Abstract
An extracellular β-amylase was produced from Aspergillus foetidus MTCC-508, and was purified 254.8-fold with 14.6 yields by precipitation with acetone and by column chromatographies with DEAE-Sephadex A-50 and Sephadex G-100. The purified enzyme was homogeneous on polyacrylamide gel electrophoresis. The band of enzyme was visible around 20 kDa on SDS-PAGE while around 80 kDa on Native-PAGE, showing its homotetrameric nature. The enzyme was optimally active at pH-6.0 and 50 °C temperature. It was fully stable at 50 °C for 2 h. The activity was strongly inhibited by Hg2+, Zn2+ and Co2+, while Mg2+ marginally enhanced the enzyme activity. The enzyme was able to hydrolyze the raw starches of potato, wheat, rice, maize, and Trapa natans, with the highest degree of saccharification of maize starch. The K m and V max values for this enzyme against boiled soluble starch were found 2.7 mg/mL and 2,100 U/mg of protein, respectively.
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Abouzeid AM (1997) Production purification and characterization of an extracellular α-amylase enzyme isolated from Aspergillus flavus. Microbios 89:55–66
Alli AI, Ogbonna CIC, Rahman ATMF (1998) Hydrolysis of certain Nigerian cereal starch using crude fungal amylase. Nig J Biotechnol 9:24–36
Bahrim GE, Scantees M, Negoitan T (2007) Biotechnological conditions of amylase and complex production and utilization involving filamentous bacteria. The Annals of the University Dunarea de jos of Gulati-, Fascicle iv- Food Technol 76–81
Bernfeld P (1955) Amylase α/β. Methods Enzymol 1:149
Chang CT, Liou HY, Tang HL, Sung HY (1996) Activation, purification and properties of β-amylase from sweet potatoes (Ipomea batatas). Biotechnol App Bichem 24:113–118
Colman P, Matthews B (1971) Symmetry, molecular weight and crystallographic Data for sweet potato beta-amylase. J Mol Biol 60:163
Dey PM, Pridham JB (1977) Biochemistry of alpha galactosidase. Adv Enzymol 15:91–130
Hyun HH, Zeikus JG (1985) General biochemical characterization of thermostable extracellular β-amylase from Clostridium thermosulfurogenes. Appl Environ Microbiol 49:1162–1167
Kwan HS, So KH, Chan KY, Cheng SC (1994) Purification and properties of β-amylase from Bacillus circulans S31. World J Microbiol Biotechnol 10:597–598
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. J Am Chem Sci 56:658–666
Lowry OH, Rosenbrough NJ, Farr Al, Randall RJ (1951) Protein estimation with the Folin-phenol reagent. J Biol Chem 193:265–275
Obi SKC, Odibo FJC (1984) Partial purification and characterization of a thermostable actinomycete β-amylase. Appl Environ Microbiol 47:571–575
Oboh G (2005) Isolation and characterization of amylase from fermented cassava (Manihot esculenta Crantz) waste water. Afr J Biotechnol 4:1117–1123
Oboh G, Ajele JO (1997) Effect of some metallic chlorides on the activity of β-amylase from sweet potatoes. Nig J Biochem Mol Biol 12:73–75
Okada y, Yoshigi N, Sahara H, Koshino S (1995) Increase in thermostability of recombinant barley beta-amylase by random mutagenesis. Biosci Biotechnol Biochem 59:1152–1153
Omemu AM, Akpan I, Bankole MO, Teniola OD (2005) Hydrolysis of raw tuber starches by amylase of Aspergillus niger AM07 isolated from the soil. Afr J Biotechnol 4:19–25
Pandey A, Nigam P, Sccol CR, Sccol VT, Singh D, Mohan R (2000) Advances in microbial amylases (Review). Biotechnol Appl Biochem 31:135–152
Rama M, Swamy MV, Seenayya G (1998) Purification and characterization of thermostable β-amylase and pullulanase from high yielding Clostridium thermosulfurogenes SV2. World J Microbiol Biotechnol 14:89–94
Schokker EP, Van Boekel AJS (1999) Kinetic of thermal inactivation of extracellular proteinase from Pseudomonas fluorescens 22F, Influence of pH, calcium and protein. J Agric Food Chem 47:1681–1686
Schuster E, Dunn-Coleman N, Frisvad JC, Van Dijck PWM (2002) On the safety of Aspergillus niger-a review. Appl Microbiol Biotechnol 59:426–435
Shen GJ, Saha BC, Lee YE, Bhatnagar L, Zeikus JG (1988) Purification and characterization of a novel thermostable β-amylase from Clostridium thermosulphurogenes, BiChemi. J 254:835–840
Swamy MV, Sairam N, Seenayya G (1994) β-amylase from Clostridium thermocelluum SS8 a thermophillic, anaerobic, cellulolytic bacterium. Lett Appl Microbiol 18:301–304
Thoma JA, Spradlin JE, Dygert S (1997) Plant and animal amylases. In: Boyer PD (ed) The enzymes, 3rd edn. Academic Press, New York, pp 115–189
Thoma JA, Spradlin JE, Dygert S (1971) Plant and animal amylases. In: Boyer PD (ed) The enzymes, 5th edn. Academic, New York, pp 115–189
Walker JM (2002) The protein protcols handbook. Humana Press Inc, New Jersey, pp 343–345
White JS, White DC (1997) Source book of enzymes. CRC Press, New York, p 572
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The support from the Department of Biotechnology, DDU Gorakhpur University, Gorakhpur is duly acknowledged.
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Mishra, S., Shivam, K., Diwakar, S., Shukla, S. (2014). Purification and Characterization of a Novel Thermostable β-Amylase from Aspergillus foetidus MTCC-508. β-Amylase from Aspergillus foetidus MTCC-508. In: Kharwar, R., Upadhyay, R., Dubey, N., Raghuwanshi, R. (eds) Microbial Diversity and Biotechnology in Food Security. Springer, New Delhi. https://doi.org/10.1007/978-81-322-1801-2_43
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DOI: https://doi.org/10.1007/978-81-322-1801-2_43
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