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Food Science and Human Wellness 2023, 12(5): 1820-1827 https://doi.org/10.1016/j.fshw.2023.02.046
Research Article | Open Access | Issue | Published: 21 March 2023

Structural and antioxidative properties of royal jelly protein by partial enzymatic hydrolysis

Show Author's Information Shanshan Lia,  Lingchen Taoa,  Shiqin Penga,  Xinyu Yua,  Xiaobin Mab( ),  Fuliang Hua( )
Key laboratory of silkworm and bee resource utilization and innovation of Zhejiang Province, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China
Teagasc Food Research Centre, Moorepark, Fermoy, Co. Cork P61 C996, Ireland

Peer review under responsibility of KeAi Communications Co., Ltd.

Keywords:
Structure, Enzymatic hydrolysis, Antioxidative activity, Royal jelly protein, Acalase
Cite this article:
Li S, Tao L, Peng S, et al. Structural and antioxidative properties of royal jelly protein by partial enzymatic hydrolysis. Food Science and Human Wellness, 2023, 12(5): 1820-1827. https://doi.org/10.1016/j.fshw.2023.02.046
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Abstract Full text About this article

The objective of this study was to investigate the structural and antioxidative properties of royal jelly protein (RJP) at different degrees of hydrolysis (DH) by partial enzymatic hydrolysis. RJP was hydrolyzed by alcalase for 0 min, 15 min, 1 h, 5 h and 8 h to obtain hydrolysates at DH of 5.34%, 11.65%, 15.19%, 21.38% and 23.91%, respectively. With the increased DH, the RJP hydrolysates showed elevated antioxidative activities. The molecular weight of RJP hydrolysates was significantly decreased but their primary backbone kept unchanged. Analysis of circular dichroism spectra revealed that the enzymolysis reduced the content of α-helix but increased the contents of β-sheet, β-turn and random coil. Meanwhile, the surface hydrophobicity and fluorescence intensity of RJP hydrolysates were decreased and a red shift occurred. As the enzymolysis continued, the surface morphology of RJP was gradually changed from a sheet-like structure into microparticles. Changes in antioxidative activities and structures generally followed a DH-dependent manner, however these changes became insignificant for samples at DH beyond 20%. Taking into consideration of both effectiveness and productivity, the optimum enzymatic duration was determined at 5 h.


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About this article

Structural and antioxidative properties of royal jelly protein by partial enzymatic hydrolysis

Show Author's information Shanshan Lia, Lingchen Taoa, Shiqin Penga, Xinyu Yua, Xiaobin Mab( ), Fuliang Hua( )
Key laboratory of silkworm and bee resource utilization and innovation of Zhejiang Province, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China
Teagasc Food Research Centre, Moorepark, Fermoy, Co. Cork P61 C996, Ireland

Peer review under responsibility of KeAi Communications Co., Ltd.

Abstract

The objective of this study was to investigate the structural and antioxidative properties of royal jelly protein (RJP) at different degrees of hydrolysis (DH) by partial enzymatic hydrolysis. RJP was hydrolyzed by alcalase for 0 min, 15 min, 1 h, 5 h and 8 h to obtain hydrolysates at DH of 5.34%, 11.65%, 15.19%, 21.38% and 23.91%, respectively. With the increased DH, the RJP hydrolysates showed elevated antioxidative activities. The molecular weight of RJP hydrolysates was significantly decreased but their primary backbone kept unchanged. Analysis of circular dichroism spectra revealed that the enzymolysis reduced the content of α-helix but increased the contents of β-sheet, β-turn and random coil. Meanwhile, the surface hydrophobicity and fluorescence intensity of RJP hydrolysates were decreased and a red shift occurred. As the enzymolysis continued, the surface morphology of RJP was gradually changed from a sheet-like structure into microparticles. Changes in antioxidative activities and structures generally followed a DH-dependent manner, however these changes became insignificant for samples at DH beyond 20%. Taking into consideration of both effectiveness and productivity, the optimum enzymatic duration was determined at 5 h.

