DE2022359A1 - Permanent nutrient preparation for the production of culture media for microorganisms and process for its production - Google Patents

Description

HANS H. PONTANI Patent attorney

1752 Kleiiiosfhelm at ÄidBÄWei
Hirschpfad 3 - Tel. 06027/325

May 5th 1970

SICALY, 49 SAINT-CYR-EN-BOWRG, France

"Permanent nutrient preparation for the production of culture media for '.' Microorganisms and process for their production ¹¹

The invention relates to new nutrients or * nutrient preparations for microbiological cultures.

Culture media must be available to the bacteriologist or microbiologist stand, their composition as well as chemical, physical and biochemical properties over one or even several Remaining constant for years · However, these conditions are with the plant-based nutrient media currently in use only »or rather difficult to meet, since the properties of plants depend on the climatic conditions, especially the irrigation and solar radiation, as well as with the culture conditions, d · h » the time of sowing, the soil conditions, the fertilization, the Treatment with insecticides, etc., fluctuate · For this reason Culture media such as malt or grain extracts rarely interact with one another identical, so that their use for microbiological cultures cannot always give the same results. Besides the preservation of such culture media represents a serious pro- *

009147/1801;

In fact, the classic method offers au i-irer Preservation of the numerous difficulties inherent in the means used to protect the culture media from extreme exposure. preserve borrowed impurities. If the Msdien in river siger form preserved or stored in a sterile state, leads The space requirements of containers and contents lead to space problems, while the use of air-dried powders increases the risk decomposition or changes during drying, during which the product undergoes oxidation processes, Maillard reactions or is exposed to other more or less negative influences. The seasonings, high-sugar products, are subject to a certain degree of caramelization and the proteins they contain decompose.

Although fresh Prucht- or grain extracts can be immediately prior to the preparation produce en3 culture medium, depending however, are thereby gen Qualitätssehwankun-the aforementioned not overcome or avoided and you need for this procedure every time at least an hour and usually even 1 to 2 1/2 hours.

The invention therefore has as its object to overcome these difficulties and a new nutrient continuous preparation .for culture media to put at your disposal _s very long time can be preserved without change in quality and therefore makes all attempts or preparations reproducible.

The subject of the invention is therefore a nutrient long-term preparation for " Production of culture media for microorganisms with a content of fermentable sugar, which is characterized in that

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ss from a powdery ^ slightly diluted with water atfea uni Easily soluble in water lyophilisate of an aqueous fruit / itocier Gctreioeliornextrakts as an essential component iD

The invention thus solves the problem of storing culture media for bacteria in a particularly economical and convenient way.

man
riological cultures. In addition, with the help of the long-life fuel preparations according to the invention, a. B. in 5 to 15 minutes, produce a culture medium.

A particularly advantageous inventive nutrient duration " preparation is characterized in that the lyophilizate from a halz wort is made and 50 to 70 wt .- ^ maltose, Contains 15 to 50% by weight of dextrin and 1 to 5% by weight of water.

The solid permanent nutrient preparations of the invention can, in addition to or instead of part of the main nutrient (igrophilioate), auxiliary substances such as, for example, zuclier, gelatinizing agents, iJinerelsalse, antibiotics or other antiseptic agents, vitamins and. possibly contain other substances

Since the permanent nutrient preparations of the invention are powdered and can be made sterile, they can be made very easily without the slightest risk of a change over a very long Ιεη ^ ε time Store kit with moderate space requirements in traditional packaging baw. kenservieren · Aufgxnmd their very little Waasergehsdts are these new tecBiiischeii products against iniltrobiologisclio, ensynatic or chemical heaktS-onsn protected.

