Succinylation of cyclodextrin glycosyltransferase from Thermoanaerobacter sp. 501 enhances its transferase activity using starch as donor.
A simple modification procedure, the succinylation of amino groups, was suitable to increase the transferase (disproportionation) activity of cyclodextrin glycosyltransferase (CGTase) from Thermoanaerobacter sp. 501 using different linear oligosaccharides as acceptors. On the contrary, the synthesis of cyclodextrins (CDs), the coupling of CDs with oligosaccharides, and the hydrolysis of starch decreased after chemical modification. The degree of succinylation of amino groups (45%) was accurately determined by MALDI-TOF mass spectrometry. The formation of CDs under industrial conditions was analyzed for native and succinylated CGTases, showing similar selectivity to alpha-, beta-, gamma-CD. The acceptor reaction with D-glucose using soluble starch as glucosyl donor was studied at 60 degrees C and pH 5.5. Malto-oligosaccharides (MOS) production was notably higher using the semisynthetic enzyme at different ratios (w/w) starch:D-glucose. Thus, more than 90% of the initial starch was converted into MOS (G2-G7) in 48 h employing a ratio donor:acceptor 1:2 (w/w).
Alcalde
M
M
Departamento de Biocatálisis, Instituto de Catálisis, C.S.I.C., Cantoblanco, 28049, Madrid, Spain.
Plou
F J
FJ
Teresa Martín
M
M
Valdés
I
I
Méndez
E
E
Ballesteros
A
A
D000215:Acylation; D001421:Bacteria, Anaerobic / Q000201:enzymology*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D003505:Cyclodextrins / Q000378:metabolism; D005964:Glucosyltransferases / Q000737:chemistry* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008320:Maltose / Q000378:metabolism; D009844:Oligosaccharides / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013213:Starch / Q000378:metabolism*; D013329:Structure-Activity Relationship; D019802:Succinic Acid / Q000737:chemistry*; D013387:Succinic Anhydrides / Q000737:chemistry
D003505:Cyclodextrins; D009844:Oligosaccharides; D013387:Succinic Anhydrides; D008320:Maltose; C031801:succinic anhydride; D013213:Starch; D019802:Succinic Acid; D005964:Glucosyltransferases; C019963:cyclomaltodextrin glucanotransferase