|
KMID : 1094720120170040729
|
|
Biotechnology and Bioprocess Engineering
2012 Volume.17 No. 4 p.729 ~ p.738
|
|
|
Characterization of a family 3 polysaccharide lyase with broad temperature adaptability, thermo-alkali stability, and ethanol tolerance
|
|
Zhou Junpei
Dong Yanyan
Gao Yajie
Tang Xianghua
Li Junjun
Yang Yunjuan
Xu Bo
Xie Zhenrong
Huang Zunxi
|
|
|
Abstract
|
|
|
|
The 774-bp pectate lyase gene plyAI4 from Bacillus sp. I4 was cloned and expressed in E. coli. The gene encodes a 257-residue polypeptide (PlyAI4, 28.3 kDa) with the highest identities of 97.3% with a putative pectate lyase from Bacillus subtilis BSn5 (ADV94306) and 60.3% with an identified pectate lyase of the polysaccharide lyase family (PL) 3 from Paenibacillus amylolyticus 27C64 (ADB78774). The purified recombinant PlyAI4 (rPlyAI4) exhibited apparently optimal activity at pH 10.5 ∼ 11.0 and 50°C. Compared with the majority of reported alkaline pectate lyases, rPlyAI4 exhibited more residual enzyme activity at 20°C (∼45%) or at 70°C (∼50%) and better thermostability at 70°C (∼60 min half-life at 70°C). In the presence of 20% (v/v) ethanol, pectate lyase activity was enhanced by 0.2 fold. After incubation in 40% (v/v) ethanol at 37°C and pH 8.5 for 1 h, the purified rPelAI4 retained more than 75% of the initial activity. Sequence analysis proposed a new signature block, A-D-G-[V/I]-H, for PL 3 pectate lyases. These properties may prove to be important with regards to PlyAI4 for basic research and industrial application.
|
|
|
KEYWORD
|
|
|
pectate lyase, temperature adaptability, thermoalkali stability, ethanol, signature block
|
|
|
FullTexts / Linksout information
|
|
|
|
|
Listed journal information
|
|
 
|