%0 Journal Article
%T Microplate for high throughput screening of 6-hydroxynicotinic acid transforming strains<br>基于微孔板的高通量筛选6-羟基烟酸转化菌方法的建立
%A Yao Yang
%A Sheng Yuan
%A Yijun Dai
%A <br>杨瑶
%A 袁生
%A 戴亦军等
%J 微生物学报
%D 2008
%I 
%X Recently, new insecticides containing 3-chloropyridylmethyl group as a versatile building block have been developed, among which imidachloprid is a promising one. The synthesis of imidachloprid can use 6-Hydroxynicotinic acid, the first intermediate of the bacterial degradation of nicotinic acid, as a starting material. It is difficult to hydroxylate nicotinic acid at C6 position in a chemical synthesis. However, biotransformation can produce 6-hydroxynicotinic acid with industrial application possibility. Therefore, methods for large scale screening of 6-hydroxynicotinic acid-producing microorganisms are urgent. A high through-put screening method for 6-hydroxynicotinic acid transforming strains was established by determining 6-hydroxynicotinic acid based on 96-well Microplate-Multiskan Spectrum. The determination wavelength and the reference wavelength of 6-hydroxynicotinic acid were 251nm and 231nm respectively. Beer's law is obeyed in the range of 0.5-11 microg/mL (R2 = 0.9999) for measuring 6-hydroxynicotinic acid. The average recovery rate was 99.11%-100.81%. The results showed that there was no apparent difference between our microplate method and previous HPLC method in the detecting 6-hydroxynicotinic acid formation. The microplate method is simple, convenient and accurate. It has the potential for large scale (about 2000 - 5000 reactions/d) screening of 6-hydroxynicotinic acid-producing microorganisms.
%K Microplate
%K High through-put screening
%K Dual-wavelength spectrum determination
%K Nicotinic acid
%K 6-hydroxynicofinic acid<br>微孔板
%K 高通量筛选
%K 双波长紫外分光光度测定
%K 烟酸
%K 6-羟基烟酸
%K 微孔板
%K 高通量筛选
%K 羟基
%K 烟酸含量
%K 转化
%K 方法
%K strains
%K transforming
%K acid
%K high
%K throughput
%K screening
%K 反应
%K 筛选量
%K 利用
%K 平均回收率
%K 比尔定律
%K 朗伯
%K 线性关系
%K 浓度范围
%K 样品
%K 参比波长
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=87C7244656E9352B91938437EE896DAC&yid=67289AFF6305E306&vid=B6DA1AC076E37400&iid=CA4FD0336C81A37A&sid=4BB057F167CF3A60&eid=EDA22B444205D04A&journal_id=0001-6209&journal_name=微生物学报&referenced_num=0&reference_num=15