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Cloning and overexpression of the xylose isomerase gene from Burkholderia sacchari and production of polyhydroxybutyrate from xylose

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A different organization for the xyl operon was found in different genomes of Burkholderia and Pseudomomas species. Degenerated primers were designed based on Burkholderia genomes and used to amplify the xylose isomerase gene (xylA) from Burkholderia sacchari IPT101. The gene encoded a protein of 329 amino acids, which showed the highest similarity (90%) to the homologous gene of Burkholderia dolosa. It was cloned in the broad host range plasmid pBBR1MCS-2, which partially restored growth and polyhydroxybutyrate production capability in xylose to a B. sacchari xyl- mutant. When xylA was overexpressed in the wild-type strain, it was not able to increase growth and polyhydroxybutyrate production, suggesting that XylA activity is not limiting for xylose utilization in B. sacchari.

L’opéron xyl est organisé de façon différente dans différents génomes des espèces Burkholderia et Pseudomonas. Des amorces dégénérées ont été conçues à partir des génomes de Burkholderia pour amplifier le gène de la xylose isomérase (xylA) de Burkholderia sacchari IPT101. Le gène codait une protéine de 329 acides aminés montrant la plus forte similarité (90 %) avec le gène homologue de Burkholderia dolosa. Il a été cloné dans le plasmide pBBR1MCS-2 possédant un large spectre d’hôtes, et a restauré partiellement la croissance et la capacité de production de polyhydroxybutyrate sur xylose du mutant xyl- de B. sacchari. Lorsque xylA était surexprimé dans la souche de type sauvage, il était incapable d’augmenter la croissance et la production de polyhydroxybutyrate, suggérant que l’activité de XylA ne soit pas limitante à l’utilisation de xylose par B. sacchari.

Document Type: Research Article

Publication date: 2009-08-01

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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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