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Detection and substrate portrayal on the serum phenoloxidase activity from the grub of rhinoceros beetle, Oryctes rhinoceros

Published online by Cambridge University Press:  31 July 2023

Balashanmuga Nehru Marieshwari
Affiliation:
Department of Zoology, University of Madras, Guindy Campus, Chennai 600025, India
Chandran Prithi
Affiliation:
Department of Zoology, University of Madras, Guindy Campus, Chennai 600025, India
Ramanathan Nivetha
Affiliation:
Department of Zoology, University of Madras, Guindy Campus, Chennai 600025, India
Sreeramulu Bhuvaragavan
Affiliation:
Department of Zoology, University of Madras, Guindy Campus, Chennai 600025, India
Janarthanan Sundaram*
Affiliation:
Department of Zoology, University of Madras, Guindy Campus, Chennai 600025, India
*
Corresponding author: Janarthanan Sundaram; E-mail: janas_09@yahoo.co.in

Abstract

Phenoloxidase (PO) is a significant biomolecule involved in humoral defence mechanism of invertebrates. Spontaneous melanization of insect haemolymph is the major hinderance for studying PO activity, as haemolymph was collected devoid of phenylthiourea. In the study, no visible melanization was observed in crude serum from the grub of Oryctes rhinoceros up to 30 min of incubation amongst crude haemolymph, diluted haemolymph, crude serum and diluted serum that were subjected to visual observation for spontaneous melanization reaction. Accordingly, crude serum was taken for evaluating PO activity. At the same time, as PO substrates tend to auto-oxidize and provide false optical density value, tris-buffered saline devoid of any substrates were used as blank for PO assays. The ideal wavelength at which maximum PO activity occurred for each substrate, namely, tyrosine, tyramine, dopamine, L-dopa, DL-dopa, catechol, protocatechuic acid and pyrogallol was determined as 407, 410, 429, 465, 403, 466, 428 and 400 nm, respectively. Additionally, time course of oxidation for each phenolic substrate by the serum PO were examined and DL-dopa was identified as the specific substrate for serum PO in the grub of O. rhinoceros. Furthermore, maximum PO activity was observed at 5 min of incubation for 10 mM of DL-dopa that was considered as optimum concentration. The ideal pH and temperature for serum PO activity was observed as 7.5 and 20°C, respectively. These results suggested that standardizing a suitable substrate is an essential prerequisite to evaluate the real PO activity of serum which might significantly fluctuate in each insect model.

Type
Research Paper
Copyright
Copyright © The Author(s), 2023. Published by Cambridge University Press

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References

Ajamhassani, M, Sendi, JJ, Farsi, MJ and Zibaee, A (2012) Purification and characterization of phenoloxidase from the hemolymph of Hyphantria cunea (Lepidoptera: Arctiidae). Invertebrate Survival Journal 9, 6471.Google Scholar
Andersen, SO (2010) Insect cuticular sclerotization: a review. Insect Biochemistry and Molecular Biology 40, 166178.CrossRefGoogle ScholarPubMed
Arumugam, G, Sreeramulu, B, Paulchamy, R, Thangavel, S and Sundaram, J (2017) Purification and functional characterization of lectin with phenoloxidase activity from the hemolymph of cockroach, Periplaneta americana. Archives of Insect Biochemistry and Physiology 95, 2139021393.CrossRefGoogle ScholarPubMed
