Characteristics of an organic solvent-tolerant β-fructofuranosidase from Arthrobacter arilaitensis NJEM01 and efficient synthesis of prebiotic kestose.
An organic solvent-tolerant β-fructofuranosidase (β-FFase) from Arthrobacter arilaitensis NJEM01 was purified, characterized, cloned, and overexpressed in Escherichia coli. The mature β-FFase contained 495 amino acid residues with an estimated molecular mass of 55 kDa. The purified β-FFase from strain NJEM01 was very stable in the buffer systems (pH 5.0-9.5) and showed high stability below 45 °C. Furthermore, the enzyme exhibited relatively high solvent stability in various aqueous organic mixtures and retained nearly 100% of its initial activity after incubation for 10 days in 20% (v/v) DMSO. In addition, the β-FFase exhibited high transfructosylation activity, synthesized prebiotic products of mainly 6-kestose (up to 476 g/L), and showed fructosyl receptor specificity to C-glucosyl flavone. A relatively high yield of FOS was achieved by the β-FFase from bacterium with a high concentration of sucrose. It made the β-FFase an exploitable biocatalyst for the production of glycosides of natural products and prebiotic kestose.
College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology , 30 Puzhunan Road, Nanjing 211816, People's Republic of China.
Jianlin
J
Chu
Xueming
X
Wu
Bin
B
Wu
Rui
R
Wang
Bingfang
B
He
D001173:Arthrobacter / Q000201:enzymology*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D009930:Organic Chemicals / Q000737:chemistry; D056692:Prebiotics; D012997:Solvents / Q000737:chemistry; D013379:Substrate Specificity; D014312:Trisaccharides / Q000096:biosynthesis*; D043324:beta-Fructofuranosidase / Q000235:genetics / Q000378:metabolism*
D009930:Organic Chemicals; D056692:Prebiotics; D012997:Solvents; D014312:Trisaccharides; C073432:6-kestose; D043324:beta-Fructofuranosidase