Synthesis of Thermostable Amylase Enzyme from Bacillus licheniformis and it's Optimization to Different Sources

Abstract:

S: - Among the six isolated amylase producing strains, Bacillus licheniformis (B1), the thermostable strain was selected from the soil of a paddy field. Its enzyme productivity and activity were evaluated. The activity of enzyme was calculated as 27.77 IU/ml. Effects of various carbon and organic nitrogen sources, and C/N ratio on enzyme production were examined. Maximum α - amylase production was obtained in medium containing 1% starch. Fructose supported the maximum amylase production among all the sugar studied. Of the organic nitrogen sources tested, peptone was found to be the best organic nitrogen source for excess yield of the enzyme. The optimum C/N ratio was found to be 1:1. The α amylase exhibited activity at a wide pH and temperature range and activity were found to be optimal at pH 6 and 40 °C respectively. The molecular weight of α amylase was calculated by sodium dodecyl sulphate gel electrophoresis and found to be around 29,000 Daltons.

• Periodical: Materials Science Forum (Volume 760)
• Pages: 73-78
• DOI: https://doi.org/10.4028/www.scientific.net/MSF.760.73
• Online since: July 2013
• Authors: Anima Nanda, T. Sudhakar, B.K. Nayak, J. Prem Kumar
• Keywords: Bacillus licheniformis, Thermostable α-Amylase

References

[1] Abouzied M.and Reddy M.1986. Direct fermentation of Potato Starch to ethanolby cocultures of Aspergillus niger and Saccharmyces cerevisiae. Appl. Environ. Microbiol. 52:1055-1059.

DOI: 10.1128/aem.52.5.1055-1059.1986

[2] Aiyer P.V. D. 2004. Effect of C:N ratio on alpha amlase production by Bacillus licheniformis spt 27. African Journal of Biotechnology. 3:519-522

DOI: 10.5897/ajb2004.000-2103

[3] Bergman F.W., Abe J, Hizukurji S. 1988. Selection of microorganism which produce raw starch degrading amylase. Appl. Microbiol. Biotechno. 27:443-446.

DOI: 10.1007/bf00451610

[4] Burbidge E. and Collier B. 1968. Production of bacterial amylases. Process Biochem. 3:53-56.

[5] Dobreva Elena. 1994. Thermostabile -amylase production by immobilized Bacillus licheniformis cells in agar gel and on acrylonitrile/acrylamide membranes Appl. Мicrobiol. Biotechnol., 41: 517-523.

DOI: 10.1007/bf00178482

[6] Ekunsumi Toye 2002. Laboratory Production and assay of amylase by bacteria and fungi.1:1-8

[7] Gangadharan D., Sivaramakrishnan S., Madhavan K. N. and Pandey A. 2006 Alpha- Amylase production by B. amyloliquefaciens. Food Techno Biotechnol 44: 269-274.

[8] Guchte Van de, Kodde M.J., Vander Vossen J.M.B.M., Kok J. and Venema G. 1990. Heterologous gene expression in Lactococcus lactis sub sp. lactis: synthesis, secretion and processing of the Bacillus subtilis neutral protease. Appl. Environ, Microbiol. 56:2606-2611.

DOI: 10.1128/aem.56.9.2606-2611.1990

[9] Hols P.T., Ferain D., Garmyn N., Bernard, A. and Delcour J. 1994. Use of homologous expression. Secretion signals and vector – free stable chromosomal integration 'n' engineering of Lactobacillus plantarum for Alpaamylase and levanase expression. Appl.Environ-Microbiol. 60:1401- 1413.

DOI: 10.1128/aem.60.5.1401-1413.1994

[10] Krishnan T. and Chandra A. K. 1983. Purification and Characterization of -Amylase from Bacillus licheniformis CUMC 305. Appl Environ Microbiol. 46(2): 430-437

DOI: 10.1128/aem.46.2.430-437.1983

[11] Lealem F.and Gashe B.A. 1994. Amylase Production by gram Positive bacteria isolated from fermenting tef (Eragrostis tef) Journal of Applied Microbiology. 86:557-560

DOI: 10.1111/j.1365-2672.1994.tb03084.x

[12] Mamo G., Gashe B.A. and Gessesse A. 2001. A highly thermostable amylase from a newly isolated thermophilic Bacillus sp. WN 11. Journal of Applied Microbiology. 86:557-560.

DOI: 10.1046/j.1365-2672.1999.00685.x

[13] Medda S. and Chandra A. K. 2008. New Strains of Bacillus licheniformis and Bacillus coagulans producing Thermostable α-Amylase Active at Alkaline pH.Journal of Applied Microbiology. 48 (1): 47 – 58

DOI: 10.1111/j.1365-2672.1980.tb05205.x

[14] Nigam. P. and Singh D. 1995. Enzyme and microbial system involved in starch processing. Enzyme Microbe Techno. 17:770-778.

DOI: 10.1016/0141-0229(94)00003-a

[15] Rao D. M., Purnima A. , Ramesh D.V. and Ayyanna C. 2002. Purification of α- amylase from Bacillus licheniformis by chromatofocusing and gel filtration chromatography. World Journal of Microbiology and Biotechnology. 18: 959- 993.

[16] Rothstein D. M., Devlin P. E., and Cate R. L. 1986. Expression of alphaamylase in Bacillus licheniformis. J. Bacteriol. 168(2): 839–842.

DOI: 10.1128/jb.168.2.839-842.1986

[17] Saito N.K. Yamamoto.1975.Regulatory factors affecting Alpha-amylase production in Bacillus licheniformis. Journal of Bacteriology. 121:848-856.

DOI: 10.1128/jb.121.3.848-856.1975

[18] Thaddeus C., Ezeji and Hubert Bahl. 2006. Purification, characterization and synergistic action of phytate-resistant α-amylase and α-glucosidase from Geobacillus thermodenitrificans HRO10. Journal of Biotechnology. 125: 27-38

DOI: 10.1016/j.jbiotec.2006.02.006

[19] Yamane K, Hirata Y., Furusato J., Yamazaki H. and Nakayama A. 1984. Changes in the properties and molecular weight of Bacillus subtilis M-type and N-type Alpha-amylases resulting from a spontaneous deletion. J. Biochem. 96:1849-1858.

DOI: 10.1093/oxfordjournals.jbchem.a135019