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001-es BibID:BIBFORM018946
Első szerző:Mótyán János András (biokémikus, molekuláris biológus)
Cím:Transglycosylation by barley alfa-amylase 1 / Mótyán János A., Fazekas Erika, Mori Haruhide, Svensson Birte, Bagossi Péter, Kandra Lili, Gyémánt Gyöngyi
Dátum:2011
ISSN:1381-1177
Megjegyzések:The transglycosylation activity of barley alfa-amylase 1 (AMY1) and active site AMY1 subsite mutant enzymes was investigated. We report here the transferase ability of the V47A, V47F, V47D and S48Y single mutants and V47K/S48G and V47G/S48D double mutant AMY1 enzymes in which the replaced amino acids play important role in substrate binding at subsites at-3 through-5. Although mutation increases the transglycosylation activity of enzymes, in the presence of acceptors the difference between wild type and mutants is not so significant. Oligomer transfer reactions of AMY1 wild type and its mutants were studied using maltoheptaose and maltopentaose donors and different chromophore containing acceptors. The conditions for the chemoenzymatic synthesis of 4-methylumbelliferyl-alfa-d-maltooligosaccharides (MU-alfa-d-MOSs) were optimized using 4-methylumbelliferyl-alfa-d-glucoside as acceptor and maltoheptaose as donor. 4-Methylumbelliferyl-alfa-d-maltoside, -maltotrioside, -maltotetraoside and -maltopentaoside have been synthesized. Products were identified by MALDI-TOF MS. 1H and 13C NMR analyses showed that AMY1 V47F preserved the stereo- and regioselectivity. The produced MU-alfa-d-MOSs of degree of polymerization DP 2, DP 3 and DP 5 were successfully applied to detect activity of Bacillus stearothermophilus maltogenic alfa-amylase, human saliv
Tárgyszavak:Természettudományok Biológiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
Barley alpha-amylase 1
Transglycosylation
Methylumbelliferyl-glycosides
Chemoenzymatic synthesis
Amylase assay
Megjelenés:Journal Of Molecular Catalysis B-Enzymatic. - 72 : 3-4 (2011), p. 229-237. -
További szerzők:Fazekas Erika (1985-) (kémikus) Mori, Haruhide Svensson, Birte Bagossi Péter (1966-2011) (biokémikus, vegyész) Kandra Lili (1943-) (biokémikus) Gyémánt Gyöngyi (1960-) (vegyész)
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