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http://dx.doi.org/10.4014/jmb.0910.10014

Isolation, Optimization, and Partial Purification of Amylase from Chrysosporium asperatum by Submerged Fermentation  

Sanghvi, Gaurav V. (Department of Botany, Faculty of Science, The Maharaja Sayajirao University of Baroda)
Koyani, Rina D. (Department of Botany, Faculty of Science, The Maharaja Sayajirao University of Baroda)
Rajput, Kishore S. (Department of Botany, Faculty of Science, The Maharaja Sayajirao University of Baroda)
Publication Information
Journal of Microbiology and Biotechnology / v.21, no.5, 2011 , pp. 470-476 More about this Journal
Abstract
A potent fungus for amylase production, Chrysosporium asperatum, was isolated from among 30 different cultures obtained from wood samples collected in the Junagadh forest, India. All of the isolated cultures were screened for their ability to produce amylase by submerged fermentation. Among the selected cultures, C. asperatum (Class Euascomycetes; Onygenales; Onygenaceae) gave maximum amylase production. In all of the different media tested, potato starch was found to be a good substrate for production of amylase enzyme at $30^{\circ}C$ and pH 5.0. Production of enzyme reached the maximum when a combination of starch and 2% xylose, and organic nitrogen (1% yeast extract) and ammonium sulfate were used as carbon and nitrogen sources, respectively. There was no significant effect of metal ions on enzyme activity. The enzyme was relatively stable at $30^{\circ}C$ for 20 min, and no inhibitory effect of $Ca^{+2}$ ions on amylase production was observed.
Keywords
Amylase; Chrysosporium asperatum; submerged fermentation; optimization;
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1 Abe, J., F. W. Bergman, K. Obeta, and S. Hizukuri. 1988. Production of the raw starch degrading amylase of Aspergillus sp. K-27. Appl. Microbiol. Biotechnol. 27: 447-450.
2 Abou Zeid, A. M. 1997. Production, purification and characterization of an extracellular alpha-amylase enzyme isolated from Aspergillus flavus. Microbios 89: 55-66.
3 Adebiyi, C. A. B. and J. A. Akinyanju. 1998. Thermophilic amylase producers from the soil. Nigerian J. Sci. Technol. 11: 30-38.
4 Akpan, I., M. O. Bankole, A. M. Adesemowo, and G. O. Latunde Dada. 1999. Production of amylase by A. niger in a cheap solid medium using rice bran and agricultural materials. Trop. Sci. 39: 77-79.
5 Alva, S., J. Anupama, J. Savla, Y. Y. Chiu, P. Vyshali, M. Shruti, et al. 2007. Production and characterization of fungal amylase enzyme isolated from Aspergillus sp. JGI 12 in solid state culture. Afr. J. Biotechnol. 6: 576-581.
6 Ariff, A. B. and C. Web. 1998. Effect of initial carbon and nitrogen sources concentrations on growth of Aspergillus awamori and glucoamylase production. Asia Pacific J. Mol. Biol. 6: 161-169.
7 Asgher, M., M. Javaid Asad, S. U. Rahman, and R. L. Legge. 2007. A thermostable $\alpha$-amylase from a moderately thermophilic Bacillus subtilis strain for starch processing. J. Food Eng. 79: 950-955.   DOI   ScienceOn
8 Beguin, P. and J. P. Aubert. 1994. The biological degradation of cellulose. FEMS Microbiol. Rev. 13: 25-58.   DOI   ScienceOn
9 Borchert, T. V., S. F. Lassen, A. Svedsen, and H. B. Frantzen. 1995. Oxidation stable amylases for detergents, pp. 175-179. In S. B. Petersen, B. Svensson, and S. Pedersen. (eds.). Progress in Biotechnology. Carbohydrate Bioengineering. Elsevier, The Netherlands. 10: 175-179.
