Main content area

Identification of a New 1,4-beta-D-xylosidase Pae1263 from the Whole Genome Sequence of Paenibacillus terrae HPL-003

Kim, DalRye, Lim, HeeKyung, Lee, KeeIn, Hwang, InTaek
Biotechnology and bioprocess engineering 2018 v.23 no.2 pp. 168-175
Escherichia coli, Lactococcus raffinolactis, Paenibacillus peoriae, bacteria, enzyme activity, genes, genetic vectors, high performance liquid chromatography, open reading frames, pH, sequence analysis, soil, thermal stability, xylose, Korean Peninsula
The gene of Pae1263 (2,196 bp, 732 aa) was found from the full-length sequence analysis of bacterium Paenibacillus terrae HPL-003 isolated from soil on Gara Mountain in Korea (CP003107, our previous study). Among the 20 open reading frames (ORFs) related with the xylose substrate, only the recombinant enzyme of ORF Pae1263 showed a 1,4-beta-D-xylosidase activity when all of the ORFs were transformed into E. coli. This gene is considered to be a new 1,4-beta-D-xylosidase because it has up to 93% similarity with other genes of ZP_10240221.1 from Lactococcus raffinolactis 4877 and ZP_11237858.1 from Paenibacillus peoriae in the GenBank blast search. The enzyme activity was confirmed by HPLC in which xylose was produced from xylobiose as a substrate by this recombinant enzyme. Mass production of the recombinant enzyme was done with the construction of the pET22(+)- Pae1263-6H expression vector system from E. coli. This new 1,4-beta-D-xylosidase was highly active at 50°C in a pH range between 6.0 and 8.0 and had thermo-stability for at least 24 h at 50°C and a K ₘ and V ₘₐₓ of 6.42 mg/mL and 75.76 U/mg on a xylobiose substrate, respectively.