DE1442089A1 - Process for the production of high protein foods - Google Patents

Description

Bbbo Research and Eng. Co, description

ESSO HESBARCH AKD ENGINEERING COMPANY, Elizabeth, New Jersey, V ₀ StοΑ.

Process for the production of high protein foods.

For this filing, the priorities of Shark 21, 1963 from US patent application Serial No. 281 895 and dated April 15, 1964 from U.S. Patent Application Serial No. 360 141

taken in Anapruoh

The invention relates to improved and economical methods sub culturing bacteria old aid of inexpensive hydrocarbons as nutrients and proteinreiohen for the direct recovery of an excellent ", Z _0, 70 to 75 J" protein containing food or feed additive for fours or or Hanschen · Genes of the invention is continuously a liquid or gaseous hydrocarbon nutrient, a liquid, aqueous, mineral-containing substance

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Medium and a gas containing excess oxygen are fed into a reaction vessel with stirring, which nourishes the hydrocarbon, contains the aqueous medium and bacteria that are able to grow by converting the hydrocarbon, -eich in the logarithmic growth phase (in which they multiply exponentially ) and are present in a concentration which is equal to or greater than that at which the exponential bacterial multiplication is equal to the continuously withdrawn amount, while part of the reaction mixture is continuously withdrawn at such a rate that critical liquid residence times are maintained, whereupon the microorganisms separated from the withdrawn mixture and dried sohliesslioh, whereby the desired, protein-rich bacterial food additive is immediately obtained. In a preferred embodiment of the invention, critical amounts of the hydrocarbon nutrient are obtained on the aqueous, saline medium supplied. In a particularly preferred embodiment of the invention, the bacterial nutrient used is a selected fraction of paraffinic C ₁₁ to G ^ Q petroleum hydrogen which has been purified to reduce its aromatic content to less than 0.5% by weight, with bacteria from Mlcrococcus oerifloans can be used and preferably optimal, non-growth reaction media containing inorganic salts are used in order to obtain a high selectivity for bacterial formation and as few esters as possible as by-products.

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le was found to be an excellent protein heron food additive in an extremely economical way from inexpensive petroleum hydrocarbons according to the present Process can be produced. The first and foremost requirement for the procedure is the application of critical short residence times in the reaction vessel. Ss was namely Surprisingly found »that a strong shift in, .the selectivity in favor of bacterial formation takes place, if you use liquid dwell times from 1.5 to 4 times, preferably from 2 to 3 times the minimum development time of the microorganism in question, e.g. Hioroooocus cerifleans, works in the following way:

1. at 2.7 times the minimum development time:

77 kg η-paraffins + 89 kg O ₂ + 22 kg inorganic salts, NH, etc. —j> 100 kg dried cells + 62 kg 0O ₂ + 26 kg H ₂ O

2. at 5.3 times the minimum development time

89 kg η-paraffins + 127 kg Og + 22 kg inorganic salts, NH ^ uswo - $ ► 100 kg dried cells + 97 kg CO ₂ + 41 kg H ₂ O.

Under the "minimum development time" is the shortest period of time to understand the bacteria in the logarithmic growth phase (i.e. the ones on a logarithmic diagram gives a straight line) at intermittent under the most favorable conditions, i.e. temperature, pH and non-limiting growth Supply (see »strong agitation of the medium) of

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Hydrocarbons, oxygen and the most beneficial aqueous salt medium for the bacteria Need to double their consentration. If at the continuous Proceedings an Ifaohetumsbeeohenschwiger sugesstst is, the "Mindeetentwioklungezeit * with addition of this Acceleratorβ can be measured. The child development side for the respective bacteria can be obtained from the literature or from the literature can be determined empirically in the laboratory, preferably using a Feraentatlonsgefässe working absatsweise

with a rotating at a speed of 1500 rpm Sehauf elbeater used.

