CLONING AND EXPRESSION OF CHAETOMIUM THERMOPHILUM XYLANASE 11-A GENE IN PICHIA PASTORIS
DOI:
https://doi.org/10.71330/thenucleus.2008.1001Abstract
The various thermophilic fungi like Chaetomium thermophile has potential to secrete xylanase and cellulase enzymes. In
the present study eukaryotic expression system of Pichia pastoris (yeast) was used to express xylanase gene. The
xylanase (Xyn 11-A) gene was isolated from C. thermophile strain NIBGE-1. Primers were designed to amplify the gene,
ligated into P. pastoris pPIC3.5K vector, the resultant recombinant clone pSSZ810 was transformed into the genome of
P. pastoris GS115 strain through electroporation. Transformants were selected on yeast peptone dextrose medium
(YPD) plates containing antibiotic geneticin (100 mg/mL) upto final concentration of 0.75 mg/mL. The maximum activity
of xylanase 2.04 U/mL after incubation of 2 hrs at 50ºC was observed in the presence of 100% methanol inducer upto
final concentration of 30μL (0.5%) as compared to control. HPLC analysis represented high peak of xylose as compared
to control. SDS-PAGE indicated approx. 28 kDa protein of expressed xylanase gene.
References
M.P. Coughtan and G.P. Hazlewood,
Biotechnol. Appl. Biochem. 17 (1993) 259.
J. Buchert. Appl. Microbiol. Biotechnol., 37
(1992) 825.
3 Alam, Gomes, G. Mohiuddin and M. Haq.
J. Enzyme. Microb. Technol., 16 (1994) 298.
L.P. Christov and B.A Prior, Enzyme and
Molecular Technology, 18 (1996) 244.
K.K.Y. Wong and J.N. Saddler, J. Critical
Reviews in Biochem., 12, No. 5/6 (1992)
C. Daneault. A Review. Tappi J., 77, No. 6
(1994) 125.
F. Latif, M.I. Rajoka, K.A. Malik, W.J.
Microbiol. Biotechnol., 11 (1995) 347.
8 J.M. Cregg, J. F. Tschopp, C. Stillman,R.
Siegel, M. Akong, W.S. Craig, R.G.
Buckholz, L.R. Madden, P.A. Kellaris, G.R.
Davis,
B.L. Smiley, J. Cruze, R. Torregrossa,
G. Velicelebi and G.P. Thill., Nucleic Acids
Res., 15, No. 9 (1987) 3859.
R.A. Brierley, C. Bussineau, R. Kosson, A.
Melton and R. S. Siegel. Annals New York
Academy of Sciences (1992) pp. 350-362.
J. F. Tschopp, G. Sverlow, R. Kosson, W.
Craig and L. Grinna., Bio Tech., 5 (1987)
Y. Chen, J. Krol, J. Cino, D. Freedman, C.
White and E. Komives. J. Chem. Tech.
Biotech., 67 (1996) 143.
K.N. Faber, W.Harder and M. Veenhuis,
Yeast, 11 (1995) 1331.
J.L. Cereghino and J.M. Cregg, FEMS
Microbial. Rev., 24, No. 1 (2000) 45.
F. Latif, M.I. Rajoka and K.A. Malik,
Bioresource Technol., 50 (1994) 107.
S.A. Khan, Z. Mukhtar, A. Akram, A. Ghaffar,
Y. Zafar, and F. Latif., Report No. AY366479,
National Institute for Biotechnology and
Genetic Engineering (NIBGE) 2003.
S.N. Cohen, A.C.Y. Chang and L. Hsu, Proc.
Natl. Acad. Sci. USA, 69 (1972) 2110.
M. Tuncer, A.S. Ball, A. Rob and M.T. J
Wilson., Enzyme Microbe. Tech., 25 (1999)
T. B. Li, Y. Hu, Y.G. Wang and H. Z. Xia.
Sheng Wu Hong Cheng Xue Bao., 21 No. 1
(2005) 25-9.
X. L. Li , Z. Q. Zhang , J.F. Dean , K. E.
Eriksson and L. G. Ljungdahl . Appl. Environ.
Microbiol., 59, No. 10 (1993) 3212.
M. Bottner and C. Lang. Mol. Bio., 267
(2004) 277.
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