The use of the new, püI.verförEigsn nutrient duration preparations for microbiological culture is extremely easy. It follows the production of an Ivah ^ bov-ilio'i, which: *: sv. ^; imsch '^:; € I'erRc

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Pul before dilute with water au, Burch corresponding Av.iv;,; .. ^y l the-

ta

With this amount you can adjust the nutrient concentration of the liährijc 1131 lor exactly to the desired value; to adjust. On the other hand, strongly concentrated culture media can in any case not be longer, which was by no means possible with conventional liquid culture media without inconvenient or complicated work processes for narrowing or dissolving.

l) ic permanent food preparations of the invention can advantageously be clolled on the basis of a wort or an extract; h from a flüsr.igen, natural Ausgxanjirn-.l? rirji _o Kan, this liquid Ausgangsiünterial zv / ar directly dui-cl · lyophilization in a puiveriörraiges product .u & v / ar.deln. According to the invention, r'c-dooJv is permissible to subject it to a nox · /orlMil'ion.dlung beforehand. This Vorbbt.gjafllung kaim ro in plrjr-ikalircV rj.; Ir ·. or a chemical treatment and / or the addition of various other substances. "With the help of such a pre-shredder you can produce a permanent nutrient preparation that is intended for a serial culture medium that the microbiological log no longer needs to modify when it ends.

The physical pretreatments of the starting point are ins general treatments such as filtering, decanting, Ausi ^ .le.i, Centrifugation and sterilization

Others, in AEET Rule B <shv zwe ^, -;? '' / URL ^ e treatments bestcOien since: cin, the Äußgangsmaterial 'rsr ü.z ^ -> i "-; ^ lilii £ S3ren versshiltdenc. iiinbc · »the above mentioned substances to be incorporated. Kan can "also for the development of the Staonaes to be cultivated necessary or cheap sugars> such as- Slökose, Rohrsucksr, iiannit uncl * äex» gl ·

009847/1101. bad original

to add.

If you want to get long-term nutritional supplements that can be used in the cold one can produce more or less festive bouillons in itself add known gelling agents. For the growth of the microorganisms or for the identification of certain ones during cultivation Mineral salts necessary for the compounds formed are mostly salts such as magnesium or iron sulfate and the like, phosphates etc. It can also be useful to use the liquid medium ■ add vitamins before lyophilization. "

It can also be recommended to use the used as starting material Add antiseptic agents to liquid, for example, to control their effectiveness or to develop favoring certain tribes at the expense of the development of others. Antibiotics in particular can be used for this purpose.

The liquid starting material can, ^ e after,

which special properties the culture medium to be produced should have, other substances are also incorporated. ■ ■ Ä

The lyophilization is carried out in a known manner, wherein the first process step, ie the freezing preferably takes place at a temperature of -25 ° to 0 or below, particularly in the range of -25 to -3O ⁰ O. It is advisable to carry out the freezing process quickly in order to avoid precipitation and decomposition reactions. When the selected freezing temperature has been reached, the apparatus containing the starting material is evacuated. During the sublimation which then takes place, precautions are taken to avoid the one

009847/11 (H.

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Part of the material is carried along by the steam. During the sublimation, the temperature in the treated material is usually between about -23 and about -25 ⁰ O and the prevailing pressure fluctuates between 0.4 and 0.3 Torr.

The desorption, ie the last phase of the lyophilization, is usually carried out at +30 to +50 ⁰ O and under a pressure of about 0.2 to 0.3 Torr. When removing the negative pressure, it is advisable to introduce an inert dry gas into the apparatus, for example nitrogen or COg »in order to avoid possible oxidation by air.

The specific conditions listed below are preferred, but are in no way to be understood as limiting.

The lyophilized powders or permanent nutrient preparations according to the invention can be among those known for this type of product Conditions conditioned and stored.

By mixing the lyophilized powder with a certain amount of water, solutions identical to the extracts used as starting material can be obtained. In the case of malt, the mixing ratios are generally between 200 and 250 g of dried powder / liter. distilled water. The characteristics of some samples of malt wort restored to this VJsise are given below:

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009847/1101

Sample 1 (made by dissolving 234 ε lyophilizate powder

in 1 liter of distilled water):

Density at 20 ^o / 20 ° C .... 1.06655 (. Decrease (Grentert) 5.50

P ^H ··· 3.55 («.. (Actually.) 7i2O

Kaitose in g / ltr. ..... 140 lbs isohumulone <1 og / ltr.

