Samain E, Chazalet V, Geremia R A
Centre de Recherche sur les Macromolécules Végétales (CERMAV-CNRS), Grenoble, France.
J Biotechnol. 1999 Jun 11;72(1-2):33-47. doi: 10.1016/s0168-1656(99)00048-6.
High cell density cultivation of recombinant Escherichia coli strains harboring the nodBC genes (encoding chitooligosaccharide synthase and chitooligosaccharide N-deacetylase, respectively) from Azorhizobium caulinodans has been previously described as a practical method for the preparation of gram-scale quantities of penta-N-acetyl-chitopentaose and tetra-N-acetylchitopentaose (Samain, E., Drouillard, S., Heyraud, A., Driguez, H., Geremia, R.A., 1997. Carbohydr. Res. 30, 235-242). We have now extended this method to the production of sulfated and O-acetylated derivatives of these two compounds by coexpressing nodC or nodBC with nodH and/or nodL that encode chitooligosaccharide sulfotransferase and chitooligosaccharide O-acetyltransferase, respectively. In addition, these substituted chitooligosaccharides were also obtained as tetramers by using nodC from Rhizobium meliloti instead of nodC from A. caulinodans. These compounds should be useful precursors for the preparation of Nod factor analogues by chemical modification.
此前已描述了对携带来自茎瘤固氮根瘤菌的nodBC基因（分别编码壳寡糖合酶和壳寡糖N - 脱乙酰酶）的重组大肠杆菌菌株进行高细胞密度培养，这是一种制备克级数量的五 - N - 乙酰 - 壳五糖和四 - N - 乙酰壳五糖的实用方法（Samain, E., Drouillard, S., Heyraud, A., Driguez, H., Geremia, R.A., 1997. Carbohydr. Res. 30, 235 - 242）。我们现在将此方法扩展到通过分别与编码壳寡糖磺基转移酶和壳寡糖O - 乙酰转移酶的nodH和/或nodL共表达nodC或nodBC来生产这两种化合物的硫酸化和O - 乙酰化衍生物。此外，通过使用来自苜蓿根瘤菌的nodC而非来自茎瘤固氮根瘤菌的nodC，这些取代的壳寡糖也以四聚体形式获得。这些化合物应是通过化学修饰制备Nod因子类似物的有用前体。
