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

• Published in: Biotechnology and bioprocess engineering 2018, 23(2), , pp.168-175
• Main Authors: Kim, Dal Rye, Lim, Hee Kyung, Lee, Kee In, Hwang, In Taek
• Format: Article
• Language: English
• Subjects: Biotechnology; Chemistry; Chemistry and Materials Science; Industrial and Production Engineering; Research Paper; 생물공학
• ISSN: 1226-8372; 1976-3816
• DOI: 10.1007/s12257-017-0246-5

The gene of Pae 1263 (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 Pae 1263 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(+)- Pae 1263-6H 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.