10 Burhan, A., U. Nisa, C. Gokhan, C. Omer, A. Ashabil, and G. Osman. 2003. Enzymatic properties of a novel thermostable, thermophilic, alkaline and chelator resistant amylase from an alkaliphilic Bacillus sp. isolate ANT-6. Process Biochem. 38: 1397-1403.   DOI   ScienceOn
11 Aberle, T. H., W. Burchard, W. Vorwerg, and S. Radosta. 1994. Conformation, contributions of amylase and amylopectin to the structural properties of starches from various sources. Starke 46: 329-335.   DOI   ScienceOn
12 Duochuan, L., Y. Yijun, P. Youliang, S. Chongyao, Z. Peijin, and H. Yicun. 1997. Purification and properties of a thermostable $\alpha$-amylase from the thermophilic fungus Thermomyces lanuginosus. Acta Microbiol. Sin. 37: 107-114.
13 Buzzini, P. and A. Martini. 2002. Extracellular enzymatic activity profiles in yeast and yeast-like strains isolated from tropical environments. J. Appl. Microbiol. 93: 1020-1025.   DOI   ScienceOn
14 Chakraborty, K., B. K. Bhattacharyya, and S. K. Sen. 2000. Purification and characterization of a thermostable $\alpha$-amylase from Bacillus stearothermophilus. Folia Microbiol. 45: 207-210.   DOI   ScienceOn
15 Cruz, R., L. Souza, E. H. E. Hoffmann, M. Z. Bellini, V. A. Cruz, and C. R. Vieira. 1997. Relationship between carbon source, production and pattern action of $\alpha$-amylase from Rhizopus sp. Rev. Microbiol. 28: 101-105.
16 Dutta, T. K., M. Jana, P. R. Pahar, and T. Bhattacharya. 2006. The effect of temperature, pH and salt on amylase in Heliodiaptomus viduus (Gurney) (Crustacia: Copepoda: Calanoida). Turk. J. Zool. 30: 187-195.
17 Ellaiah, P., K. Adinarayana, Y. Bhavani, P. Padmaja, and B. Srinivasula. 2002. Optimization of process parameters for glucoamylase production under solid state fermentation by a newly isolated Aspergillus species. Process Biochem. 38: 615-620.   DOI   ScienceOn
18 El-Safey, E. M. and M. S. Ammar. 2004. Purification and characterization of $\alpha$-amylase isolated from Aspergillus falvus var. columnaris. Ass. Univ. Bull. Environ. Res. 7: 93-100.
19 Kunamneni, A. 2005. Amylase production by solid state fermentation by the thermophilic fungus Thermomyces lanuginosus. J. Biosci. Bioeng. 100: 168-171.   DOI   ScienceOn
20 Kirk, T. K., E. Schultz, W. J. Connors, L. F. Lorenz, and J. G. Zeikus. 1978. Influence of culture parameters on lignin metabolism by Phanerochaete chrysosporium. Arch. Microbiol. 117: 277-285.   DOI   ScienceOn
21 Laderman, K. A., B. R. Davis, H. C. Krutzsch, M. S. Lewis, Y. V. Griko, P. L. Privalov, and C. B. Anfinsen. 1993. The purification and characterization of extremely thermostable alpha-amylase from the hyperthermophilic archaebacterium Pyrococcus furiosus. J. Biol. Chem. 268: 394-401.
22 Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685.   DOI   ScienceOn
23 Lange, N. K. 1993. Application of cellulases in the textile industry, pp. 263-272. In P. Suominen and T. Reinikainen. (eds.). Trichoderma reesei Cellulases and Other Hydrolases. Foundation for Biotechnical and Industrial Fermentation Research, Helsinki.
24 Lineback, D. R., C. E. Georgi, and K. L. Doty. 1996. Glucoamylase production by Aspergillus niger as influenced by medium composition. J. Gen. Appl. Microbiol. 12: 27-34.
