High-level expression of a xylanase gene from the thermophilic fungus Paecilomyces thermophila in Pichia pastoris.
A xylanase gene from Paecilomyces thermophila was functionally expressed in Pichia pastoris. The recombinant xylanase (xynA) was predominantly extracellular; in a 5 l fermentor culture, the total extracellular protein was 8.1 g l(-1) with an activity of 52,940 U ml(-1). The enzyme was purified to homogeneity with a recovery of 48 %. The recombinant xynA was optimally active at 75 °C, as measured over 10 min, and at pH 7. The enzyme was stable up to 80 °C for 30 min. It hydrolyzed birchwood xylan, beechwood xylan and xylooligosaccharides to produce xylobiose and xylotriose as the main products.
Department of Biotechnology, College of Food Science and Nutritional Engineering, China Agricultural University, No. 17 Qinghua Donglu, Haidian District, Beijing, China. fgs-ln@163.com
Guangsen
G
Fan
Priti
P
Katrolia
Huiyong
H
Jia
Shaoqing
S
Yang
Qiaojuan
Q
Yan
Zhengqiang
Z
Jiang
D004591:Electrophoresis, Polyacrylamide Gel; D043364:Endo-1,4-beta Xylanases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D004795:Enzyme Stability; D005285:Fermentation; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D005965:Glucuronates / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D009844:Oligosaccharides / Q000378:metabolism; D010143:Paecilomyces / Q000201:enzymology* / Q000235:genetics; D010843:Pichia / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D013696:Temperature; D014990:Xylans / Q000378:metabolism
D005656:Fungal Proteins; D005965:Glucuronates; D009844:Oligosaccharides; D011994:Recombinant Proteins; D014990:Xylans; C570991:xylooligosaccharide; D043364:Endo-1,4-beta Xylanases