The result is the limitation of the liquid dwell time Improvement is surprising! because it was not expected that the selectivity would improve the lingering pages would, and the skilled artisan normally would not have done without this improvement in selectivity tries * to work with such short liquid lingering pages Bb is (i) theoretically unsuitable for a continuous Processes with liquid dwell sides of less than 1.45 times the minimum development rate su work, i.e. at a reaction in which the rate of formation of the product is proportional to the amount of product present, is the minimum residence time for a continuous process 1 / ln 2 or 1.45 times the minimum development since fermentation was carried out in absent amounts, and (2) since the fermentation is continuous Procedure for malfunctions and serious deviations

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If conditions occur under the most favorable conditions, there must be a margin of slope of at least the simple, preferably the double, the minimum settling time for a continuous process (i.e. 1.45 times the minimum development time) so that the bacterial culture is not exhausted, i.e. it is flushed out of the reaction vessel. It should be noted that the only cost for this safety factor is the enlargement of the reaction vessel, and this is a subordinate cost factor compared to the geeant costs of the procedure »

It was not to be expected that the short residence times would significantly improve the selectivity, and that The technical progress achieved by the use of short residence times is very important, as can be seen from the following Considerations result

¹ · cooling costs . All bacterial growth processes are strongly exothermic, in that they develop from 2 770 to 5550 koal / kg cells, and are carried out at temperatures close to room temperature. Saher 1st usually nioht water cooling _r but refrigeration required. In a typical process, which works with short dwell times (equation 1), the costs of this deep freezing are around 40 Jt of the #ermentation costs (ie the total costs of the ronstoff »* of the fomentation and the product extraction) ο increase with longer dwell times (equation 2) this cost by about £ 6from that is proportional to the additional amount

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Oxygen, which leads to the formation of carbon dioxide and water is consumed.

2. Energy costs . In all current bacterial breeding processes, the hydrocarbons are in low concentration dissolved in an aqueous waohetum medium containing mineral salts. Since the solubility of oxygen in water is very low, there is a very high energy requirement (stirrers and blowers) in order to achieve the transfer of oxygen to the microorganisms. This energy requirement is not reduced by measures such as emulsification, but can be reduced by using oxygen-enriched air instead of ordinary air. The cost of procurement of oxygen-enriched air are but reoht significantly · In any case, be in use short residence times (Equation 1) energy costs about 40 $> the Permentationekoßten, and increase it by about 4-0 # or more if you with long residence time works (equation 2).

3ο hydrocarbon costs . The Kohleuwaeseretoffkosten be reduced by applying short reaction times by 15 ^ ♦ These costs are, however, in comparison sin the costs discussed above low.

4. Cost of the remote control system . The size of the reaction vessel * is also reduced by the use of Itura reactionaries by £ 15. These costs are, however, likewise

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klein ia Tezkältnis eu those discussed in paragraphs 1 and 2 Costs.

Venn nan naoh a bevorBugten Ausfuhrungsfora the Brfindung as Bakteriennähretoff C ₆ - to C ^ hydrocarbons used, so that cleaning of these Kohlenwaseerstoffe as has herauegeeteilt such that from them ia substantially all Aroaaten be removed extremely sweekaassig or necessary. It was also surprisingly found that the aonooyclic and polycyclic aromas normally contained in petroleum are not consumed by many bacteria, but are adsorbed on or in the bacteria, even if the formation of ebbs as meoen products and the amount of unconverted oil by the process according to the invention to be teed up on a mininua. So these aromas can be left out. Remove the final product, and cattle feeders tend to have a tendency to impair the growth of the animals if the feed contains more than 0.05 Gev the aonooyolisohen Aroaaten sioh iaaer nooh ftuseerst sohädlioh effect on the Vaohstua of the animals · 39a «inventive method thus an empty economic extraction of the bacteria possible, while it practiioh all aroaat from has removed, because the heat did not show that the outgoing material was Starting hydrocarbons and the Beter formed as by-products

