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PLANT PHYSIOLOGY , Vol 109, Issue 4 1199-1205, Copyright © 1995 by American Society of Plant Biologists
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GENE REGULATION AND MOLECULAR GENETICS |
Stable Accumulation of Aspergillus niger Phytase in Transgenic Tobacco Leaves
T. C. Verwoerd, P. A. van Paridon, AJJ. van Ooyen, JWM. van Lent, A. Hoekema and J. Pen
MOGEN International N.V., Einsteinweg 97, 2333 CB Leiden, The Netherlands (T.C.V., J.P.)
Phytase from Aspergillus niger increases the availability of phosphorus
from feed for monogastric animals by releasing phosphate from the substrate
phytic acid. A phytase cDNA was constitutively expressed in transgenic
tobacco (Nicotiana tabacum) plants. Secretion of the protein to the
extracellular fluid was established by use of the signal sequence from the
tobacco pathogen-related protein S. The specific phytase activity in
isolated extracellular fluid was found to be approximately 90-fold higher
than in total leaf extract, showing that the enzyme was secreted. This was
confirmed by use of immunolocalization. Despite differences in
glycosylation, specific activities of tobacco and Aspergillus phytase were
identical. Phytase was found to be biologically active and to accumulate in
leaves up to 14.4% of total soluble protein during plant maturation.
Comparison of phytase accumulation and relative mRNA levels showed that
phytase stably accumulated in transgenic leaves during plant growth.
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