Asano, T and Ashida, M (2001) Cuticular pro-phenoloxidase of the silkworm, Bombyx mori: purification and demonstration of its transport from hemolymph. Journal of Biological Chemistry 276, 1110011112.CrossRefGoogle ScholarPubMed
Ashida, M (1971) Purification and characterization of pre-phenoloxidase from hemolymph of the silkworm Bombyx mori. Archives of Biochemistry and Biophysics 144, 749762.CrossRefGoogle ScholarPubMed
Ashida, M and Brey, PT (1995) Role of the integument in insect defense: pro-phenol oxidase cascade in the cuticular matrix. Proceedings of National Academy of Sciences, USA 92, 1069810702.CrossRefGoogle ScholarPubMed
Ashida, M and Brey, PT (1998) Recent advances on the research of the insect prophenoloxidase cascade. In Molecular Mechanisms of Immune Responses in Insects, pp. 135172.Google Scholar
Ashida, M and Yamazaki, H (1990) Biochemistry of the phenoloxidase system in insects: with special reference to its activation, in molting and metamorphosis Ohnishi, E and Ishizaki, H (eds), Molting and Metamorphosis pp. 239265.Google Scholar
Bae, S and Kim, Y (2004) Host physiological changes due to parasitism of a braconid wasp, Cotesia plutellae, on diamondback moth, Plutella xylostella. Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology 138, 3944.CrossRefGoogle ScholarPubMed
Baruah, GS, Sarma, HK, Bardoloi, S and Bora, D (2019) Purification and characterization of phenoloxidase from the hemolymph of healthy and diseased Antheraea assamensis Helfer (Lepidoptera: Saturniidae): effects of certain biological components and chemical agents on enzyme activity. Archives of Insect Biochemistry and Physiology 100, 2153121534.CrossRefGoogle ScholarPubMed
Beck, M, Theopold, U and Schmidt, O (2000) Evidence for serine protease inhibitor activity in the ovarian calyx fluid of the endoparasitoid Venturia canescens. Journal of Insect Physiology 46, 12751283.CrossRefGoogle ScholarPubMed
Benesova, J, Dobes, P and Hyrsl, P (2009) Developmental changes in phenol-oxidizing activity in the greater wax moth Galleria mellonella. Bulletin of Insectology 62, 237243.Google Scholar
Bhuvaragavan, S, Sruthi, K, Nivetha, R, Ramaraj, P, Hilda, K, Meenakumari, M and Janarthanan, S (2023) Insect galectin stimulates the human CD4+ T cell proliferation by regulating inflammation (T cell and monocyte) through Th2 immune response. Process Biochemistry 125, 1535.CrossRefGoogle Scholar
Bidla, G, Hauling, T, Dushay, MS and Theopold, U (2009) Activation of insect phenoloxidase after injury: endogenous versus foreign elicitors. Journal of Innate Immunity 1, 301308.CrossRefGoogle ScholarPubMed
Brunet, PCJ (1980) The metabolism of the aromatic amino acids concerned in the cross-linking of insect cuticle. Insect Biochemistry 10, 467500.CrossRefGoogle Scholar
Cerenius, L, Lee, BL and Soderhall, K (2008) The proPO system: pros and cons for its role in invertebrate immunity. Trends in Immunology 29, 263271.CrossRefGoogle ScholarPubMed
Chase, MR, Raina, K, Bruno, J and Sugumaran, M (2000) Purification, characterization and molecular cloning of prophenoloxidases from Sarcophaga bullata. Insect Biochemistry and Molecular Biology 30, 953967.CrossRefGoogle ScholarPubMed
Clark, KD (2015) Altered tyrosine metabolism and melanization complex formation underlie the developmental regulation of melanization in Manduca sexta. Insect Biochemistry and Molecular Biology 58, 6675.CrossRefGoogle ScholarPubMed
Da Silva, C, Dunphy, GB and Rau, ME (2000) Interaction of hemocytes and prophenoloxidase system of fifth instar nymphs of Acheta domesticus with bacteria. Developmental & Comparative Immunology 24, 367379.CrossRefGoogle ScholarPubMed
