Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process...
Patent
1997-11-17
1999-12-07
Naff, David M.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
435176, 435395, 435402, 435410, 4352891, C12P 100, C12N 1114, C12N 500, C12N 504
Patent
active
059981628
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention relates to a process for continuously or discontinuously producing secondary metabolites of viable plant cells in porous and inorganic matrices.
The invention further relates to an article suitable for entrapping and immobilizing the plant cells in such a condition to maintain their viability for production of secondary metabolites.
The production of secondary metabolites on an industrial scale constitutes the main but not the only field of application of the present invention, since the process according to the invention and the immobilization obtained, as described in detail hereafter, can be advantageously used in any other equivalent field in which immobilized plant cells are used.
BACKGROUND OF THE INVENTION
The most commonly used immobilization matrix is algal polysaccharide alginate, cross-linked with calcium ions as described in FEBS Lett. 103: 93-97 (1979) and 122: 312-316 (1980), Plant Cell Rep. 5: 302-305 (1986), and Appl. Microbiol. Biotechnol. 30: 475-481 (1989). Other methods include entrapment with polyurethane foam, described in Biotechnol. Bioeng. 33: 293-299 (1989), 35: 660-667 (1990), Appl. Microbiol. Biotechnol. 33: 36-42 (1990), 37: 397-403 (1991) and 35: 382-392 (1991).
The use of porous and amorphous sol-gel derived silica for entrapment of viable cells has been described in cases of non-plant cells, reported in J. Biotechnol. 30: 197 (1993), J. Ceram. Soc. Jpn. 100: 426 (1992), Biochim. Biophys. Acta 276: 323 (1972), Chemistry of Materials 6: 1605-1613 (1994) and Angew, Chem, Int. Engl. 34: 301-303 (1995). These latter cannot be considered as methods actually suitable for applications to higher plant cells, which are severely poisoned under the experimental conditions reported for immobilization of bacteria and yeast cells. As for processes applied to, and claimed for, plant cell immobilization, they entail some problems: first of all, processes based on simple adhesion to the surface cannot properly be considered as immobilization, since cell reproduction and the increase in biological mass unavoidably cause release of cells in solution.
Polyurethane foaming matrices may cause severe transport limitations to and from immobilized cells. One important drawback of these host matrices is their poor mechanical stiffness, so that prolonged use for industrial production in practice does not appear feasible.
Immobilization in alginate beads allows direct contact of cells with the gel matrix, so that the cells are inevitably subjected to a high concentration of a variety of ions and organic compounds, causing negative physiological effects.
SUMMARY OF THE INVENTION
A main object of the present invention is therefore to overcome the above described disadvantages by means of a process which allows plant cells to be immobilized while maintaining their viability and avoiding the release of cells from the matrices, with free transport between the immobilized phase and the culture medium.
A further object of the present invention is to provide a process which may be repeated in standard conditions with constant results.
Another object of the invention is to provide a host matrix with stiffness suitable to tolerate stress and share strain during production.
Still another object is to provide immobilization articles or matrices which, allowing free exchange of organic species and nutrients through the open pores, ensure increased intercellular contacts and consequently the potential for biochemical communications.
A further object of the invention is to provide a process which may be performed with industrial-scale devices and relevant production equipment.
These objects are achieved by a process comprising the steps of: matrix of inorganic material having a tensile strength of at least 500 MPa; matrix; colloidal suspension not interfering with the cell viability; including a carrier gas saturated with volatile SiO.sub.2 or organic modified SiO.sub.2 precursors.
The employment of culture systems in which plant cells are immobilized on an inert sup
REFERENCES:
patent: 4845054 (1989-07-01), Mitchener
patent: 5041138 (1991-08-01), Vacanti et al.
Journal of Biotechnology, 30 (1993) 197-210.
Cappelletti Elsa Mariella
Carturan Giovanni
Piovan Anna
Biosil AG
Coleman Henry D.
Naff David M.
Sudol R. Neil
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