Production and Characterization of Glucoamylase by Aspergillus niger
Applied Food Biotechnology,
Vol. 4 No. 1 (2017),
Background and Objective: Glucoamylase is a potent starch degrading enzyme whose cheap production has been an area of research. Its production by Aspergillus niger in solid-state fermentation was studied using dried garden pea peel as a substrate, which enormously reduced the production cost. The current study intended to produce glucoamylase by a cost-effective strategy and exhaustively characterize the enzyme.
Material and Methods: Garden pea peel was used as a substrate in solid state fermentation by Aspergillus niger for the production of glucoamylase under process parameters. Response surface methodology, a statistical tool for optimization, was applied to setup the experimental design for glucoamylase production. Characterization studies of the enzyme were carried out with temperature, pH, metal salts and elemental composition analysis.
Results and Conclusion: The process parameters were temperature, amount of substrate and time of fermentation. Glucoamylase production was highest in the pH range of 5.4-
6.2, was stable at pH 3.8, and maintained its maximum activity even at 70°C for 30 min. It showed higher catalytic efficiency when incubated with metal ions Fe2+, Cu2+, Mg2+, and Pb2+. Km and Vmax for glucoamylase were 0.387 mg of soluble starch ml-1 and 35.03 U μl-1 min-1, respectively. Glycogen was also used as a substrate, which gave an increased Km by 2.585, whose KI was found to be 0.631. Energy-dispersive X-ray spectroscopy was performed for obtaining composition of the pea peel. C, N, and O were found to be 12.53%, 29.9%, and 55.27% by atomic weights, respectively. Cost- and time-effective production of glucoamylase was achieved by utilizing dried garden pea peel (a vegetable residue) powder as the substrate for production. Its high stability ensures efficient utilization under industrial conditions. This work provides a very good platform for the enzyme immobilization studies and scale up production in future.
Conflict of interest: The authors declare that there is no conflict of interest.
- ▪Garden pea peal ▪Glucoamylase ▪Energy-dispersive X-ray spectroscopy ▪Response surface methodology ▪Solid state fermentation
How to Cite
Montibeller VW, de Souza Vandenberghe LP, Amore A,
Soccol CR, Birolo L, Vinciguerra R, Salmon DNX, Spier
MR, Faraco V. Characterization of hemicellulolytic enzymes
produced by Aspergillus niger NRRL 328 under solid state
fermentation on soybean husks. BioRes. 2014; 9(4):7128-
Akhtaruzzaman M, Mozumder NR, Jamal R, Rahman A,
Rahman T. Isolation and characterization protease enzyme
from leguminous seeds. Agric. Sci. Res. J. 2012; 2:434-440.
Pandey A, Webb C, Soccol C, LArroche C. Rice bran as a
substrate for proteolytic enzyme production. Enzyme
technology. New Delhi: Asiatech Publishers, Inc; 2005.
Prajapati VS, Trivedi UB, Patel KC. Optimization of
glucoamylase production by Colletotrichum sp. KCP1 using
statistical methodology. Food Sci. Biotechnol. 2013; 22(1):
Weill C, Burch R, Vandyk J. An alpha-amyloglucosidase that produces beta-glucose. Cereal Chem. 1954; 31(2): 150-158.
Jonathan MC, van Brussel M, Scheffers MS, Kabel MA.
Characterisation of branched gluco-oligosaccharides to study the mode-of-action of a glucoamylase from Hypocrea jecorina. Carbohydr. Polym. 2015;132:59-66. doi
Koshland DE. Stereochemistry and the mechanism of
enzymatic reactions. Biol Rev. 1953; 28(4): 416-436.
McCarter JD, Withers SG. Mechanisms of enzymatic
glycoside hydrolysis. Curr Opin Struct Biol. 1994; 4(6): 885-
Zhang YH, Lynd LR. Toward an aggregated understanding of enzymatic hydrolysis of cellulose: noncomplexed cellulase systems. Biotechnol. Bioeng. 2004; 88(7): 797-824. doi 10.1002/bit.20282.
Sanchez OJ, Cardona CA. Trends in biotechnological
production of fuel ethanol from different feedstocks.
Bioresour. Technol. 2008;99(13):5270-5295.
Sauer J, Sigurskjold BW, Christensen U, Frandsen TP,
Mirgorodskaya E, Harrison M, Roepstorff P, Svensson B.
Glucoamylase: structure/function relationships, and protein engineering. Biochim Biophys Acta. 2000; 1543(2): 275-293.
Ellaiah P, Adinarayana K, Bhavani Y, Padmaja P, Srinivasulu B. Optimization of process parameters for glucoamylase production under solid state fermentation by a newly isolated Aspergillus species. Process Biochem. 2002; 38(4):615-620.
Malhotra R, Noorwez S, 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(5):378-384.
Pandey A. Solid-state fermentation. Biochem. Eng. J. 2003; 13(2): 81-84.
Reguly J. Biotecnologia dos processos fermentativos: fundamentos, matérias primas agrícolas, produtos e processos. Pelotas: UFPel. 1996; 12-21.
Anto H, Trivedi U, Patel K. Glucoamylase production by
solid-state fermentation using rice flake manufacturing waste products as substrate. Bioresour Technol. 2006; 97(10): 1161-1166.
Zambare V. Solid state fermentation of Aspergillus oryzae for glucoamylase production on agro residues. Int J Lif Sc. 2010; 4: 16-25.
Ominyi M, Ogbonna J, Nwoba E, Nwagu K, Ukachi R.
Isolation, and screening of α-amylase and glucoamylase
producing fungi and their glucomaylase producing fungi and their application in bioethanol, production. Int J Sci Nat 2013; 4(1): 44-50.
Nahid P, Vossoughi M, Azad RR, Ahmadi M, Zarrabi A,
Hosseini S. The production of glucoamylase by Aspergillus
niger under solid state conditions (research note). Int J Engg-Trans B: Appl. 2011; 25(1): 1-2.
Puri S, Arora M, Sarao L. Production and optimization of amylase and glucoamylase using Aspergillus oryzae under solid state fermentation. Int J Res Pur Appl Microbiol. 2013;3:83-88.
Miller GL. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem. 1959; 31(3): 426-428.
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