Feng, C, Song, Q, , W and Lu, J (2008) Purification and characterization of hemolymph prophenoloxidase from Ostrinia furnacalis (Lepidoptera: Pyralidae) larvae. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 151, 139146.CrossRefGoogle ScholarPubMed
Ganesh, A, Bhuvaragavan, S, Ramaraj, P and Janarthanan, S (2021) Grubs of rhinoceros beetle Oryctes rhinoceros with enormous potential for bioconversion of organic solid waste into value-added compost. Journal of Entomological Research 45, 179185.CrossRefGoogle Scholar
Gholami, T, Ghadamyari, M, Oliaee, AO and Ajamhasani, M (2013) Effects of inhibitors on haemolymph phenoloxidase from rosaceous branch borer, Ospheranteria coerulescens (Coleoptera: Cerambycidae). Journal of Plant Protection Research 53, 324332.CrossRefGoogle Scholar
González-Santoyo, I and Córdoba-Aguilar, A (2012) Phenoloxidase: a key component of the insect immune system. Entomologia Experimentalis et Applicata 142, 116.CrossRefGoogle Scholar
Goonewardena, HF (1958) The rhinoceros beetle (Oryctes rhinoceros Linn.) in Ceylon Introduction, distribution and life history. Tropical Agricultuist Ceylon 114, 3960.Google Scholar
Gressitt, JL (1953) The coconut rhinoceros beetle (Oryctes rhinoceros) with particular reference to the Palau Islands. The Coconut Rhinoceros Beetle (Oryctes rhinoceros) with particular Reference to the Palau Islands. Bulletins of Bishop Museum 212, 1157.Google Scholar
Halwani, AE, Niven, DF and Dunphy, GB (2000) Apolipophorin-III and the interactions of lipoteichoic acids with the immediate immune responses of Galleria mellonella. Journal of Invertebrate Pathology 76, 233241.CrossRefGoogle ScholarPubMed
Huger, AM (1966) A virus disease of the Indian rhinoceros beetle, Oryctes rhinoceros (Linnaeus), caused by a new type of insect virus, Rhabdion virus Oryctes gen. n., sp. n. Journal of Invertebrate Pathology 8, 3851.CrossRefGoogle Scholar
Huger, AM (2005) The Oryctes virus: its detection, identification, and implementation in biological control of the coconut palm rhinoceros beetle, Oryctes rhinoceros (Coleoptera: Scarabaeidae). Journal of Invertebrate Pathology 89, 7884.CrossRefGoogle Scholar
Hung, SY and Boucias, DG (1996) Phenoloxidase activity in hemolymph of naive and Beauveria bassiana-infected Spodoptera exigua larvae. Journal of Invertebrate Pathology 67, 3540.CrossRefGoogle Scholar
Jeyaraj, R, Thangaraj, T, Vasuki, CA and Aruchami, M (1986) Characteristics of phenoloxidases in larval cuticle of the coconut pest, Oryctes rhinoceros. Proceedings: Animal Sciences 95, 739744.Google Scholar
Kamalanathan, T, Rajeswari, IT and Sundaram, J (2020) Detection and characterization of phenoloxidase from the larval haemolymph of blow fly, Hemipyrellia tagaliana in response to non-self-molecules. Uttar Pradesh Journal of Zoology 41, 128142.Google Scholar
Kim, MS, Baek, MJ, Lee, MH, Park, JW, Lee, SY, Söderhäll, K and Lee, BL (2002) A new easter-type serine protease cleaves a masquerade-like protein during prophenoloxidase activation in Holotrichia diomphalia larvae. Journal of Biological Chemistry 277, 3999940004.CrossRefGoogle Scholar
Laughton, AM and Siva-Jothy, MT (2011) A standardised protocol for measuring phenoloxidase and prophenoloxidase in the honey bee, Apis mellifera. Apidologie 42, 140149.CrossRefGoogle Scholar