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j that are retained on the bacteria are not poisonous aind and in any other way do not impair the growth of the animals. However, these substances retained on the bacteria offer no advantage, and their amount decreases the anticipated execution forms of the invention deliberately reduced to a mini-space "

Bb it has been found that an improved selectivity can be achieved if, during the fermentation, large amounts of air in the range from 0.5 to 4.0, preferably from 0.8 to 2.5 parts by volume of the air / volume supplied to the reaction vessel are used the liquid in the reaction vessel / minute per weight <, -? C dry bacterial concentration in the reaction vessel / liquid retention time in hours. According to another embodiment, oxygen-enriched air is used. In this case, the amount of air required is reduced proportionally with the increase in its oxygen content. According to this embodiment, the enriched air contains more than $ 70 and preferably more than 90 tons of oxygen in view of the savings resulting from the reduced cost of the compression plant. So there is an initial conversion of the oxygen into fatty acids, which are then used by the bacteria for growth with the formation of carbon dioxide and water as by-products * If there is not enough air, the initial reaction of fatty acid formation continues, es but there is not enough air sur available so that the bacteria can use the fatty acids for their Waohetutt

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ηβη, and the fatty acids are converted into ebbs, which are anchor in the ropes.

Se was also found that working bit on Saueretoff enriched air the use of very high levels of hydrocarbons, s.B. “More than 2 wt. 56, based on the die aqueous containing inorganic salts (including BH, 0H) Medina, with full use of these hydrocarbons «Öglioh« eight, so that in the continuously sucked off Very low hydrocarbon concentrations are included. Surprisingly, the high selectivity remains even when using these large amounts of hydrocarbons au in favor of the bacterial growth in contrast to Prayer education received in the course of the procedure. If aan old works in oxygen-enriched air »the preventive hydrocarbon quantities are 1 to 5» in particular 1 up to 3% by weight of the aqueous containing the inorganic salts Mediuaso

8ohlies8llch are preferred by using a aqueous Z & ohtungsaedluas, which is a critical coabination of inorganic salts, the selectivities are greatly increased, so that the generation of esters is reduced by a factor of 10, i.e. the Gleiobgewiohtsgenisoh after drying only 2 Jt ester, absorbed on the bacteria, instead of 20% with only a slight change in the reaction medium containing the mineral salts.

By combining all of the measures described above

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the cost of producing feed from hydrocarbons is significantly reduced. So z. B. the energy requirement sum stirring the Reaktiönsgefäes and the air supply is greatly reduced and also achieved a very significant improvement in that it is possible to separate the bacteria by a simple process, which does not use the extremely expensive cleaning methods such as ZoB. repeated washing with surfactants, extraction, etc., requires.

Coagulants »e» B are preferably used. katlonisohe surfactants to reduce the cost of initial separation such as centrifugation or settling Reduce.

To further explain the invention, reference is made to the drawing, which shows a preferred device for continuous Carrying out the process using the preferred, aromatic-containing Cg to CUQ raw material ο The hydrocarbons are released through line 1 of the cleaning system 2, from which non-straight-chain components and impurities are drawn off through line 3. This cleaning of the raw material is preferably carried out by adsorption of the straight-chain hydrocarbons, preferably Paraffins »with the help of 5 ^ molecular sieves, attached Desorption and purification of the desorbed straight-chain hydrocarbons with the help of 1 ^ molecular sieves or silica gel for the purpose of adsorption of the nooh impurities contained therein elnsohliesslioh the aromatics. The cleaned,

