DAO (Diamine Oxidase) Purification from Natural Sources and Enzyme Enriched Food Production
Project no. 22256 · Status: Kapanmış (Yayın bekleniyor) · 2021
Summary
<p>Histamine intolerance occurs in approximately 1-3% of the population. DAO is a group of enzymes that oxidize diamines such as histamine and some primary monoamines. If DAO activity is low, the consumption of foods containing high amounts of histamine can cause intolerance reactions. The first aim of the research was to purify a DAO enzyme with high specific activity from wheat. The second aim of the research was to enrich pickles with DAO enzyme during the production process.</p><p> </p><p>Wheatgrass was obtained from registered wheat from the Aegean Agricultural Research Directorate. DAO enzyme activity was determined by spectrophotometric method and LC/MS-MS method. Ammonium sulfate protein precipitation, dialysis, column chromatography and purification processes were applied to obtain DAO enzyme. DAO enzyme was added to sauerkraut at the beginning of the fermentation stage to obtain 6 different samples. Samples without DAO were used as control. Histamine levels of sauerkraut were determined by spectrophotometric method and LC MS/MS method at the end of 3 weeks.</p><p> </p><p>We determined that the optimum pH of the DAO we obtained was 6.5 and the optimum enzyme working time was 10 min in the method we applied. The specific enzyme activity of wheatgrass homogenate analyzed by spectrophotometric method was 21.37 U/mg, while it increased to 206.46 U/mg after dialysis and 4138.07 U/mg after column chromatography.</p><p>We found that the pickle samples without added DAO had the highest histamine content at 6% salinity. Compared to the control without added DAO enzyme, the dialysis sample decreased the histamine content by 28% and the chromatography sample by 34%. 25 U of commercial DAO enzyme reduced the histamine content in pickle juice by 92% after 3 weeks compared to the control, whereas the DAO enzyme obtained by dialysis (25 U/L) reduced it by 87%. With the widespread use of this application, we anticipate that people with histamine intolerance will be able to safely use foods that cause intolerance and their quality of life will improve.</p>