Keywords: Structure, Enzymatic hydrolysis, Antioxidative activity, Royal jelly protein, Acalase

References(54)

[1]

A. Balan, M.A. Moga, L. Dima, et al., Royal jelly-a traditional and natural remedy for postmenopausal symptoms and aging-related pathologies, Molecules 25(14) (2020) 3291. https://doi.org/10.3390/molecules25143291.
DOI Google Scholar
[2]

S. Ahmad, M.G. Campos, F. Fratini, et al., New insights into the biological and pharmaceutical properties of royal jelly, Int. J. Mol. Sci. 21(2) (2020) 382. https://doi.org/10.3390/ijms21020382.
DOI Google Scholar
[3]

A.N.K.G. Ramanathan, A.J. Nair, V.S. Sugunan, A review on royal jelly proteins and peptides, J. Funct. Foods 44 (2018) 255-264. https://doi.org/10.1016/j.jff.2018.03.008.
DOI Google Scholar
[4]

M. Kamakura, Royalactin induces queen differentiation in honeybees, Nature 473(7348) (2011) 478-483. https://doi.org/10.1038/nature10093.
DOI Google Scholar
[5]

P. Fan, B. Han, M. Feng, et al., Functional and proteomic investigations reveal major royal jelly protein 1 associated with anti-hypertension activity in mouse vascular smooth muscle cells, Sci. Rep. 6 (2016) 30230. https://doi.org/10.1038/srep30230.
DOI Google Scholar
[6]

M. Feng, Y. Fang, C. Ma, et al., Mechanistic insight into royal protein inhibiting the gram-positive bacteria, Biomolecules 11(1) (2021) 64. https://doi.org/10.3390/biom11010064.
DOI Google Scholar
[7]

K. Kohno, I. Okamoto, O. Sano, et al., Royal jelly inhibits the production of proinflammatory cytokines by activated macrophages, Biosci. Biotech. Bioch. 68(1) (2004) 138-145. https://doi.org/10.1271/bbb.68.138.
DOI Google Scholar
[8]

I. Okamoto, Y. Taniguchi, T. Kunikata, et al., Major royal jelly protein 3 modulates immune responses in vitro and in vivo, Life Sci. 73(16) (2003) 2029-2045. https://doi.org/10.1016/s0024-3205(03)00562-9.
DOI Google Scholar
[9]

S. Takaki-Doi, K. Hashimoto, M. Yamamura, et al., Antihypertensive activities of royal jelly protein hydrolysate and its fractions in spontaneously hypertensive rats, Acta. Med. Okayama. 63(1) (2009) 57-64. https://doi.org/10.18926/amo/31859.
DOI Google Scholar
[10]

A. Sultana, A.H. Nabi, K.B. Biswas, et al., A peptide YY inhibits the human renin activity in a pH dependent manner, Front. Biosci. (Landmark Ed) 14 (2009) 3286-3291. https://doi.org/10.2741/3451.
DOI Google Scholar
[11]

H. Guo, Y. Kouzuma, M. Yonekura, Isolation and properties of antioxidative peptides from water-soluble royal jelly protein hydrolysate, Food. Sci. Technol. Res. 11(2) (2005) 222-230. https://doi.org/10.3136/fstr.11.222.
DOI Google Scholar
[12]

H. Guo, A. Ekusa, K. Iwai, et al., Royal jelly peptides inhibit lipid peroxidation in vitro and in vivo, J. Nutr. Sci. Vitaminol. (Tokyo) 54(3) (2008) 191-195. https://doi.org/10.3177/jnsv.54.191.
DOI Google Scholar
[13]

H. Guo, Y. Kouzuma, M. Yonekura, Structures and properties of antioxidative peptides derived from royal jelly protein, Food. Chem. 113(1) (2009) 238-245. https://doi.org/10.1016/j.foodchem.2008.06.081.
DOI Google Scholar
[14]

T. Nagai, R. Inoue, N. Suzuki, et al., Antioxidant properties of enzymatic hydrolysates from royal jelly, J. Med. Food. 9(3) (2006) 363-367. https://doi.org/10.1089/jmf.2006.9.363.
DOI Google Scholar
[15]

M. Barati, F. Javanmardi, S.M.H.M. Jazayeri, et al., Techniques, perspectives, and challenges of bioactive peptide generation: a comprehensive systematic review, Compr. Rev. Food. Sci. Food. Saf. 19(4) (2020) 1488-1520. https://doi.org/10.1111/1541-4337.12578.
DOI Google Scholar
[16]

