Exploration of Amylases Producing Competency of Helicoverpa armigera Gut Bacterial Strain, Bacillus subtilis RTSBA6 6.00

  • Ashok A. Shinde Department of Biotechnology, MGM’s Institute of Biosciences and Technology, Aurangabad, Maharashtra, India
  • Faiyaz K. Shaikh (1) Department of Biotechnology, MGM’s Institute of Biosciences and Technology, Aurangabad, Maharashtra, India; (2) Department of Biochemistry, Dr. Babasaheb Ambedkar Marathwada University, Aurangabad, Maharashtra, India
  • Sarwan W. Bradosty Department of Biology, Cihan University-Erbil, Kurdistan region, Iraq https://orcid.org/0000-0002-4659-4081
  • Manvendra S. Kachole Department of Biotechnology, MGM’s Institute of Biosciences and Technology, Aurangabad, Maharashtra, India
Keywords: Amylase, Bacillus subtilis RTSBA6 6.00, Helicoverpa armigera, zymography

Abstract

The Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae) is a polyphagous insect pest of agriculturally important crops. The alkaline gut of this insect pest possesses diverse bacterial communities which may assist in digestive physiology. As part our investigations of understanding the role of gut bacterial communities in insect gut, here amylase producing competency of earlier identified H. armigera gut bacterial strain, i.e., Bacillus subtilis RTSBA6 6.00 is reported. Initial screening for amylase activity was assessed by starch agar plate. Upon 7% sodium dodecyl sulfate polyacrylamide gel electrophoresis amylase zymography, bacterial culture supernatant produced seven amylase bands on the gel. The observed molecular weights of amylases were 191.2 KDa, 158.0 KDa, 131.7 KDa, 54.0 KDa, 31.3 KDa, 67.2 KDa, and 44.6 KDa, respectively. Considerable amylase activity was observed in neutral to alkaline pH with optimum at pH 6.8. The optimal activity temperature of amylases was found to be 50°C, and the activity decreased dramatically at temperatures above 75°C.

Downloads

Download data is not yet available.

References

N. A. Moran, P. H. Degnan, S. R. Santos, H. E. Dunbar and H. Ochman. “The players in a mutualistic symbiosis: Insects, bacteria, viruses, and virulence genes.” Proceedings of the National Academy of Sciences of the United States of America, vol. 102, pp. 16919-16926, 2005.

J. C. Clemente, L. K. Ursell, L. W. Parfrey and R. Knight. “The impact of the gut microbiota on human health: An integrative view”. Cell, vol. 148, pp. 1258-1270, 2012.

R. Dillon and V. Dillon. “The gut bacteria of insects: Nonpathogenic interactions”. Annual Review of Entomology, vol. 49, pp. 71-92, 2004.

F. M. Pilon, L. E. Visotto, R.N.C. Guedes and M. G. A. Oliveira. “Proteolytic activity of gut bacteria isolated from the velvet bean caterpillar Anticarsia gemmatalis”. Journal of Comparative Physiology B: Biochemical, Systemic, and Environmental Physiology, vol. 183, pp. 735-747, 2013.

J. A. Breznak and A. Brune. “Role of microorganisms in the digestion of lignocellulose by termites”. Annual Review of Entomology, vol. 39, pp. 453-487, 1994.

V. D. Parde, H. C. Sharma and M. S. Kachole. “In vivo inhibitions of Helicoverpa armigera gut pro-proteinase activation by nonhost plant protease inhibitors”. Journal of Insect Physiology, vol. 56, no 9, pp. 1315-1324, 2010.

A. A. Shinde, F. K. Shaikh, P. P. Gadge, M. V. Padul, S. P. Govindwar and M. S. Kachole. “Conserved nature of Helicoverpa armigera gut bacterial flora on different host plants and in vitro interactions with PI proteins advocates role in host digestive physiology”. Journal of the Saudi Society of Agricultural Sciences, vol. 18, pp. 141-149, 2017.

