A rapid enzymatic assay for high-throughput screening of adenosine-producing strains
메타 데이터
바이오화학분류
바이오정밀화학
화학제품
연료
논문
A rapid enzymatic assay for high-throughput screening of adenosine-producing strains
학술지
Microbial biotechnology
저자명
Dong, Huina; Zu, Xin; Zheng, Ping; Zhang, Dawei
초록
<P>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 NH<SUB>3</SUB>, 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.</P>