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求助糖部分酸水解实验详细步骤
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我在一篇文献上看到的一个部分酸水解的实验流程，可是他写的对于我这个从未接触过的人来说太简略了（所用的仪器，容器，100℃用水浴锅烘箱还是马弗炉，试剂具体用量等都弄不清楚），希望有这方面的专家来帮我看看，能不能指导我一下详细是要怎么做，非常感谢。 文献：A. Inoue, K. Takadono, R. Nishiyama, K. Tajima, T. Ojima, Characterization of an alginate lyase, FlAlyA, from Flavobacterium sp. strain UMI-01 and its expression in Escherichia coli, Mar. Drugs 2 8 (2014) 4693e4712. 原文：Briefly, 50 g of sodium alginate was suspended in 1 L of 0.3 N HCl and partially hydrolyzed at 100 °C for 20 min. Poly(MG) block released to the supernatant was collected by centrifugation, neutralized, and precipitated with ethanol in a final concentration of 60%. Insoluble materials after the partial hydrolysis were again hydrolyzed with 0.3 N HCl for 24 h and collected by centrifugation. The precipitates were suspended in 500 mL of distilled water, dissolved by the addition of small amount of 1 M NaOH, and adjusted to pH 2.85 with 1 N HCl. The precipitates formed (poly(G) block) were collected by centrifugation, dissolved in 10 mM Na2CO3, and precipitated with ethanol. While the supernatant (poly(M) block) was neutralized with 0.1 M Na2CO3 and precipitated with ethanol. By this procedure, 4.6 g, 17.9 g, and 12.4 g of poly(MG), poly(M) and poly(G) blocks, respectively, were obtained. 翻译：1、将50 g海藻酸钠悬浮于1 L 0.3N HCl中，并在100°C下部分水解20min。 2、离心收集上清液（pMG），调pH至7，用乙醇沉淀（使上清液最终乙醇浓度为60%，即加上清液1.5倍体积的乙醇）。 3、部分水解后的不溶物用0.3N HCl再次水解24h，离心收集沉淀。 4、将沉淀物悬浮在500 mL蒸馏水中，通过添加少量1 M NaOH溶解，并用1 N HCl调节至pH 2.85。 5、离心收集沉淀物（pG），溶解于10mM（1mmol/L） Na2CO3中，用乙醇沉淀。 6、上清液（pM）用0.1 M Na2CO3中和，用乙醇沉淀。 7、通过该方法，分别得到了4.6g、17.9g和12.4g聚（MG）、聚（M）和聚（G）嵌段。 发自小木虫Android客户端 |
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