Research Article | Volume: 6, Issue: 3, May-June, 2018

Characterization of extracellular amylase from Bacillus sp. strain RU1

Aleem Basha Pinjari Vijayalakshmi Kotari   

Open Access   

Published:  Apr 05, 2018

DOI: 10.7324/JABB.2018.60305

The present study reveals the isolation of amylolytic bacteria from the industrially contaminated soils collected around Kurnool district, Andhra Pradesh, India. A bacterium belonging to genus Bacillus was isolated by serial dilution method. The bacterium was designated as Bacillus sp. strain RU1. Compared to others, strain RU1 has shown maximum amylase activity with starch as substrate. Various parameters have been optimized for enhancing the production of amylase which includes carbon sources, nitrogen sources, substrate concentration, and inoculum size. The results have shown fructose and ammonium carbonate are the best carbon and nitrogen sources for amylase production. Crude amylase enzyme studies revealed the optimum temperature and pH as 45°C and 10.0. Therefore, the amylase enzyme isolated in the present study will find applications in the industry as the enzyme is active at moderately thermophilic and at alkaline pH.

Keyword:     Bacillus sp. Extracellular amylase Characterization Carbon sources Temperature pH.


Pinjari AB, Kotari V. Characterization of extracellular amylase from Bacillus sp. strain RU1. J App Biol Biotech. 2018;6(3):29-34. DOI: 10.7324/JABB.2018.60305

Copyright: Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license.

HTML Full Text

1. Pandey A, Nigam P, Soccol CR, Soccol VT, Singh D, Mohan R, et al. Advances in microbial amylases. Biotechnol Appl Biochem 2000;31:135-52.

2. Schmidt S, Martin A, Schadt C, Lipson D, Meyer A, Costello E, et al. Profound Seasonal Changes in Microbial Diversity and Function in an Alpine Environment. New York: Mc-Graw Hill; 2005.

3. Sindhu MK, Singh BK, Prased T. Changes in starch content of anhar seed due to fungal attack. Phytopathology 1997;34:269-71.

4. Rao MB, Tanksale AM, Ghatge MS, Deshpande VV. Molecular and biotechnological aspects of microbial proteases. Microbiol Mol Biol Rev 1998;62:597-635.

5. Ashwini K, Gaurav K, Karthik L, Bhaskara Rao KV. Optimization, production and partial purification of extracellular α-amylase from Bacillus sp. Marini. Arch Appl Sci Res 2011;3:33-42.

6. Lévêque E, Janeček Š, Haye B, Belarbi A. Thermophilic archaeal amylolytic enzymes. Enzyme Microb Technol 2000;26:3-14.

7. Vijayalakshmi, Sushma K, Abha S, Chander P. Isolation and characterization of Bacillus subtilis KC3 for amylolytic activity. Int J Biosci Biochem Bioinf 2012;2:336.

8. Gupta R, Gigras P, Mohapatra H, Goswami VK, Chauhan B. Microbial α-amylases: A biotechnological perspective. Process Biochem 2003;38:1599-616.

9. Nielsen JE, Borchert TV. Protein engineering of bacterial alpha-amylases. Biochim Biophys Acta 2000;1543:253-74.

10. Sivaramakrishan S, Gangadharan D, Nampoothiri KM, Soccol CR, Pandey A. α-amylases from microbial sources-an overview on recent developments. Food Technol Biotechnol 2006;44:173-84.

11. Mishra S, Behera N. Amylase activity of a starch degrading bacteria isolated from soil receiving kitchen wastes. Afr J Biotechnol 2008;7:3326-31.

12. Holt JG. Gram-positive cocci. Bergey's Manual of Determinative Bacteriology. Maryland: Williams and Wilkins; 1994:527-58.

13. Bernfeld P. Amylase, α and β. Methods Enzymol 1955;1:149-51.

14. Van Der Maarel MJ, Van Der Veen B, Uitdehaag JC, Leemhuis H, Dijkhuizen L. Properties and applications of starch-converting enzymes of the α-amylase family. J Biotechnol 2002;94:137-55.

15. Grata K, Nabrdalik M, Latała A. Effect of different environmental factors on amylolytic activity of Bacillus cereus and Bacillus mycoides. Proc ECOpole 2010;4:49-53.

16. Salva TD, Moraes IO. Effect of the carbon source on alpha-amylase production by Bacillus subtilis BA-04. Rev Microbiol 1995;26:46-51.

17. Aqeel BM, Umar DM. Effect of alternative carbon and nitrogen sources on production of alpha-amylase by Bacillus megaterium. J Biotechnol 2008;136:S293.

18. Suman S, Ramesh K. Production of a thermostable extracellular amylase from thermophilic Bacillus species. J Pharm Sci Res 2010;2:149-54.

