Regulation and Characterization of Amylase Enzyme Secretion In the Digestive Tract of Antheraea assamensis Helfer
Keywords:Antheraea assamensis, Pebrine Disease, Starvation, Feeding, Temperature, pH, Ca ion, Larval Stage, Amylase Activity, Zymography, SDS PAGE, Enzyme Purification.
Antheraea assamensis Helfer is an endemic non-mulberry lepidopteran species of North East India with high commercial demand for its golden hued silk. Antheraea assamensis suffers from a protozoan disease pebrine which lead to death of the silkworm. In this study, the effect of various factors (starvation, feeding, temperature, pH, Ca ion and larval stage) in the regulation of amylase secretion is compared in healthy and diseased muga silkworm. Moreover, enzyme zymography and purification of amylase from the midgut is also performed. Result shows that, the secretion of amylase in Antheraea assamensis in starved condition, significantly decreases. Refeeding experiments (after 2 days of starvation) suggest that amylase secretion sharply increases due to feeding stimulus. The purified enzyme molecular weight is confirmed as 56 kDa by SDS PAGE. The purification fold of purified enzyme is 34 times higher than the crude enzyme extract.
How to Cite
Abd El-latif A O. 2020. Partial characterization of the digestive proteases and α-amylase of the larvae of the red palm weevil, Rhynchophorus ferrugineus. Arthropods 9(1): 7-14.
Abraham E G, Nagaraju J, Datta R K. 1992. Biochemical studies of amylases in the silkworm, Bombyx mori L.: Comparative analysis in diapausing and nondiapausing strains. Insect Biochemistry and Molecular Biology 228: 867-873.
Bernfeld P. 1955. Amylases, α andβ. In: Colowick S P, Kaplan N. (Eds.), Methods in enzymology, Vol. I. New York, Academic Press, pp. 149-158.
Bhide A J, Channale S M, Patil S S, Gupta V S, Ramasamy S, Giri A P. 2015. Biochemical, structural and functional diversity between two digestive α-amylases from Helicoverpa armigera. Biochimica etBiophysica Acta - General Subjects 1850: 1719-1728.
Blakemore D, Williams S, Lehane M J. 1995. Protein stimulation of trypsin secretion from the opaque zone midgut cells of Stomoxyes calcitrans. Comparative Biochemistry and Physiology Part B: Biochemisrty and Molecular Biology 110: 301-307.
Chapman R F. 1985. Coordination of digestion. In: Kerkut G A, Gilbert L I (Eds.), Comprehensive insect physiology, biochemistry and pharmacology, Vol. 5. UK: Pergamon Press. pp. 213-240.
Cristofoletti P T, Ribeiro A F, Terra W R. 2001. Apocrine secretion of amylase and exocytosis of trypsin along the midgut of Tenebrio molitor larvae. Journal of Insect Physiology 47: 143-155.
Da Lage J L. 2018. The amylases of insects.International Journal of Insect Science10: 1179543318804783. dos Santos D D G, e Silva R R, de Barros C M, Nepomuceno-Silva J L, Nunes-da-Fonseca R, da Silva J R. 2022. Regeneration of the midgut during larval stages of the red flour beetle Tribolium castaneum(Herbst, 1797) upon fasting. Journal of Stored Products Research 98: 102003.
El-Didamony G, Ismaiel A A, El-Sheikh A, El-Gendy R, Fekry D. 2022. Digestive enzymes of Greater wax moth (Galleria mellonella) (Lepidoptera: Pyrilidae) through some strains of endophytic fungi infection. Journal of Pharmaceutical Negative Results 13(8): 2614-2619.
Ferreira C, Terra W R. 1989. Spatial organization of digestion, secretory mechanisms and digestive enzyme properties in Pheropsophus aequinoctialis (Coleoptera:Carabidae). Insect Biochemistry 19: 383-391.
Franco O L, Rigden D J, Melo F R, Bloch Jr C, Silva C P, Grossi-de-Sa M F. 2000. Activity of wheat a-amylase inhibitors towards bruchid a-amylases and structural explanation of observed specificities. European Journal of Biochemistry 267: 2166-2173.
García-Carreño F L, Dimes L E, Haard N F. 1993. Substrate gel electrophoresis for composition and molecular weight of proteinases and proteinaceous proteinase inhibitors. Analytical Biochemistry 214: 65-69.
