Production and Characterization of Pectic Enzymes of Xanthomonas campestris pv. sesami
Abstract
The bacterium Xanthomonas campestris pv. sesami, is the causal organism of the leaf spot disease (Marad eldum) in sesame plants. Pectic enzymes which were reported to be produced by many plant pathogenic bacteria, were found to play an important role in pathogenesis. There are three main types of pectic enzymes (Polygalacturonase, PG, Polygalacturonate trans-eliminase PTE and Pectin methylesterase PME). The present study was conducted to investigate the capacity of the bacterium X. campestris pv. sesami, to produce the different types of pectic enzymes. A salt medium supplemented with different pectic materials as carbon sources were tested for the production of the enzymes. The produced enzymes were characterized using different selective tests, both the viscosity and reducing group methods were used to measure the enzyme activity. The results showed that no pectin methylestrease (PME) was produced and only the Polygalacturonase trans-eliminase (PTE) was produced by the bacterium. Sodium polypectate and polygalacturonic acid were found as the best substrates giving 26.9 and 19.8 viscosity enzyme units, respectively. The enzyme activity was maximum at the 6th day (9.3 viscosity units), then the activity was decreasing with further incubation time. The test of the effect of the pH indicated that the pH optimum of the enzyme activity was at pH 9.0. The effects of different metallic ions to the enzyme activity showed that,= a higher activity was found with Ca++ which gave 12.2 viscosity units. When different concentrations of Ca++ were tested, the maximum activity of the enzyme was found at 50 mg/ ml concentration, which gave 18.0 viscosity units. It could be recommended that the tested bacterium should be investigated for the production of the cellulases and hemi-cellulases which also have a role in the pathogenesis process.
References
sesame (Sabet and Dowson). Ph.D. thesis, Department of Botany and Plant
Technology, Imperial Collage of Science and Technology. London.
Acuna, A. M.; Gutierrez, R. M.; Viniegra, G. G. and Favela, T. E. (1995). Production
and properties of three pectinolytic Activities produced by Asperigillus niger
in submerged and solid state fermentation. Journal of Applied Microbiology
and Biotechnology, 43:808-814.
Azanaw, A., Belay, Y., Ebabuye, Y., & Ayizengaw, G. (2018). Screening of Sesame
Germplasms Against Bacterial Blight (Xanthomonas campestris pv.sesami) at
Metema Ethiopia. Academic Journal of Life Sciences, 4(3), 15 -17.
Bhat, M. K. and Bhat, S. (1997). Cellulose degrading enzymes and their potential
industrial application. Biotechnology Advances, 15: 583-620.
Birch,G. G., Blakebrough, N. and Parker, J. (1981). Enzyme and Food processing.
Applied Science Publisher LTD. England. Pp 5- 13.
Braverman, J. B. F. (1974). Introduction to Biochemistry of food. 3rd. elsevier
publishing company ed.. London. Pp 31.
Chapin, F. S., Matson, P. A. and Mooney, H. A. (2002). Principles of Terrestrial
Ecosystem Ecology. Springer- Verlag, New York. Pp 150.
Coral, G.; Arican, B. M. Nisa, O. and Hatice, G. (2001). Some properties of crude
carboxymethyl cellulase of Asperigillus niger. Turkish Journal of Biology.
26: 209 – 213.
Fogarty, W. M. and Kelly, C. J. (1979). Development of Microbial extracellular
enzymes. in: enzyme and fermentation biotechnology. (Alan Wiseman.ed)
Ellis Harwood LTD. Publishers. England, pp 239.
Golla, W. N., Kebede, A. A., & Kindeya, Y. B. (2020). Evaluation of sesame
genotypesfor seed yield and bacterial blight (Xanthomonas campestris pv.
sesami) disease resistance in optimum moisture areas of Western Tigray,
Ethiopia. Cogent Food & Agriculture , 6(1)
Gupta, V. K. and Ayyachamy, M. (2012). Biotechnology of Fungal Genes. Science
publisher. England. Pp 53- 71.
Hasunuma, T.; Fukusaki, E. and Kobayashi, A. (2003). A Methanol Production is
enhanced by expression of an Asperigillus niger Pectinmethylesterase in
tabacco cells. Journal of Biotechnology. 106: 45-52.
Production and Characterization of Pectic Enzymes of
Xanthomonas campestris pv. sesami
45
Laurent, F.; Kotoujansky, A. and Bertheau, U. (2000). Over Production in
Escherichia coli of the Pectinmethylesterase. Canadian Journal of
Microbiology. 46: 474-480.
Maria, S.M.; Celestino, C.; Maria de Freitas, S.; Javier, F. M.; Valle desouse, M.;
Ximenes, E. and Ferrira, F. (2006). Purification and Characterization of a
Novel Pectinase from Acrophialophora nainiana with Emphasis on
Physiochemical Properties. Journal of Biochemistry, 123 (1): 33-42
Mirjana, G. A., Antov, D. M. and Stana, N. P. (2004). Pectinases partitioning in
aqueous two- phase system: an integration of the systems poly (ethylene
glycol)/ crude dextrin and poly (ethylene glycol) ammonium sulphate.
Journal of Chemical Society. 69 (4): 299- 307.
Mustafa, A.A. (2008). Production and charactterazation of pectic enzymes. M. Sc.
Thesis University of Gezeira
Ranveer,S. J.; shivalika, S. and Reena, G. (2005). Microbial Pectinolytic Enzymes.
Elsevier publishing company. London and New York. Pp 34- 50.
Rombouts, F. M. and Pinlnik, W. (1980). Pectic enzymes In: microbial enzymes and
bioconversion. ( Rose A. H.ed). Academic press, London. 5: 227- 270.
Sabet , K ,A. and Dowson, W.T (1960). Abacterial leaf spot disease of sesame
(Sesamun orientel). Phytobathological Z. 37:252-256.
Scawen, M. D. and Milling, J. (1985). Practical Aspect of large –scale protein
purification in: hand book of enzyme biotechnology ( A. A Wiseman.ed)
John Willy and Sons. New York. Pp36 – 70.
Timoth, Y. D. (2001). Enzymes: General Properties. John Wiley and Sons. Ltd. New
York and London. Pp 151- 167.
Urmila, P.; Vikram, D.; Shobhna, S. and Chadha, B. S. (2005). Pectinase and
Polygalacturonase production by A thermophilic A. fumigates isolated from
orange peels. Brazilian Journal of Microbiology, 36: 63 – 69.
Vinoth ,B. Manivasagaerumal, R. and Belamurugan, S. (2012). Phytochemical
analysis and antibacterial activity of Moringa oleifera (lam), India.
International J. for research in Biological Sciences, 2012; 2(3):98-102