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Purification and characterization of trypsin produced by gut bacteria from Anticarsia gemmatalis.
Arch Insect Biochem Physiol. 2017 Oct; 96(2)AI

Abstract

Purification of active trypsin in the digestive process of insects is essential for the development of potent protease inhibitors (PIs) as an emerging pest control technology and research into insect adaptations to dietary PIs. An important aspect is the presence of proteolytic microorganisms, which contribute to host nutrition. Here, we purified trypsins produced by bacteria Bacillus cereus, Enterococcus mundtii, Enterococcus gallinarum, and Staphylococcus xylosus isolated from the midgut of Anticarsia gemmatalis. The trypsins had a molecular mass of approximately 25 kDa. The enzymes showed increased activity at 40°C, and they were active at pH values 7.5-10. Aprotinin, bis-benzamidine, and soybean Kunitz inhibitor (SKTI) significantly inhibited trypsin activity. The l-1-tosyl-amido-2-phenylethylchloromethyl ketone (TPCK), pepstatin A, E-64, ethylenediamine tetraacetic acid, and calcium ions did not affect the enzyme activity at the concentrations tested. We infer the purified trypsins do not require calcium ions, by which they differ from the trypsins of other microorganisms and the soluble and insoluble trypsins characterized from A. gemmatalis. These data suggest the existence of different isoforms of trypsin in the velvetbean caterpillar midguts.

Authors+Show Affiliations

Departamento de Bioquímica e Biologia Molecular, Instituto de Biotecnologia Aplicada a Agropecuária-BIOAGRO, Universidade Federal de Viçosa, Viçosa, MG, Brazil.Instituto de Ciências Agrárias, Universidade Federal de Viçosa Campus Rio Paranaíba, MG, Brazil.Instituto de Ciências Biológicas e da Saúde, Universidade Federal de Viçosa Campus Rio Paranaíba, MG, Brazil.Departamento de Bioquímica e Biologia Molecular, Instituto de Biotecnologia Aplicada a Agropecuária-BIOAGRO, Universidade Federal de Viçosa, Viçosa, MG, Brazil.Departamento de Bioquímica e Biologia Molecular, Instituto de Biotecnologia Aplicada a Agropecuária-BIOAGRO, Universidade Federal de Viçosa, Viçosa, MG, Brazil.Departamento de Engenharia de Biossistemas, Universidade Federal de São João Del Rei, São João Del Rei, MG, Brazil.Departamento de Bioquímica e Biologia Molecular, Instituto de Biotecnologia Aplicada a Agropecuária-BIOAGRO, Universidade Federal de Viçosa, Viçosa, MG, Brazil.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

28762531

Citation

Pilon, Franciny Martins, et al. "Purification and Characterization of Trypsin Produced By Gut Bacteria From Anticarsia Gemmatalis." Archives of Insect Biochemistry and Physiology, vol. 96, no. 2, 2017.
Pilon FM, Silva CDR, Visôtto LE, et al. Purification and characterization of trypsin produced by gut bacteria from Anticarsia gemmatalis. Arch Insect Biochem Physiol. 2017;96(2).
Pilon, F. M., Silva, C. D. R., Visôtto, L. E., Barros, R. A., da Silva Júnior, N. R., Campos, W. G., & de Almeida Oliveira, M. G. (2017). Purification and characterization of trypsin produced by gut bacteria from Anticarsia gemmatalis. Archives of Insect Biochemistry and Physiology, 96(2). https://doi.org/10.1002/arch.21407
Pilon FM, et al. Purification and Characterization of Trypsin Produced By Gut Bacteria From Anticarsia Gemmatalis. Arch Insect Biochem Physiol. 2017;96(2) PubMed PMID: 28762531.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Purification and characterization of trypsin produced by gut bacteria from Anticarsia gemmatalis. AU - Pilon,Franciny Martins, AU - Silva,Camila da Rocha, AU - Visôtto,Liliane Evangelista, AU - Barros,Rafael de Almeida, AU - da Silva Júnior,Neilier Rodrigues, AU - Campos,Wellington Garcia, AU - de Almeida Oliveira,Maria Goreti, Y1 - 2017/08/01/ PY - 2017/8/2/pubmed PY - 2017/10/5/medline PY - 2017/8/2/entrez KW - bacteria KW - pest control KW - protease inhibitor KW - trypsin JF - Archives of insect biochemistry and physiology JO - Arch Insect Biochem Physiol VL - 96 IS - 2 N2 - Purification of active trypsin in the digestive process of insects is essential for the development of potent protease inhibitors (PIs) as an emerging pest control technology and research into insect adaptations to dietary PIs. An important aspect is the presence of proteolytic microorganisms, which contribute to host nutrition. Here, we purified trypsins produced by bacteria Bacillus cereus, Enterococcus mundtii, Enterococcus gallinarum, and Staphylococcus xylosus isolated from the midgut of Anticarsia gemmatalis. The trypsins had a molecular mass of approximately 25 kDa. The enzymes showed increased activity at 40°C, and they were active at pH values 7.5-10. Aprotinin, bis-benzamidine, and soybean Kunitz inhibitor (SKTI) significantly inhibited trypsin activity. The l-1-tosyl-amido-2-phenylethylchloromethyl ketone (TPCK), pepstatin A, E-64, ethylenediamine tetraacetic acid, and calcium ions did not affect the enzyme activity at the concentrations tested. We infer the purified trypsins do not require calcium ions, by which they differ from the trypsins of other microorganisms and the soluble and insoluble trypsins characterized from A. gemmatalis. These data suggest the existence of different isoforms of trypsin in the velvetbean caterpillar midguts. SN - 1520-6327 UR - https://www.unboundmedicine.com/medline/citation/28762531/Purification_and_characterization_of_trypsin_produced_by_gut_bacteria_from_Anticarsia_gemmatalis_ L2 - https://doi.org/10.1002/arch.21407 DB - PRIME DP - Unbound Medicine ER -