Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Using tissue cell culture to make a protein or polypeptide
Patent
1994-07-18
1996-11-05
Lilling, Herbert J.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Using tissue cell culture to make a protein or polypeptide
424 59, 424 69, 424 706, 424 74, 424115, 435 41, 514415, C12P 2100, C12P 1700, A61K 700
Patent
active
055717004
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to a process for preparing melanin pigments by bioconversion, to the pigments obtained and also to their use in the field of cosmetics.
It is known to use colored pigments in the cosmetics field, and these are essentially inorganic pigments or alternatively pigments derived from synthetic direct dyes and, in the case of black pigments, pure carbon.
It is, in particular, known to produce yellow-brown dyes by oxidation of 5-hydroxyindole. It is known, moreover, to prepare melanin pigments enzymatically, by oxidative polymerization of 5,6-dihydroxyindole derivatives such as 5,6-dimethoxyindole or 5,6-methylenedioxyindole.
The Applicant has now discovered that melanin pigments could be obtained rapidly by bioconversion of simple substrates, in particular using plant cells cultured in vitro in a standard manner.
In effect, the processes proposed to date employ conventional precursors of melanin biosynthesis, which prove expensive.
In contrast, according to the invention, the materials employed are common precursors and biological compounds which are modestly priced and easy to use. The mechanism of enzymatic bioconversion according to the invention does not employ polyphenol oxidases.
Thus, the present invention leads to a process for preparing melanin pigments by enzymatic bioconversion, employing a melanin precursor substrate and plant cells. According to the invention:
a) poppy (Papaver somniferum) plant cells previously cultured, are separated from their culture medium and subcultured in a culture medium at a concentration ranging from 10 to 100 g/l,
b) the cells are at least partially ground before or at the end of the latency time,
c) the at least partially ground cells are brought into contact, in a bioconversion medium, with a melanin precursor substrate chosen from indole, indoline, dihydroxyphenylalanine, tyramine and tyrosine,
d) the precipitated melanin pigment is recovered.
In a first step, undifferentiated poppy cells are cultured in vitro in a standard culture medium. After a sufficient growth time, the cells are separated from their medium, for example by filtration. The duration of the growth time of the cells is of no importance to the process, and depends in particular on the quantity of cells which it is desired to obtain.
As a standard culture medium, Heller's medium may be used. This medium can optionally contain vitamins and/or hormones in addition.
Undifferentiated poppy cells are understood to mean cell cultures derived from fragments of whole plants, decontaminated and placed on solid culture media chosen in such a way that the cells multiply thereon without giving rise to organized tissue forms. These cells are then transferred to liquid media, where their growth takes place by simple cell multiplication. These cultures are comparable to those of bacteria.
In a second step, the cells obtained are subcultured in a standard culture medium, at a concentration of fresh cellular matter of 10 to 100 g/l. As culture medium for the subculturing, a new culture medium is used. For the subculturing, a Heller's medium optionally containing vitamins and/or hormones may also be used. The cells are maintained in this medium for a period not exceeding the latency time.
The latency time is the time needed for a freshly subcultured cell to adapt to the medium, and during which the cell concentration remains constant before growing therein. By taking periodic samples, it is thus possible to establish that the cell concentration remains substantially identical to the initial concentration--the cells are in the latency period--then the concentration grows significantly when the latency time has ended; the cells have then passed into the growth step. Depending on the strain used, the latency time can vary between 6 hours and 7 days.
It is preferable to subculture the cells at a concentration of fresh cellular matter of 20 to 50 g/l.
In a third step, before or at the end of the latency time, the cells are preferably ground to obtain a ground cell preparation.
Cell grinding may
REFERENCES:
patent: Re33993 (1992-07-01), Grollier et al.
patent: 4459285 (1984-10-01), Grollier et al.
patent: 4569839 (1986-02-01), Grollier et al.
patent: 4581230 (1986-04-01), Grollier et al.
patent: 4746510 (1988-05-01), Grollier et al.
patent: 4767618 (1988-08-01), Grollier et al.
patent: 4880621 (1989-11-01), Grollier et al.
patent: 4933177 (1990-06-01), Grollier et al.
patent: 4961754 (1990-10-01), Grollier
patent: 5205837 (1993-04-01), Andrean et al.
Chemical Abstracts, No. 43036h, vol. 102, No. 5, 4 Feb. 1985.
Hilaire Pascal
Junino Alex
Martin Richard
"L'Oreal"
Lilling Herbert J.
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