Optimization of a rapid enzymatic assay for adenosine in fermentation broth based on adenosine deaminase by response surface methodology
Author(s): ZU Xin, DONG Hui-na, LI Ning, ZHANG Da-wei, SUN Yu-mei, School of Biological Engineering, Dalian Polytechnic University, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, School of Basic Medical Sciences, Lanzhou University
Pages: 81-85+91
Year: 2015 Issue: 14
Journal: Science and Technology of Food Industry
Keyword: adenosine; adenosine deaminase; high-throughput screening; response surface methodology;
Abstract: This study attempted to establish a new rapid assay for adenosine combined adenosine deaminase(ADA) with indophenol method. Based on the single factor tests,the optimal reaction conditions were explored by response surface methodology with four variables of sodium hydroxide,salicylic acid,sodium nitroferricyanide and sodium hypochlorite. And a verification experiment using the optimum composition( sodium hydroxide25.85g/L,salicylic acid 68.05g/L,sodium nitroferricyanide 2.19g/L,sodium hypochlorite 40.9m L/L) was performed.A adenosine standard curve was established under the optimum reaction conditions in range of 0.01~10mmol/L.The recovery of adenosine by ADA assay were between 95.5%~103.8% and RSD were between 0.9%~3.3%.This assay showed high consistency with HPLC method when determine the concentration of adanosine in fermentaton broth. Then the reliability of the ADA assay was further supported by spike and recovery test. The results showed that the method could rapidly and high-throughput determine the adenosine concentration in fermentation broth.
Pages: 81-85+91
Year: 2015 Issue: 14
Journal: Science and Technology of Food Industry
Keyword: adenosine; adenosine deaminase; high-throughput screening; response surface methodology;
Abstract: This study attempted to establish a new rapid assay for adenosine combined adenosine deaminase(ADA) with indophenol method. Based on the single factor tests,the optimal reaction conditions were explored by response surface methodology with four variables of sodium hydroxide,salicylic acid,sodium nitroferricyanide and sodium hypochlorite. And a verification experiment using the optimum composition( sodium hydroxide25.85g/L,salicylic acid 68.05g/L,sodium nitroferricyanide 2.19g/L,sodium hypochlorite 40.9m L/L) was performed.A adenosine standard curve was established under the optimum reaction conditions in range of 0.01~10mmol/L.The recovery of adenosine by ADA assay were between 95.5%~103.8% and RSD were between 0.9%~3.3%.This assay showed high consistency with HPLC method when determine the concentration of adanosine in fermentaton broth. Then the reliability of the ADA assay was further supported by spike and recovery test. The results showed that the method could rapidly and high-throughput determine the adenosine concentration in fermentation broth.
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