Goat dung as a feedstock for hyper-production of amylase from Glutamicibacter arilaitensis strain ALA4

Abstract

Background

In modern times, bacteria-associated products; especially enzymes are gaining immense interest among worldwide researchers. Among several enzymes, amylases are of great significance in bioprocess engineering. This investigation was aimed to optimize the amylase production from Glutamicibacter arilaitensis strain ALA4 using goat dung as an inexpensive substrate in solid-state fermentation.

Results

Amylase production was initially improved by optimizing physical factors and medium components by one factor at a time method. Two-level full factorial design (2⁵ factorial matrix) was applied to screen the selected variables using first-order polynomial model. Parameters such as incubation temperature, moisture, starch, and yeast extract affected the amylase activity significantly (P < 0.05). Central composite design at N = 30 was further employed to evaluate the optimum levels of these variables by a second-order polynomial equation. Maximal amylase activity of 4572.53 ± 41.71 U/g was estimated in the goat dung medium supplemented with 100% moisture, 1% (w/w) starch, and 1% (w/w) yeast extract, being incubated at 40 °C. The optimized parameters revealed approximately twofold increment in the amylase yield (R ² 0.9169) with respect to the original medium. The amylase showed stability at high pH and temperature up to 4 h of incubation with residual activities of 52.32 ± 2.2 and 46.12 ± 3.3%, respectively. Additionally, the enzyme revealed resistant property not only towards various metal ions and organic solvents but also surfactants and inhibitors. Most importantly, the amylases obtained from strain ALA4 depicted remarkable tolerance to commercially available various detergents.

Conclusions

This study reports first reference on the hyper-production of amylase from G. arilaitensis using goat dung as the low-cost agro-waste medium. G. arilaitensis strain ALA4 may be utilized for wide spread applications in several bioprocess industries due to the high stability of its amylase towards diversified pH, temperature, solvents, surfactants, and detergents.

Keywords

Amylases / 2⁵ factorial design, solid-state fermentation / Stability, central composite design

References

[1] Abd-Elhalem BT, El-Sawy M, Gamal RF, Abou-Taleb KA. Production of amylases from Bacillus amyloliquefaciens under submerged fermentation using some agro-industrial by-products. Ann Agric Sci, 2015, 60: 193-202.

[2] Awodun MA, Omonijo LI, Ojeniyi SO. Effect of goat dung and NPK fertilizer on soil and leaf nutrient content, growth and yield of pepper. Int J Soil Sci, 2007, 2: 142-147.

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

[4] Azad MAK, Bae JH, Kim JS, Lim JK, Song KS, Shin BS, . Isolation and characterization of a novel thermostable -amylase from Korean pine seeds. N Biotechnol, 2009, 26: 143-149.

[5] Bajpai B, Chaudhary M, Saxena J. Production and characterization of α-amylase from an extremely halophilic archaeon, Haloferax sp. HA10. Food Technol Biotechnol, 2015, 53: 11-17.

[6] Blanco AS, Durive OP, Perez SB, Montes ZD, Guerra NP. Simultaneous production of amylases and proteases by Bacillus subtilis in brewery wastes. Braz J Microbiol, 2016, 47: 665-674.

[7] Bradford MM. A rapid and sensitive for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analyt Biochem, 1976, 72: 248-254.

[8] Coronado M, Vargas C, Hofemeister J, Ventosa A, Nieto JJ. Production and biochemical characterization of α-amylase from the moderate halophile Halomonas meridian. FEMS Microbiol Lett, 2000, 183: 67-71.

[9] Das K, Doley R, Mukherjee AK. Purification and biochemical characterization of a thermostable, alkaliphilic, extracellular -amylase from Bacillus subtilis DM-03, a strain isolated from the traditional fermented food of India. Biotechnol Appl Biochem, 2004, 40: 291-298.

[10] 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.

