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飛雪_2019新蟲 (初入文壇)
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[求助]
求助糖部分酸水解實(shí)驗(yàn)詳細(xì)步驟
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我在一篇文獻(xiàn)上看到的一個(gè)部分酸水解的實(shí)驗(yàn)流程，可是他寫的對(duì)于我這個(gè)從未接觸過的人來說太簡(jiǎn)略了（所用的儀器，容器，100℃用水浴鍋烘箱還是馬弗爐，試劑具體用量等都弄不清楚），希望有這方面的專家來幫我看看，能不能指導(dǎo)我一下詳細(xì)是要怎么做，非常感謝。 文獻(xiàn)：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. 翻譯：1、將50 g海藻酸鈉懸浮于1 L 0.3N HCl中，并在100°C下部分水解20min。 2、離心收集上清液（pMG），調(diào)pH至7，用乙醇沉淀（使上清液最終乙醇濃度為60%，即加上清液1.5倍體積的乙醇）。 3、部分水解后的不溶物用0.3N HCl再次水解24h，離心收集沉淀。 4、將沉淀物懸浮在500 mL蒸餾水中，通過添加少量1 M NaOH溶解，并用1 N HCl調(diào)節(jié)至pH 2.85。 5、離心收集沉淀物（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、通過該方法，分別得到了4.6g、17.9g和12.4g聚（MG）、聚（M）和聚（G）嵌段。 發(fā)自小木蟲Android客戶端 |
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