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The improvement of lipase secretion and stability by addition of inert compounds into Acinetobacter calcoaceticus cultures

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Abstract:



Acinetobacter calcoaceticus BD413 produces variable amounts of an exocellular lipase that becomes rapidly inactivated upon secretion. To achieve high yield and protect the enzyme, we assayed the addition of several inert compounds to cell-free supernatants, cell fractions, and whole cultures. Glass beads, poly(ethylene glycol) 600, Triton X-100, saccharose, gum arabic, and -cyclodextrin were among the compounds tested. -Cyclodextrin and gum arabic (and saccharose to a lesser extent) were effective enzyme stabilizers in cell-free supernatants, while gum arabic, glass beads, and Triton X-100 improved lipase secretion from cells, and, therefore, total lipase yield (30–50%, according to the additive). In whole cultures, -cyclodextrin was the most effective additive, particularly in combination with glass beads or gum arabic. Indeed, cultures containing -cyclodextrin plus gum arabic were able to maintain 95% (±1.5%) of the initial lipase activity for more than 16 h, while control cultures with no additives maintained only 10% (±4%) of the enzyme activity after the same period. In conclusion, the addition of inert compounds in cultures may be considered a useful approach for achieving increased yield and lipase stabilization, amenable for downstream processing.Key words: Acinetobacter calcoaceticus, lipase, secretion, stabilization, inert additives.

Acinetobacter calcoaceticus BD413 peut produire des quantités variables d'une lipase exocellulaire qui est rapidement inactivée à la suite de sa sécrétion. Afin d'obtenir un rendement élevé et de protéger l'enzyme, nous avons fait des tentatives d'ajout d'une variété de composés inertes aux surnageants acellulaires, aux fractions cellulaires et aux cultures entières. Parmi les composés testés se retrouvaient des billes de verre, du polyéthylène glycol 600, du Triton X-100, du saccharose, de la gomme arabique et de la -cyclodextrine. La cyclodextrine et la gomme arabique (et le saccharose dans une moindre mesure) furent d'efficaces stabilisateurs enzymatiques, alors que la gomme arabique, les billes de verre et le Triton X-100 ont amélioré la sécrétion de lipase par les cellules, et donc le rendement total de lipase (de 30 à 50 %, selon l'additif). Dans les cultures entières, la -cyclodextrine se sont avérées les additifs les plus efficaces, surtout en combinaison avec des billes de verre ou de la gomme arabique. En effet, les cultures contenant de la -cyclodextrine et de la gomme arabique furent capables de conserver 95 % (± 1,5 %) de l'activité lipase initiale pendant plus de 16 heures, alors que les culture témoins sans additifs n'ont conservé que 10 % (± 4 %) de leur activité lipase à la fin de cette même période. En conclusion, l'ajout de composés inertes dans les cultures pourrait être considéré comme une approche utile dans le but d'obtenir un rendement supérieur et une stabilisation de la lipase, qui pourrait se prêter à des traitements subséquents.Mots clés : Acinetobacter calcoaceticus, lipase, sécrétion, stabilisation, additifs inertes

Keywords: Acinetobacter calcoaceticus; additifs inertes; inert additives; lipase; secretion; stabilisation; stabilization; sécrétion

Document Type: Research Article

Publication date: December 1, 2002

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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