| 查看: 632 | 回复: 1 | ||
| 本帖产生 1 个 翻译EPI ，点击这里进行查看 | ||
[求助]
求翻译，谢谢，站内信求大神帮忙
|
||
|
A novel glyA gene from the marine bacterium Alcanivorax sp. was cloned and expressed in Escherichia coli BL21(DE3). The recombinant glyA encodes a polypeptide of 418 amino acids, which was designated as AdSHMT that shows the highest identity (70%) with a SHMT from Shewanella algae. The purified enzyme showed a single band at about 45 kDa by SDS-PAGE analysis. It was found that AdSHMT exhibited the maximal activity at 50 ◦ C and pH 7.0. The K m , V max , and K cat values of AdSHMT against dl-threo-3-phenylserine were calculated to be 0.097 mol/L, 3.255 ?mol/min/mg and 2.451/s, respectively. More importantly, RP-HPLC detection showed that the AdSHMT achieved an 88.37% molecular conversion rate in catalyzing glycine to l-serine, with the final concentration of l-serine being 353.15 mM in the reaction at 35 ◦ C and 22nd hour when the initial concentration of the substrate (glycine) was 0.399 M. The molecular conversion rate of the AdSHMT from the Alcanivorax sp. was 1.26-fold that of the EcSHMT from the E. coli,which is currently applied in industrial production. Therefore, AdSHMT has the potential for industrial applications due to its high enzymatic conversion rate. |
回复此楼» 猜你喜欢
博士延得我，科研能力直往上蹿
已经有7人回复
-
退学或坚持读
已经有27人回复
-
面上基金申报没有其他的参与者成吗
已经有5人回复
-
有70后还继续奋斗在职场上的吗？
已经有5人回复
-
遇见不省心的家人很难过
已经有22人回复
-
多组分精馏求助
已经有6人回复
UTKProgress
木虫 (小有名气)
- 翻译EPI: 11
- 应助: 7 (幼儿园)
- 金币: 3605.5
- 散金: 7
- 红花: 3
- 帖子: 280
- 在线: 236.4小时
- 虫号: 3500502
- 注册: 2014-10-27
- 性别: GG
- 专业: 高分子合成化学
【答案】应助回帖
★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★
bxzc123: 金币+20, 翻译EPI+1, ★★★很有帮助 2014-11-25 22:11:57
bxzc123: 金币+20, 翻译EPI+1, ★★★很有帮助 2014-11-25 22:11:57
| 一种来自海洋细菌Alcanivorax的新型的glyA基因被克隆，并在大肠杆菌BL21（DE3）上表达。重组的glyA 编码了一个由418个氨基酸组成的多肽，被命名为 AdSHMT， 它显示出与来自Shewanella藻的SHMT最高70％的相似。在SDS-PAGE分析中纯化的酶为45 kDa的单链 (多肽)。结果发现，在50◦C和pH7.0时，AdSHMT表现出最大的活性。AdSHMT against dl-threo-3-phenylserine (这里的一个专业知识点，我不懂。应该是一生化反应)的 Km, Vmax, 和 Kcat 数值分别为0097摩尔/升，3255摩尔/分钟/毫克和2451/秒。更重要的是反相高效液相色谱法检测表明，在35 ◦ C时（AdSHMT）催化glycine为l-serine反应中当glycine的初始浓度为0.399 M时在第22小时AdSHMT取得了88.37%的转化率。从Alcanivorax SP到AdSHMT的分子转化率是目前在工业生产中应用的从E. coli到EcSHMT的转化率的1.26倍。因此，AdSHMT因其高的酶转化率可能在工业上得到应用。 |
2楼2014-11-25 10:06:15