A. Sultana, A.H.M.N. Nabi, U.M. Nasir, et al., A dipeptide YY derived from royal jelly proteins inhibits renin activity, Int. J. Mol. Med. 21(6) (2008) 677-681. https://doi.org/10.3892/ijmm.21.6.677.
DOI Google Scholar
[17]

N. Ahmadifard, J.H. Murueta, A. Abedian-Kenari, et al., Comparison the effect of three commercial enzymes for enzymatic hydrolysis of two substrates (rice bran protein concentrate and soy-been protein) with SDS-PAGE, J. Food. Sci. Technol. 53(2) (2016) 1279-1284. https://doi.org/10.1007/s13197-015-2087-6.
DOI Google Scholar
[18]

V.G. Tacias-Pascacio, R. Morellon-Sterling, E.H. Siar, et al., Use of alcalase in the production of bioactive peptides: a review, Int. J. Biol. Macromol. 165 (2020) 2143-2196. https://doi.org/10.1016/j.ijbiomac.2020.10.060.
DOI Google Scholar
[19]

W. Chansuwan, M. Khamhae, Z. Yang, et al., Hydrolase-treated royal jelly attenuates LPS-induced inflammation and IgE-antigen-mediated allergic reaction, Funct. Foods Health D. 10(3) (2020) 127-142. https://doi.org/10.31989/ffhd.v10i3.694.
DOI Google Scholar
[20]

W. Tian, M. Li, H. Guo, et al., Architecture of the native major royal jelly protein 1 oligomer, Nat. Commun. 9(1) (2018) 3373. https://doi.org/10.1038/s41467-018-05619-1.
DOI Google Scholar
[21]

P.M. Nielsen, D. Petersen, C. Dambmann, Improved method for determining food protein degree of hydrolysis, J. Food Sci. 66(5) (2001) 642-646. https://doi.org/10.1111/j.1365-2621.2001.tb04614.x.
DOI Google Scholar
[22]

T.T. Bu, M.J. Zhou, J.X. Zheng, et al., Preparation and characterization of a low-phenylalanine whey hydrolysate using two-step enzymatic hydrolysis and macroporous resin adsorption, LWT-Food Sci. Technol. 132 (2020) 109753. https://doi.org/10.1016/j.lwt.2020.109753.
DOI Google Scholar
[23]

W.J. Chen, X.B. Ma, W.J. Wang, et al., Preparation of modified whey protein isolate with gum acacia by ultrasound maillard reaction, Food Hydrocoll. 95 (2019) 298-307. https://doi.org/10.1016/j.foodhyd.2018.10.030.
DOI Google Scholar
[24]

A. Kato, S. Nakai, Hydrophobicity determined by a fluorescence probe method and its correlation with surface properties of proteins, Biochim. Biophys. Acta. 624(1) (1980) 13-20. https://doi.org/10.1016/0005-2795(80)90220-2.
DOI Google Scholar
[25]

T.H. Yan, S.J. Lim, A.S. Babji, et al., Sarbini, enzymatic hydrolysis: sialylated mucin (SiaMuc) glycoprotein of edible swiftlet's nest (ESN) and its molecular weight distribution as bioactive ESN SiaMuc-glycopeptide hydrolysate, Int. J. Biol. Macromol. 175 (2021) 422-431. https://doi.org/10.1016/j.ijbiomac.2021.02.007.
DOI Google Scholar
[26]

N. Akbari, J.M. Milani, P. Biparva, Functional and conformational properties of proteolytic enzyme-modified potato protein isolate, J. Sci. Food Agr. 100(3) (2020) 1320-1327. https://doi.org/10.1002/jsfa.10148.
DOI Google Scholar
[27]

A.M. Ghribi, I.M. Gafsi, A. Sila, et al., Effects of enzymatic hydrolysis on conformational and functional properties of chickpea protein isolate, Food Chem. 187 (2015) 322-330. https://doi.org/10.1016/j.foodchem.2015.04.109.
DOI Google Scholar
[28]

P.R. Roberts, J.D. Burney, K.W. Black, et al., Effect of chain length on absorption of biologically active peptides from the gastrointestinal tract, Digestion 60(4) (1999) 332-337. https://doi.org/10.1159/000007679.
DOI Google Scholar
[29]

