Cloning and expression of a novel, moderately thermostable xylanase-encoding gene (Cflxyn11A) from Cellulomonas flavigena.
The Cfl xyn11A gene, encoding the endo-1,4-beta-xylanase Cfl Xyn11A from Cellulomonas flavigena, was isolated from a genomic DNA library. The open reading frame of the Cfl xyn11A gene was 999 base pairs long and encoded a polypeptide (Cfl Xyn11A) of 332 amino acids with a calculated molecular mass of 35,110Da. The Cfl xyn11A gene was expressed in Escherichia coli and the recombinant enzyme, with an estimated molecular weight of 31kDa was purified and xylanase activity was measured. Cfl Xyn11A showed optimal activity at pH 6.5 and 55 degrees C. The enzyme demonstrated moderate thermal stability as Cfl Xyn11A maintained 50% of its activity when incubated at 55 degrees C for 1h or at 45 degrees C for 6h. This is the first report describing the cloning, expression and functional characterization of an endo-1,4-beta-xylanase-encoding gene from C. flavigena. Cfl Xyn11A may be suitable for industrial applications in the food and feed industries, or in the pre-treatment of lignocellulosic biomass required to improve the yields of fermentable sugars for bioethanol production.
Amaya-Delgado
Lorena
L
Departamento de Biotecnología y Bioingeniería, CINVESTAV, Av Instituto Politécnico Nacional No 2508, CP 07360, México DF, Mexico.
Mejía-Castillo
Teresa
T
Santiago-Hernández
Alejandro
A
Vega-Estrada
Jesús
J
Amelia
Farrés-G-S
FG
Xoconostle-Cázares
Beatriz
B
Ruiz-Medrano
Roberto
R
Montes-Horcasitas
María Del Carmen
Mdel C
Hidalgo-Lara
María Eugenia
ME
D018533:Biomass; D040143:Cellulomonas / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry*; D005786:Gene Expression Regulation; D015964:Gene Expression Regulation, Bacterial; D015723:Gene Library; D016680:Genome, Bacterial; D006863:Hydrogen-Ion Concentration; D007218:Industrial Microbiology / Q000379:methods; D008031:Lignin / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D013696:Temperature; D013997:Time Factors
C036909:lignocellulose; D008031:Lignin; D004247:DNA; D043364:Endo-1,4-beta Xylanases