Lee, KM, Lee, KY, Choi, HW, Cho, MY, Kwon, TH, Kawabata, SI and Lee, BL (2000) Activated phenoloxidase from Tenebrio molitor larvae enhances the synthesis of melanin by using a vitellogenin-like protein in the presence of dopamine. European Journal of Biochemistry 267, 36953703.CrossRefGoogle ScholarPubMed
Liu, W, Xue, C, Zhang, J, Yu, J and Luo, W (2010) Inhibitory effect of tannic acid on growth, development and phenoloxidase activity of Spodoptera exigua larva. Journal of Plant Resources and Environment 19, 3237.Google Scholar
Longankumar, K, Thangaraj, T, Manimegalai, M, Aruchami, M and Vinayakam, A (1996) Latent larval cuticular phenoloxidase in the coconut pest, Oryctes rhinoceros. Archives of Insect Biochemistry and Physiology 33, 2738.3.0.CO;2-U>CrossRefGoogle Scholar
Lowry, OH (1951) Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193, 265275.CrossRefGoogle ScholarPubMed
Mullen, L and Goldsworthy, G (2003) Changes in lipophorins are related to the activation of phenoloxidase in the haemolymph of Locusta migratoria in response to injection of immunogens. Insect Biochemistry and Molecular Biology 33, 661670.CrossRefGoogle Scholar
Nappi, AJ, Frey, F and Carton, Y (2005) Drosophila serpin 27A is a likely target for immune suppression of the blood cell-mediated melanotic encapsulation response. Journal of Insect Physiology 51, 197205.CrossRefGoogle ScholarPubMed
Radha, S, Mullainadhan, P and Arumugam, M (2013) Detection of two distinct types of hemolymphatic prophenoloxidase and their differential responses in the black tiger shrimp, Penaeus monodon, upon infection by white spot syndrome virus. Aquaculture 376, 7684.CrossRefGoogle Scholar
Rafiei, B, Ghadamyari, M, Imani, S, Hosseininaveh, V and Ahadiyat, A (2018) Characterization and inhibition studies of hemolymph phenoloxidase from Dociostaurus maroccanus. Toxin Reviews 37, 4451.CrossRefGoogle Scholar
Ratcliffe, NA, Rowley, AF, Fitzgerald, SW and Rhodes, CP (1985) Invertebrate immunity: basic concepts and recent advances. International Review of Cytology 97, 183350.CrossRefGoogle Scholar
Ryazanova, AD, Alekseev, AA and Slepneva, IA (2012) The phenylthiourea is a competitive inhibitor of the enzymatic oxidation of DOPA by phenoloxidase. Journal of Enzyme Inhibition and Medicinal Chemistry 27, 7883.CrossRefGoogle ScholarPubMed
Sadawarte, AK, Moharil, MNM and Satpute, NS (2019) Phenoloxidase activity in hemolymph of naïve and HaNPV infected larvae of Helicoverpa armigera (HB), its characterization and inhibition. Journal of Pharmacognosy and Phytochemistry 8, 17561763.Google Scholar
Saul, SJ and Sugumaran, M (1987) Protease mediated prophenoloxidase activation in the hemolymph of the tobacco hornworm, Manduca sexta. Archives of Insect Biochemistry and Physiology 5, 111.CrossRefGoogle Scholar
Schmid-Hempel, P (2005) Evolutionary ecology of insect immune defenses. Annual Review of Entomology 50, 529.CrossRefGoogle ScholarPubMed
Schmid, MR, Brockmann, A, Pirk, CW, Stanley, DW and Tautz, J (2008) Adult honeybees (Apis mellifera L.) abandon hemocytic, but not phenoloxidase-based immunity. Journal of Insect Physiology 54, 439444.CrossRefGoogle Scholar
Sharifi, M, Ghadamyari, M, Sajedi, RH and Mahmoodi, NO (2015) Effects of 4-hexylresorcinol on the phenoloxidase from Hyphantria cunea (Lepidoptera: Arctiidae): in vivo and in vitro studies. Insect Science 22, 639650.CrossRefGoogle ScholarPubMed
Shelby, KS and Popham, HJ (2008) Cloning and characterization of the secreted hemocytic prophenoloxidases of Heliothis virescens. Archives of Insect Biochemistry and Physiology 69, 127142.CrossRefGoogle ScholarPubMed
