High-level heterologous expression of Bacillus halodurans putative xylanase xyn11a (BH0899) in Kluyveromyces lactis.
The putative xyn11A structural gene (BH0899) encoding a family-11 xylanase from alkaliphilic Bacillus halodurans strain C-125 was heterologously expressed in the yeast Kluyveromyces lactis CBS 1065 and secreted to a level of 156 microg/ml under selective culture conditions in shake flasks. The Xyn11A production level in shake flask cultures of K. lactis CBS 1065 was higher than that reported for other xylanase genes placed under the control of the regulated LAC4 promoter on a plasmid containing an entire sequence of pKD1 from Kluyveromyces drosophilarium. Recombinant Xyn11A was highly active over pH range from 3 to 10, with maximal activity around pH 7. The enzyme showed a specific activity of 628 U/mg-protein on birchwood xylan as substrate, but no cellulase or beta-xylosidase activity.
Wamalwa
Benson Munyali
BM
Applied Microbiology Laboratory, Graduate School of Bioresources, Mie University, Tsu, Japan.
Zhao
Guangshan
G
Sakka
Makiko
M
Shiundu
Paul Mwanza
PM
Kimura
Tetsuya
T
Sakka
Kazuo
K
D001407:Bacillus / Q000201:enzymology*; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D007716:Kluyveromyces / Q000201:enzymology*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014995:Xylosidases / Q000096:biosynthesis* / Q000235:genetics
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D014995:Xylosidases