19. Lin LL, Chyau CC, Hsu WH. Production and properties of a raw-starch-degrading amylase from the thermophilic and alkaliphilic Bacillus sp. TS-23. Biotechnol Appl Biochem 1998;28:61-8.

20. Forgaty W, Kelly C. Amylase, amyloglucosidase and related gluconases. Microbial Enzymes and Bioconversions Economic Microbiology. Vol. 5. London: Academic Press; 1980. p. 115-70.

21. van Hille RP, Bromfield LV, Botha SS, Jones G, van Zyl AW, Harrison ST. The effect of nutrient supplementation on growth and leaching performance of bioleaching bacteria. Adv Mater Res 2009;71:413-6.

22. Rasooli I, Astaneh SD, Borna H, Barchini KA. A thermostable α-amylase producing natural variant of Bacillus spp. isolated from soil in Iran. Am J Agric Biol Sci 2008;3:591-6.

23. Dash BK, Rahman MM, Sarker PK. Molecular identification of a newly isolated Bacillus subtilis BI19 and optimization of production conditions for enhanced production of extracellular amylase. Biomed Res Int 2015;2015:859805.

24. Ikram-Ul-Haq NS, Shamim N, Ashraf H, Ali S, Qadeer M. Effect of surfactants on the biosynthesis of alpha amylase by Bacillus subtilis GCBM-25. Pak J Bot 2005;37:373-9.

25. Raza SK. Effect of Pigment Volume Concentration on the Enzyme Activity of Bioactive Coatings. Karlstad: Master of Science in Engineering, Faculty of Technology and Science, Karlstads University; 2011. p. 66.

26. Swain MR, Kar S, Padmaja G, Ray RC. Partial characterization and optimization of production of extracellular alpha-amylase from Bacillus subtilis isolated from culturable cow dung microflora. Polish J Microbiol 2006;55:289-96.

27. Malhotra R, Noorwez SM, Satyanarayana T. Production and partial characterization of thermostable and calcium-independent α-amylase of an extreme thermophile Bacillus thermooleovorans NP54. Lett Appl Microbiol 2000;31:378-84.

28. Alk A, Ibrahim HM. A potential new isolate for the production of a thermostable extracellular-amylase. Afr J Bacteriol Res 2011;3:129-37.

29. Burhan A, Nisa U, Gökhan C, Ömer C, Ashabil A, Osman G. Enzymatic properties of a novel thermostable, thermophilic, alkaline and chelator resistant amylase from an alkaliphilic Bacillus sp. Isolate ANT-6. Process Biochem 2003;38:1397-403.

30. Vaseekaran S, Balakumar S, Arasaratnam V. Isolation and identification of a bacterial strain producing thermostable a-amylase. Trop Agric Res 2010;22:1-11.

Article Metrics
307 Views 95 Downloads 402 Total



Related Search

By author names

Similar Articles

Optimization production of thermo active levansucrase from Bacillus subtilis Natto CCT 7712

Bruna Caroline Marques Gonçalves, Janaina Mantovan, Mara Lúcia Luis Ribeiro, Dionísio Borsato, Maria Antonia Pedrine Colabone Celligoi

Molecular Characterization of some Date palms in Nigeria Using RAPD Markers

Beatrice O. Emoghene, Omorefe Asemota, MacDonald Idu, Chukwuemeka R. Eke

Submerged fermentation, purification, and characterization of L-asparaginase from Streptomyces sp. isolated from soil

Savitha S. Desai, Basavaraj S. Hungund

Purification and characterization of tannase from the local isolate of Aspergillus niger

Saja Taha Yaseen Al-Mraai, Dhia Falih Al-Fekaiki, Alaa Jabbar Abd Al-Manhel

Kinetic and thermodynamic parameters of four acid phosphatases from edible seeds of oleaginous cucurbit Lagenaria siceraria (Molina) Standl.

Djary Michel Koffi, Jean Bedel Fagbohoun

Partial purification and characterization of antimicrobial peptide from the hemolymph of cockroach Periplaneta americana

Reymend Encily Martin, Yogita Raosaheb Channe

Purification and characterization of an iron-activated alkaline phosphatase produced by Rhizopus microsporus var. microsporus under submerged fermentation using rye flour

Pedro Henrique de Oliveira Ornela, Luis Henrique Souza Guimarães

Purification and biochemical characterization of protease from the seeds of Cyamopsis tetragonoloba

Rajesh Kumar Rawaliya, Preeti Patidar, Sadhana Sharma, Krishnan Hajela

Cloning and expression of a GH11 xylanase from Bacillus pumilus SSP-34 in Pichia pastoris GS115: Purification and characterization

