title: Molecular cloning and expression of the trichoderma harzianum C4 endo-beta-1,4-Xylanase Gene in Saccharomyces cerevisiae.
abstract: An endo-beta-1,4-xylanase (beta-xylanase) from Trichoderma harzianum C4 was purified without cellulase activity by sequential chromatographies. The specific activity of the purified enzyme preparation was 430 units/mg on D-xylan. The complementary DNA (cDNA) encoding beta-xylanase (xynII) was amplified by PCR and isolated from cDNA PCR libraries constructed from T. harzianum C4. The nucleotide sequence of the cDNA fragment contained an open reading frame of 663 bp that encodes 221 amino acids, of which the mature protein is homologous to several beta- xylanases II. An intron of 63 bp was identified in the genomic DNA sequence of xynII. This gene was expressed in Saccharomyces cerevisiae strains under the control of adh1 (alcohol dehydrogenase I) and pgk1 (phosphoglycerate kinase I) promoters in 2 mu-based plasmids, which could render recombinants able to secrete beta-xylanase into the media.
authors.affiliation: Skin Biotechnology Center, Graduate School of Biotechnology and Institute of Life Science and Resources, Kyung Hee University,Yongin 446-701, Korea.
authors.forename: Jung Min
authors.initials: JM
authors.lastname: Lee
authors.forename: Ji-Won
authors.initials: JW
authors.lastname: Shin
authors.forename: Jae-Kook
authors.initials: JK
authors.lastname: Nam
authors.forename: Ji-Young
authors.initials: JY
authors.lastname: Choi
authors.forename: Choon-Soo
authors.initials: CS
authors.lastname: Jeong
authors.forename: In-Seob
authors.initials: IS
authors.lastname: Han
authors.forename: Soo-Wan
authors.initials: SW
authors.lastname: Nam
authors.forename: Yun-Jaie
authors.initials: YJ
authors.lastname: Choi
authors.forename: Dae Kyun
authors.initials: DK
authors.lastname: Chung
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis / Q000235:genetics; D004271:DNA, Fungal / Q000032:analysis / Q000235:genetics; D043364:Endo-1,4-beta Xylanases / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D007218:Industrial Microbiology; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D014242:Trichoderma / Q000201:enzymology* / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D004271:DNA, Fungal; D011993:Recombinant Fusion Proteins; D043364:Endo-1,4-beta Xylanases