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直链淀粉在体外通过支链淀粉的延伸和裂解来合成。

Amylose is synthesized in vitro by extension of and cleavage from amylopectin.

作者信息

van de Wal M, D'Hulst C, Vincken J P, Buléon A, Visser R, Ball S

机构信息

Department of Plant Breeding, Graduate School of Experimental Plant Sciences, Agricultural University of Wageningen, P. O. Box 386, 6700 AJ Wageningen, The Netherlands.

出版信息

J Biol Chem. 1998 Aug 28;273(35):22232-40. doi: 10.1074/jbc.273.35.22232.

Abstract

Amylose synthesis was obtained in vitro from purified Chlamydomonas reinhardtii starch granules. Labeling experiments clearly indicate that initially the major granule-bound starch synthase extends glucans available on amylopectin. Amylose synthesis occurs thereafter at rates approaching or exceeding those of net polysaccharide synthesis. Although these results suggested that amylose originates from cleavage of a pre-existing external amylopectin chain, such transfer of chains from amylopectin to amylose was directly evidenced from pulse-chase experiments. The structure of the in vitro synthesized amylose could not be distinguished from in vivo synthesized amylose by a variety of methods. Moreover high molecular mass branched amylose synthesis preceded that of the low molecular mass, suggesting that chain termination occurs consequently to glucan cleavage. Short pulses of synthesis followed by incubation in buffer with or without ADP-Glc prove that transfer requires the presence of the glucosyl-nucleotide. Taken together, these observations make a compelling case for amylopectin acting as the in vivo primer for amylose synthesis. They further prove that extension is followed by cleavage. A model is presented that can explain the major features of amylose synthesis in plants. The consequences of intensive amylose synthesis on the crystal organization of amylopectin are reported through wide angle x-ray analysis of the in vitro synthesized polysaccharides.

摘要

直链淀粉的合成是在体外从莱茵衣藻纯化的淀粉颗粒中获得的。标记实验清楚地表明,最初主要的颗粒结合淀粉合酶延伸支链淀粉上可用的葡聚糖。此后直链淀粉的合成速率接近或超过净多糖合成速率。尽管这些结果表明直链淀粉起源于预先存在的外部支链淀粉链的裂解,但通过脉冲追踪实验直接证明了链从支链淀粉向直链淀粉的这种转移。通过多种方法无法区分体外合成的直链淀粉与体内合成的直链淀粉的结构。此外,高分子量支链直链淀粉的合成先于低分子量直链淀粉,这表明链终止发生在葡聚糖裂解之后。在含有或不含有ADP-葡萄糖的缓冲液中孵育之前进行短时间的合成脉冲证明转移需要葡糖基核苷酸的存在。综上所述,这些观察结果有力地证明了支链淀粉作为直链淀粉合成的体内引物。它们进一步证明延伸之后是裂解。提出了一个可以解释植物中直链淀粉合成主要特征的模型。通过对体外合成多糖的广角X射线分析报告了大量直链淀粉合成对支链淀粉晶体组织的影响。

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