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Optimal conditions for levan formation by an overexpressed recombinant levansucrase
Authors:Hafedh Belghith  Ki-Bang Song  Chul-Ho Kim  Sang-Ki Rhee
Affiliation:(1) Applied Microbiology Research Division, Korea Research Institute of Bioscience and Biotechnology, KIST, P.O.Box 115, 305-600 Yusong, Taejon, Korea
Abstract:Summary A genetically modified levansucrase, which contained His-affinity tag in its C-terminal, was constructed by PCR reaction using two synthetic primers. This modified protein was produced up to 30 % in total cell protein of E. coli, and was purified by a one-step affinity chromatography. The optimum pH for levan production was pH 5 and the optimum temperature was 0 °C. The higher velocity of levan formation within shorter enzyme reaction times was achieved by increasing the levels of enzyme concentration. The optimal sucrose concentration for levan production was around 20 %. Under these conditions, more than 50 g levan/l was produced.
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