Abstract
Canola seed oil-bodies were investigated as a production vehicle and immobilization matrix for xylanases. A recombinant xynC gene from Neocallimastix patriciarum encoding a xylanase (XynC) was fused to an oleosin coding sequence suitable for targeting the xylanase to the oil-body membrane. This fusion gene was introduced into Brassica napus using Agrobacterium-mediated transformation. Transgenic Canola plants were obtained expressing xylanase which was targeted to the oil-bodies of seeds as shown by analysis with XynC-specific antibodies. Oil-bodies extracted from transgenic seeds exhibited xylanase activity, indicating the immobilization of XynC on the surface of oil bodies and the functioning of the xylanase as a fusion protein. The immobilized XynC retained its optimal temperature, Km value and specificity. However, it exhibited reduced sensitivity to pH. Furthermore, it was shown that the enzyme immobilized on oil-bodies could be recycled by flotation several times without loss of activity.
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Liu, JH., Selinger, L.B., Cheng, KJ. et al. Plant seed oil-bodies as an immobilization matrix for a recombinant xylanase from the rumen fungus Neocallimastix patriciarum. Molecular Breeding 3, 463–470 (1997). https://doi.org/10.1023/A:1009604119618
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DOI: https://doi.org/10.1023/A:1009604119618