Volume 4, Issue 4 (3-2018)                   nbr 2018, 4(4): 288-298 | Back to browse issues page

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Afrisham S, Badoei-delfard A, Namaki Shoushtari A, Karami Z, Malek-abadi S. Isolation and identification of Bacillus producing thermophilic alpha amylase: production optimization and investigation of the activity and stability of enzyme.. nbr 2018; 4 (4) :288-298
URL: http://nbr.khu.ac.ir/article-1-3023-en.html
Isolation and identification of Bacillus producing thermophilic alpha amylase: production optimization and investigation of the activity and stability of enzyme.
Saide Afrisham * , Arastoo Badoei-delfard , Abdolhamid Namaki Shoushtari , Zahra Karami , Saeid Malek-abadi
Abstract:   (6874 Views)
Alpha-amylases are the most important amylases used in industry. Among them, thermophilic alpha-amylases are of particular importance, which is due to their activity and stability in high temperatures. These enzymes produced by thermophile micro-organisms including bacteria. These thermophilic alpha-amylases are used in various industries such as processing of starch as well as production of detergents and biofuels. In this research, the bacteria which produce the thermophilic alpha-amylases were isolated and characterized in hot springs of Gorooh village in Kerman province. According to the results of screening on the specific liquid and solid media, AT59 was selected as the best strain. Morphological and biochemical characterization of the isolated strain indicated that it belonged to Bacillus sp. and was gram-positive, catalase positive, casein hydrolyzing and acid producing from lactose and sucrose. The results obtained from the optimization of the enzyme production medium showed that among the carbon, nitrogen and ion sources investigated, starch (1 gr/l), gelatin (2 g/l) and magnesium sulfate (1 g/l) had the most increasing effect on the production of AT59 alpha-amylase. Moreover, the highest enzyme production was obtained at pH 5. This enzyme also demonstrated the highest degree of activity and stability in 80 and 70 ℃, respectively. These findings suggested that this enzyme has a considerable potential for use in starch industry.
 

 
 
Keywords: hot spring, production, screening, thermophilic enzymes
Full-Text [PDF 497 kb]   (3728 Downloads)    
Type of Study: Original Article | Subject: Microbiology
Received: 2016/08/19 | Revised: 2021/05/23 | Accepted: 2017/11/25 | Published: 2018/03/20 | ePublished: 2018/03/20
References
1. Abdel-Fattah, Y.R., Soliman, N.A., El-Toukhy, N.M., El- Gendi, H. and Ahmed, R.S. 2012. Production, purification, and characterization of thermostable alpha-amylase produced by Bacillus licheniformis isolate AI20. – J. Chem. 13: 1-11.
3. Afrisham, S., Badoei-Dalfard, A., Namaki-Shoushtari, A. and Karami, Z. 2016. Characterization of a thermostable, CaCl2-activated and raw-starch hydrolyzing alpha-amylase from Bacillus licheniformis AT70: Production under solid statef-ermentation. – J. Mol. Catal. B: Enzym. 132: 98-106.
4. Ahmadi, A., Ghobadi, S., Khajeh, K., Nomanpour, B. and Badoei-Dalfard, A. 2010. Purification of α-Amylase from Bacillus sp. GHA1 and its partial characterization. – J. Iran Chem. Soc. 7: 432-440.
5. Aiyer, P.D. 2005. Effect of C: N ratio on alpha-amylase production by Bacillus licheniformis SPT 27. – African J. Biotech. 3: 519-522.
6. Amoozegar, M.A., Samareh-Abolhasani, B., Shafiei, M., Didari, M. and Hamedi, J. 2013. Production of halothermotolerant alpha-amylase from a moderately halophilic bacterium, Nesterenkonia Strain F. – Progress in Biological Sciences. 2: 85-97.
7. Antranikian, G. and Egorova, K. 2007. Extremophiles, a unique resource of biocatalysts for industrial biotechnology. Extremophiles. – ASM Press, Washington 361-406.
8. Asgher, M., Asad, M.J., Rahman, S.U. and Legge, R.L. 2007. A thermostable α-amylase from a moderately thermophilic Bacillus subtilis strain for starch processing. – J. Food Eng. 79: 950-955.
