Effect of temperature, pH and metal ions on amylase produced from selected indigenous extremophile bacteria in Pakistan

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Research Paper 01/09/2018
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Effect of temperature, pH and metal ions on amylase produced from selected indigenous extremophile bacteria in Pakistan

Farkhanda Kalsoom, Faisal Rasheed Anjum, Sidra Anam, Samiullah Khan, Fariha Hasan
Int. J. Biosci.13( 3), 262-275, September 2018.
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There is a burgeoning demand for amylase production due to wide range applications of amylase in different industrial processes like saccharification of starchy materials, food, detergents and textile industries. But high cost of fermentation media is one of the technical barriers in amylase production from microbial sources. Extremophiles microorganisms (thermophilic and halophilic) could be potential source for thermostable amylase. Present study deals with isolation of extremophilic amylase-producing bacteria from soil samples in starch agar medium and their subsequent identification through 16sRNA analysis. Different parameters like pH, temperature, and metal ions concentration (Ca+2, Mn+2, Fe+2, Zn+2) were optimized for amylase production. Five thermophilic strains and one halophilic strain were found positive for amylase production. Phylogenetic analysis showed that the amylase producing thermophilic strains includes Bacillus spp., Rheinheimera spp., Alishewanella spp., Pseudomonas spp., Microbacterium spp. while halophilic strain includes Bacillus spp. Thermophilic strains showed optimum amylase production at pH of 8 & 60°C, while maximum amylase activity for halophilic strains was observed at pH 7 and 40°C. Divalent ions Ca+2 and Mn+2 enhanced the amylase production while Zn and Fe did not a have any significant effect. Current research revealed that use of extremophile bacteria could be an important step towards the development of environmental friendly and cost effective process for thermostable amylase production.


Afifi AF, Kamel EM, khalil AA,  Foaad MA, Fawziand EM, Houseny M. 2008. Purification and characterization of a-amylase from penicillium olsonii under the effect of some antioxidant vitamins. Global Journal of Biotechnology and Biochemistry 3, 14-21. https://doi.org/10.1155/2013/527570

Asad W, Asif M, Ajaz RAS. 2011. Extracellular enzyme production by indigenous the rmophilic bacteria: Partial purification and characterization of α-amylase by Bacillus sp. Wa21. Pakistan Journal of Botany 43, 1045–1052. https://doi.org/10.1016/j.jgeb.2015.09.004

Autha K, Priya KJ. 2011. Effect of pH, temperature and metal ions on amylase activity Bacillus subtilis IJPBS.

Božic N, Ruiz J, López-Santín J, Vujˇci´c Z. 2011. Production and properties of the highly effic ient raw starch digesting α-amylase from a Bacillus licheniformis ATCC9945a. Biochemical Engineering Journal 53, 203–209. https://doi.org/10.1016/j.bej.2010.10.014

Burhan A, Nisa U, Gokhan C, Omer C, Ashabil A, Osman G. 2003. Enzymatic properties of a novel thermostable, thermophilic, alkaline and chelator resistant amylase from an alkaliphilic Bacillus spp. isolate ANT-6. Process Biochemistry38, 1397-03. https://doi.org/10.1208/s12249-011-9586-1

Coronado M, Vargas C, Hofemeister J, Ventosa A, Nieto JJ. 2000. Production and biochemical characterization of an alpha-amylase from the moderate  halophile Halomonas meridiana. FEMS Microbiology Letters 183, 67-71. https://doi.org/10.17113/ftb.

Dar GH, Kamili AN, Nazir R, Bandh SA, Jan TR, Chishti MZ. 2015. Enhanced production of α-amylase by penicillium chrysogenum in liquid culture by modifying the process parameters. Microbial Pathogenesis 88, 10–15. https://doi.org/10.1016/j.micpath.2015.07.016

Enache M, Kamekura M. 2010. Hydrolytic enzymes of halophilic microorganisms and their economic values. Romanian Journal of Biochemistry 47, 46–59.