[11] Deb P, Talukdar SA, Mohsina K, Sarker PK, Abu Sayem SM. Production and partial characterization of extracellular amylase enzyme from Bacillus amyloliquefaciens P-001. SpringerPlus, 2013, 2: 154.

[12] Dey U, Chatterjee S, Mondal NK. Isolation and characterization of arsenic-resistant bacteria and possible application in bioremediation. Biotechnol Rep, 2016, 10: 1-7.

[13] Finore I, Donato PD, Poli A, Kirdar B, Kasavi C, Toksoy EO, . Use of agro waste biomass for α-amylase production by Anoxybacillus amylolyticus: purification and properties. J Microb Biochem Technol, 2014, 6: 320-326.

[14] Ghasemi Y, Rasoul-Amini S, Ebrahiminezhad A, Zarrini G, Kazemi A, Mousavi-Khorshidi S, . Halotolerant amylase production by a novel bacterial strain, Rheinheimera aquimaris. Res J Microbiol, 2010, 5: 144-149.

[15] Haq IU, Hameed U, Mahmood Z, Javed MM. Solid state fermentation for the production of α-amylase by Paenibacillus amylolyticus. Pak J Bot, 2012, 44: 341-346.

[16] Hasan MM, Marzan LW, Hosna A, Al-Hakim Azad AK. Optimization of some fermentation conditions for the production of extracellular amylases by using Chryseobacterium and Bacillus isolates from organic kitchen wastes. J Genet Eng Biotechnol, 2017

[17] Hernandez MS, Rodriguez MR, Guerra NP, Roses RP. Amylase production by Aspergillus niger in submerged cultivation on two wastes from food industries. J Food Eng, 2006, 73: 93-100.

[18] Khusro A, Aarti C. Molecular identification of newly isolated Bacillus strains from poultry farm and optimization of process parameters for enhanced production of extracellular amylase using OFAT method. Res J Microbiol, 2015, 10: 393-420.

[19] Khusro A, Barathikannan K, Al-Dhabi NA, Arasu MV, Agastian P. Statistical optimization of thermo-alkali stable xylanase production from Bacillus tequilensis strain ARMATI. Electron J Biotechnol, 2016, 22: 16-25.

[20] Khusro A, Barathikannan K, Aarti C, Agastian P. Optimization of thermo-alkali stable amylase production and biomass yield from Bacillus sp. under submerged cultivation. Fermentation, 2017, 3: 7.

[21] Kikani BA, Singh SP. Single step purification and characterization of a thermostable and calcium independent α- amylase from Bacillus amyloliquefaciens TSWK1-1 isolated from Tulsi Shyam hot spring reservoir, Gujarat (India). Int J Biol Macromol, 2011, 48: 676-681.

[22] Kumar SS, Sangeeta R, Soumya S, Ranjan RP, Bidyut B, Pradee DM. Characterizing novel thermophilic amylase producing bacteria from Taptapani hot spring, Odisha, India. Jundishapur J Microbiol, 2014, 7: 1-7.

[23] Li X, Yu HY. Purification and characterization of novel organic-solvent tolerant β-amylase and serine protease from a newly isolated Salimicrobium halophilum strain LY20. FEMS Microbiol Lett, 2012, 329: 204-211.

[24] Maity S, Mallik S, Basuthakur R, Gupta S. Optimization of solid state fermentation conditions and characterization of thermostable α-amylase from Bacillus subtilis (ATCC 6633). J Bioprocess Biotech, 2015, 5: 1-7.

[25] Miller GL. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem, 1959, 31: 426-428.

[26] Mitidieri S, Martinelli AHS, Schrank A, Vainstein MH. Enzymatic detergent formulation containing amylase from Aspergillus niger: a comparative study with commercial detergent formulations. Bioresour Technol, 2006, 97: 1217-1224.

[27] Mnkeni PNS, Austin LM. Fertilizer value of human manure from pilot urine-diversion toilets. Water SA, 2009, 35: 133-138.