S.Y. Lee, S.J. Hur, Antihypertensive peptides from animal products, marine organisms, and plants, Food Chem. 228 (2017) 506-517. https://doi.org/10.1016/j.foodchem.2017.02.039.
DOI Google Scholar
[30]

J.J. Gorman, T.P. Wallis, J.J. Pitt, Protein disulfide bond determination by mass spectrometry, Mass. Spectrom. Rev. 21(3) (2002) 183-216. https://doi.org/10.1002/mas.10025.
DOI Google Scholar
[31]

J. Chen, X.Y. Chen, Q.J. Zhu, et al., Determination of the domain structure of the 7S and 11S globulins from soy proteins by XRD and FTIR, J. Sci. Food Agr. 93(7) (2013) 1687-1691. https://doi.org/10.1002/jsfa.5950.
DOI Google Scholar
[32]

R. Gnanasambandam, A. Proctor, Determination of pectin degree of esterification by diffuse reflectance Fourier transform infrared spectroscopy, Food Chem. 68(3) (2000) 327-332. https://doi.org/10.1016/S0308-8146(99)00191-0.
DOI Google Scholar
[33]

L. Wang, M. Xiao, S.H. Dai, et al., Interactions between carboxymethyl konjac glucomannan and soy protein isolate in blended films, Carbohyd. Polym. 101 (2014) 136-145. https://doi.org/10.1016/j.carbpol.2013.09.028.
DOI Google Scholar
[34]

X.B. Ma, W.J. Chen, T.Y. Yan, et al., Comparison of citrus pectin and apple pectin in conjugation with soy protein isolate (SPI) under controlled dry-heating conditions, Food Chem. 309 (2020). https://doi.org/10.1016/j.foodchem.2019.125501.
DOI Google Scholar
[35]

X. Ma, W. Wang, D. Wang, et al., Degradation kinetics and structural characteristics of pectin under simultaneous sonochemical-enzymatic functions, Carbohydr. Polym. 154 (2016) 176-185. https://doi.org/10.1016/j.carbpol.2016.08.010.
DOI Google Scholar
[36]

R.L.J. Almeida, T. Dos Santos Pereira, V. de Andrade Freire, et al., Influence of enzymatic hydrolysis on the properties of red rice starch, Int. J. Biol. Macromol. 141 (2019) 1210-1219. https://doi.org/10.1016/j.ijbiomac.2019.09.072.
DOI Google Scholar
[37]

M.Y. Zhang, A.M. Liao, K. Thakur, et al., Modification of wheat bran insoluble dietary fiber with carboxymethylation, complex enzymatic hydrolysis and ultrafine comminution, Food Chem. 297 (2019) 124983. https://doi.org/10.1016/j.foodchem.2019.124983.
DOI Google Scholar
[38]

N.J. Greenfield, Using circular dichroism spectra to estimate protein secondary structure, Nat. Protoc. 1(6) (2006) 2876-2890. https://doi.org/10.1038/nprot.2006.202.
DOI Google Scholar
[39]

G.C.N. Cruz, L. Garcia, A.J. Silva, et al., Calcium effect and pH-dependence on self-association and structural stability of the Apis mellifera major royal jelly protein 1, Apidologie 42(3) (2011) 252-269. https://doi.org/10.1007/s13592-011-0025-9.
DOI Google Scholar
[40]

J. Kocot, M. Kielczykowska, D. Luchowska-Kocot, et al., Antioxidant potential of propolis, bee pollen, and royal jelly: possible medical application, Oxid. Med. Cell. Longev. 2018 (2018) 7074209. https://doi.org/10.1155/2018/7074209.
DOI Google Scholar
[41]

C.T. Wen, J.X. Zhang, J. Zhou, et al., Effects of slit divergent ultrasound and enzymatic treatment on the structure and antioxidant activity of arrowhead protein, Ultrason. Sonochem. 49 (2018) 294-302. https://doi.org/10.1016/j.ultsonch.2018.08.018.
DOI Google Scholar
[42]

X.B. Ma, F.R. Hou, H.H. Zhao, et al., Conjugation of soy protein isolate (SPI) with pectin by ultrasound treatment, Food Hydrocoll. 108 (2020) 106056. https://doi.org/10.1016/j.foodhyd.2020.106056.
DOI Google Scholar
[43]