9. Asoodeh, A., Alemi, A., Heydari, A. and Akbari, J. 2013. Purification and biochemical characterization of an acidophilic amylase from a newly isolated Bacillus sp. DR90. – Extremophiles 17: 339-348.
10. Aullybux, A.A. and Puchooa, D. 2013. Alpha-amylase production on low-cost substrates by Naxibacter sp. isolated from Mauritian soils. – Br. Microbiol. Res. J. 3: 478-491.
11. Azadian, F., Badoei-Dalfard, A., Namaki-Shoushtari, A. and Hassanshahian, M. 2016. Purification and biochemical properties of a thermostable, haloalkaline cellulase from Bacillus licheniformis AMF-07 and its application for hydrolysis of different cellulosic substrates to bioethanol prod-uction. – Mol. Bio. Res. Comm. 5: 143-155.
12. Babu, K.R. and Satyanarayana, T. 1995. Alpha-amylase production by thermophilic Bacillus coagulans in solid state fermentation. – Process Biochem. 30: 305-309.
13. Badoei-Dalfard, A., Karami, Z., Ramezani-Pour, N. 2016. Bench scale production of nicotinic acid using a newly isolated Stenotrophomonas maltophilia AC-21 producing highly-inducible and versatile nitrilase. – J. Mol. Catal. B: Enzym. 133: 552-559.
14. Badoei-Dalfard, A., Karami, Z., Ramezani-pour, N. 2016. Production and characterization of a nitrilase from Pseudomonas aeruginosa RZ44 and its potential for nitrile biotransformation. – Iranian J. Biotechnol. 14: 142-153.
15. Baysal, Z., Uyar, F. and Aytekin, Ç. 2003. Solid state fermentation for production of alpha-amylase by a thermotolerant Bacillus subtilis from hot-spring water. – Process Biochem. 38: 1665-1668.
16. Bernfeld, P. 1955. Amylases, α and β. – Methods Enzy-mol. 1: 149-158.
17. Bouzas, T.M., Barros-Velázquez, J. and Gonzalez Villa, T. 2006. Industrial applications of hyperthermophilic enzymes: a review. – Protein Pept. Lett. 13: 645-651.
18. Bozic, N., Ruiz, J., Santin, J., and Vujcic, Z. 2011. Optimization of the growth and alpha-amylase production of Bacillus subtilis IP 5832 in shake flask and laboratory fermenter batch cultures. – J. Serb. Chem. Soc. 76: 965-972.
19. Burhan, A., Nisa, U., Gökhan, C., Ömer, C., Ashabil, A., and Osman, G. 2003. Enzymatic properties of a novel thermostable, thermophilic, alkaline and chel-ator resistant amylase from an alkaliphilic Bacillus sp. isolate ANT-6. – Process Biochem. 38: 1397-1403.
20. Chandra, M.S., Mallaiah, K.V., Sreenivasulu, P. and Choi, Y.L. 2010. Purification and characterization of highly thermostable alpha-amylase from thermophilic Alicyclobacillus acidocaldarius. Biotechnol. – Bioprocess Eng. 15: 435-440.
21. Fooladi, J. and Sajjadian, A. 2010. Screening the thermophilic and hyperthermophilic bacterial popu-lation of three Iranian hot-springs to detect the therm-ostable alpha-amylase producing strain. – Iran J. Med. Microbiol. 2: 46-50.
22. Gangadharan, D., Sivaramakrishnan, S., Nampoothiri, K.M. and Pandey, A. 2006. Solid culturing of Baci-llus amyloliquefaciens for alpha-amylase production. – Food Technol. Biotechnol. 44: 269-274.
23. Haki, G.D. and Rakshit, S.K. 2003. Developments in industrially important thermostable enzymes: a revi-ew. – Bioresour. Technol. 89: 17-34.
24. Karakaş, B., İnan, M. and Certel, M. 2010. Expression and characterization of Bacillus subtilis PY22 alpha-amylase in Pichia pastoris. – J. Mol. Catal. B: Enzym. 64: 129-134.
