title: Gene cloning, overexpression, and characterization of a xylanase from Penicillium sp. CGMCC 1669.
abstract: A xylanase-encoding gene, xyn11F63, was isolated from Penicillium sp. F63 CGMCC1669 using degenerated polymerase chain reaction (PCR) and thermal asymmetric interlaced (TAIL)-PCR techniques. The full-length chromosomal gene consists of 724 bp, including a 73-bp intron, and encodes a 217 amino acid polypeptide. The deduced amino acid sequence of xyn11F63 shows the highest identity of 70% to the xylanase from Penicillium sp. strain 40, which belongs to glycosyl hydrolases family 11. The gene was overexpressed in Pichia pastoris, and its activity in the culture medium reached 516 U ml(-1). After purification to electrophoretic homogeneity, the enzyme showed maximal activity at pH 4.5 and 40 degrees C, was stable at acidic buffers of pH 4.5-9.0, and was resistant to proteases (proteinase K, trypsin, subtilisin A, and alpha-chymotrypsin). The specific activity, K (m), and V (max) for oat spelt xylan substrate was 7,988 U mg(-1), 22.2 mg ml(-1), and 15,105.7 micromol min(-1) mg(-1), respectively. These properties make XYN11F63 a potential economical candidate for use in feed and food industrial applications.
authors.affiliation: Key Laboratory of Feed Biotechnology of Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing 100081, People's Republic of China.
authors.forename: Wanli
authors.initials: W
authors.lastname: Liu
authors.forename: Pengjun
authors.initials: P
authors.lastname: Shi
authors.forename: Qiang
authors.initials: Q
authors.lastname: Chen
authors.forename: Peilong
authors.initials: P
authors.lastname: Yang
authors.forename: Guozeng
authors.initials: G
authors.lastname: Wang
authors.forename: Yaru
authors.initials: Y
authors.lastname: Wang
authors.forename: Huiying
authors.initials: H
authors.lastname: Luo
authors.forename: Bin
authors.initials: B
authors.lastname: Yao
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D015870:Gene Expression; D006868:Hydrolysis; D007221:Industry; D007700:Kinetics; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010407:Penicillium / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D010447:Peptide Hydrolases / Q000378:metabolism; D010843:Pichia / Q000235:genetics; D013379:Substrate Specificity
chemical_list: D043364:Endo-1,4-beta Xylanases; D010447:Peptide Hydrolases