Hyper expression of kojibiose phosphorylase gene and trehalose phosphorylase gene from Thermoanaerobacter brockii ATCC35047 in Bacillus subtilis and selaginose synthesis utilizing two phosphorylases.
The kojibiose phosphorylase (KP) gene and trehalose phosphorylase (TP) gene from Thermoanaerobacter brockii ATCC35047 were intracellularly hyper-expressed under the control of the Bacillus amyloliquefaciens alpha-amylase promoter in Bacillus subtilis. The production yields were estimated to be 2.1 g of KP and 4.9 g of TP per liter of medium. Selaginose, non-reducing trisaccharide, was synthesized from trehalose utilizing the recombinant KP and TP from B. subtilis. Selaginose was not hydrolyzed by salivary amylase, artificial gastric juice, pancreatic amylase, or small intestinal enzymes.
Yamamoto
Takuo
T
Amase Institute, Hayashibara Biochemical Laboratories, Inc., 7-7 Amase Minami-machi, Okayama 700-0834, Japan. amaseken@hayashibara.co.jp
Mukai
Kazuhisa
K
Maruta
Kazuhiko
K
Watanabe
Hikaru
H
Yamashita
Hiroshi
H
Nishimoto
Tomoyuki
T
Kubota
Michio
M
Chaen
Hiroto
H
Fukuda
Shigeharu
S
D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D001709:Biotechnology / Q000379:methods*; D003001:Cloning, Molecular; D004187:Disaccharides / Q000378:metabolism; D015870:Gene Expression; D005964:Glucosyltransferases / Q000235:genetics* / Q000378:metabolism; D006005:Phosphorylases / Q000235:genetics* / Q000378:metabolism; D046968:Thermoanaerobacter / Q000201:enzymology* / Q000235:genetics; D014312:Trisaccharides / Q000096:biosynthesis*
D004187:Disaccharides; D014312:Trisaccharides; C505610:selaginose; C008237:kojibiose; D005964:Glucosyltransferases; D006005:Phosphorylases; C023181:alpha,alpha-trehalose phosphorylase