Inhibitory activity of protein hydrolysates from rice bran on mushroom tyrosinase


  • Thaniya Wunnakup Drug and Herbal Product Research and Development Center, College of Pharmacy, Rangsit University, Patumthani 12000, Thailand
  • Chaowalit Monton Medicinal Cannabis Research Institute, College of Pharmacy, Rangsit University, Patumthani 12000, Thailand
  • Laksana Charoenchai Drug and Herbal Product Research and Development Center, College of Pharmacy, Rangsit University, Patumthani 12000, Thailand


inhibition, kinetic, mushroom tyrosinase, protein hydrolysates, rice bran protein


Rice bran protein hydrolysates (RBPHs) obtained by Alcalase® have attracted attention because of their bioactivity properties (antioxidant, anti-acetylcholinesterase and anti-butyrylcholinesterase).  However, there has been no report on their improved inhibition of melanogenesis.  This study aimed to investigate the inhibitory tyrosinase activity of RBPH.  The defatted Khao Hom Mali RD15 rice bran protein was extracted into water (RBP1), 2% NaCl (RBP2) and 0.1 N NaOH (RBP3) fractions.  All protein fractions were hydrolyzed with Alcalase® to produced RBPH1, RBPH2 and RBPH3.  The protein content, % yield, degree of hydrolysis (DH) and molecular weight patterns of each fraction were investigated.  The RBPH2 and RBPH3 showed high potential inhibition on mushroom tyrosinase activity, the IC50 values were determined to be 1.92 mg/ml and 0.46 mg/ml, respectively.  The inhibition kinetics showed that RBPH2 was an uncompetitive mechanism, with inhibition constants (Ki and Kis) were 6.1 mg/ml and 4.5 mg/ml, respectively.  Moreover, the RBPH3 displayed a non-competitive mechanism and Ki and Kis were 2.8 mg/ml and 2.5 mg/ml, respectively.  Therefore, the RBPHs could be potential candidates for use in the cosmetics and food industries.


Adler-Nissen, J. (1986). Enzymic hydrolysis of food proteins. Elsevier applied science publishers.

Aguilar-Toalá, J. E., & Liceaga, A. M. (2020). Identification of chia seed (Salvia hispanica L.) peptides with enzyme inhibition activity towards skin-aging enzymes. Amino Acids, 52(8), 1149-1159. DOI:

Burnett, C., Bergfeld, W. F., Belsito, D. V., Hill, R. A., Klaassen, C. D., Liebler, D. C., ... & Heldreth, B. (2018). Safety assessment of hydrolyzed wheat protein and hydrolyzed wheat gluten as used in Cosmetics. International journal of toxicology, 37(1_suppl), 55S-66S. DOI:

Boonloh, K., Kukongviriyapan, V., Kongyingyoes, B., Kukongviriyapan, U., Thawornchinsombut, S., & Pannangpetch, P. (2015). Rice bran protein hydrolysates improve insulin resistance and decrease pro-inflammatory cytokine gene expression in rats fed a high carbohydrate-high fat diet. Nutrients, 7(8), 6313-6329. DOI:

Braspaiboon, S., Osiriphun, S., Peepathum, P., & Jirarattanarangsri, W. (2020). Comparison of the effectiveness of alkaline and enzymatic extraction and the solubility of proteins extracted from carbohydrate-digested rice. Heliyon, 6(11), e05403. DOI:

Brunelle, E., Le, A. M., Huynh, C., Wingfield, K., Halámková, L., Agudelo, J., & Halámek, J. (2017). Coomassie brilliant blue G-250 dye: an application for forensic fingerprint analysis. Analytical chemistry, 89(7), 4314-4319. DOI:

Chai, W. O.-Y., Ou-Yang, C., Huang, Q., Lin, M. Z., Wang, Y. X., Xu, K. L., Huang, W. Y., & Pang, D. D. (2018). Antityrosinase and antioxidant properties of mung bean seed proanthocyanidins: Novel insights into the inhibitory mechanism. Food chemistry, 260, 27-36. DOI:

Chanput, W., Theerakulkait, C., & Nakai, S. (2009). Antioxidative properties of partially purified barley hordein, rice bran protein fractions and their hydrolysates. Journal of Cereal Science, 49(3), 422-428.