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Wheel-chain paraffins are conducted through line 4 to the storage container 5 '. The straight-chain paraffins pass through line 6 into the reaction vessel 7, which contains the bacteria and is equipped with a stirrer 8. Phosphoric acid is sucked through line 9t inorganic salts through line 10 and water through line 11 of the milk container 12 provided with agitator 13. The boiled salt solution passes through line 14 into the reaction vessel. Aawmiuahydroxyd is sucked into the reaction vessel through line 15. Air or air enriched with oxygen is introduced into the reaction vessel through line 16. Air, carbon dioxide and water vapor are discharged from the reaction vessel through line 17 to the outside air. "Sin Seelsoh from product see Ilen, Salslöeung and small amounts of unset hydrocarbons is continuously withdrawn through line 18, and part of this Geeiechee is through line 19, cooling tower 20 and line 21 ia cycle out, among other things in the reaction vessel to exert a cooling effect. Additionally or alternatively, a cooling coil 22 can also be used in the reaction vessel. The best of the product reaches the centrifuges or settling vessels 24 through line 23, where the product sources are separated off. The supernatant liquid is sucked off through line 25 and the circuit is conducted into the reaction vessel office. The separated concentrated cells pass through line 26 into the dryers 27, which are preferably atomization dryers, and to which hot air is fed through line 28. The dried product bells are

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device 29 deducted. The separation in the centrifuge is preferably carried out in such a way that the pulp sucked out of the centrifuge contains 7 to 28 GeW ₀ -Jt, preferably 10 to 15 ow.-jt srockengel, whereby the intra-eellial water is not included carried out economically, and because of the critical Reaktionsbedingiangen contain obtained as a final product cells NaOH drying only 7 to 10 Qew.-jt Ctesaatfett, of which about 75 i »ester omitted and 5 inorganic to 9 9EW ₉ ~ # Salsa ο the loss © on the last "which are not particularly advantageous as a component of cattle feed, are therefore not excessive" Instead of separation in the centrifuge, other separation methods, such as filtration or settling with or without a filter aid or coagulating agent, can be used, etc. to cancel the initial separation of the cells , and of course you can also use different drying methods. In any case, this process is extremely economical, since it does not require any cumbersome separation processes and no washing out of old surface-active agents to remove oil and by-products (although these processes can also be used in other ways of the invention), an end product with an extremely high protein content being obtained which does not contain any impurities harmful to animals or humans.

AIb serve as starting material for the process according to the invention Petroleum hydrocarbons with 1 to 30 carbonatoses, voreugswelee Gaeöle with a boiling range of 190 to 400 ° 0,

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In particular τοη 190 tie 320 ° C. Other preferred starting materials are gsradkettige and isoparaffins, CyοIoparaffins, Monoolefins, diolefins and aromatics old 1 bio 30 carbon atoms 1 »Molecule and mixtures thereof. Special preferred straight-chain paraffins * are available in large quantities Standing and particularly suitable starting materials are straight-chain paraffins from gas oils or light petrol old 11 to 30 carbon atoms as well as normally gaseous Hydrocarbons, such as methane, ethane and propane, as well as oils of the same, such as hydrogen gas When starting material is used, it is preferably made of the reaction vessel in Gaeforia through distribution pipes supplied «

Key to the success of this preferred one The embodiment of the invention is the cleaning of any used Qg to 0 «Q carbons for the purpose of reduction their content of polynuclear and mononuclear aromatics less than 0.5 wt.-p.t, preferably less than 0.1 wt.-p.t., especially less than 0.01 Qeν.-36. This is necessary because sioh has surprisingly found that aromatics of the microorganisms are not attacked, and if the economic separation process genäse the invention without one If such a coating of the starting material are used, the product is extremely harmful to animals and humans. she Harmfulness manifests itself in the fact that these substances the wholesomeness the old fed the feed in question

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Belittle animals. This is completely independent of the minor Amounts of polynuclear aromatics that are considered carcinogens and should preferably not be included in the end product. These latter substances can selectively after several processes are eliminated from the initial material} but it is surprising that also the single-core. Aroma- ' which accounted for a considerable part, i.e. £ 20 to £ 50, of the total Represent aromatic content, are also removed must if the process is to lead to a valuable end product. A number of others, in the literature as teohnisoh feasible described procedure makes of extremely expensive washing methods with surface-active agents, Extraction methods and other procedures that use the End product has to be subjected in order to obtain good fodder «

