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s090604054鐵桿木蟲 (著名寫手)
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[求助]
發(fā)酵方面的翻譯
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一、方法： 將2ml 0.02mol/l PH值5.0醋酸鹽緩沖液配制成的3%右旋糖酐70KDa溶液置于35°C保溫10min,再加入適當(dāng)稀釋的酶液0.5ml保溫1hr后，利用3，5二硝基水楊酸（DNS）法測(cè)定生成的還原糖。在上述條件下每分鐘產(chǎn)生1umol還原糖（葡萄糖當(dāng)量）所需要的酶量為一個(gè)酶活單位（u）。 二、檢測(cè)步驟： （一）、對(duì)照組樣品處理 1、發(fā)酵液處理：發(fā)酵液于4000rpm條件離心10min，吸取1ml上清液，用純化水稀釋20-400倍，記作1#溶液。 2、取2mlPH值5.0的醋酸鹽緩沖液配制的3%右旋糖酐70KDa試液于10ml離心管中，35°C水浴10min備用，記做2#溶液。 3、用1000 ul移液器吸取375ul 3,5二硝基水楊酸（DNS）試液于20ml比色皿中，扣塞備用，記做3#溶液。 4、吸取1#溶液0.5ml加入2#溶液中、試液快速振勻，記作4#溶液。 5、吸取4#溶液0.5ml加入3#溶液中、試液記作5#溶液。 6、4#剩余溶液放回35°C水浴60min，記作6#溶液。 7、5#溶液搖勻后放入100°C水浴，計(jì)時(shí)，5min酶液滅活處理后，立即放入冷水冷卻后，再加入5375ul純化水混勻，記作7#溶液，作為零點(diǎn)校對(duì)組備用。 （二）、發(fā)酵樣品處理 1、 用P1000移液器吸取375ul 3,5二硝基水楊酸（DNS）試液于20ml比色皿中，扣塞備用（記作8#溶液）。 2、 吸取6#溶液0.5ml加入8#溶液，搖勻后放入100°C水浴，計(jì)時(shí)5min酶液滅活處理后，立即放入冷水充分冷卻后，再加入5375ul 純化水混勻（記作9#溶液）。 （三）、樣品檢測(cè) 分光光度計(jì)提前開啟20min后，將波長(zhǎng)旋轉(zhuǎn)到540nm，7#溶液作對(duì)照、9#溶液作樣品，檢測(cè)，記錄樣品吸光值，每一樣品測(cè)定三次吸光值，取平均值。 （四）、酶活性計(jì)算 計(jì)算公式：酶活（u/ml）=（吸光值/0.6023）*稀釋倍數(shù)/60/180*1000*10 （五）、計(jì)算公式解釋：稀釋倍數(shù)是指發(fā)酵業(yè)稀釋倍數(shù)，稀釋倍數(shù)多少取決于酶活單位且不能讓酶解液吸光值大于1；0.6023是計(jì)算常數(shù)；60是酶液反應(yīng)時(shí)間；180是葡萄糖分子量；1000是摩爾與微摩爾之間轉(zhuǎn)換；10是0.1ml酶液與1ml酶液之間轉(zhuǎn)換。 |
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s090604054
鐵桿木蟲 (著名寫手)
- 應(yīng)助: 17 (小學(xué)生)
- 金幣: 6279.9
- 散金: 99
- 紅花: 5
- 帖子: 1169
- 在線: 595.4小時(shí)
- 蟲號(hào): 879306
- 注冊(cè): 2009-10-21
- 專業(yè): 能源化工
2樓2012-07-12 22:38:10
s090604054
鐵桿木蟲 (著名寫手)
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3樓2012-07-13 21:02:36
s090604054
鐵桿木蟲 (著名寫手)
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4樓2012-07-16 23:17:12
a86267397
銅蟲 (初入文壇)
- 應(yīng)助: 3 (幼兒園)
- 金幣: 188.3
- 帖子: 23
- 在線: 14.7小時(shí)
- 蟲號(hào): 1752041
- 注冊(cè): 2012-04-12
- 性別: GG
- 專業(yè): 食品加工技術(shù)
【答案】應(yīng)助回帖
book2005593: 請(qǐng)注意版規(guī)，本版禁止使用機(jī)器翻譯，謝謝合作！ 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樓2012-07-17 10:44:41
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