Sagar Krishna Bhat,, Kavya Purushothaman, Appu Rao Gopala Rao Appu Rao, K Ramachandra Kini

Media optimization for the production of alkaline protease by Bacillus cereus PW3A using response surface methodology

Gururaj B. Tennalli, Soumya Garawadmath, Lisa Sequeira, Shreya Murudi, Vaibhavi Patil, Manisha N. Divate, Basavaraj S. Hungund

Isolation and partial characterization of serine proteases from jellyfish of the Antarctic region

Nataliia Raksha, Teyiana Halenova, Tetiana Vovk, Tetiana Beregova, Tetiana Maievska, Victor Tomchuk, Olexiy Savchuk, Ludmila Ostapchenko

The physicochemical and biological properties of novel silver nanoparticles synthesized by the extract of Holigarna ferruginea

Kumbar Mudakappa Manjunath, Y. L. Krishnamurthy

Synthesis of bio-nanofiber from pectin/polyvinyl alcohol for therapeutic application

Nagula Harshith, Palleti Sai Yogesh, Kuttikattu Narayanan Kutty Bibin, Ramachandran Rabin, Ganesan Anusiya, Rengarajan Jai Ganesh

Assessment of bioactivity of the novel exopolysaccharide secreted by Bacillus subtilis isolated from the gut of marine anchovies

Thejaswi Bhandary, Paari Kuppusamy Alagesan

Characterization of an alkaline endo-α-1-4-amylase from alkaliphilic Bacillus halodurans LBW 5117 and demonstration of its desizing potential

Kevin Raymond Oluoch, Edward Kirwa Muge, Yvonne Wanjiku Mwangi, Francis Jakim Mulaa

Thermal treatment and inhibition study with non-toxic product of polyphenol oxidase from Mediterranean Palm heart (Chamaerops humilis L.)

Abdelaziz Bouchaib, Abdellatif Ben Abdellah, Tarik Chafik, Amine Laglaoui, Abdelhay Arakrak, Mohamed Bakkali, Abdelaziz Benjouad, Amal Maurady

Phenotypic characterization and genetic diversity of the Khiew-Phalee chicken (Gallus gallus): A fighting cock originating from Uttaradit, Thailand

Siriwadee Phromnoi, Preeda Lertwatcharasarakul, Wallaya Phongphaew, Pisit Poolprasert

Agronomic and disease responses of three watermelon (citrilus lanatus l.) varieties to fungicide spraying regimes in a tropical environment

Nathaniel Dauda , Sandra Nneamaka Ugwuagu, Patience Ukamaka Ishieze, Kevin Ugwuoke, Vivian Ogechi Osadebe, Solomon Oluwaseyi Adewuyi, Uchenna Noble Ukwu

Amplification and cloning of mcrAgene from enriched consortia of Methanogens

Rachana Chandragupta Acharya, Usha Mukundan

Effect of drought stress on epicuticular wax load in peanut genotypes

Jayant K Solanki, S. K. Sarangi

Production and Optimization of Protease by Filamentous Fungus Isolated from Paddy Soil in Thiruvarur District Tamilnadu

Sudarkodi Chandrasekaran, Sundar Sundaram Pillai Kumaresan, Murugan Manavalan

The effects of diet and temperature on enzymes, hormones, and fecundity of the African Catfish Clarias gariepinus (Burchell 1822)

Waleed Abdul-Aziz A. Al-Deghayem, El Amin Mohamed Suliman

Partial purification and characterization of lectin from serum of American cockroach, Periplaneta americana

Reymend Encily Martin, Yogita Raosaheb Channe

Isolation and partial characterization of amylase produced by fungal isolates from the agro-industrial waste source

Soumya Nandi, Twinkle Bose, Susmita Mahato, Somashree Chatterjee, Annalakshmi Chatterjee

Cowpea (Vigna unguiculata L. Walp.): A choice crop for sustainability during the climate change periods

Tewodros Ayalew, Tarekegn Yoseph

Isolation and characterization of starch degrading bacteria from disparate soil samples

Vidhyutha Srivathsan, Mahima Bhandari, Priya Swaminathan

Assessing the role of temperature as an elicitor for indole-3-acetic acid production in cyanobacterial species

Priyanka Behera, Dhanesh Kumar,, Shubhashree Mahalik

Screening and identification of heat tolerance in Indian wheat genotypes using generalized regression neural network (GRNN) model

Anil Kumar, Sadaf Fatima, Iffat Azim, Anshika Negi, Shalini Bhadola, Pooja Jha, Suboot Hairat

Effects of liquid storage of buck semen at refrigeration temperatures on sperm viability and fertility to develop ready to use goat semen diluent

Manish Kumar,, Ravi Ranjan, Alok Bhardwaj