A rapid enzymatic assay for high-throughput screening of adenosine-producing strains.
A rapid enzymatic assay for high-throughput screening of adenosine-producing strains.
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用于高通量筛选腺苷生产菌株的快速酶测定
DOI：
10.1111/1751-7915.12189
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发表时间：
2015-03
影响因子：
5.7
通讯作者：
Zhang D
中科院分区：
文献类型：
--
作者：
Dong H;Zu X;Zheng P;Zhang D
腺苷是组织功能的主要局部调节剂，在工业上用作生产药用核苷物质的前体。腺苷高产菌株的高通量筛选对工业微生物育种具有重要意义。将腺苷脱氨酶(ADA)与吲哚酚结合，建立了腺苷的酶法测定方法。ADA催化腺苷裂解成肌苷和NH3，后者可用吲哚酚法准确测定。该检测系统经过优化，具有良好的性能，可以容忍无机盐和多种营养成分添加到检测混合物中。用96孔微孔板可以准确地测定腺苷。加标和回收试验表明，该方法可以准确和重复性地测定发酵液中腺苷的增加，而无需任何去除蛋白质和潜在干扰低分子分子的预处理。该方法还可用于腺苷高产菌株的高通量筛选。ADA分析的高选择性和准确性提供了大量样品中腺苷的快速和高通量分析。
Adenosine is a major local regulator of tissue function and industrially useful as precursor for the production of medicinal nucleoside substances. High-throughput screening of adenosine overproducers is important for industrial microorganism breeding. An enzymatic assay of adenosine was developed by combined adenosine deaminase (ADA) with indophenol method. The ADA catalyzes the cleavage of adenosine to inosine and NH3, the latter can be accurately determined by indophenol method. The assay system was optimized to deliver a good performance and could tolerate the addition of inorganic salts and many nutrition components to the assay mixtures. Adenosine could be accurately determined by this assay using 96-well microplates. Spike and recovery tests showed that this assay can accurately and reproducibly determine increases in adenosine in fermentation broth without any pretreatment to remove proteins and potentially interfering low-molecular-weight molecules. This assay was also applied to high-throughput screening for high adenosine-producing strains. The high selectivity and accuracy of the ADA assay provides rapid and high-throughput analysis of adenosine in large numbers of samples.
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影响因子：
7.4
作者：
Li, Le-Le;Ge, Pinghua;Selvin, Paul R.;Lu, Yi
通讯作者：
Lu, Yi
影响因子：
6.4
作者：
Dong H;Zhang D
通讯作者：
Zhang D
一种新型无酶、无标记荧光适体传感器，用于腺苷的放大检测
影响因子：
12.6
作者：
Fu, Bo;Cao, Jichao;Wang, Lei
通讯作者：
Wang, Lei
影响因子：
6.2
作者：
González-Rodríguez, J;Pérez-Juan, P;de Castro, MDL
通讯作者：
de Castro, MDL
影响因子：
7.4
作者：
NGO, TT;PHAN, APH;LENHOFF, HM
通讯作者：
LENHOFF, HM