Gomes I, Gomes J, Steiner W. 2003. Highly thermostable amylase and pullulanase  of the extreme thermophilic eubacterium Rhodothermus marinus: production and partial characterization. Bioresource Technology 90, 207-14. https://doi.org/10.1016/S0960-8524(03)00110-X

Goyal N, Gupta J, Soni S. 2005. A novel raw starch digesting thermostable α-amylase from Bacillus sp. I-3 and its use in the direct hydrolysis of raw potato starch. Enzyme and Microbial Technology 37, 723–734. https://doi.org/10.1016/j.enzmictec.2005.04.017

Gupta R, Gigras P, Mohapatra H, Goswami VK, Chauhan B. 2003. Microbial α-amylases: a biotechnological perspective. Process Biochemistry 38, 1599-1616. https://doi.org/10.1016/S0032-9592(03)00053-0.

Jaspreet SJ, Kaur L, McCarthy OJ. 2007. Factors influencing the physico-chemical, morphological, thermal and rheological properties of some chemically modified starches for food application. Food Hydrocolloids 21, 1-22. https://doi.org/10.1016/j.foodhyd.2006.02.006

Kamekura M. 2011. Physiology of halophilic eubacteria. Journal of Biochemistry 36, 83–92.

Leveque E, Janeek S, Haye B, Belarbi A. 2000. Thermophilic Archaeal Amylolytic EnzymesEnzyme and Microbial Technology 26, 314. https://doi.org/10.1016/S0141-0229(99)00142-8

Muralikrishna G, Nirmala M. 2005. Cereal alpha-amylases (overview).Carbohydrate Polymers 60, 163-173. https://doi.org/10.1016/j.carbpol.2004.12.002.

Nouadri T, Meraihi Z, Shahrazed D, Leila B. 2010. Purification and characterization of the α-amylase isolated from Penicillium camemberti PL21. African Journal of Biochemical Research 4,155-162.

Prakash O, Jaiswal N. 2009. Alpha-Amylase: An Ideal Representative of Thermostable Enzymes. Applied Biochemistry and Biotechnology 160, 2401-2414. https://doi.org/10.1007/s12010-009-8735-4.

Qureshi AS, Bhutto MA, Chisti Y, Khushk I, Dahot MU, Bano S. 2012. Production of pectinase by Bacillus subtilisfrl 01 in a date syrup medium. African Journal of Biotechnology 11, 12563–12570.

Qureshi AS, Dahot MU, Panhwar SI. 2010. Biosynthesis of alkaline phosphatase by Escherichia coliEFRL 13 in submerged fermentation. World Applied Sciences Journal 8, 50–56. https://doi.org/10.1016/j.enzmictec.2004.11.017

Qureshi AS, Dahot MU. 2009. Production of proteases by staphylococcus epidermidis EFRL 12 using cost effective substrate (molasses) as a carbon source. Pakistan Journal of Biotechnology 6, 55–60. https://doi.org/10.5897/AJB2015.15174

Ramesh MV, Lonsane BK. 1989. Solid state fermentation for production of highertiters of thermostable alpha-amylase with two peaks for pH optima by Bacillus licheniformis M27. Biotechnology letters 11, 49-52.

Simair AA, Khushk I, Qureshi AS, Bhutto MA, Chaudhry HA, Ansari KA, Lu C. 2017. Amylase Production from Thermophilic Bacillus sp. BCC 021-50 Isolated from a Marine Environment. Fermentation 3, 25. d https://doi.org/10.3390/fermentation3020025

Stamford TL, Stamford NP, Coelho LC, Araujo JM. 2001. Production and characterization of a thermostable alpha-amylase from Nocardiopsis sp. endophyte of yam bean. Bioresource Technology 76, 137-141. https://doi.org/10.1016/S0960-8524(00)00089-4

Sterner R, Liebl W. 2001. Thermophilic adaptation of proteins. Thermophilic adaptation of proteins. Critical Reviews in Biochemistry and Molecular Biology 36, 39-106. https://doi.org/10.1080/20014091074174

Tapan KD, Malabendu J, Priti R, Pahari T. 2006. The Effect of Temperature, pH, and Salt on  Amylase in Heliodiaptomus viduus. Turkish Journal of Zoology 30, 187-195. https://doi.org/10.1080/09291010802402196

Whitcomb DC, Lowe ME. 2007. Human  pancreatic digestive enzymes. Digestive  Diseases and Sciences 52, 1-17. https://doi.org/10.1007/s10620-006-9589-z

Yasser RASoliman NA, Nabil ME, El-Gendi H, Rania SA. 2013. Production, Purification, and Characterization of Thermos table α-Amylase produced by Bacillus licheniformis Isolate AI20. Journal of Chemistry 1, 10-21.