Potocki-Veronese Gabrielle, Putaux Jean-Luc, Dupeyre Danielle, Albenne Cécile, Remaud-Siméon Magali, Monsan Pierre, Buleon Alain
Laboratoire Biotechnologie-Bioprocédés, UMR INRA 792, UMR CNRS 5504, INSA DGBA, 135 avenue de Rangueil, 31077 Toulouse Cedex 4, France.
Biomacromolecules. 2005 Mar-Apr;6(2):1000-11. doi: 10.1021/bm049326g.
The recombinant amylosucrase from Neisseria polysaccharea was used to synthesize in vitro amylose from sucrose as unique substrate. The morphology and structure of the insoluble residue were shown to depend only on the initial sucrose concentration (100, 300, or 600 mM), which controlled both the chain length and concentration at the precipitation stage. The average degree of polymerization (DP) in the precipitated product varied from 58 for the lowest initial sucrose concentration (100 mM) to 45 and 35 for higher sucrose concentrations (300 and 600 mM, respectively). The shorter chains (DP 35 and 45), produced in high yields (54 and 24 g/L respectively), precipitated as polycrystalline aggregates with exceptional crystallinity, without optimization of the reaction medium for crystallization. The longer chains (DP 58), produced in lower amount (2.9 g/L), formed networks similar to those observed for amylose gels. All synthesized products displayed a B-type crystal structure. Their melting behavior was also studied, the thermostability being higher for the precipitate containing the longer chains. Further thermal treatments were shown to still improve the crystallinity and yield substrates usable as new standards for the determination of the relative crystallinity of starchy products. The kinetics of chain elongation and aggregation were thoroughly investigated in order to explain how the action of amylosucrase resulted in such different amylose structures. These results emphasize the potentiality of amylosucrase in the design of amylodextrins with controlled morphology, structure, and physicochemical properties.
来自多糖奈瑟氏菌的重组淀粉蔗糖酶被用于以蔗糖作为唯一底物在体外合成直链淀粉。结果表明,不溶性残余物的形态和结构仅取决于初始蔗糖浓度(100、300或600 mM),这一浓度在沉淀阶段既控制了链长又控制了浓度。沉淀产物中的平均聚合度(DP)从最低初始蔗糖浓度(100 mM)时的58变化到较高蔗糖浓度(分别为300和600 mM)时的45和35。较短的链(DP 35和45),高产率生成(分别为54和24 g/L),以具有优异结晶度的多晶聚集体形式沉淀,无需对反应介质进行结晶优化。较长的链(DP 58),产量较低(2.9 g/L),形成类似于直链淀粉凝胶中观察到的网络结构。所有合成产物均呈现B型晶体结构。还研究了它们的熔化行为,含较长链的沉淀物热稳定性更高。进一步的热处理显示仍可提高结晶度并产生可用作测定淀粉类产品相对结晶度新标准的底物。为了解释淀粉蔗糖酶的作用如何导致形成如此不同的直链淀粉结构,对链伸长和聚集的动力学进行了深入研究。这些结果强调了淀粉蔗糖酶在设计具有可控形态、结构和物理化学性质的支链淀粉方面的潜力。