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Biochemical characterization a digestive trypsin in the midgut of large cabbage white butterfly, Pieris brassicae L. (Lepidoptera: Pieridae)

Published online by Cambridge University Press:  07 November 2017

A. Sharifloo ,
A. Zibaee ,
J. Jalali Sendi  and
K. Talebi Jahroumi
A. Sharifloo
Affiliation:
Department of Plant Protection, Faculty of Agricultural Sciences, University of Guilan, Rasht 416351314, Iran
A. Zibaee*
Affiliation:
Department of Plant Protection, Faculty of Agricultural Sciences, University of Guilan, Rasht 416351314, Iran
J. Jalali Sendi
Affiliation:
Department of Plant Protection, Faculty of Agricultural Sciences, University of Guilan, Rasht 416351314, Iran
K. Talebi Jahroumi
Affiliation:
Department of Plant Protection, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
*
*Author for correspondence Phone:+98-013-33690485 Fax:+98-013-33690281 E-mail: [email protected], [email protected]
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Abstract

A comprehensive study on digestive trypsin was undertaken in the larval midgut of Pieris brassicae L. Results of enzymatic compartmentalization showed a significantly higher activity of crude trypsin in the anterior larval midgut rather than posterior-midgut. Using Diethylaminoethyl cellulose fast flow column chromatography a purified trypsin was obtained by specific activity of 21 U mg−1 protein, recovery of 22%, purification fold of 28-fold and molecular weight of 25 kDa. This purified enzyme showed the highest activity at pH 8 and the corresponding temperature of 40°C. However, the specific inhibitors used including 4-(2-Aminoethyl) benzenesulfonyl fluroride hydrochloride, N-p-Tosyl-L-lysine methyl ester hydrochloride and Soybean Trypsin Inhibitor significantly lowered the activity of the purified enzyme in vitro. Moreover, the activity of trypsin and likewise the nutritional indices were significantly altered in the larval midgut feeding upon the leaves treated by 1 mM concentration of each inhibitor in comparison with control. Determination of enzymatic characteristics of insect trypsins is crucial in paving the path for controlling pests by potential natural compounds via transgenic plants.

Keywords

Digestive trypsincharacterizationenzymatic inhibitionPieris brassicae
Type
Research Papers
Information
Bulletin of Entomological Research , Volume 108 , Issue 4 , August 2018 , pp. 501 - 509
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Copyright
Copyright © Cambridge University Press 2017 

