Recombinant expression of soluble gonad inhibiting hormone (GIH) from Penaeus monodon in a bacterial expression system

Reynold Peter mangottil; K. K. Vijayan; M. A. Pradeep; C. P. Balasubramanyan

doi:10.21077/ijf.2025.72.4.173307-10

Authors

Reynold Peter icar
K. K. Vijayan
M. A. Pradeep
C. P. Balasubramanyan

https://doi.org/10.21077/ijf.2025.72.4.173307-10

Keywords:

Penaeus monodon, Gonad inhibiting hormone, recombinant GIH protein, eyestalk ablation, Escherichia coli expression system

Abstract

Successful reproduction of captive shrimp has been the primary goal of commercial breeding program. Gonad inhibiting hormone (GIH), a member of CMG neuropeptide hormone family present within the neural tissue of the eyestalk of crustaceans, exert profound influence on reproductive maturation in shrimps. Present study reports the recombinant expression and purification of the GIH gene, using an Escherichia coli expression system. This is the first report of expressing the recombinant GIH of P. monodon using E. coli expression system in its native soluble form secreted to the media. The purified recombinant peptide is useful in structure and function analysis, which can be used for identifying its interacting partners and elucidating its mode of action. Production of an antibody/antagonist for the GIH would facilitate the development of an immunological tool for induced maturation in shrimp industry. This procedure could replace the physiologically destructive eyestalk ablation technique.

Keywords: Escherichia coli expression system, Eyestalk ablation, Immunological tool, Recombinant GIH protein

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References

Aguilar M B, Quackenbush L S, Hunt D T, Shabanowitz J and Huberman A. 1992. Identification, purification and initial characterization of the vitellogenesis-inhibiting hormone from the Mexican crayfish Procambarus bouvieri (Ortmann). Comparative Biochemistry and Physiology -- Part B: Biochemistry and 102(3): 491–498. DOI: https://doi.org/10.1016/0305-0491(92)90039-T

Anand P S S, Balasubramanian C P, Aravind R, Biju I F, Rajan R V, Vinay T N, Panigrahi A, Sudheer N S, Rajamanickam S, Kumar S, Antony J, Ambasankar K, Jithendran K P and Vijayan K K. 2023. Reproductive performance of captive-reared Indian white shrimp, Penaeus indicus, broodstocks over two generations. Frontiers in Marine Science 10. DOI: https://doi.org/10.3389/fmars.2023.1101806

Bhassu S, Shama M, Tiruvayipati S, Soo T C C, Ahmed N and Yusoff K. 2024. Microbes and pathogens associated with shrimps - implications and review of possible control strategies. Frontiers in Marine Science 1397708. DOI: https://doi.org/10.3389/fmars.2024.1397708

Choi J H and Lee S Y. 2004. Secretory and extracellular production of recombinant proteins using Escherichia coli. Applied Microbiology and Biotechnology 625–635. DOI: https://doi.org/10.1007/s00253-004-1559-9

Chung J S and Webster S G. 2003. Moult cycle-related changes in biological activity of moult-inhibiting hormone (MIH) and crustacean hyperglycaemic hormone (CHH) in the crab, Carcinus maenas: From target to transcript. European Journal of Biochemistry 270(15): 3280–3288. DOI: https://doi.org/10.1046/j.1432-1033.2003.03720.x

DeKleijn D P V, Janssen K P C, Waddy S L, Hegeman R, Lai W Y, Martens G J M and Van Herp F. 1998. Expression of the crustacean hyperglycaemic hormones and the gonad- inhibiting hormone during the reproductive cycle of the female American lobster Homarus americanus. Journal of Endocrinology 156(2): 291–298. DOI: https://doi.org/10.1677/joe.0.1560291

Edomi P, Azzoni E, Mettulio R, Pandolfelli N, Ferrero E A and Giulianini P G. 2002. Gonad-inhibiting hormone of the Norway lobster (Nephrops norvegicus): cDNA cloning, expression, recombinant protein production, and immunolocalization. Gene 284(1–2): 93–102. DOI: https://doi.org/10.1016/S0378-1119(02)00389-X

Esposito D and Chatterjee D K. 2006. Enhancement of soluble protein expression through the use of fusion tags. Current Opinion in Biotechnology 353–358. DOI: https://doi.org/10.1016/j.copbio.2006.06.003