(The decrease is the change in density due to fermentation).

Sample 2 (from 246 g powder per liter of water):

Density 20 ° / 20 ^o C ............. 1.08925 C Maltose ..... 167 g / ltr.

Isohumulone: <1 mg / Ltr.

) neuter) 7.44

Sample 3 (from powder made from beer wort mixed with 10% Haltodextrin 65/35 before lyophilization, 246 g of which is dissolved in 1 liter of water):

Degree Balling ................ 19.13 C Maltose # ia dry ^ Pul-

) Powder ......... 29.2

Sample 4 (from 231 g of powder per liter of Wasuer) ».

Balling degree ................ 20.51 (decrease 1.01752 ... 4.46

pH ·. ·. «·. ··. · ..« ·. ···. 3.30 ("(limit value) .. 78.3

Kai tose $ in dry powder ... »66.44 Dextrin Ji ^B ·" ..... 15 _s O3

The examples illustrate the invention.

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example 1

A cube is made from a light, coarsely ground corn using the traditional brewing method. When the malt is completely sucked out and the starch has disappeared, the wort is separated from the pomace by filtration, whereupon its density is adjusted to the desired value by adding washing water from the! Drester. The liquid obtained in this manner is' given in large Pyrex glass flask and then heated for 20 minutes 12O ⁰ C in an autoclave, to aussufallen coagulable products, after which they are filtered and then the clear lyophilieiert FiItrat. ".

For this purpose, the wort is distributed in an amount of 10 l / m on Sdbalen and then frozen by cooling to -28 ⁰ G * where-

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The apparatus is evacuated to a final pressure of 0.4! Dorr.

Then, sublimed at a product temperature of -25 ⁰ C, wherein the temperature of -38 and -42 ⁰ Wasseröampffalle C interlocutory fluctuates.

To compensate for the amount of heat consumed in the sublimation, a source of scattering is used, the temperature of which is 80 ⁰ at the beginning, and which then gradually cools to + 40 °. The pressure is then 0.4 Torr. During the duration of the desorption, the product is kept at a temperature of + 4Q ⁰ C and under a bridge of 0.55 SJorr.

Then the prevailing vacuum is created by introducing dry,

The product that is solid in your «Weis © ΐ:% α3.ΐο» ©; contains 2 bits. 3 . Moisture wvl 2x% w & rn ea la eine yasserdiclbitiiii? ©: ·: · χ · Γο: υ.

009847 / ifOI.

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2022358

is stored, well-preserved, 2 To divide culture broth from this product, it is sufficient to ^{warm the powder in about 22 0} G -

mem, distilled water to dissolve. Let the Hährbouillons obtained in this way I _»9 after they have been previously sterilized 10 minutes at 120 ⁰ G, auageseichnet used as culture media.

Example 2 ■

Analogous to example 1, another powdery nutrient duration preparation produced »with (however, the wort after filtering» sterilized and the lyophilization under sterile conditions is carried out * The product obtained in this way leaves directly without sterilization for the production of nutrient bouillons use·

Example ?

Using malt as the starting material is worked ia analogously to Example 1, operating however, the clear EiXtrat 5 $ glucose added "As in Example 1 ma" leg taping with 10 Iitr "Elüssigköit / m, but freezes up to JEndtemperatur from -32 ⁰ O and evacuates Ms to 0.3

During sublimation the product temperature is -28 ⁰ G and in the water vapor trap it fluctuates between -44 and «48 ^ 0» "■ L

The required to compensate for the consumed during sublimation Wärsie calories are supplied to the product by a 0trablungs ~ heat source, the temperature during the first 'four stood 6Q ⁰ O? and then gradually decreased ^{to JO 0 O.} The pressure in the sublimation apparatus is? then about '; -

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0.25 to 0 _f 3 tea · For desorption, the product is then ^{held at 30 0} O for 4 hours under a “pressure ψοη ®tw® 0.2 to stm 0.3 ^ ©”.