The hydrocarbons, in particular the gas oil feed, are preferably cleaned with the aid of molecular sieves * Here, the straight-chain hydrocarbons are selectively adsorbed on the molecular sieve and consequently are practically completely purified from aromatics. A preferred method of purifying gas oils in this sense is in the. US Patent 3,070,542 described and this method is the preferred Reinigungemethode for the output product of the invention vegetables. In essentially is carried out in said patent that the A'dsorptionegesohwindlfkeit increased and the naohfolgende desorption, special when it SiOH, inter alia, substances having high molecular weight is important

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can be made easier "if the molecular sieve is old as a" drainage agent, voraugeweiee Aaaoniak, preloaded and that, TerdrSngangsaittel susaaaen old the Auegangegut is euegangegut. As a precautionary measure, aan eich also serves a sus & tsliohen journey, as it is in the Belgian patents

637 262 or 637 315 is described.

Suitable bacteria for the method of the present invention Invention binds all bacteria that are produced using coal waoheen, non-toxic for animals and off of the class of the Sdhisooyoeten. These include the following comments

1. Peeudoaonadalee,

2, Ohlaaydrobacteriales,
3 hyphoaicrobials,

. 4 <Jtoibaoterialee »

5. Aotinoayoetalee,

6 »Oaryophonalee,

7 »Beggiatoalee,

8 «Kyxobaoteralee,

9ο Bpirochaetaaee,
, 10 "Ifyooplaeaatalee.

The bacteria preferred for the method according to the invention are the following: Mioroooocue oerifioane, Pseudoaonas aeroginoea, Paeudonooae fluoreeoene, Vocardia opaoa, Sooardla rubra, Xooardia ooralina, Pseudoaonas aethanioa, Peeudcwonae deeauaolytlefyool. ÄLoroooooue oerifloans _f the Dr. B »3« Kallio et al., "Iottmal of Bacteriology", Volume 78, Vr. 3, pp. 441-448 (September 1959) was isolated and identified. Cultures the microorganisms and in the "Aaerioan Type Culture Oil Lotion", 212 H Street, IV, Washington 7, B ₀ C ,, V.St.A.,

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laid down under 14,987. Di® complete identification this material "is the following

Morphology The ropes are small, spherical and tend to form an elliptical shape in ancient cultures and in media with hot nitrogen content. The barks au® defined media have medium surehnease? from 0.5 to 1.0 μ, which should have office ohms from 1.0 to 2.0 μ in complicated susamase-set media. The cells are found individually or in clumps. Immobile, BetBohroaatiseh © grains and sudanophilic grains are not observed <.

Gram reaction negative.

The colonies on defined agar are small (1 sam), circular, convex and have a full hand · you Colonies on nutrient agar are larger (2 to 5 ms), raised muooid, "generally round

Pigmentation Ee can be found white, beige or brownish variants "

Necessarily aerobic. Bas growth takes place on a large number of substances, ZoB _e yeast extract, casein hydro-IyBat, long-chain alcohols and acids, long-chain, straight-chain alkanes and olefins.,

Fermentation of Carbohydrates * Carbohydrates are not fermented. Many carbon hydrates are aerobically assimilated. to these include glucose, maltose, mannitol, sucrose, lactose, arabinose, bhamnose, sorbitol, duoitol and inulin. Becomes aerobic Glucose processed with acid formation. Bs became Gluooneaura identified.

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Iitrate reductionj negative.

Gelatin liquefaction generally negative. Some strains may liquefy slowly · Urea hydrolysis negative.
Be katalaee is produced »
Hydrogen is not grooved.
Best temperature 25 ° E.

Optimal pH value for growth 7.0 to 8.5 · Fundeteller soil from Iowa.
Locations »Soils.