Grève P, Sorokine O, Berges T, Lacombe C, Van Dorsselaer A and Martin G. 1999. Isolation and amino acid sequence of a peptide with vitellogenesis inhibiting activity from the terrestrial isopod Armadillidium vulgare (Crustacea). General and Comparative Endocrinology 115(3): 406–414. DOI: https://doi.org/10.1006/gcen.1999.7330

Hopkins P M. 2012. The eyes have it: A brief history of crustacean neuroendocrinology. General and Comparative Endocrinology 175(3): 357–366. DOI: https://doi.org/10.1016/j.ygcen.2011.12.002

Jeong K J and Lee S Y. 2001. Secretory production of human granulocyte colony-stimulating factor in Escherichia coli. Protein Expression and Purification 23(2): 311–318. DOI: https://doi.org/10.1006/prep.2001.1508

Katayama H, Ohira T, Aida K and Nagasawa H. 2002. Significance of a carboxyl-terminal amide moiety in the folding and biological activity of crustacean hyperglycemic hormone. Peptides 23(9): 1537–1546. DOI: https://doi.org/10.1016/S0196-9781(02)00094-3

Kotzsch A, Vernet E, Hammarström M, Berthelsen J, Weigelt J, Gräslund S and Sundström M. 2011. A secretory system for bacterial production of high-profile protein targets. Protein Science 20(3): 597–609. DOI: https://doi.org/10.1002/pro.593

Lorenzon S. 2005. Hyperglycemic stress response in Crustacea. Invertebrate Survival Journal 132–141.

Makrides S C. 1996. Strategies for achieving high-level expression of genes in Escherichia coli. Microbiological Reviews 60(3): 512–538. DOI: https://doi.org/10.1128/mr.60.3.512-538.1996

Mandal A and Singh P. 2025. Global scenario of shrimp industry: Present status and future prospects. Shrimp Culture Technology: Farming, Health Management and Quality Assurance, pp1–24. Springer Nature. DOI: https://doi.org/10.1007/978-981-97-8549-0_1

Mettulio R, Giulianini P G, Ferrero E A, Lorenzon S and Edomi P. 2004. Functional analysis of crustacean Hyperglycemic Hormone by in vivo assay with wild-type and mutant recombinant proteins. Regulatory Peptides 119(3): 189–197. DOI: https://doi.org/10.1016/j.regpep.2004.02.002

Morera Y, Lugo J M, Ramos L, Rodríguez-Ramos T, Huberman A and Estrada M P. 2013. A chimeric recombinant crustacean hyperglycemic hormone from Litopenaeus schmitti (Burkenroad) obtained as C-terminus fusion protein boost hemolymph glucose concentration in Litopenaeus vannamei (Boone). Aquaculture Research 44(7): 1066–1075. DOI: https://doi.org/10.1111/j.1365-2109.2012.03110.x

Mosco A, Zlatev V, Guarnaccia C, Pongor S, Campanella A, Zahariev S and Giulianini P G. 2012. Novel Protocol for the Chemical Synthesis of Crustacean Hyperglycemic Hormone Analogues — An Efficient Experimental Tool for Studying Their Functions. PLOS ONE 7(1): e30052. DOI: https://doi.org/10.1371/journal.pone.0030052

Moss S M, Moss D R, Arce S M, Lightner D V. and Lotz J M. 2012. The role of selective breeding and biosecurity in the prevention of disease in penaeid shrimp aquaculture. Journal of Invertebrate Pathology 247–250. DOI: https://doi.org/10.1016/j.jip.2012.01.013

Ohira T, Okumura T, Suzuki M, Yajima Y, Tsutsui N, Wilder M N and Nagasawa H. 2006. Production and characterization of recombinant vitellogenesis-inhibiting hormone from the American lobster Homarus americanus. Peptides 27(6): 1251–1258. DOI: https://doi.org/10.1016/j.peptides.2005.10.019

Ohira T, Tsutsui N, Nagasawa H and Wilder M N. 2006. Preparation of two recombinant crustacean hyperglycemic hormones from the giant freshwater prawn, Macrobrachium rosenbergii, and their hyperglycemic activities. Zoological Science 23(4): 383–391. DOI: https://doi.org/10.2108/zsj.23.383

Okumura T. 2004. Perspectives on hormonal manipulation of shrimp reproduction. Japan Agricultural Research Quarterly 49–54. DOI: https://doi.org/10.6090/jarq.38.49

Pasteur C. 1959. FISHERIES RESEARCH BOARD OF CANADA Translation Series No. 2798 PRELIMINARY NOTE ON THE ROLE OF THE X ORGAN IN THE CONTROL OF MOLTING IN THE SHRIMP PALAEMON (= LEANDER) SERRATUS (PENNANT) title: Note préliminaire sur le rôle de l’organe X dans le contrôle. Comptes Rendus des Séances Mensuelles de la Société des Sciences Naturelles et Physiques du Maroc 25: 123–124.