25 McMahon, E. M., C. T. Kelly, and W. M. Fogarty. 1999. High maltose-producing amylolytic system of a Streptomyces sp. Biotechnol. Lett. 21: 23-26.   DOI   ScienceOn
26 Miller, G. L. 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem. 31: 426-429.   DOI
27 Hema, A., U. B. Trivedi, and K. C. Patel. 2006. Glucoamylase production by solid-state fermentation using rice flask manufacturing waste products as substrate. Bioresour. Technol. 97: 1161-1166.   DOI   ScienceOn
28 Gold, M. H., M. B. Mayfield, T. M. Cheng, K. Krisnangkura, A. Enoki, M. Shimada, and J. K. Glenn. 1982. A Phanerochaete chrysosporium mutant defective in lignin degradation as well as several other secondary metabolic functions. Arch. Microbiol. 132: 115-122.   DOI   ScienceOn
29 Goto, C. E., E. P. Barbosa, L. C. L. Kistner, R. F. Gandra, V. L. Arrias, and R. M. Peralta. 1998. Production of amylase from Aspergillus fumigatus. Rev. Microbiol. 29: 99-103.
30 Haq, I., H. Ashraf, S. Rani, and M. A. Qadeer. 2002. Biosynthesis of alpha amylase by chemically treated mutant of Bacillus subtilis GCBU-20. Pak. J. Biol. Sci. 2: 73-75.   DOI
31 Hoover, R. 2001. Composition, molecular structure, and physicochemical properties of tuber and root starches: A review. Carbohydr. Polym. 45: 253-267.   DOI   ScienceOn
32 Kennedy, J. F., V. M. Cabalda, and C. A. White. 1988. Enzymatic starch utilization and genetic engineering. TIBTECH 6: 184-189.   DOI   ScienceOn
33 Khoo, S. L., A. A. Amirul, M. Kamaruzaman, M. Nazalan, and M. N. Azizan. 1994. Purification and characterization of alphaamylase from Aspergillus flavus. Folia Microbiol. 39: 392-398.   DOI   ScienceOn
34 Pommier, J. C., G. Goma, J. L. Fuentes, C. Rousset, and O. Jokinen. 1990. Using enzymes to improve the process and the product quality in the recycled paper industry. Part 2: Industrial applications. Tappi J. 73: 197-202.
35 Nahas, E. and M. M. Waldermarin. 2002. Control of amylase production and growth characteristics of Aspergillus ochraceus. Microbiologia 44: 5-10.
36 Omemu, A. M., I. Akpan, M. O. Bankole, and O. D. Teniola. 2005. Hydrolysis of raw tuber starches by amylase of Aspergillus niger AM07 isolated from soil. Afr. J. Biotechnol. 4: 19-25.
37 Pandey, A., P. Nigam, V. T. Soccol, D. Singh, and R. Mohan. 2000. Advances in microbial amylases. Biotechnol. Appl. Biochem. 31: 135-152.   DOI   ScienceOn
38 Prasad, D. Y., J. A. Heitmann, and T. W. Joyce. 1993. Enzymatic deinking of colored offset newsprint. Nordic Pulp Paper Res. J. 8: 284-286.   DOI   ScienceOn
39 Reyed, R. M. 2007. Biosynthesis and properties of extracellular amylase by encapsulation Bifidobatrium bifidum in batch culture. Aust. J. Basic Appl. Sci. 1: 7-14.
40 Sambrook, J. and D. W. Russell. 2001. Molecular Cloning: A Laboratory Manual, 3rd Ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
41 Schokker, E. P. and A. J. S. van Boekel. 1999. Kinetics of thermal inactivation of the extracellular proteinase from Pseudomonas fluorescens 22F: Influence of pH, calcium and protein. J. Agric. Food Chem. 47: 1681-1686.   DOI   ScienceOn
42 Teotia, S., R. Lata, S. K. Khare, and M. N. Gupta. 2001. Onestep purification of glucoamylase by affinity precipitation with alginate. J. Mol. Recognit. 14: 295-299.   DOI   ScienceOn
43 Van der Maarel, M. J., B. van der Veen, J. C. Uitdehaag, H. Leemhuis, and L. Dijkhuizen. 2002. Properties and applications of starch-converting enzymes of the $\alpha$-amylase family. J. Biotechnol. 94: 137-155.   DOI   ScienceOn