Eggs relaxed mm looked at the vacuum Introduction of oxygen-free, dry nitrogen *

The pulvertonaige Iteodukt (nutrient permanent preparation) obtained in this way contains Z to $ $! Moisture and lets Aich well preserve bsw · Keep "Aeugi? ItBd eeines Glulrosegehsltes can aan it aur B ^ potellung voa litabötiillöiis use that to, investigation of femtatierung hzw® the breakdown of glucose by microbial strains should serve "

By dissolving the powder in distilled water can be easily KultnrMGtien produce * can sterilize mm _s -indea they durcli filtered through a membrane üteiliniöiisacle "

Example 4

Example 5 is like that, which deviates from this. IiI-entered sterilized and lyophilized under sterile conditions.

BeJBPiel ^

Example 3 is repeated, starting with a beer wort and deviating from this, instead of eex glucose, 2% gelose is added to the filtered wort, after which the pH of the liquid is adjusted to 6.5.

The SubliiÄtifJifindtt held at -26 ⁰ O. -

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The powder obtained is a culture medium concentrate from which one can dissolve in distilled water under weak erosion. "warm up" nutrient media containing gelatin in one step in a water bath can share.

009847/1801

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Claims (10)

t ~ \ * t- C. Vf \ J 1) Long-term nutrient preparation for the production of Ki-iltusrae-diGii for microorgarismeii with a Gol10.lt of fermentierlisrem sucker, thereby gekcssseicli & et, it is made from a powdery!., With water easily "/ epduinibaren m & in VJ easy löölichen lijophilisat an i? isei ^l sodium Fruclit- odei- Gct-rei-icorn as an essential part of the Bostaxid. 2) -iiähi? Tofi '(3rd preparation according to claim 1, thereby. gekennae refuses, cJiiß es ssusätsXieli mindostsas W Mineralaals and / or at least one Sslierungsmittel imd / ocis:. * rainaestene another zucli; © ?? vmä / oäm? mitidfestens contains a vitamin and / or at least one antiseptic win » 5) permanent nutrient pi'iipapat according to claim 1 or C-, d so "rc! 1, characterized 'that cUa x-yophilisate is made from one: -:' - malt wort and from 50 to 70% by weight of Kaltoce, 15 cowardly JO Gew · - ^ dextrin as well as 1 "to 5 # Ifeucfctigkeit ecth ^ lt. 4) nutrient long-term preparation according to claim 3, d a d u r c Jj characterized in that it contains 0.5 to 10 6cw .- ^ gelose Contains P or gelatin 5) Kührstoff duration preparation according to claim 'J or 4 _f gekennaeichnet that it contains 1 Ms 20 wt. -Ja glucose · 6) A process for producing τοη tßlhrstoffdauexpräpasben üaoh least one of claims 1 to 5, characterized in _t that an aqueous fruit CDEI. " Cereal grain extract 00 lyophilized to make a powdery egg! ·, 009847/1801 BATH ORIGINAL dry-looking lyophilisate that is packaged waterproof will. . - 7) The method according to claim 6, d a d u r c k s e k 3 η η ~ draws that the aqueous extract is lyophilized, by cooling it to a temperature of -25 to -3Ö ° C freezes, then the water contained therein to -23 C and under a pressure of about 0.3 to 0.4 Sorr abdUbli- miert, whereupon the lyophilisate is finally adsorbed by holding it at +30 to +50 ⁰ O under a pressure of about 0.2 to 0.3 Torr. 8) Method according to claim 6 or 7, characterized thereby e η η ζ gauges that nan as a starting material is one Beer sausages used. 9) Method according to claim 6 or 7 »characterized in that the && ß aqueous extract is mixed with about 0.5 to 10 wt .- # glucose. 10) Method according to claim 6 or 7 »d · d u r c h g e» indicates that one is the aqueous extract 0.25 Ms 5% by weight of gelose or gelatin added. BATH ORIGINAL