title: Cloning of a novel xylanase gene from a newly isolated Fusarium sp. Q7-31 and its expression in Escherichia coli.
abstract: A strain of Q7-31 was isolated from Qinghai-Tibet Plateau and was identified as Fusarium sp. based on its morphological characteristics and ITS rDNA gene sequence analysis. It has the highest capacity of degrading cell wall activity compared with other 11 strains. To do research on its xylanase activity of Fusarium sp. Q7-31 while the degrading the rice cell walls, the complete gene xyn8 that encodes endo-1, 4-β-xylanase secreted by Fusarium sp. Q7-31 was cloned and sequenced. The coding region of the gene is separated by two introns of 56bp and 55bp. It encodes 230 amino acid residues of a protein with a calculated molecular weight of 25.7 kDa. The animo acids sequence of xyn8 gene has higher similarity with those of family 11 of glycosyl hydrolases reported from other microorganisms. The nature peptide encodeing cDNA was subcloned into pGEX5x-1 expression vector. The recombinant plasmid was expressed in Escherichia coli BL21-CodonPlus (DE3)-RIL, and xylanase activity was measured. The expression fusion protein was identified by SDS-PAGE and Western blotting, a new specific band of about 52kDa was identified when induced by IPTG. Enzyme activity assay verified the recombinants proteins as a xylanase. A maxium activity of 2.34U/ mg, the xylanase had optimal activity at pH 6.0 and temperature 40℃.
authors.affiliation: Gansu Agricultural University , Lanzhou 730000 , China ; Qinghai University , Xining 810016 , China.
authors.forename: Zhan-Ling
authors.initials: ZL
authors.lastname: Xie
authors.forename: Hai-Yan
authors.initials: HY
authors.lastname: Gao
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhang
authors.forename: Huan
authors.initials: H
authors.lastname: Wang
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
keywords: Fusarium sp.; Gene cloning; xylanase
references.citation: Carbohydr Res. 1997 Aug 7;302(3-4):191-5
references.pmid: 9291571
references.citation: Antonie Van Leeuwenhoek. 2010 Aug;98(2):213-23
references.pmid: 20361256
references.citation: J Biotechnol. 2002 May 9;95(2):109-31
references.pmid: 11911922
references.citation: Mycopathologia. 2007 Jul;164(1):19-25
references.pmid: 17554639
references.citation: Mol Gen Genet. 1999 Apr;261(3):530-6
references.pmid: 10323234
references.citation: Can J Microbiol. 2007 Aug;53(8):1000-10
references.pmid: 17898857
references.citation: FEMS Microbiol Lett. 1994 May 15;118(3):305-10
references.pmid: 8020753
references.citation: Plant Physiol. 1994 Apr;104(4):1113-1118
references.pmid: 12232152
references.citation: Fungal Genet Biol. 2002 Apr;35(3):213-22
references.pmid: 11929211
references.citation: J Bacteriol. 1992 Sep;174(18):5848-53
references.pmid: 1522062
references.citation: Med Mycol. 1998 Oct;36(5):299-303
references.pmid: 10075499
references.citation: Nature. 1970 Aug 15;227(5259):680-5
references.pmid: 5432063
references.citation: Appl Microbiol Biotechnol. 2000 Jun;53(6):732-5
references.pmid: 10919335
references.citation: Curr Genet. 2001 Dec;40(4):268-75
references.pmid: 11795847
references.citation: Anal Biochem. 1976 May 7;72:248-54
references.pmid: 942051
references.citation: FEMS Microbiol Lett. 1996 Dec 1;145(2):295-9
references.pmid: 8961570
references.citation: Antonie Van Leeuwenhoek. 2005 Jul;88(1):48-59
references.pmid: 15928976
references.citation: J Biotechnol. 1996 Nov 1;51(2):181-9
references.pmid: 8987884