H. Sharma, M. Dhillon and R. Arora. “Effects of Bacillus thuringiensis dendotoxin-fed Helicoverpa armigera on the survival and development of the parasitoid Campoletis chlorideae”. Entomologia Experimentalis et Applicata, vol. 126, pp. 1-8, 2008.

P. K. Mishra and S. M. Tandon. “Gut bacterial flora of Helicoverpa armigera (Lepidoptera, Noctuidae)”. Indian Journal of Microbiology, vol. 43, no. 1, pp. 55-56, 2003.

A. A. Shinde, F. K. Shaikh, M. V. Padul and M. S. Kachole. “Bacillus subtillis RTSBA6 6.00, a new strain isolated from gut of Helicoverpa armigera (Lepidoptera: Noctuidae) produces chymotrypsin-like proteases”. Saudi Journal of Biological Sciences, vol. 19, pp. 317-323, 2012.

P. N. Gayatri, A. Ojha, M. K. Kajla, A. Raj and R. Rajagopal. “Host plant induced variation in gut bacteria of Helicoverpa armigera”. PLoS One, vol. 7, pp. e30768, 2012.

L. E. Visoˆtto, M. G. A. Oliveira, A. O. B. Ribon, T. R. MaresGuia and R. N. C. Guedes. “Characterization and identification of proteolytic bacteria from the gut of the velvetbean caterpillar (Lepidoptera: Noctuidae)”. Environmental Entomology, vol. 38, no. 4, pp. 1078-1085, 2009.

R. Kaur, N. Kaur and A. K. Gupta. “Structural features, substrate specificity, kinetic properties of insect α-amylase and specificity of plant α-amylase inhibitors”. Pesticide Biochemistry and Physiology, vol. 116, pp. 83-93, 2014.

J. S. Buyer. “Improved fast gas chromatography for FAME analysis of bacteria”. Journal of Microbiological Methods, vol. 54, pp. 117120, 2003.

S. Sharmin, M. D. Towhid and M. N. Anwar. “Isolation and characterization of a protease producing bacteria Bacillus amoviv-orus and optimization of some factors of culture conditions for protease production”. Journal of Biological Sciences, vol. 5, no. 3, pp. 358-362, 2005.

O. H. Lowry, N. J. Rosebrough, A. L. Farr and R. J. Randall. “Protein measurement with the folin phenol reagent”. Journal of Biological Chemistry, vol. 193, pp. 265, 1951.

U. K. Laemmli. “Cleavage of structural proteins during the assembly of the head of bacteriophage T4”. Nature, vol. 227, pp. 680-685, 1970.

E. H. Fisher and E. A. Stein. “Alpha amylase from human saliva”. Biochemical Preparations, vol. 8, pp. 27-32, 1961.

G. G. Priest. “Extracellular enzyme synthesis in the genus Bacillus”. Bacteriological Reviews, vol. 41, no. 3, pp. 711-753, 1977.

A. Andrades and L. M. Contreras. Amylase zymography”. Methods in Molecular Biology, vol. 1626, pp. 301-308, 2017.

F. Zeng and A. C. Cohen.“Partial characterization of a-amylase in the salivary glands of Lygus hesperus and Lygus lineolaris”. Comparative Biochemistry and Physiology Part B, vol. 126, pp. 9-16, 2000.

P. C. Aryati, A. Pangastuti and S. L. A. Sari. “Amylase Production Potentials of Bacterial Isolates Obtained from the Gut of Oryctes rhinoceros Larvae”. IOP Conference Series: Materials Science and Engineering, 2017. 23. T. M. Clark. “Evolution and adaptive significance of larval midgut alkalinization in the insect superorder Mecopterida”. Journal of Chemical Ecology, vol 25, pp. 1945-1960, 1999.

Published
2019-06-30
How to Cite
1.
Shinde A, Shaikh F, Bradosty S, Kachole M. Exploration of Amylases Producing Competency of Helicoverpa armigera Gut Bacterial Strain, Bacillus subtilis RTSBA6 6.00. cuesj [Internet]. 30Jun.2019 [cited 27Jan.2022];3(1):75-9. Available from: https://journals.cihanuniversity.edu.iq/index.php/cuesj/article/view/97
Section
Research Article