Abstract
A soil isolate, Bacillus sp. DT7 has been found to produce significant amounts of an extracellular pectinase subsequently characterized as pectin lyase (EC 4.2.2.10). By optimizing growth conditions, Bacillus sp. DT7 produced higher amount of pectin lyase (53 units/ml) than that has been reported in the literature. Using gel filtration and ion exchange chromatography, this enzyme was purified and found to have a molecular mass of 106 kDa. The purified enzyme exhibited maximal activity at a temperature of 60 ∘C and pH 8.0. The presence of 100 mM concentrations of CaCl2 and mercaptoethanol significantly enhanced pectinase activity of the purified enzyme. This pectinase has tremendous applications in textile industry, plant tissue maceration and fruit juice wastewater treatments.
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References
Acuna-Arguelles, M.E., Gutierrez-Rajas, M., Viniegra-Gonzalez, G. & Favela-Toress, E. 1995 Production and properties of three pectinolytic activities produced by A. niger in submerged and solid state fermentation. Applied Microbiology and Biotechnology 43, 808-814.
Aguillar, G. & Huitron, C. 1990 Constitutive exopectinase produced by Aspergillus sp. CH-Y-1043 on different carbon sources. Biotechnology Letters 12, 655-660.
Borin, M.D.F., Said, S. & Fonseca, M.J.V. 1996 Purification and biochemical characterization of an extracellular endopolygalacturonase from Penicillium frequentans. Journal of Agriculture and Food Chemistry 44, 1616-1620.
Cao, J., Zheng, L., & Chen, S. 1992 Screening of pectinase producer from alkalophilic bacteria and study on its potential application in degumming of rammie. Enzyme and Microbial Technology 14, 10131016.
Dave, B.A. & Vaughn, R.H. 1971 Purification and properties of a polygalaturonic acid trans eliminase produced by Bacillus pumilus. Journal of Bacteriology 108, 166-174.
Fanseca, M.J.V. & Said, S. 1995 Sequential production of pectinase by Penicillium frequentans. World Journal of Microbiology and Biotechnology 11, 174-177.
Fogarty, M.V. & Kelly, C.T. 1983 In Microbial Enzymes and Biotechnology. ed. Fogarty, M.W. pp. 131-182. London & New York: Elsevier Applied Science Publishers. ISBN 1-85166486-6
Gainvors, A. & Belarbi, A. 1995 Detection methods for polygalactu-ronase producing strains of Saccharomyces cerevisae. Yeast 10, 1311-1319.
Garibaldi, A. & Bateman, D.F. 1971 Pectic enzymes produced by Erwinia chrysanthemi and their e.ect on plant tissue. Physiological Plant Pathology 1, 25-40.
Horikoshi, K. 1972 Production of alkaline enzymes by alkalophilic microorganisms Part III. Alkaline pectinase of Bacillus No P-4-N. Agricultural and Biological Chemistry 36, 285-293.
Karbassi, A. & Vaughn, R.H. 1980 Purification and properties of poygalacturonic acid trans-eliminase from Bacillus stearothermophilus. Canadian Journal of Microbiology 26, 377-384.
Kelly, C.T. & Fogarty, W.M. 1978 Production and properties of polygalacturonate lyase by an alkalophilic microorganism Bacillus sp. RK9. Canadian Journal of Microbiology 29, 1164-1172.
Laemmli, U.K. 1970 Cleavage of structural proteins during the assembly of head bacteriophage T4. Nature 22, 680-685.
Martinez, M.J., Alconda, M.T., Guillrn, F., Vazquez, C. & Reyes, F. 1991 Pectic activity from Fusarium oxysporium f. sp. melonis: purification and characterization of an exopolygalacturonase. FEMS Microbiology Letters 81, 145-150.
Merril, C.R., Goldman, D. & Ibert, M.H. 1981 Ultrasensitive stain for proteins in polyacrylamide gel shows regional variations in cerebrospinal fluid proteins. Science 211, 1437-1438.
Miller, L.G. 1959 Use of dinitrosalicylic acid reagent for the determination of reducing sugars. Analytical Chemistry 31, 426-428.
Moran, F. & Starr, M.P. 1969 Metabolic regulation of polygalacturonic acid trans eliminase in Erwinia. European Journal of Biochemistry 11, 291-295.
Nagel, C.W. & Vaughn, R.H. 1961 The characteristic of a polygalacturonase produced by Bacillus polymyxa. Archives of Biochem-istry and Biophysics 93, 344-352.
Nagel, C.W. & Hasegawa, S. 1968 The isolation and characterization of an 1,4 (4,4-dihydrogalacturonosyl) galacturonate hydrolase. Journal of Food Science 33, 378-382.
Nasumo, S. & Starr, M.P. 1967 Polygalacturonic acid tran eliminase of Xanthomonas compestris. Biochemical Journal 104, 178-184.
Pitt, O. 1988 Pectic lyase from Rhoma medicabinis var. pinodella. Methods in Enzymology 161, 350-365.
Rombouts, F.M. & Pilnik, W. 1986 Pectinases and other cell-wall degrading enzymes of industrial importance. Symbiosis 2, 79-89.
Siessere, V. & Said, S. 1989 Pectic enzyme production in solid state fermentation using citrus pulp pellets by Talaromyces flavus, Tubercularia vulgaris and Penicillium charlessi. Biotechnology Letters 11, 343-344.
Sharma, H.S.S. 1987 Enzymatic degradation of residual non cellulosic polysaccharides present on dew retted flax fibers. Applied Microbiology and Biotechnology 26, 358-362.
Starr, M.P. & Nasumo, S. 1967 Proteolytic activity of phytopathogenic Xanthomonads. Journal of General Microbiology 46, 425-433.
Tanabe, H., Yoshihara, K., Tamura, K., Kobayashi, Y., Akamatsu, I., Niyomwan, N., & Footrakul, P. 1987 Pretreatment of pectic waste water from orange canning process by an alkalophilic Bacillus sp. Journal of Fermentation Technology 65, 243-246.
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Kashyap, D., Chandra, S., Kaul, A. et al. Production, purification and characterization of pectinase from a Bacillus sp. DT7. World Journal of Microbiology and Biotechnology 16, 277–282 (2000). https://doi.org/10.1023/A:1008902107929
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DOI: https://doi.org/10.1023/A:1008902107929