Dhalleine, C., & Delepierre, S. (2015). Method for preparing alkaline hydrolysates of plant proteins. (U.S. Patent No. 9149063B2). U.S. Patent and Trademark Office. DOI:

Deng, Y., Huang, L., Zhang, C., Xie, P., Cheng, J., Wang, X., & Liu, L. (2020). Skin-care functions of peptides prepared from Chinese quince seed protein: Sequences analysis, tyrosinase inhibition and molecular docking study. Industrial Crops and Products, 148, 112331. DOI:

Dogra, S., & Sarangal, R. (2014). Pigmentary disorders: An insight. Pigment International, 1(1), 5. DOI: 10.4103/2349-5847.135429.

Doucet, D., Otter, D. E., Gauthier, S. F., & Foegeding, E. A. (2003). Enzyme-induced gelation of extensively hydrolyzed whey proteins by Alcalase: peptide identification and determination of enzyme specificity. Journal of Agricultural and Food Chemistry, 51(21), 6300-6308. DOI:

Dupont, C., Bocquet, A., Tomé, D., Bernard, M., Campeotto, F., Dumond, P., ... & Kalach, N. (2020). Hydrolyzed Rice Protein-Based Formulas, a Vegetal Alternative in Cow’s Milk Allergy. Nutrients, 12(9), 2654. DOI:

Fabian, C., & Ju, Y. H. (2011). A review on rice bran protein: its properties and extraction methods. Critical reviews in food science and nutrition, 51(9), 816-827. DOI:

Feng, B., Ma, L., Yao, J., Fang, Y., Mei, Y., & Wei, S. (2013). Protective effect of oat bran extracts on human dermal fibroblast injury induced by hydrogen peroxide. Journal of Zheijang University SCIENCE B, 14, 97-105. DOI:

Ferri, M., Graen-Heedfeld, J., Bretz, K., Guillon, F., Michelini, E., Calabretta, M. M., ... & Tassoni, A. (2017). Peptide fractions obtained from rice by-products by means of an environment-friendly process show in vitro health-related bioactivities. PloS one, 12(1), e0170954. DOI:

Gou, L. L. (2017). The effect of oxaloacetic acid on tyrosinase activity and structure: Integration of inhibition kinetics with docking simulation. International journal of biological macromolecules, 260, 59-66. DOI:

Guan, J., Takai, R., Toraya, K., Ogawa, T., Muramoto, K., Mohri, S., ... & Cho, S. J. (2017). Effects of alkaline deamidation on the chemical properties of rice bran protein. Food Science and Technology Research, 23(5), 697-704. DOI:

Ishikawa, S., Sasaki, Y. F., Kawaguchi, S., Mochizuki, M., & Nagao, M. (2006). Characterization of genotoxicity of kojic acid by mutagenicity in Salmonella and micronucleus induction in rodent liver. Genes and Environment, 28(1), 31-37. DOI:

Jan-On, G., Sangartit, W., Pakdeechote, P., Kukongviriyapan, V., Senaphan, K., Boonla, O., ... & Kukongviriyapan, U. (2020). Antihypertensive effect and safety evaluation of rice bran hydrolysates from Sang-Yod rice. Plant Foods for Human Nutrition, 75(1), 89-95. DOI:

Jiamyangyuen, S., Srijesdaruk, V., & Harper, W. J. (2005). Extraction of rice bran protein concentrate and its application in bread. Extraction, 27(1), 55-64.

Kim, J. K., Park, K. T., Lee, H. S., Kim, M., & Lim, Y. H. (2012). Evaluation of the inhibition of mushroom tyrosinase and cellular tyrosinase activities of oxyresveratrol: comparison with mulberroside A. Journal of enzyme inhibition and medicinal chemistry, 27(4), 495-503. DOI:

Kooyers, T. J., & Westerhof, W. (2006). Toxicology and health risks of hydroquinone in skin lightening formulations. Journal of the European academy of Dermatology and Venereology, 20(7), 777-780. DOI:

Kubglomsong, S., Theerakulkait, C., Reed, R. L., Yang, L., Maier, C. S., & Stevens, J. F. (2018). Isolation and Identification of Tyrosinase-Inhibitory and Copper-Chelating Peptides from Hydrolyzed Rice-Bran-Derived Albumin. Journal of agricultural and food chemistry, 66(31), 8346-8354. DOI:

Kumagai, T., Kawamura, H., Fuse, T., Watanabe, T., Saito, Y., Masumura, T., ... & Kadowaki, M. (2006). Production of rice protein by alkaline extraction improves its digestibility. Journal of nutritional science and vitaminology, 52(6), 467-472. DOI:

Logarušić, M., Slivac, I., Radošević, K., Bagović, M., Redovniković, I. R., & Srček, V. G. (2019). Hempseed protein hydrolysates’ effects on the proliferation and induced oxidative stress in normal and cancer cell lines. Molecular biology reports, 46(6), 6079-6085. DOI:

Muyan, C., Xiumei, Z., Tianxiang, G., & Chao, C. (2006). Effects of temperature, pH and NaCl on protease activity in digestive tract of young turbot, Scophthalmus maximus. Chinese Journal of Oceanology and Limnology, 24(3), 300-306. DOI:

Ota, Y., Imai, T., Onose, J. I., Takami, S., Cho, Y. M., Hirose, M., & Nishikawa, A. (2009). A 55-week chronic toxicity study of dietary administered kojic acid (KA) in male F344 rats. The Journal of toxicological sciences, 34(3), 305-313. DOI:

Phantuwong, N., Thongraung, C., & Yupanqui, C. T. (2017). Enzymatic hydrolysis on protein and β-glucan content of Sang-yod rice bran hydrolysates and their anti-inflammatory activity on RAW 264.7 cells. Functional Foods in Health and Disease, 7(12), 958-971. DOI: 10.31989/ffhd.v7i12.388.

Phongthai, S., Homthawornchoo, W., & Rawdkuen, S. (2017). Preparation, properties and application of rice bran protein: A review. International Food Research Journal, 24(1), 25-34.

Prakash, J., & Ramaswamy, H. S. (1996). Rice bran proteins: properties and food uses. Critical Reviews in Food Science & Nutrition, 36(6), 537-552. DOI:

Saran, A. S. (2004). Loss of tyrosinase activity confers increased skin tumor susceptibility in mice. Oncogene, 23(23), 4130-4135. DOI:

Sanguigno, L., Casamassa, A., Funel, N., Minale, M., Riccio, R., Riccio, S., Boscia, F., Brancaccio, P., Pollina, L.E., Anzilotti, S., & Di Renzo, G. (2018). Triticum vulgare extract exerts an anti- inflammatory action in two in vitro models of inflammation in microglial cells. PLoS One, 13(6), e0197493. DOI:

Su, N. W., Wang, M. L., Kwok, K. F., & Lee, M. H. (2005). Effects of temperature and sodium chloride concentration on the activities of proteases and amylases in soy sauce koji. Journal of agricultural and food chemistry, 53(5), 1521-1525. DOI:

Thamnarathip, P., Jangchud, K., Jangchud, A., Nitisinprasert, S., Tadakittisarn, S., & Vardhanabhuti, B. (2016). Extraction and characterisation of Riceberry bran protein hydrolysate using enzymatic hydrolysis. International Journal of Food Science & Technology, 51(1), 194-202. DOI:

Uraipong, C., & Zhao, J. (2016). Rice bran protein hydrolysates exhibit strong in vitro α‐amylase, β‐glucosidase and ACE‐inhibition activities. Journal of the Science of Food and Agriculture, 96(4), 1101-1110. DOI:

Vandenplas, Y., De Greef, E., Hauser, B., & Paradice Study Group. (2014). Safety and tolerance of a new extensively hydrolyzed rice protein-based formula in the management of infants with cow’s milk protein allergy. European journal of pediatrics, 173(9), 1209-1216. DOI: 10.1007/s00431-014-2308-4.

Zhao, Q., Xiong, H., Selomulya, C., Chen, X.D., Zhong, H., Wang, S., Sun, W., & Zhou, Q. (2012). Enzymatic hydrolysis of rice dreg protein: effects of enzyme type on the functional properties and antioxidant activities of recovered proteins. Food chemistry, 134(3), 1360-1367. DOI:




How to Cite

Thaniya Wunnakup, Chaowalit Monton, & Laksana Charoenchai. (2023). Inhibitory activity of protein hydrolysates from rice bran on mushroom tyrosinase. Journal of Current Science and Technology, 11(2), 324–333. Retrieved from



Research Article