Schägger H. 2006. Tricine-SDS-PAGE. Nature Protocols 1(1): 16–22. DOI: https://doi.org/10.1038/nprot.2006.4

Soyez D, Van Deijnen J E and Martin M. 1987. Isolation and characterization of a vitellogenesis‐inhibiting factor from sinus glands of the lobster, Homarus americanus. Journal of Experimental Zoology 244(3): 479–484. DOI: https://doi.org/10.1002/jez.1402440314

Swartz J R. 2001. Advances in Escherichia coli production of therapeutic proteins. Current Opinion in Biotechnology 195–201. DOI: https://doi.org/10.1016/S0958-1669(00)00199-3

Tamura K, Stecher G and Kumar S. 2021. MEGA11: Molecular Evolutionary Genetics Analysis Version 11. Molecular Biology and Evolution 38(7): 3022–3027. DOI: https://doi.org/10.1093/molbev/msab120

Taranger G L, Carrillo M, Schulz R W, Fontaine P, Zanuy S, Felip A, Weltzien F A, Dufour S, Karlsen Ø, Norberg B, Andersson E and Hansen T. 2010. Control of puberty in farmed fish. General and Comparative Endocrinology 165(3): 483–515. DOI: https://doi.org/10.1016/j.ygcen.2009.05.004

Terpe K. 2006. Overview of bacterial expression systems for heterologous protein production: From molecular and biochemical fundamentals to commercial systems. Applied Microbiology and Biotechnology 211–222. DOI: https://doi.org/10.1007/s00253-006-0465-8

Treerattrakool S, Boonchoy C, Urtgam S, Panyim S and Udomkit A. 2014. Functional characterization of recombinant gonad-inhibiting hormone (GIH) and implication of antibody neutralization on induction of ovarian maturation in marine shrimp. Aquaculture 428–429: 166–173. DOI: https://doi.org/10.1016/j.aquaculture.2014.03.009

Treerattrakool S, Panyim S, Chan S M, Withyachumnarnkul B and Udomkit A. 2008. Molecular characterization of gonad-inhibiting hormone of Penaeus monodon and elucidation of its inhibitory role in vitellogenin expression by RNA interference. FEBS Journal 275(5): 970–980. DOI: https://doi.org/10.1111/j.1742-4658.2008.06266.x

Treerattrakool S, Panyim S and Udomkit A. 2011. Induction of Ovarian Maturation and Spawning in Penaeus monodon Broodstock by Double-Stranded RNA. Marine Biotechnology 13(2): 163–169. DOI: https://doi.org/10.1007/s10126-010-9276-0

Tsutsui N, Ohira T, Okutsu T, Shinji J, Bae S H, Kang B J and Wilder M N. 2013. Molecular cloning of a cDNA encoding vitellogenesis-inhibiting hormone in the whiteleg shrimp Litopenaeus vannamei and preparation of its recombinant peptide using an E. coli expression system. Fisheries Science 79(3): 357–365. DOI: https://doi.org/10.1007/s12562-013-0603-z

Vaca A A and Alfaro J. 2000. Ovarian maturation and spawning in the white shrimp, Penaeus vannamei, by serotonin injection. Aquaculture 182(3–4): 373–385. DOI: https://doi.org/10.1016/S0044-8486(99)00267-7

Vijayan K K, Reynold P, Mohanlal D L and Balasubramanian C P. 2013. Preliminary characterisation of gonad inhibiting hormone (GIH) gene and its expression pattern during vitellogenesis in giant tiger shrimp, Penaeus monodon Fabricius, 1798. Indian Journal of Fisheries 60(2).

Wilder M, Okumura T and Tsutsui N. 2010. Reproductive Mechanisms in Crustacea Focusing on Selected Prawn Species: Vitellogenin Structure, Processing and Synthetic Control. Aqua-BioScience Monographs 3(3): 73–110. DOI: https://doi.org/10.5047/absm.2010.00303.0073

Wurts W A and Stickney R R. 1984. An hypothesis on the light requirements for spawning penaeid shrimp, with emphasis on Penaeus setiferus. Aquaculture 41(2): 93–98. DOI: https://doi.org/10.1016/0044-8486(84)90086-3