Shelomi, M, Lin, SS and Liu, LY (2019) Transcriptome and microbiome of coconut rhinoceros beetle (Oryctes rhinoceros) larvae. BMC Genomics 20, 113.CrossRefGoogle ScholarPubMed
Strand, MR (2008) The insect cellular immune response. Insect Science 15, 114.CrossRefGoogle Scholar
Sugumaran, M (2002) Comparative biochemistry of eumelanogenesis and the protective roles of phenoloxidase and melanin in insects. Pigment Cell Research 15, 29.CrossRefGoogle ScholarPubMed
Sugumaran, M and Nellaiappan, K (1990) On the latency and nature of phenoloxidase present in the left colleterial gland of the cockroach Periplaneta americana. Archives of Insect Biochemistry and Physiology 15, 165181.CrossRefGoogle ScholarPubMed
Sugumaran, M and Nellaiappan, K (2000) Characterization of a new phenoloxidase inhibitor from the cuticle of Manduca sexta. Biochemical and Biophysical Research Communications 268, 379383.CrossRefGoogle ScholarPubMed
Sugumaran, M, Tan, S and Sun, HL (1996) Tyrosinase-catalyzed oxidation of 3,4-dihydroxyphenylglycine. Archives of Biochemistry and Biophysics 329, 175180.CrossRefGoogle ScholarPubMed
Sugumaran, M, Nellaiappan, K, Amaratunga, C, Cardinale, S and Scott, T (2000) Insect melanogenesis: III. Metabolon formation in the melanogenic pathway—regulation of phenoloxidase activity by endogenous dopachrome isomerase (decarboxylating) from Manduca sexta. Archives of Biochemistry and Biophysics 378, 39.CrossRefGoogle ScholarPubMed
Tsukamoto, T, Ishiguro, M and Funatsu, M (1986) Isolation of latent phenoloxidase from prepupae of the housefly, Musca domestica. Insect Biochemistry 16, 573581.CrossRefGoogle Scholar
Valadez-Lira, JA, Alcocer-Gonzalez, JM, Damas, G, Nunez-Mejia, G, Oppert, B, Rodriguez-Padilla, C and Tamez-Guerra, P (2012) Comparative evaluation of phenoloxidase activity in different larval stages of four lepidopteran pests after exposure to Bacillus thuringiensis. Journal of Insect Science 12, 80.CrossRefGoogle ScholarPubMed
Vavricka, CJ, Christensen, BM and Li, J (2010) Melanization in living organisms: a perspective of species evolution. Protein and Cell 1, 830841.CrossRefGoogle ScholarPubMed
Xue, CB, Luo, WC, Chen, QX, Wang, Q and Ke, LN (2006) Enzymatic properties of phenoloxidase from Pieris rapae (Lepidoptera) larvae. Insect Science 13, 251256.CrossRefGoogle Scholar
Yousefi-Lardeh, L and Zibaee, A (2020) Biochemical characterization of a hemolymph phenoloxidase and its endogenous inhibitor in the larvae of an invasive moth, Cydalima perspectalis Walker (Lepidoptera: Crambidae). Journal of Asia Pacific Entomology 23, 890900.CrossRefGoogle Scholar
Zdybicka-Barabas, A, Mak, P, Jakubowicz, T and Cytryńska, M (2014) Lysozyme and defense peptides as suppressors of phenoloxidase activity in Galleria mellonella. Archives of Insect Biochemistry and Physiology 87, 112.CrossRefGoogle ScholarPubMed
Zibaee, A, Bandani, AR and Malagoli, D (2011) Purification and characterization of phenoloxidase from the hemocytes of Eurygaster integriceps (Hemiptera: Scutelleridae). Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 158, 117123.CrossRefGoogle ScholarPubMed
Zufelato, MS, Lourenço, AP, Simões, ZL, Jorge, JA and Bitondi, MM (2004) Phenoloxidase activity in Apis mellifera honey bee pupae, and ecdysteroid-dependent expression of the prophenoloxidase mRNA. Insect Biochemistry and Molecular Biology 34, 12571268.CrossRefGoogle ScholarPubMed
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