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References

Ahmad, Z., Saleemuddin, M. & Siddiqui, M. (1980) Purification and characterisation of three alkaline proteases from the larvae of army worm Spodoptera litura. Insect Biochemistry 10, 667–673.Google Scholar
Anwar, A. & Saleemudin, M. (2002) Purification and characterization of a digestive alkaline protease from the larvae of Spilosoma oblique. Archives of Insect Biochemistry and Physiology 51, 1–12.Google Scholar
Bernardi, R., Tedeschi, G., Ronchi, S. & Palmieri, S. (1996) Isolation and some molecular properties of a trypsin-like enzyme from larvae of European corn borer Ostrinia nubilalis Hubner (Lepidoptera: Pyralidae). Insect Biochemistry and Molecular Biology 26, 883–889.CrossRefGoogle Scholar
Buller, A.R. & Townsend, C.A. (2013) Intrinsic evolutionary constraints on protease structure, enzyme acylation, and the identity of the catalytic triad. Proceedings of the National Academy of Sciences of the United States of America (PNAS) 110, 653–661.Google ScholarPubMed
de Oliveira, C.F.T., Luz, L.A., Paiv, P.M.G., Coelho, L.C.B.B., Marangoni, S. & Macedo, M.L.R. (2011) Evaluation of seed coagulant Moringa oleifera lectin (cMoL) as a bioinsecticida l tool with potential for the control of insects. Process Biochemistry 46, 498–504.CrossRefGoogle Scholar
Frugoni, J.A.C. (1957) Tampone universal di Brittona Robinson a forzaionica costante. Gazzetta chimica Italiana 87, 403–407.Google Scholar
Graham, J.S. & Ryan, C.A. (1997) Accumulation of metallocarboxy-peptidase inhibitor in leaves of wounded potato plants. Biochemical and Biophysics Research Communication 101, 1164–1170.Google Scholar
Grover, S., Kaur, S., Gupta, A.K., Kumar, G. & Kaur, T.J. (2016) Characterization of Trypsin Like Protease from Helicoverpa armigera (Hubner) and its potential inhibitors. Proceedings of the National Academy of Sciences, India Section B: Biological Sciences 1–8, doi:10.1007/s40011-016-0732-0.Google Scholar
Hedstrom, L. (2002) Serine protease mechanism and specificity. Chemical Reviews 102, 4501–4523.Google Scholar
Kanost, M.R. & Clem, R.J. (2012) Insect proteases. pp. 346–364. in Gilbert, L. (Ed.) Insect Molecular Biology and Biochemistry. New York, Elsevier.Google Scholar
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of bacteriophage T4. Nature 227, 680–685.Google Scholar
Lawrence, P., Nirmala, K., Jayaveeramuthu, A. & Koundal, K.R. (2001) Nucleotide sequence of a genomic clone encoding a cowpea (Vigna unguiculata L.) trypsin inhibitor. Electronic Journal of Biotechnology 4, 1. doi: 10.2225/vol4-issue1-fulltext-4.Google Scholar
Lazarevic, J. & Jankovic-Tomanic, M. (2015) Dietary and phylogenetic correlates of digestive trypsin activity in insect pests. Entomologia Experimentalis et Applicata 157, 123–151.CrossRefGoogle Scholar
Lee, S.I., Lee, S.H., Koo, J.C., Chaun, H.J., Lim, C.O., Mun, J.H., Song, Y.H. & Cho, M.J. (1999) Soybean Kunitz trypsin inhibitor (SKTI) confers resistance to the brown planthopper (Nilaparvata lugens Stal) in transgenic rice. Molecular Breeding 5, 1–9.Google Scholar
Lopes, A.R., Juliano, M.A., Marana, S.R., Juliano, L. & Terra, W.R. (2006) Substrate specificity of insect trypsins and the role of their subsites in catalysis. Insect Biochemistry and Molecular Biology 36, 130–140.CrossRefGoogle ScholarPubMed
Lowry, O.H., Rosebrough, N.J., Farr, A.L. & Randall, R.J. (1951) Protein measurement with the Folin phenol reagent. Journal of Biology and Chemistry 193, 265–275.CrossRefGoogle ScholarPubMed
Marchetti, S., Chiaba, C., Chiesa, F., Bandiera, A. & Pitotti, A. (1998) Isolation and partial characterization of two trypsins from the larval midgut of Spodoptera littoralis (Boisduval). Insect Biochemistry and Molecular Biology 28, 449–458.Google Scholar
Novillo, C., Castanera, P. & Ortego, F. (1999) Isolation and characterization of two digestive trypsin-like proteinases from larvae of the stalk corn borer, Sesamia nonagrioides. Insect Biochemistry and Molecular Biology 29, 177–184.Google Scholar
Oliveira, M.G.A., Simone, S.G., Xavier, L.P. & Geudes, R.N.C. (2005) Partial purification and characterization of digestive trypsinlike proteases from the velvet bean caterpillar, Anticarsia gemmatalis. Comparative Biochemistry and Physiology Part B 140, 369–380.Google Scholar
Ranjbar, M., Zibaee, A. & Sendi, J.J. (2014) A trypsin-like proteinase in the midgut of Ectomyeloise ceratoniae Zeller purification, characterization and host plant inhibitors. Archives of Insect Biochemistry and Molecular Biology 85, 1–12.Google Scholar
Scriber, J.M. & Slansky, F. (1981) The nutritional ecology of immature insects. Annual Review of Entomology 26, 183–211.CrossRefGoogle Scholar
Sharifloo, A., Zibaee, A., Sendi, J.J. & Talebi Jahroumi, K.H. (2016) Characterization of a Digestive α-Amylase in the Midgut of Pieris brassicae L. (Lepidoptera: Pieridae). Frontiers in Physiology 7, 96.Google Scholar
Shaw, E., Mares-Guia, M. & Cohen, W. (1965) Evidence for an active center histidine in trypsin through use of a specific reagent 1-chloro-3-tosylamido-7-amino-2-heptanona, the chloromethyl ketone derived from N-α-tosyl-l-lysine. Biochemistry 4, 2219–2224.Google Scholar
Stryer, L. (2000) Biochemistry. 5th edn. New York City: WH Freeman & Co.Google Scholar
Telang, M.A., Giri, A.P., Sainani, M.N. & Gupta, V.S. (2005) Elastase like proteinase of Helicoverpa armigera is responsible for inactivation of a proteinase inhibitor from chickpea. Journal of Insect Physiology 51, 513–522.Google Scholar
Terra, W.R. & Ferriera, C. (2012) Biochemistry of digestion. pp. 365–418. in Gilbert, L.I. (Ed.) Insect Molecular Biology and Biochemistry. Oxford, UK: Elsevier.Google Scholar
Terra, W.R., Ferreira, C., Jordao, B.P. & Dillon, R.J. (1996) Digestive enzymes. pp. 171–224. in Lehance, M.J., & Billingsley, P.F. (Eds) Biology of the Insect Midgut. London: Chapman and Hall.Google Scholar
Walsmann, P., Richter, M. & Markwardt, F. (1972) Inactivation of trypsin and thrombin by 4-amidinobenzolsulfofluoride and 4-(2-aminoethyl)-benzolsulfofluoride. Acta Biologia Medico Germanica 28, 577–585.Google Scholar
Zibaee, A. (2012) A digestive lipase of Pieris brassicae L. (Lepidoptera: Pieridae): Purification, Characterization and host plants effects. Archive of Insect Biochemistery and Physiology 81, 1–19.CrossRefGoogle ScholarPubMed
Zibaee, A., Bandani, A.R., Fazeli-Dinan, M., Zibaee, I., Sendi, J.J. & Maleki, F.A. (2011) A trypsin-like protease in rice green semi-looper, Naranga aenescens Moore (Lepidoptera: Noctuidae): purification and characterization. Archives of Insect Biochemistry and Physiology 78, 1–16.Google Scholar