A novel role of L-Asparaginase enzyme production from fungal species

S. Vivekanandha, M. Sankareswaran, P. Prabhavathi

Abstract


The soil samples were collected from oil factory in Salem, Vellore and Chennai. About 12 organisms were isolated from these soil samples. The strain was then screened using MM9 media for the L-asparaginase enzyme production. Aspergillus niger which showed pink colour around the colony on selected media. The screened isolate was optimized at different substrates (wheat bran, gingelly oil cake, cotton seed, rice bran, rice bran), temperature (30°C, 40°C, 50°C, 60°C and 70°C), moisture content (10%, 20%, 30%, 40% and 50%), pH (5, 6, 7, 8 and 9), incubation days (3, 4, 5, 6 and 7), carbon sources (glucose, maltose, sucrose, lactose and mannitol), nitrogen sources (urea, peptone, casein and yeast extract) and metal ions (ZnCl2, FeSO4, MgSO4, NaCl, and CaCl2). A.niger produced maximum L-asparaginase enzyme in gingelly oil cake at temperature 40°C, in 30 % of moisture content, at pH 7 in 4 days. The maximum enzyme activity was observed in mannitol as carbon source, yeast extract as the nitrogen source and ZnCl2 as the metal ion source. The L-asparaginase enzyme was purified by ammonium sulphate precipitation, dialysis and DEAE-cellulose column chromatography. Purification by DEAE-cellulose column chromatography has increased the specific activity to 4166.66 (U/g dry substrate) with a purification fold of 27.7 and a yield of 55.5% in Aspergillus niger.

 

Keywords: Fungal isolates, L- Aspergine, Anion exchange column chromatography.


Full Text:

PDF

References


A Pandey, P Selvakumar, R Carlos Soccol, P Nigam, 1994, Solid state fermentation for the production of industrial enzymes, Wiley Eastern Publishers, New Delhi, 33–37.

E A Abu, S A Ado, D B James, 2005, African J. Biotechnol, 4, 785 – 790.

J D Broome, 1961, Nature, 191, 1114-1115.

, 9] L T Mashburn, J C Wriston, 1964, Arch Biochem Biophys, 105, 450-452.

J C Gilman, 1971, A Manual of Soil Fungi, 2nd Ed. Iowa State College Press, Ames, Iowa, 450.

R Gulati, R K Saxena, R Gupta, 1997, Lett Appl Microbiol, 24, 23-26.

H A Lechevalier, S A Waksman, 1962, The actinomycetes. III. Antibiotics of Actinomycetes, Williams & Wilkins, Baltimore, 430.

, 15] M Raimbault, D Alazard, 1980, Eur, J, Appl Microbiol Biotechnol, 199–209.

T Theantana, K D Hyde, S Lumyong, 2007, KMITL Sci .Tech. J, 7, 13-18.

O H Lowry, N J Rosebrough, A L Farr, 1951, Randall RJ. J. Biol. Chem., 48, 17-25.

F Beisson, A Tiss, C Rivière, R Verger, 2000, Eur J Lipid Technol., 133-153, A comprehensive and detailed review about all lipase assays involving carboxylic ester hydrolysis.

A Benny, Ayyakkannu, 1992, Phuket Mar Biol Spec, 11, 137-140.

A Ashraf, E I Bessoumy, M Sarhan, J Mansour, 2003, Journal of Biochemistry and Molecular Biology, 37(4), 387-393.

S A Gaffer, Y I Shethna, 1975, J Appl Microbiol, 44, 727-729.

K R Babu, T Satyanarayana, 1995, J. Sci. Ind. Res, 55, 464- 467.

, 21] C K Venil, P Lakshmanaperumalsamy, 2009, Iranian Journal of Biotechnology, 7(1).

H A Campbell, T L Marshburn, E A Boyse, L J Old, 1967, Biochem Genet, 8, 721-730.

H Anto, U P Trivedi, K C Patel, 2006, Bioresour. Technol, 1997, 1161-1166.

N Saleem Basha, R Rekha, M Komala, S Ruby, 2009, Tropical Journal of Pharmaceutical Research, 8 (4), 353-360.

P K Praseetha, V S Sugunan, R Shabi, S Jegadambikadevi, 2012, J. Microbiol. Biotech. Res, 2(5), 657- 664.




Copyright (c) 2017 Journal of Microbiology and Biotechnology Research




<Microbiology+Biotechnology+Research>Journal of Microbiology and Biotechnology Research