Reaction of phosphorylase-a with α-D-glucose 1-phosphate and maltodextrin acceptors to give products with degree of polymerization 6-89.
A series of linear glucan saccharides (GS) with defined quantity and degree of polymerization (DP) were synthesized from α-d-glucose 1-phosphate (α-d-Glc 1-P) by phosphorylase-a. The GS product fractions with average DP 11, 22, 38, 52, 60, 70, and 79 were measured by HPSEC-ELSD system. Then the same seven fractions were resolved into individual peaks with DP: 6-14, 10-32, 27-55, 37-67, 44-75, 49-83 and 53-89 by HPAEC-PAD system. Results showed that measurement of α-d-Glc 1-P amount consuming during GS synthesis by both systems enable calculation of reaction yield. The reaction yield for the 24h biosynthesis of the GS product was 25.3% (measured by HPSEC-ELSD) or 29.1% (measured by HPAEC-PAD). The HPSEC-ELSD and HPAEC-PAD systems were also successfully used for phosphorylase-a activity measurement in order to perform its kinetic characterization. This study established feasible systems for preparation of various sizes of the GS with defined DP and quantity as well as characterization of phosphorylase-a kinetics.
Department of Food Science, Biotechnology Division, College of Life Sciences, National Taiwan Ocean University, 2 Pei-Ning Road, Keelung, 20224, Taiwan, ROC. Electronic address: bart_kazlowski@hotmail.com.
Bartosz
B
Kazłowski
Department of Food Science, Biotechnology Division, College of Life Sciences, National Taiwan Ocean University, 2 Pei-Ning Road, Keelung, 20224, Taiwan, ROC. Electronic address: irisko@ntou.edu.tw.
Yuan-Tih
YT
Ko
D000818:Animals; D002138:Calibration; D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange / Q000379:methods; D005936:Glucans / Q000138:chemical synthesis* / Q000302:isolation & purification; D005958:Glucosephosphates / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular; D010762:Phosphorylase a / Q000737:chemistry*; D058105:Polymerization; D011108:Polymers / Q000138:chemical synthesis*; D011134:Polysaccharides / Q000737:chemistry*
D005936:Glucans; D005958:Glucosephosphates; D011108:Polymers; D011134:Polysaccharides; C008315:maltodextrin; C031590:glucose-1-phosphate; D010762:Phosphorylase a
Degree of polymerization; Glucan saccharides; HPAEC-PAD; HPSEC-ELSD; Phosphorylase