Synthesis of novel fructooligosaccharides by substrate and enzyme engineering
Synthesis of novel fructooligosaccharides by substrate and enzyme engineering
复制标题
DOI：
10.1016/j.jbiotec.2008.07.1998
复制
发表时间：
2008-11-06
影响因子：
4.1
通讯作者：
Seibel, Juergen
中科院分区：
文献类型：
--
作者：
Beine, Rafael;Moraru, Roxana;Seibel, Juergen
低聚果糖(FOSS)和多果苷(PSS)因其作为益生素的有益作用而受到特别关注。本文报道了利用底物和酶工程技术合成一类新的低聚果糖。使用改良的左旋蔗糖酶(枯草芽孢杆菌的SACB)和蔗糖类似物(α-木基-1，2-β-果糖或α-半乳糖-1，2-β-果糖)，产品图谱从果聚糖(Levan)聚合物转变为一系列新的更高的低聚糖(低聚木糖)，或多糖(半乳多聚果糖苷)，大小不一。此外，该酶还被随机诱变，用于合成短链低聚果糖，以产生不能产生聚合物的变体A5(N242H)。它将其产品模式转变为短链低聚糖和水解，并首次与蔗糖类似物木糖果糖一起直接合成6-酮糖类似物(α-木糖-1，2-β-果糖-2，6-β-果糖)。覆盖低聚果糖的不同糖苷残基(即半乳糖和木糖)可能改变益生菌和生化性质。(C)2008爱思唯尔B.V.保留所有权利。
Fructooligosaccharides (FOSs) and polyfructosides (PSs) have received particular attention due to its beneficial effects as prebiotics. Here we report the synthesis of a new class of fructooligosaccharides by substrate and enzyme engineering. Using an engineered levansucrase enzyme (SacB of Bacillus subtilis), and sucrose analogues (alpha-Xyl-1,2-beta-Fru or alpha-Gal-1,2-beta-Fru), the product profile shifted from the fructan (levan) polymer to a range of new higher oligosaccharides (xylooligofructosides), or polysaccharides (galactopolyfructosides), of varying size. Further the enzyme was tailored by random mutagenesis, for the synthesis of short-chain fructooligosaccharides to yield variant A5 (N242H), which is unable to produce polymers. It shifts its product pattern to short-chain oligosaccharides and hydrolysis and enabled in combination with the sucrose analogue Xyl-Fru for the first time the direct synthesis of a 6-kestose analogue (alpha-Xyl-1,2-beta-Fru-2,6-beta-Fru). The different glycopyranosyl-residues (i.e. galactose and xylose) that cap fructooligosaccharides may alter prebiotic and biochemical properties. (C) 2008 Elsevier B.V. All rights reserved.