Characterization of an acidophilic α-amylase from Aspergillus niger RBP7 and study of catalytic potential in response to nutritionally important heterogeneous compound


  • Riddha Mukherjee
  • Tanmay Paul
  • Suman Kumar Halder
  • Jyoti Prakash Soren
  • Amrita Banerjee
  • Keshab Chandra Mondal
  • Bikash Ranjan Pati
  • Pradeep Kumar Das Mohapatra



characterization, digestion of food material, molecular weight determination, purification


An acidophilic α-amylase from Aspergillus niger RBP7 was purified after solid state fermentation on potato peel substrate. Molecular mass of the purified α-amylase was 37.5 kDa and it exhibited 1.4 mg/ml and 0.992 μ/mol/min Km and Vmax values, respectively. The enzyme was stable in the pH range from 2.0 to 6.0, at high NaCl concentration (3 M) and at temperatures between 40 °C and 70 °C. The enzyme showed an optimal activity at pH 3.0 and at 45 °C. The enzyme was inhibited by Hg2+ and was stable in the presence of different surfactants (Tween 60, Tween 80, and SDS at 1% level) and different inhibitory reagents (β-mercaptoethanol, phenylmethylsulfonyl fluoride, and sodium azide). This acidophilic amylase enzyme can digest heterogeneous food materials, i.e. the mixture of rice, fish, bread and curry with comparable activity to the commercial diastase enzymes available.


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Aygan A, Arikan B, Korkmaz H, Dinçer S, Colak O (2008) Highly thermostable and alkaline α-amylase from a halotolerant-alkaliphilic Bacillus sp. AB68. Braz J Microbiol 39(3):547-553.

Gavrilescu M, Chisti Y (2005) Biotechnology - a sustainable alternative for chemical industry. Biotechnol Adv 23(7-8):471-499.

Ghorai, S, Banik SP, Verma D, Chowdhury S, Mukherjee S, Khowala S (2009) Fungal biotechnology in food and feed processing. Food Res Int 42(5):577-587.

Gupta R, Gigras P, Mohapatra H, Goswami VK, Chauhan B (2003) Microbial α-amylases: a biotechnological prospective. Process Biochem 38:1599-1616.

Halder SK, Jana A, Paul T, Das AP, Ghosh K, Pati BR, Mondal KC (2016) Purification and biochemical characterization of chitinase of Aeromonas hydrophila SBK1 biosynthesized using crustacean shell. Biocat Agri Biotechnol 5:211-218.

Haq I, Ali S, Javed MM, Hameed U, Saleem A, Adnan F, Qadeer MA (2010) Production of alpha amylase from a randomly induced mutant strain of Bacillus amyloliquefaciens and its application as a desizer in textile industry. Pak J Bot 42(1):473-484.

Haq IU, Javed MM, Hameed U, Adnan F (2010) Kinetic and thermodynamic studies of alpha amylase from Bacillus licheniformis mutant. Pak J Bot 42(5):3507-3516.

Jana M, Pati B (1997) Thermostable, salt-tolerant α-amylase from Bacillus sp. MD 124. J Basic Microbiol 37(5):323-326.

Khan JA, Priya R (2011) A study on partial purification and characterization of extracellular amylases from Bacillus subtilis. Adv Appl Sci Res 2(3):509-519.

Konsula Z, Liakopoulou-Kyriakides M (2004) Hydrolysis of starches by the action of an α-amylase from Bacillus subtilis. Process Biochem 39:1745-1749.

Krishnan T, Chandra AK (1983) Purification and characterization of α-amylase from Bacillus licheniformis CUMC 305. Appl Environ Microbiol 46:430-437.

Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680-685.

Lineweaver H, Burk D (1934) The determination of enzyme dissociation constants. Acc Chem Res 56(3):658-666.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with Folin phenol reagent. J Biol Chem 193:265-275.

Maity C, Samanta S, Halder SK, Das Mohapatra PK, Pati BR, Jana M, Mondal KC (2010) Isozymes of α-amylases from newly isolated Bacillus thuringiensis CKB 19: production from immobilized cells. Biotechnol Bioprocess Eng 16(2):312-319.

Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31(3):426-428.

Mukherjee R, Paul T, Soren JP, Halder SK, Mondal KC, Pati BR, Das Mohapatra PK (2017) Acidophilic α-amylase production from Aspergillus niger RBP7 using potato peel as substrate: a waste to value added. Waste Biomass Valor, DOI:

Pandey A, Nigam P, Soccol CR, Soccol VT, Singh D, Mohan R (2000) Advances in microbial amylases. Biotechnol Appl Biochem 31(2):135-152.

Parker K, Salas M, Nwosu VC (2010) High fructose corn syrup: production, uses and public health concerns. Biotechnol Mol Biol Rev 5(5):71-78.

Raghu HS, Rajeshwara NA (2015) Immobilization of α-amylase (1,4-α-D-glucanglucanohydralase) by calcium alginate encapsulation. Int Food Res J 22(2):869-871.

Samanta S, Das A, Halder SK, Jana A, Kar S, Das Mohapatra PK, Pati BR, Mondal KC (2014) Thermodynamic and kinetic characteristics of an α-amylase from Bacillus licheniformis SKB4. Acta Biol Szeged 58(2):147-156.

Sethi BK, Nanda PK, Sahoo S, Sena S (2016) Characterization of purified α- amylase produced by Aspergillus terreus NCFT 4269.10 using pearl millet as substrate. Cogent Food Agric 2(1).

Singh R, Kumar V, Kapoor V (2014) Partial purification and characterization of heat stable α-amylase from a thermophilic actinobacteria, Streptomyces sp. MSC 702. Enzyme Res 2014:8, ID 106363.

Souza PMD, Magalhães PDOE (2010) Application of microbial α-amylase in industry - a review. Braz J Microbiol 41:850-861.

Tallapragada P, Dikshit R, Jadhav A, Sarah U (2017) Partial purification and characterization of amylase enzyme under solid state fermentation from Monascus sanguineus. Genet Eng Biotechnol J 15(1):95-101.

Uchino F (1982) A thermophilic unusually acidophilic amylase produced by a thermophilic acidophilic Bacillus sp. Agric Biol Chem 46(1):7-13.

Varalakshmi KN, Kumudini BS, Nandini BN, Solomon J, Suhas R, Mahash B, Kavitha AP (2009) Production and characterization of α-amylase from Aspergillus niger JGI 24 isolated in Bangalore. Pol J Microbiol 58(1):29-36.

Yandri, Susanti D, Suhartati T, Hadi S (2012) Immobilization of α-amylase from locale bacteria isolate Bacillus subtilis ITBCCB148 with carboxymethyl cellulose (CMCellulose). Modern Appl Sci 6(3).




How to Cite

Mukherjee, R., Paul, T., Halder, S. K., Soren, J. P., Banerjee, A., Mondal, K. C., Pati, B. R. and Mohapatra, P. K. D. (2018) “Characterization of an acidophilic α-amylase from Aspergillus niger RBP7 and study of catalytic potential in response to nutritionally important heterogeneous compound”, Acta Biologica Szegediensis, 62(1), pp. 75–82. doi: 10.14232/abs.2018.1.75-82.