Fu D T, Robyt J F
Department of Biochemistry and Biophysics, Iowa State University, Ames 50011.
Carbohydr Res. 1991 Sep 18;217:201-11. doi: 10.1016/0008-6215(91)84130-7.
The maltodextrin (maltose through maltoheptaose) acceptor reactions of two Streptococcus mutans 6715 glucosyltransferases (GTF-I and GTF-S) were studied. The acceptor product structures were determined by comparing them with the known structures of the acceptor products of Leuconostoc mesenteroides B-512FM dextransucrase (EC 2.4.1.5) and L. mesenteroides B-1355 alternansucrase (EC 2.4.1.140). When reacted with maltose (G2), both GTF-I and GTF-S transferred a D-glucopyranose from sucrose to the nonreducing glucosyl residue to give panose (6(2)-alpha-D-glucopyranosyl maltose). Panose then served as an acceptor to give two further acceptor products, 6(2)-alpha-isomaltosyl maltose and 6(2)-alpha-nigerosyl maltose. 6(2)-alpha-Isomaltosyl maltose then went on to serve as an acceptor to give a series of homologous acceptor products with isomaltodextrin chains attached to C-6 of the nonreducing-end residue of maltose, while 6(2)-alpha-nigerosyl maltose did not further react. When reacted with other maltodextrins (G3-G7), both GTF-I and GTF-S transferred a D-glucopyranose to C-6 of either the nonreducing-end or the reducing-end residues of the maltodextrins, forming alpha(1----6) linkages. When D-glucopyranose was transferred to the nonreducing-end residue by GTF-I or GTF-S, the first product was also an acceptor to give the second product, which then served as an acceptor to give the third product, etc., to give a homologous series of products. When D-glucopyranose was transferred to the reducing-end residue, the acceptor product that formed did not readily serve as an acceptor, or served only as a very poor acceptor, to give a small amount of the next homologue, as was the case for G7 with GTF-S. In addition, GTF-I also transferred D-glucopyranose to the reducing-end or to the nonreducing-end residue of maltotriose, forming alpha(1----3) linkages, to give 3(3)-alpha-D-glucopyranosyl maltotriose and 3(1)-alpha-D-glucopyranosyl maltotriose. Neither of these acceptor products further served as acceptors to give a homologous series. Under equivalent conditions of equimolar amounts of acceptor and sucrose, maltose and maltotriose are much better acceptors with GTF-I than they are with GTF-S, which is better than L. mesenteroides B-512FM dextransucrase. The three enzymes display significantly different efficiencies for the different maltodextrin acceptor reactions, GTF-I and GTF-S having much higher efficiencies than L. mesenteroides B-512FM dextransucrase.
研究了两种变形链球菌6715葡糖基转移酶(GTF-I和GTF-S)的麦芽糊精(麦芽糖至麦芽七糖)受体反应。通过将受体产物结构与肠膜明串珠菌B-512FM葡聚糖蔗糖酶(EC 2.4.1.5)和肠膜明串珠菌B-1355交替蔗糖酶(EC 2.4.1.140)的受体产物已知结构进行比较来确定其结构。当与麦芽糖(G2)反应时,GTF-I和GTF-S均从蔗糖转移一个D-吡喃葡萄糖至非还原葡糖基残基,生成潘糖(6(2)-α-D-吡喃葡萄糖基麦芽糖)。然后潘糖作为受体生成另外两种受体产物,即6(2)-α-异麦芽糖基麦芽糖和6(2)-α-黑曲霉糖基麦芽糖。6(2)-α-异麦芽糖基麦芽糖接着作为受体生成一系列同源受体产物,其异麦芽糊精链连接至麦芽糖非还原端残基的C-6,而6(2)-α-黑曲霉糖基麦芽糖不再进一步反应。当与其他麦芽糊精(G3-G7)反应时,GTF-I和GTF-S均将一个D-吡喃葡萄糖转移至麦芽糊精非还原端或还原端残基的C-6,形成α(1→6)键。当GTF-I或GTF-S将D-吡喃葡萄糖转移至非还原端残基时,第一个产物也是受体,生成第二个产物,然后第二个产物作为受体生成第三个产物等,从而生成一系列同源产物。当D-吡喃葡萄糖转移至还原端残基时,形成的受体产物不易作为受体,或仅作为非常差的受体,生成少量下一个同系物,如GTF-S与G7反应的情况。此外,GTF-I还将D-吡喃葡萄糖转移至麦芽三糖的还原端或非还原端残基,形成α(1→3)键,生成3(3)-α-D-吡喃葡萄糖基麦芽三糖和3(1)-α-D-吡喃葡萄糖基麦芽三糖。这两种受体产物均不再进一步作为受体生成同源系列产物。在受体和蔗糖等摩尔量的等效条件下,麦芽糖和麦芽三糖作为GTF-I的受体比作为GTF-S的受体要好得多,而GTF-S又比肠膜明串珠菌B-512FM葡聚糖蔗糖酶要好。这三种酶对不同麦芽糊精受体反应显示出显著不同的效率,GTF-I和GTF-S的效率远高于肠膜明串珠菌B-512FM葡聚糖蔗糖酶。