25. Kolcuoğlu, Y., Colak, A., Faiz, O. and Belduz, A.O. 2010. Cloning, expression and characterization of highly thermo-and pH-stable maltogenic amylase from a thermophilic bacterium Geobacillus caldoxyl-osilyticus TK4. – Process Biochem. 45: 821-828.
26. Kristjanson K.J. 1989. Thermophilic organisms as sour-ce of thermostable enzymes. – Trends Biotechnol. 7: 49-53.
27. Mahdavi, A., Hassan Sajedi, R., Rassa, M. and Jafarian, V. 2010. Characterization of an alpha-amylase with broad temperature activity from an acid-neutralizing Bacillus cereus strain. – Iran J. Biotechnol. 8: 103-111.
28. Miller, G.L. 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. – Anal. Chem. 31: 426-428.
29. Oziengbe, E.O. and Onilude, A.A. 2012. Production of a thermostable alpha-amylase and its assay using Bacillus licheniformis isolated from excavated land sites in Ibadan, Nigeria. – Bayero Journal of Pure and Applied Sciences 5: 132-138.
30. Pandey, A., Soccol, C.R., Rodriguez-Leon, J.A. and Nigam, P. 2001. Solid state fermentation in biotec-hnolog'y. – J. Microbiol. Methods 34: 405-423.
31. Prakash, B., Vidyasagar, M., Madhukumar, M.S., Muralikrishna, G. and Sreeramulu, K. 2009. Produ-ction, purification, and characterization of two extremely halotolerant, thermostable, and alkali-stable apha-amylases from Chromohalobacter sp. TVSP 101. – Process Biochem. 44: 210-215.
32. Ramachandran, S., Patel, A.K., Nampoothiri, K.M., Francis, F., Nagy, V., Szakacs, G. and Pandey, A. 2004. Coconut oil cake-a potential raw material for the production of alpha-amylase. – Bioresour. Tech-nol. 93: 169-174.
33. Ramezani-Pour, N., Badoei-Dalfard, A., Namaki-Shou-shtari, A. and Karami, Z. 2015. Nitrile-metabolizing potential of Bacillus cereus strain FA12; Nitrilase production, purification, and characterization. – Bioc-atal. Biotransform. 33: 156-166
34. Samie, N., Noghabi, K.A., Gharegozloo, Z., Zahiri, H.S., Ahmadian, G., Sharafi, H., Behrozi, R. and Vali, H. 2012. Psychrophilic alpha-amylase from Aeromonas veronii NS07 isolated from farm soils. – Process Bi-ochem. 47: 1381-1387.
35. Sen, S.K., Raut, S., Satpathy, S., Rout, P.R., Bandyopadhyay, B. and Mohapatra, P.K.D. 2014. Characterizing novel thermophilic amylase producing bacteria from Taptapani hot spring, Odisha, India. – Jundishapur. J. Microbiol. 7: 1-7.
36. Shafiei, M., Ziaee, A.A. and Amoozegar, M.A. 2012. Purification and characterization of a halophilic alpha-amylase with increased activity in the presence of organic solvents from the moderately halophilic Nesterenkonia sp. strain F. – Extremophiles 16: 627-635.
37. Sneath, P.H.A., Mair, N.S., Sharpe, M.E. and Holt, J.G. 1986. Bergy’s manual of systematic bacteriology, Vol. 2. – William and Wilkins, Baltimore, USA.
38. Sodhi, H.K., Sharma, K., Gupta, J.K. and Soni, S.K. 2005. Production of a thermostable alpha-amylase from Bacillus sp. PS-7 by solid state fermentation and its synergistic use in the hydrolysis of malt starch for alcohol production. – Process Biochem. 40: 525-534.
39. Souza, P.M.D. 2010. Application of microbial alpha-amylase in industry-a review. – Braz. J. Microbiol. 41: 850-861.
40. Tamura, K., Dudley, J., Nei, M. and Kumar, S. 2007. MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. – Mol. Biol. Evol. 24: 1596-1599.
41. Zeikus, J.G., Vieille, C. and Savchenko, A. 1998. Thermozymes: biotechnology and structure–function relationships. – Extremophiles 2: 179-183.
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