, Saini RK, Keum YS. Extraction of antioxidants and flavonoids from yuzu (Citrus junos Sieb ex Tanaka) peels: a response surface methodology study. Journal of Food Measurement and Characterization. 11: 364-379 (2017)
Galaverna G, Dall'Asta C. Production processes of orange juice and effects on antioxidant components. In: Processing and Impact on Antioxidants in Beverages, Preedy V (ed), Academic Press, Cambridge, MA, USA. pp. 203-214 (2014)
Gil-Chávez G, Villa JA, Ayala-Zavala JF, Heredia JB, Sepulveda D, Yahia EM, González-Aguilar GA. Technologies for extraction and production of bioactive compounds to be used as nutraceuticals and food ingredients: an overview. Comprehensive Reviews in Food Science and Food Safety. 12: 5-23 (2013)
Hollman PC, Bijsman MN, van Gameren Y, Cnossen EP, de Vries JH, Katan MB. The sugar moiety is a major determinant of the absorption of dietary flavonoid glycosides in man. Free Radical Research. 31: 569-573 (1999)
Huynh NT, Smagghe G, Gonzales GB, Camp JV, Raes K. Enzyme-assisted extraction enhancing the phenolic release from cauliflower (Brassica oleracea L. var. botrytis) outer leaves. Journal of Agricultural and Food Chemistry. 62: 7468-7476 (2014)
Hyun JM, Park KJ, Kim SS, Park SM. Lee YJ, An HJ. Antioxidant and anti-inflammatory effects of solvent fractions from the peel of the native Jeju citrus ‘Hongkyool’ and ‘Pyunkyool’. Journal of Life Science. 25: 1132-1138 (2015)
Iglesias-Carres L, Mas-Capdevila A, Bravo FI, Aragonav G, Muguerza B, Arola-Arnal A. Optimization of a polyphenol extraction method for sweet orange pulp (Citrus sinensis L.) to identify phenolic compounds consumed from sweet oranges. PLoS One. 14: e0211267 (2019)
Kawaii S, Ikuina T, Hikima T, Tokiwano T, Yoshizawa Y. Relationship between structure and antiproliferative activity of polymethoxyflavones towards HL60 cells. Anticancer Research. 32: 5239-5244 (2012)
Lee JH, Cho BK. Effects of red grape, wild grape and black raspberry wines on ground pork during refrigerated storage. Acta Alimentaria. 43: 553-563 (2014)
Lee MH, Huh D, Jo DJ, Lee GD, Yoon SR. Flavonoids components and functional properties of citrus peel hydrolysate. Journal of the Korean Society of Food Science and Nutrition. 36: 1358-1364 (2007)
Ma YQ, Chen JC, Liu DH, Ye XQ. Effect of ultrasonic treatment on the total phenolic and antioxidant activity of extracts from citrus peel. Journal of Food Science. 73: T115-T120 (2008)
Montero-Calderon A, Cortes C, Zulueta A, Foglia A, Esteve MJ. Green solvents and ultrasound-assisted extraction of bioactive orange (Citrus sinensis) peel compounds. Scientific Reports. 9: 16120 (2019)
Nishad J, Saha S, Kaur C. Enzyme‐and ultrasound‐assisted extractions of polyphenols from Citrus sinensis (cv. Malta) peel: A comparative study. Journal of Food Processing and Preservation. 43: e14046 (2019)
Pinelo M, Arnous A, Meyer AS. Upgrading of grape skins: Significance of plant cell-wall structural components and extraction techniques for phenol release. Trends in Food Science & Technology. 17: 579-590 (2006)
Ribeiro DS, Henrique SMB, Oliveira LS, Macedo GA, Fleuri LF. Enzymes in juice processing: a review. International Journal of Food Science & Technology. 45: 635-641 (2010)
Sharma S, Sandhu DK, Bagga PS. Physical characterization of isozymes of endo-β,1,4-glucanase and β-1,4-glucosidase from Aspergillus species. FEMS Microbiology Letters. 79: 99-104 (1991)
Shin KS, Chang HJ, Lee JH. Effect of commercial carbohydrases on the hesperetin and narigenin contents of citrus fruits. Korean Journal of Food Preservation. 27: 446-456 (2020)
Song EY, Choi YH, Kang KH, Koh JS. Free sugar, organic acid, hesperidin, naringenin and inorganic elements changes of Cheju citrus fruits according to harvest date. Korean Journal of Food Science and Technology. 30: 306-312 (1998)
Trinh TK, Kang LS. Application of response surface method as an experimental design to optimize coagulation tests. Environmental Engineering Research. 15: 63-70 (2010)
Tripoli E, Guardia ML, Giammanco S, Majo DD, Giammanco M. Citrus flavonoids: Molecular structure, biological activity and nutritional properties: A review. Food Chemistry. 104: 466-479 (2007)
Yang YC, Li J, Zu YG, Fu YJ, Luo M, Wu N, Liu XL. Optimisation of microwave-assisted enzymatic extraction of corilagin and geraniin from Geranium sibiricum Linne and evaluation of antioxidant activity. Food Chemistry. 122: 373-380 (2010)
Yao X, Zhu X, Pan S, Fang Y, Jiang F, Phillips GO, Xu X. Antimicrobial activity of nobiletin and tangeretin against Pseudomonas. Food Chemistry. 132: 1883-1890 (2012)
This work (Grants No S2602087) was supported by project for Cooperative R&D between Industry, Academy, and Research Institute funded Korea Ministry of SMEs and Startups in 2018.
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Shin, KS., Lee, JH. Optimization of enzymatic hydrolysis of immature citrus (Citrus unshiu Marcov.) for flavonoid content and antioxidant activity using a response surface methodology. Food Sci Biotechnol 30, 663–673 (2021). https://doi.org/10.1007/s10068-021-00897-w
- Antioxidant activity
- Enzymatic hydrolysis
- Immature citrus
- Response surface methodology