[28] Narang S, Satyanarayana T. Thermostable -amylase production by an extreme thermophile Bacillus thermooleovorans. Lett Appl Microbiol, 2001, 32: 31-35.

[29] Nusrat A, Rahman SR. Comparative studies on the production of extracellular -amylase by three mesophilic Bacillus isolates. Bangladesh J Microbiol, 2007, 24: 129-132.

[30] Oberoi R, Beg QK, Puri S, Saxena RK, Gupta R. Characterization and wash performance analysis of an SDS stable alkaline protease from a Bacillus sp. World J Microbiol Biotechnol, 2001, 17: 493-497.

[31] Pal A, Khanum F. Production and extraction optimization of xylanase from Aspergillus niger DFR-5 through solid-state fermentation. Bioresour Technol, 2010, 101: 7563-7569.

[32] Prajapati VS, Trivedi UB, Patel KC. A statistical approach for the production of thermostable and alklophilic α-amylase from Bacillus amyloliquefaciens KCP2 under solid-state fermentation. 3 Biotech, 2015, 5: 211-220.

[33] Prakasham RS, Rao CS, Sarma PN. Green gram husk—an inexpensive substrate for alkaline protease production by Bacillus sp. in solid-state fermentation. Bioresour Technol, 2005, 97: 1449-1454.

[34] Qureshi AS, Khushk I, Ali CH, Chisti Y, Ahmad A, Majeed H. Coproduction of protease and amylase by thermophilic Bacillus sp. BBXS-2 using open solid-state fermentation of lignocellulosic biomass. Biocatal Agric Biotechnol, 2016, 8: 146-151.

[35] Ray RC, Kar S. Statistical optimization of a-amylase production by Bacillus brevis MTCC 7521 in solid-state fermentation using cassava bagasse. Biologia, 2009, 64: 864-870.

[36] Roohi Kuddus M, Saima S. Cold-active detergent-stable extracellular α-amylase from Bacillus cereus GA6: biochemical characteristics and its perspectives in laundry detergent formulation. J Biochem Tech, 2013, 4: 636-644.

[37] Sajjad M, Choudhry S. Effect of starch containing organic substrates on alpha amylase production in Bacillus strains. Afr J Microbiol Res, 2012, 6: 7285-7291.

[38] Saxena R, Singh R. Amylase production by solid-state fermentation of agro-industrial wastes using Bacillus sp. Braz J Microbiol, 2011, 42: 1334-1342.

[39] Selvam K, Selvankumar T, Rajiniganth R, Srinivasan P, Sudhakar C, Senthilkumar B, . Enhanced production of amylase from Bacillus sp. using groundnut shell and cassava waste as a substrate under process optimization: waste to wealth approach. Biocatal Agri Biotechnol, 2016, 7: 250-256.

[40] Sharma N, Vamil R, Ahmad S, Agarwal R. Effect of different carbon and nitrogen sources on α-amylase production from Bacillus amyloliquefaciens. Int J Pharm Sci Res, 2012, 3: 1161-1163.

[41] Smitha RB, Jisha VN, Sajith S, Benjamin S. Dual production of amylase and δ-endotoxin by Bacillus thuringiensis sub sp. kurstaki during biphasic fermentation. Microbiology, 2013, 82: 794-800.

[42] Tiwari S, Shukla N, Mishra P, Gaur R. Enhanced production and characterization of a solvent stable amylase from solvent tolerant Bacillus tequilensis RG-01: thermostable and surfactant resistant. Sci World J, 2014

[43] Vijayaraghavan P, Kalaiyarasi M, Vincent SGP. Cow dung is an ideal fermentation medium for amylase production in solid-state fermentation by Bacillus cereus. J Genet Eng Biotechnol, 2015, 13: 111-117.

[44] Yazid NA, Barrena R, Komilis D, Sanchez A. Solid-state fermentation as a novel paradigm for organic waste valorisation: a review. Sustainability, 2017, 9: 224.