Like other branched-chain higher alcohols used as biofuels, isoamyl alcohol has attracted considerable attention because of its advantages, which include high energy density, low hygroscopicity, and compatibility with the current infrastructure. Previous attempts to increase the microbial production of isoamyl alcohol have yielded great progress, but the existing methods of detecting isoamyl alcohol based on gas chromatography and high-performance liquid chromatography are laborious and time-consuming. In this study, we developed a simple colorimetric assay to determine high isoamyl alcohol–producing strains. The assay was based on isoamyl alcohol oxidase and peroxidase (IAOP assay) and could be performed in microplate with high throughput and had a specific detection range of 0–20 mM. Characterization analysis revealed that the developed IAOP assay was highly specific for isoamyl alcohol relative to other branched-chain alcohols. Little interference with the assay was observed from the fermentation media, microorganisms, and fermentation byproducts (e.g., lactic acid, acetic acid). We conclude that the enzyme-based IAOP assay can be used for high-throughput monitoring of strains that produce isoamyl alcohol and could be adjusted to screen for strains that produce many other metabolites.

像其他用作生物燃料的支链高级醇一样，异戊醇由于其优点而备受关注，包括高能量密度，低吸湿性以及与现有基础设施的相容性。先前增加异戊醇的微生物产量的尝试已经取得了很大的进步，但是基于气相色谱和高效液相色谱的检测异戊醇的现有方法既费力又费时。在这项研究中，我们开发了一种简单的比色测定法来确定产生高异戊醇的菌株。该测定基于异戊醇氧化酶和过氧化物酶（IAOP测定），可以在微量培养板中以高通量进行，检测范围为0–20 mM。表征分析表明，相对于其他支链醇，已开发的IAOP分析对异戊醇具有高度特异性。从发酵培养基，微生物和发酵副产物（例如乳酸，乙酸）中观察到的对测定的干扰很小。我们得出结论，基于酶的IAOP分析可用于高通量监测产生异戊醇的菌株，并可进行调整以筛选产生许多其他代谢产物的菌株。