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s090604054铁杆木虫 (著名写手)
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[求助]
发酵方面的翻译
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一、方法： 将2ml 0.02mol/l PH值5.0醋酸盐缓冲液配制成的3%右旋糖酐70KDa溶液置于35°C保温10min,再加入适当稀释的酶液0.5ml保温1hr后，利用3，5二硝基水杨酸（DNS）法测定生成的还原糖。在上述条件下每分钟产生1umol还原糖（葡萄糖当量）所需要的酶量为一个酶活单位（u）。 二、检测步骤： （一）、对照组样品处理 1、发酵液处理：发酵液于4000rpm条件离心10min，吸取1ml上清液，用纯化水稀释20-400倍，记作1#溶液。 2、取2mlPH值5.0的醋酸盐缓冲液配制的3%右旋糖酐70KDa试液于10ml离心管中，35°C水浴10min备用，记做2#溶液。 3、用1000 ul移液器吸取375ul 3,5二硝基水杨酸（DNS）试液于20ml比色皿中，扣塞备用，记做3#溶液。 4、吸取1#溶液0.5ml加入2#溶液中、试液快速振匀，记作4#溶液。 5、吸取4#溶液0.5ml加入3#溶液中、试液记作5#溶液。 6、4#剩余溶液放回35°C水浴60min，记作6#溶液。 7、5#溶液摇匀后放入100°C水浴，计时，5min酶液灭活处理后，立即放入冷水冷却后，再加入5375ul纯化水混匀，记作7#溶液，作为零点校对组备用。 （二）、发酵样品处理 1、 用P1000移液器吸取375ul 3,5二硝基水杨酸（DNS）试液于20ml比色皿中，扣塞备用（记作8#溶液）。 2、 吸取6#溶液0.5ml加入8#溶液，摇匀后放入100°C水浴，计时5min酶液灭活处理后，立即放入冷水充分冷却后，再加入5375ul 纯化水混匀（记作9#溶液）。 （三）、样品检测 分光光度计提前开启20min后，将波长旋转到540nm，7#溶液作对照、9#溶液作样品，检测，记录样品吸光值，每一样品测定三次吸光值，取平均值。 （四）、酶活性计算 计算公式：酶活（u/ml）=（吸光值/0.6023）*稀释倍数/60/180*1000*10 （五）、计算公式解释：稀释倍数是指发酵业稀释倍数，稀释倍数多少取决于酶活单位且不能让酶解液吸光值大于1；0.6023是计算常数；60是酶液反应时间；180是葡萄糖分子量；1000是摩尔与微摩尔之间转换；10是0.1ml酶液与1ml酶液之间转换。 |
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s090604054
铁杆木虫 (著名写手)
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2楼2012-07-12 22:38:10
s090604054
铁杆木虫 (著名写手)
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3楼2012-07-13 21:02:36
s090604054
铁杆木虫 (著名写手)
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4楼2012-07-16 23:17:12
a86267397
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【答案】应助回帖
book2005593: 请注意版规，本版禁止使用机器翻译，谢谢合作！ 2012-07-18 07:48:51
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1.method: Will 2 ml 0.02 mol/l PH 5.0 acetic acid salt buffer makes up to 3% dextran 70 KDa 35 ° C heat preservation solution in 10 min, add the liquid enzymes diluted properly 0.5 ml insulation 1 hr later, using 3, 5 two nitro salicylic acid (DNS) method for the determination of the generation of reducing sugar. In the above conditions ,a Enzymes living unit (u) means every minute produce 1 umol reducing sugar (glucose equivalent) need enzyme quantity. 2.Detecting steps: (1), sample with the control group 1, fermented liquid treatment: fermented liquid in condition 4000 RPM conditions centrifugal 10 min, draw on 1 ml clear liquid, with purified water dilute 20-400 times, remember as 1 # solution. 2, take 2 ml PH value of 5.0 acetic acid salt buffer made 3% dextran 70 KDa solution in 10 ml centrifugal liquid pipe, 35 ℃ water bath 10 min, remember as 2 # solution. 3,draw 375 ul 3, 5 two nitro salicylic acid (DNS) solution in the 20 ml than color dish with 1000 ul pipettor, buckle plug to spare, remember as 3# solution. 4,draw 0.5 ml liquor from 1# solution add to 2# solution, fast vibration well, remember as 4 # solution. 5, draw 0.5 ml liquor from 4# solution add to 3# solution, remember as #5 solution. 6, 4# solution remaining at 35 ℃ water bath 60 min, written as #6 solution. 7, 5# solution shake well and put into 100 ℃ water bath, timing, 5 min enzyme liquid inactivated after processing,cooling in cold water immediately, then add 5375 ul purification water blending, and remember as 7# solution, as zero proofreading group. Set aside. (2), fermentation sample processing 1,draw 375 ul 3, 5 two nitro salicylic acid (DNS) solution in the 20 ml than color dish with 1000 ul pipettor, buckle plug spare (written for 8# solution). 2,draw 0.5 ml liquid from 6# solution into 8# solution, shake well and put into 100℃ water bath, timing 5 min enzyme liquid inactivated after processing, immediately into cold water sufficient cooling, add 5375 ul purification water blending (written for 9# solution). (3), sample testing Spectrophotometer open after 20 min in advance, will be rotated to 540 nm wavelength, 7# solution as control, 9# solution as samples, detection, record sample absorb light value, each sample determine three time absorb light value, take average. (4), enzyme activity calculation Formula: the enzyme live (u/ml) = (absorb light value / 0.6023) * diluted times / 60/180 * 1000 * 10 (5) formula explanation: diluted times is refers to the fermentation industry diluted times, diluted times depends on how much enzyme unit and can't let live enzyme solution liquid absorb light a value greater than 1; 0.6023 is a calculation constant; 60 is enzyme liquid reaction time; 180 is glucose molecular weight; 1000 is Moore and micro switch between Moore; 10 is 0.1 ml enzyme solution and 1 ml enzyme convert between liquid. |
5楼2012-07-17 10:44:41
