Copyright (c) 2016 AJC
This work is licensed under a Creative Commons Attribution 4.0 International License.
Random Mutagenesis Methods for Hyper-Production of Laccase and its Effect on Azo Dye Decolourization
Corresponding Author(s) : Mani Arunkumar
Asian Journal of Chemistry,
Vol. 28 No. 3 (2016): Vol 28 Issue 3
Abstract
This work focuses on the improvement of the wild strain for the hyper production of laccase through random mutagenesis. Maximum laccase production (46956 U/L) was observed in mutant T. hirsuta, when rice bran (5 g) at 75 % moisture (w/v) was used with 1 mL inoculum at pH 6 and 25 °C in the presence of 1 % (w/v) glucose as a carbon source, 0.1 % (w/v) sodium nitrate as a nitrogen source, 2 % of (v/v) glycerol as an inducer in submerged fermentation under static conditions. Laccase purification was performed by precipitating the enzyme with 90 % ammonium sulphate followed by ion exchange chromatography. The puried laccase had an apparent molecular mass of 49.9 kDa with monomeric structure estimated by SDS-PAGE. The three azo dyes could be efficiently decolourized by the purified laccase in the presence of a HBT as redox mediators after 3 h of incubation at pH 5. Laccase mediated degradation of poneaceau was monitored by UV-visible, IR-spectroscopy, HPLC and GCMS. Furthermore, the decolourization products of poneaceau were found to be aromatic amines and less phytotoxic nature than the untreated dye.
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References
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L. Tan, H. Li, S. Ning and B. Xu, Bioresour. Technol., 158, 321 (2014); doi:10.1016/j.biortech.2014.02.063.
H. Zollinger, Colour Chemistry-Synthesis, Properties and Application of Organic Dyes and Pigment, VCH Publishers, New York, pp. 92-102 (1987).
C.M. Carliell, S.J. Barclay, N. Naidoo, C.A. Buckley, D.A. Mulholland and E. Senior, Water SA, 21, 61 (1995).
K.C. Chen, J.Y. Wu, D.J. Liou and S.C. Hwang, J. Biotechnol., 101, 57 (2003); doi:10.1016/S0168-1656(02)00303-6.
H.S. Lade, T.R. Waghmode, A.A. Kadam and S.P. Govindwar, Int. Biodeter. Biodegr., 72, 94 (2012); doi:10.1016/j.ibiod.2012.06.001.
F.P. van der Zee, G. Lettinga and J.A. Field, Chemosphere, 44, 1169 (2001); doi:10.1016/S0045-6535(00)00270-8.
M.S. Khehra, H.S. Saini, D.K. Sharma, B.S. Chadha and S.S. Chimni, Dyes Pigments, 70, 1 (2006); doi:10.1016/j.dyepig.2004.12.021.
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P. Verma and D. Madamwar, World J. Microbiol. Biotechnol., 19, 615 (2003); doi:10.1023/A:1025115801331.
G. McMullan, C. Meehan, A. Conneely, N. Kirby, T. Robinson, P. Nigam, I.M. Banat, R. Marchant and W.F. Smyth, Appl. Microbiol. Biotechnol., 56, 81 (2001); doi:10.1007/s002530000587.
S. Mishra Salony and V.S. Bisaria, J. Sci. Ind. Res. (India), 66, 684 (2007).
S. Rodriguez-Couto, M.A. Sanroman and G.M. Gubitz, Chemosphere, 58, 417 (2005); doi:10.1016/j.chemosphere.2004.09.033.
C. Lopez, I. Mielgo, M.T. Moreira, G. Feijoo and J.M. Lema, J. Biotechnol., 99, 249 (2002); doi:10.1016/S0168-1656(02)00217-1.
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G. Diamantidis, A. Effosse, P. Potier and R. Bally, Soil Biol. Biochem., 32, 919 (2000); doi:10.1016/S0038-0717(99)00221-7.
H. Jung, F. Xu and K. Li, Enzyme Microb. Technol., 30, 161 (2002); doi:10.1016/S0141-0229(01)00485-9.
U.K. Laemmli, Nat. Sci, 227, 680 (1970); doi:10.1038/227680a0.
G.D. Saratale, S.D. Kalme and S.P. Govindwar, Int. J. Biotechnol., 5, 407 (2006).
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Y. Fukushima and T.K. Kirk, Appl. Environ. Microbiol., 61, 872 (1995).
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I.K. Kapdan and F. Kargi, Enzyme Microb. Technol., 67, 195 (2002); doi:10.1016/S0141-0229(01)00468-9.
M.S. Revankar and S.S. Lele, World J. Microbiol. Biotechnol., 22, 921 (2006); doi:10.1007/s11274-006-9136-2.
D.A. Wood, J. Gen. Microbiol., 117, 327 (1980).
S. Ryan, W. Schnitzhofer, T. Tzanov, A. Cavaco-Paulo and G.M. Gübitz, Enzyme Microb. Technol., 33, 766 (2003); doi:10.1016/S0141-0229(03)00162-5.
G.M. Soares, M.T. De Amorim and M. Costa-Ferreira, J. Biotechnol., 89, 123 (2001); doi:10.1016/S0168-1656(01)00302-9.
A. Pich, S. Bhattacharya, H.J. Adler, T. Wage, A. Taubenberger, Z. Li, K.H. Van Pee, U. Böhmer and T. Bley, Macromol. Biosci., 6, 301 (2006); doi:10.1002/mabi.200500192.
K.N. Niladevi and P. Prema, Bioresour. Technol., 99, 4583 (2008); doi:10.1016/j.biortech.2007.06.056.
E.P. Chagas and L.P. Durrant, Enzyme Microb. Technol., 29, 473 (2001); doi:10.1016/S0141-0229(01)00405-7.
M. Stredansky and E. Conti, Process Biochem., 34, 581 (1999); doi:10.1016/S0032-9592(98)00131-9.
J.F.O. Cruz, M.S. Thesis, Banana Skin: A Novel Material for a Low-Cost Production of Laccase, Universitat Rovira I Virgili, Spain (2008).
G.F. Leatham and M.A. Stahmann, J. Gen. Microbiol., 125, 147 (1981).
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