Ganga G. 2003. Comprehensive sericulture, silkworm rearing and silk reeling, Oxford and IBH Publishing house co. pvt. ltd., New Delhi, 2 Hemmati S A, Shishehbor P, Stelinski L L. 2022. Life table parameters and digestive enzyme activity of Spodoptera littoralis(Boisd) (Lepidoptera: Noctuidae) on selected legume cultivars. Insects 13(7): 661.
Hori K. 1970. Some properties of amylase in thesalivary gland of Lygusdisponsi (Hemiptera). Journal of Insect Physiology 16: 373-386.
Kaur R, Kaur N, Gupta A K. 2014. Structural features, substrate specificity, kinetic properties of insect α-amylase and specificity of plant α-amylase inhibitors. Pesticide Biochemistry and Physiology 116: 83-93.
Kotkar M H, Sarate J P, Tamhane A V, Gupta S V, Giri P A. 2009. Responses of midgut amylases of Helicoverpa armigerato feeding on various host plants. Journal of Insect Physiology 55: 663-670.
Laemmli U K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685.
Lehane M J, Blakemore D, Williams S, Moffatt M R. 1995. Regulation of digestive enzyme levels in insects. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 110: 285-289.
Lowry O, Rosebrough N, Farr A L, Randall R. 1951. Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193: 265-275.
Ma R J, Wang C Y, Liu Y W, Sivakumar T R, Ren Z X, Fang Y, Jia J Q, Gui Z Z. 2014. Identification and characterization of a novel endoglucanase (CMCase) isolated from the larval gut of Bombyx mori. Journal of AsiaPacific Entomology 17: 67-71.
Madhusudhan K N, Manila Moorthy S, Gupta V P, Sinha A K, Sivaprasad V. 2018. Studies on biochemical changes during tropical tasar silkworm-pebrine interaction. International Journal of Advanced Research 6: 635-647.
Malone A L, Gatehouse S H. (née Edmonds) 1998. Effects of Nosema apis Infection on Honey Bee (Apis mellifera) Digestive Proteolytic Enzyme Activity. Journal of Invertebrate Pathology 71: 169-174.
Mikani A, Wang Q S, Takeda M. 2011. Brain-midgut short neuropeptide F mechanism that inhibits digestive activity of the American cockroach, Periplaneta Americana upon starvation. Peptides 34: 135-144.
Mohamed N T, Mohamed M S, El-Ebiarie A S, Gamal N. 2019. Biochemical Characterization of Some Digestive Enzymes in the Midgut of Eristalis megacephala(Diptera: Syrphidae). Egyptian Academic Journal of Biological Sciences. C, Physiology and Molecular Biology 11(3): 77-95.
Nadaf H L, Sirsat A K, Hivrale V K. 2022. Biochemical characterization of α-amylases from differently feeding pests: sap-sucking Aphis craccivoraand tissue chewing Pectinophora gossypiella. International Journal of Tropical Insect Science 42(6): 3747-3757.
Nagaraju J, Abraham E. G. 1995. Purification and characterization of digestiveamylase from the tasar silkworm, Antheraea mylitta (Lepidoptera: Saturniidae). Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 110: 201-209.
Nakonieczny M, Michalczyk K, Kedziorski A. 2006. Midgut glycosidases activities inmonophagous larvae of Apollo butterfly. Parnassius apollossp. Franken bergeri. Comptes Rendus Biologies 329: 765-774.
Nath R, Haloi K, Talukdar B, Devi D. 2013. Comparative study on tensile properties of different colour morphs and wild counterpart of muga silkworm (Antheraea assamensisHelfer) of North Eastern India. International Journal of Research in Biological Sciences 3: 141-144.
Rath S S, Prasad B C, Sinha B R R P. 2003. Food utilization efficiency in fifth instar larvae of Antheraea mylitta(Lepidoptera: Saturniidae) infected with Nosemasp. and its effect on reproductive potential and silk production. Journal of Invertebrate Pathology 83: 1-9.
Rahimi V, Bandani A R. 2014. Comparison of the effects of cereal and legume proteinaceous seed extracts on α-amylase activity and development of the Sunn pest. Journal of Asia Pacific Entomology 17(1): 7-11.
Sakai T, Satake H, Takeda M. 2006. Nutrient-induced α-amylase and protease activity is regulated by crustacean cardioactive peptide (CCAP) in the cockroach midgut. Peptides 27: 2157–2164.
Sarkar P, Sikdar S R. 2020. Characterization of α-amylase and its activity from mustard aphid Lipaphis erysimi (Kalt.)(Hemiptera) and its inhibition by Rorippa indica defensin. International Journal of Pest Management 66: 99-105.
