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Immobilized Sclerotinia sclerotiorum invertase to produce invert sugar syrup from industrial beet molasses by product

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Abstract

The fungus Sclerotinia sclerotiorum produces invertase activity during cultivation on many agroindustrial residues. The molasses induced invertase was purified by DEAE-cellulose chromatography. The molecular mass of the purified enzyme was estimated at 48 kDa. Optimal temperature was determined at 60 °C and thermal stability up to 65 °C. The enzyme was stable between pH 2.0 and 8.0; optimum pH was about 5.5. Apparent Km and Vmax for sucrose were estimated to be respectively 5.8 mM and 0.11 μmol/min. The invertase was activated by β-mercaptoethanol. Free enzyme exhibited 80 % of its original activity after two month’s storage at 4 °C and 50 % after 1 week at 25 °C. In order to investigate an industrial application, the enzyme was immobilized on alginate and examined for invert sugar production by molasses hydrolysis in a continuous bioreactor. The yield of immobilized invertase was about 78 % and the activity yield was 59 %. Interestingly the immobilized enzyme hydrolyzed beet molasses consuming nearly all sucrose. It retained all of its initial activity after being used for 4 cycles and about 65 % at the sixth cycle. Regarding productivity; 20 g/l of molasses by-product gave the best invert sugar production 46.21 g/day/100 g substrate related to optimal sucrose conversion of 41.6 %.

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Acknowledgments

This work was supported by the financial project of LIP-MB Laboratory, INSAT, Carthage University, Ministry of Higher Education and Scientific Research of Tunisia.

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Authors and Affiliations

  1. Laboratory of Protein Engineering and Bioactive Molecules (LIP-MB), National Institute of Applied Sciences and Technology, University of Carthage, B. P. 676, 1080, Tunis Cedex, Tunisia

    Refka Mouelhi, Ferid Abidi, Said Galai & M. Nejib Marzouki

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  1. Refka Mouelhi
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  2. Ferid Abidi
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  3. Said Galai
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  4. M. Nejib Marzouki
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Correspondence to Refka Mouelhi.

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Ferid Abidi and Galai Said have contributed equally to this work.

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Mouelhi, R., Abidi, F., Galai, S. et al. Immobilized Sclerotinia sclerotiorum invertase to produce invert sugar syrup from industrial beet molasses by product. World J Microbiol Biotechnol 30, 1063–1073 (2014). https://doi.org/10.1007/s11274-013-1525-8

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  • DOI: https://doi.org/10.1007/s11274-013-1525-8

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