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Establishment of an ex vivo laticifer cell suspension culture from Taraxacum brevicorniculatum as a production system for cis-isoprene

 
: Post, J.; Eisenreich, W.; Huber, C.; Twyman, R.M.; Prüfer, D.; Schulze Gronover, C.

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Journal of molecular catalysis. B, Enzymatic 103 (2014), pp.85-93
ISSN: 1381-1177
ISSN: 1873-3158
English
Journal Article
Fraunhofer IME ()

Abstract
Laticifers are highly specialized plant cells that produce latex enriched with secondary metabolites. The articulated laticifers of Taraxacum brevicorniculatum synthesize natural rubber, an industrially-valuable composite biopolymer comprising >95% high-molecular-weight (HMW) poly(cis-1,4-isoprene). Here we present a proof-of-concept approach for the cultivation of cell suspension cultures exclusively containing laticifers. We transformed T. brevicorniculatum plants with a plasmid conferring laticifer-specific hygromycin resistance. Transgenic callus tissue was used to induce a cell suspension culture under antibiotic selection to favor laticifer growth. The cultured cells appeared laticiferous in terms of morphology and expressed laticifer-specific genes. Confocal laser scanning microscopy revealed intracellular lipid accumulation in vesicle-like structures. Nuclear magnetic resonance and diffusion ordered spectroscopy indicated the presence of mid-length poly(cis-1,4 -isoprene) chains but no high HMW natural rubber in the cells. Precursor feeding with mevalonolactone increased the accumulation of poly(cis-1,4-isoprene) by 17-fold, reaching a concentration of 2.7 mg/g dry weight. Our approach could lead to the development of a production platform for the efficient conversion of isopentenyl diphosphate into poly(cis-1,4-isoprene) in an optimized cell suspension culture system. The apparent absence of HMW natural rubber is discussed in terms of our current knowledge of rubber biosynthesis.

: http://publica.fraunhofer.de/documents/N-301404.html