Schumaker T T S, Cristofoletti P T, Terra W R. 1993. Properties and compartmentalization of digestivecarbohydrases and proteases in Scaptotrigonabi punctata (Apidae: Meliponinae) larvae. Apidologie 24: 3-17.
Sharifloo A, Zibaee A, Sendi J J, Jahroum K T. 2016. Characterization of a Digestive a-Amylase in the Midgut of Pieris brassicae L. (Lepidoptera: Pieridae). Frontiers in Physiology 7: 96.
Soto-Robles L V, López M F, Torres-Banda V, Cano-Ramírez C, Obregón-Molina G, Zúñiga G. 2021. The Bark Beetle Dendroctonusrhizophagus (Curculionidae: Scolytinae) Has Digestive Capacity to Degrade Complex Substrates: Functional Characterization and Heterologous Expression of an α-Amylase. International Journal of Molecular Sciences 22(1): 36.
Syakuro M Y A, Haryadi N, Muhlison W, Sucipto I. 2022. Isolation and Characterization of α-Amylase Enzyme on Brown Planthopper (Nilaparvata lugens Stal) On Rice. The Journal of Experimental Life Science 12(3): 81-87.
Teo L H,Woodring J P. 1985. Digestive enzymes in the house cricket Acheta domesticus with special reference to amylase. Comparative Biochemistry and Physiology Part A: Physiology 82: 871-877.
Terra W R, Espinoza-Fuentes E P, Ferreira C. 1988. Midgut amylase, lysozyme, aminopeptidase,and trehalase from larvae and adults of Muscadomestica. Archives of Insect Biochemistry and Physiology 9: 283-297.
Tikader A, Rajan R K. 2012. Utilization of muga host plants for cocoon crop improvement. In Proceedings: National Seminar on Recent Trends in Research and Development in Muga culture – Ideas to action, Guwahati, India. pp. 68-71.
Tikader A, Vijayan K, Saratchandra B. 2013. Improvement of host plants of Muga silkworm (Antheraea assamensis) for higher productivity and better adaptation-A Review. Plant Knowledge Journal 2: 83-88.
Vale V F, Moreira B H, Moraes C S, Pereira M H, Genta F A, Gontijo N F. 2012. Carbohydrate digestion in Lutzomyia longipalpis’ larvae (Diptera–Psychodidae). Journal of Insect Physiology 58: 1314-1324.
Weidlich S, Müller S, Hoffmann K. H, Woodring J. 2013. Regulation of amylase, cellulase and chitinase secretion in the digestive tract of the two-spotted field cricket, Gryllus bimaculatus. Archives of Insect Biochemistry and Physiology 83: 69-85.
Willis J D, Klingeman W E, Oppert C, Oppert B S, Jurat-Fuentes J L. 2010. Characterization of cellulolytic activity from digestive fluids of Dissosteira carolina(Orthoptera: Acrididae). Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 157: 267-272.
Woodring J, Diersch S, Lwalaba D, Hoffmann K H, Meyering-Vos M. 2009. Control of the release of digestive enzymes in the caeca of the cricket G. bimaculatus. Physiological Entomology 34: 144-151.
Yan H, Liu Q, Wen F, Bai B, Wen Y, Chen W, Lu W, Lin Y, Xia Q, Wang G. 2021. Characterization and potential application of an α-amylase (BmAmy1) selected during silkworm domestication. International Journal of Biological Macromolecules 167: 1102-1112.
Yan H, Feng W, Haiying X, Yuchan W, Deli S, Anyang L, Yicheng N, Qingyou X, Genhong W. 2022. Biochemical characterization and overexpression of an α-amylase (BmAmy) in silkworm, Bombyx mori. Insect Molecular Biology 31(2): 251-259.
Zadeh V M, Pourabad R F, Ashouri S, Hosseini N. 2020. Impact of four diets on pupal and larval total protein and digestive α-amylase activity in Ephestia kuehniella(Lepidoptera: Pyralidae) Journal of Crop Protection 9 (4): 615-624.
Zeng W H, Liu R X, Li Z Q, Liu B R, Li Q J, Xiao W L, Chen L Q, Zhong J H. 2012. Comparative lignocellulase activity and distribution among selected termite (Isoptera) genera. Journal of Entomological Science 47: 01-109.
Zibaee A, Hoda H, Fazeli-Dinan M. 2012. Purification and biochemical properties of a salivary α-amylase in Andrallus spinidens Fabricius (Hemiptera: Pentatomidae). Invertebrate Survival Journal 9: 48-57.