J. Jin, O.D. Okagu, A.E.A. Yagoub, et al., Effects of sonication on the in vitro digestibility and structural properties of buckwheat protein isolates, Ultrason. Sonochem. 70 (2021) 105348. https://doi.org/10.1016/j.ultsonch.2020.105348.
DOI Google Scholar
[44]

M.M. Ai, T. Tang, L.D. Zhou, et al., Effects of different proteases on the emulsifying capacity, rheological and structure characteristics of preserved egg white hydrolysates, Food Hydrocoll. 87 (2019) 933-942. https://doi.org/10.1016/j.foodhyd.2018.09.023.
DOI Google Scholar
[45]

P. Malumba, S. Janas, O. Roiseux, et al., Comparative study of the effect of drying temperatures and heat-moisture treatment on the physicochemical and functional properties of corn starch, Carbohyd. Polym. 79(3) (2010) 633-641. https://doi.org/10.1016/j.carbpol.2009.09.013.
DOI Google Scholar
[46]

H.L. Ma, L.R. Huang, J.Q. Jia, et al., Effect of energy-gathered ultrasound on alcalase, Ultrason. Sonochem. 18(1) (2011) 419-424. https://doi.org/10.1016/j.ultsonch.2010.07.014.
DOI Google Scholar
[47]

C.S. Zhou, H.L. Ma, X.J. Yu, et al., Pretreatment of defatted wheat germ proteins (by-products of flour mill industry) using ultrasonic horn and bath reactors: effect on structure and preparation of ACE-inhibitory peptides, Ultrason. Sonochem. 20(6) (2013) 1390-1400. https://doi.org/10.1016/j.ultsonch.2013.04.005.
DOI Google Scholar
[48]

P.B. Subhedar, P.R. Gogate, Enhancing the activity of cellulase enzyme using ultrasonic irradiations, J. Mol. Catal. B.-Enzym. 101 (2014) 108-114. https://doi.org/10.1016/j.molcatb.2014.01.002.
DOI Google Scholar
[49]

N. Alizadeh-Pasdar, E.C.Y. Li-Chan, Comparison of protein surface hydrophobicity measured at various pH values using three different fluorescent probes, J. Agr. Food Chem. 48(2) (2000) 328-334. https://doi.org/10.1021/jf990393p.
DOI Google Scholar
[50]

A.G.B. Wouters, I. Rombouts, E. Fierens, et al., Relevance of the functional properties of enzymatic plant protein hydrolysates in food systems, Compr. Rev. Food Sci. Food Saf. 15(4) (2016) 786-800. https://doi.org/10.1111/1541-4337.12209.
DOI Google Scholar
[51]

A. Nisov, D. Ercili-Cura, E. Nordlund, Limited hydrolysis of rice endosperm protein for improved techno-functional properties, Food Chem. 302 (2020) 125274. https://doi.org/10.1016/j.foodchem.2019.125274.
DOI Google Scholar
[52]

D.E. Goldsack, R.C. Chalifoux, Contribution of the free energy of mixing of hydrophobic side chains to the stability of the tertiary structure of proteins, J. Theor. Biol. 39(3) (1973) 645-651. https://doi.org/10.1016/0022-5193(73)90075-1.
DOI Google Scholar
[53]

U. Uthumporn, I.S.M. Zaidul, A.A. Karim, Hydrolysis of granular starch at sub-gelatinization temperature using a mixture of amylolytic enzymes, Food Bioprod. Process. 88(C1) (2010) 47-54. https://doi.org/10.1016/j.fbp.2009.10.001.
DOI Google Scholar
[54]

K.A. Kristoffersen, N.K. Afseth, U. Bocker, et al., Average molecular weight, degree of hydrolysis and dry-film FTIR fingerprint of milk protein hydrolysates: intercorrelation and application in process monitoring, Food Chem. 310 (2020) 125800. https://doi.org/10.1016/j.foodchem.2019.125800.
DOI Google Scholar
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Received: 21 December 2021
Revised: 06 April 2022
Accepted: 13 June 2022
Published: 21 March 2023
Issue date: September 2023

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© 2023 Beijing Academy of Food Sciences.

Acknowledgements

This work was supported by the National Natural Science Foundation of China (31872431), the earmarked fund for the Modern Agroindustry Technology Research System from the Ministry of Agriculture of China (CARS-44).

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This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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