Free taxanes and the release of bound compounds having taxane antibody reactivity by xylanase in female, haploid-derived cell suspension cultures ofTaxus brevifolia

Durzan, D. J.; Ventimiglia, Frank

doi:10.1007/bf02823035

Cited by 18 publications

(13 citation statements)

References 15 publications

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“…Haploid egg-derived callus cultures from the female gametophyte of a 30-year-old T. bre6ifolia tree, maintained on modified B5 medium, in darkness, at 25°C [34], were used as experimental system.…”

Section: Plant Materialsmentioning

confidence: 99%

Nitric oxide induces cell death in Taxus cells

2000

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Section: Plant Materialsmentioning

confidence: 99%

Nitric oxide induces cell death in Taxus cells

2000

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“…After solvent extraction of taxanes~gold-labelled antibodies reaffirmed the occurrence of residual covalently bound laxanes on cell walls. Xylanase treatment released a number of taxanes that were recovered from the digest (Durzan and Ventimiglia, 1994). Chromatographic evidence with digests reaffirmed the occun·ence of bound baccatin III, 1O-deacetyI baccatin III, IO-deacety1-paclitaxel, several unidentified taxanes, but little or no paclitaxel.…”

Section: Taxane Yields From Cell Suspensionsmentioning

confidence: 85%

Metabolic Engineering of Plant Cells in a Space Environment

Durzan

2000

Biotechnology and Genetic Engineering Reviews

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“…Although protoplasts may be used for selected studies, they lack a functional cell wall and therefore are not as representative of cultured cells as intact single cells. This is of particular importance for hydrophobic secondary metabolites such as paclitaxel, which may be stored in the cell wall (Choi et al, 2001;Durzan and Ventimiglia, 1994;Roberts et al, 2003;Russin et al, 1995). Digestion of the cell wall and protoplast formation has been shown to stimulate release of paclitaxel to the extracellular medium (Aoyagi et al, 2002;Roberts et al, 2003).…”

Section: Discussionmentioning

confidence: 97%

Preparation of single cells from aggregated Taxus suspension cultures for population analysis

Naill

Roberts

2004

Biotech & Bioengineering

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A method for the isolation of single plant cells from Taxus suspension cultures has been developed for the analysis of single cells via rapid throughput techniques such as flow cytometry. Several cell wall specific enzymes, such as pectinase, pectolyase Y-23, macerozyme, Driselase(R), and cellulase were tested for efficacy in producing single cell suspensions. The method was optimized for single cell yield, viability, time, and representivity of aggregated cell cultures. The best combination for single cell isolation was found to be 0.5% (w/v) pectolyase Y-23 and 0.04% (w/v) cellulase. High viability (>95%) and high yields of single cell aggregates (>90%) were obtained following 4 hours of digestion for four separate Taxus cell lines. In addition, methyl jasmonate elicitation (200 microM) was found to have no effect on three of the four tested Taxus lines. Isolated single cells were statistically similar to untreated cell cultures for peroxidase activity (model cell wall protein) and paclitaxel content (secondary metabolite produced in Taxus cell cultures). In comparison, protoplasts showed marked changes in both peroxidase activity and paclitaxel content as compared to untreated cultures. The use of flow cytometry was demonstrated with isolated cells that were found to have > 99% viability upon staining with fluorescein diacetate. The development of a method for the isolation of single plant cells will allow the study of population dynamics and culture variability on a single cell level for the development of population models of plant cell cultures and secondary metabolism.

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Free taxanes and the release of bound compounds having taxane antibody reactivity by xylanase in female, haploid-derived cell suspension cultures ofTaxus brevifolia

Cited by 18 publications

References 15 publications

Nitric oxide induces cell death in Taxus cells

Nitric oxide induces cell death in Taxus cells

Metabolic Engineering of Plant Cells in a Space Environment

Preparation of single cells from aggregated Taxus suspension cultures for population analysis

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