title: Construction of minimum size cellulase (Cel5Z) from Pectobacterium chrysanthemi PY35 by removal of the C-terminal region.
abstract: Pectobacterium chrysanthemi PY35 secretes the endoglucanase Cel5Z, an enzyme of the glycoside hydrolase family 5. Cel5Z is a 426 amino acid, signal peptide (SP)-containing protein composed of two domains: a large N-terminal catalytic domain (CD; 291 amino acids) and a small C-terminal cellulose binding domain (CBD; 62 amino acids). These two domains are separated by a 30 amino acid linker region (LR). A truncated cel5Z gene was constructed with the addition of a nonsense mutation that removes the C-terminal region of the protein. A truncated Cel5Z protein, consisting of 280 amino acid residues, functioned as a mature enzyme despite the absence of the SP, 11 amino acid CD, LR, and CBD region. In fact, this truncated Cel5Z protein showed an enzymatic activity 80% higher than that of full-length Cel5Z. However, cellulase activity was undetectable in mature Cel5Z proteins truncated to less than 280 amino acids.
authors.affiliation: Division of Applied Life Science, Gyeongsang National University, Chinju, 660-701, Korea.
authors.forename: Woo Jin
authors.initials: WJ
authors.lastname: Lim
authors.forename: Su Young
authors.initials: SY
authors.lastname: Hong
authors.forename: Chang Long
authors.initials: CL
authors.lastname: An
authors.forename: Kye Man
authors.initials: KM
authors.lastname: Cho
authors.forename: Byoung Rock
authors.initials: BR
authors.lastname: Choi
authors.forename: Young Kyun
authors.initials: YK
authors.lastname: Kim
authors.forename: Jin Mee
authors.initials: JM
authors.lastname: An
authors.forename: Jung Mi
authors.initials: JM
authors.lastname: Kang
authors.forename: Sun Mi
authors.initials: SM
authors.lastname: Lee
authors.forename: Soo Jeong
authors.initials: SJ
authors.lastname: Cho
authors.forename: Hoon
authors.initials: H
authors.lastname: Kim
authors.forename: Han Dae
authors.initials: HD
authors.lastname: Yun
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D001483:Base Sequence; D044602:Cellulases / Q000737:chemistry* / Q000378:metabolism*; D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial; D016974:Dickeya chrysanthemi / Q000201:enzymology*; D015870:Gene Expression; D008969:Molecular Sequence Data; D011487:Protein Conformation; D015202:Protein Engineering
chemical_list: D001426:Bacterial Proteins; D044602:Cellulases