Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1989-06-01
1992-06-02
Griffin, Ronald W.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
536 27, C07H 100, C07H 2104
Patent
active
051186039
DESCRIPTION:
BRIEF SUMMARY
DESCRIPTION
The invention concerns a process for the inactivation of the biological activity of DNA, especially recombinant DNA.
According to the directions for the protection against danger due to in vitro newly combined nucleic acids, which have been published by Federal Minister for Research and Technology, wastes which contain nucleic acids, thus DNA, are to be sterilised or denatured. Therefore, in the case of working with recombinant DNA-containing micro-organisms, it is not only necessary to inactivate the organisms but also to destroy the possibly recombinant DNA in the organisms. As a rule, the killing off of micro-organisms takes place relatively simply. However, the DNA in the micro-organisms is, as a rule, not destroyed by the measures employed. Since, now, this sterilisation or denaturing must take place before the biomass is introduced into the clarification plant, measures are, furthermore, to be carried out in order that no substances get into the subsequent clarification plant with the biomass which, for example, damage or kill off the micro-organisms of the activated sludge if a biological clarification plant according to the activated sludge process is supplied, which is preferred. Thus, care must be taken that the substances used are not toxic or, before introduction into the clarification plant, are so broken down that they can cause no harm.
The methods previously known and recognized e.g. by the Federal Ministry of Health (BGA) of the Federal Republic of Germany and by the Central Commission for Biologicial Safety (ZkBS) of the Federal Republic of Germany for the sterilization and inactivation (e.g. steam sterilisation over 20 min. at 121.degree. C.) are very expensive. Especially in the case of the treatment of industrially usual fermentation amounts of 10 to 50 m.sup.3, these processes are very cost intensive not only for apparatus but also because of the high energy consumption.
Therefore, it was the task of the present invention to make available a process with which the biological activity of DNA, especially recombinant DNA, can be inactivated with certainty without undesired, especially toxic substances getting into the waste water. Furthermore, the process is to be capable of being carried out cost-favourably.
This task is solved by a process for the inactivation of the biological activity of DNA, especially recombinant DNA, which is cahracterized in that one adds to the DNA-containing biomass a percarboxylic acid with 1 to 3 carbon atoms, an alkali metal peroxide or an alkali metal peroxomonosulphate and heats the mixture then to 60.degree. to 100.degree. C.
Surprisingly, it was ascertained that by means of the use of the above-mentioned compounds and by heating of the batch, not only are still living micro-organisms killed off but that also the DNA contained in them is broken down to such an extent that no fragments capable of replication remain.
With the process according to the invention, a breakdown of the DNA to fragments smaller than/equal to 500 bp can be achieved which is also not exceeded by the waste disposal processes (autoclaving) permitted by the ZKBS. Such fragments are no longer biologically active and can, therefore, be passed without danger into the waste water.
The process according to the invention can be advantageously used for biomasses such as result in the case of the culturing and production of recombinant DNA within or outside of animal or human cells, as well as for micro-organisms. Waste waters which contain micro-organisms, plasmids and the like DNA-containing material can, therefore, before the introduction into the actual clarification plant, be subjected to the pre-treatment according to the invention which ensures that no DNA in a form capable of replication can any longer be present.
The biomass, such as for example a micro-organism-containing culture solution, is mixed with a percarboxylic acid with 1 to 3 carbon atoms, an alkali metal peroxide or an alkali metal peroxomonosulphate, since this is, on the one hand, relatively stable and, on t
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Comer Michael J.
Munster Michael J.
Popp Friedrich
Schumacher Gunther
Seydler Bodo
Boehringer Mannheim GmbH
Griffin Ronald W.
Kunz Gary L.
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