Schrader Paul S, Burrows Elizabeth H, Ely Roger L
Department of Chemical Engineering, Yale University, New Haven, Connecticut 06510, USA.
Anal Chem. 2008 Jun 1;80(11):4014-9. doi: 10.1021/ac702633q. Epub 2008 Apr 29.
This paper describes a screening assay, compatible with high-throughput bioprospecting or molecular biology methods, for assessing biological hydrogen (H2) production. While the assay is adaptable to various physical configurations, we describe its use in a 96-well, microtiter plate format with a lower plate containing H2-producing cyanobacteria strains and controls and an upper, membrane-bottom plate containing a color indicator and a catalyst. H2 produced by cells in the lower plate diffuses through the membrane into the upper plate, causing a color change that can be quantified with a microplate reader. The assay is reproducible, semiquantitative, sensitive down to at least 20 nmol of H2, and largely unaffected by oxygen, carbon dioxide, or volatile fatty acids at levels appropriate to biological systems.
本文描述了一种与高通量生物勘探或分子生物学方法兼容的筛选测定法,用于评估生物制氢。虽然该测定法可适应各种物理配置,但我们描述了其在96孔微量滴定板形式中的应用,其中下层板含有产氢蓝藻菌株和对照,上层板为膜底板,含有颜色指示剂和催化剂。下层板中的细胞产生的氢气通过膜扩散到上层板中,导致颜色变化,可使用酶标仪进行定量。该测定法具有可重复性、半定量性,对至少20 nmol氢气敏感,并且在生物系统适当的水平下,很大程度上不受氧气、二氧化碳或挥发性脂肪酸的影响。