A recent identification carried out in the laboratories of the inventors shows that the microorganism is true not a Micricocous, but an Arthrobacter let and has a strong affinity with arthrobatic ureafacenes. The following summary explains the reason for the preferred Identification of this organism as Arthrobacterx

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Mioroooooue

Χα early ermentation stage Always grim positive

Cells in irregular shapes

Never changes its size

Never comes in Stabfora before

Carbohydrates are often fermented

"M.Cerificana"

Always grim negative

As well as
Arthrobacter

As well as
Arthrobaeter

As well as
Arthrobot

As well as
Arthrobot

Arthrobot

Grief-negative or distressing

The formation of short threads old somewhat rudimentary eye formation can take place. gene

Sometimes larger appear as the usual oooooideh cells

Large, oooooid cells lead to the formation of rod-shaped ones Cells

Little or no acid formation from carbohydrates

As already mentioned, air is continuously introduced into the culture and preferably in the bottom of the fermentation vessel through a distributor pipe or some other similar device to divide the air into fine bubbles SU that the oxygen is completely distributed in the reacting culture medium. Ss BB * one or more Sohaufel stirrers with 1 to 100, preferably 5 to 30 PS per 3785 l of liquid culture medium can be used. The stirring possibilities with the bucket stirrer are in advance more than 1000, in particular more than 1500 rpm. It is preferable to work with vortex ventilation, ie if a vertical one is used

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Cylinder, which extends from a point immediately below the liquid surface to close to the bottom of the vessel, over to achieve a better mix and to focus on the fomentation forming foam away from the surface of the liquid Pull and distribute through the pipe to the bottom of the vessel.

Ale source of nitrogen can be organic or inorganic Nitrogen derivative can be used. Suitable organic compounds are proteins, hydrolyzed with acid Proteins, enzyme digested proteins, amino acids, yeast extract, asparagine and urea. For economic For reasons, asm preferably uses an inorganic compound, such as ammonia, ammonium hydroxide or salts thereof, such as ammonium phosphate, Ammonium citrate, etc. A very advantageous method of adding the nitrogen is to add ammonium hydroxide. In this way, a pH range of 5.5 to 7.5 achieved and supplied the required nitrogen. For this purpose you can also use gaseous ammonia directly initiate into the culture. If necessary, additional acid or base, such as sulfuric acid, can be added from time to time. Add. The present method does not require expensive stimulants such as vitamins and the like.

Besides energy and sources for nitrogen are in the fermentation medium in order to achieve the greatest selectivity critical amounts of selected mineral nutrients are also required. In this sense are potassium, sulfur and phosphorus

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required These elements can be added in poms of their salts «The potassium can Z ₀ Ba as chloride, phosphate,
Sulphate, citrate, acetate, nitrate etc. ₀ can be carried out. Sulfur and phosphorus can ode in the form of sulfates? Phosphates, such as alkali or ammonium phosphate, are supplied. The preferred critical amounts of minerals used to achieve the highest selectivity are the following »
where the appropriate amounts for different concentrations of cells are determined by multiplying the relevant percentage by the cell concentration?

Particularly preferred amount of salt in the supplied aqueous medium in GeWo-Jt * based on the increase in a cell concentration of
1 wt «- *

Minerals P. Preferred amount of salt
In the supplied
aqueous medium in
Weight ~ #, based on
breeding one
Cell concentration
from 1 wt H ₃ PO ₄ S + Na 0.01 to 1.0 Na ₂ SO ₄ K + Cl 0.01 * 0.5 KCl Man 0.01 "0.05 MgSO ₄ Oa 0.005 "0.5 OaOl ₉ 0.005 «0.5

0.05 to 0.25 0.025 * 0.05 0.05 "0.10 0.02 * 0.05 0 * 02> 0.05

the following minerals are required, dtren Quantities, however, are less critical

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Particularly preferred

Preferred salt tongs, salt tines, weight jfc, Gew. ^ C, based on based on the Züohdie Growing a cell concentration concentration of minerals of 1 weight per cent

FeSO ₄ Ve 0.001 to 0.1 0.005 to 0.02 Rn 0.001 "0.1 0.005" 0.02

The ions given above can also be compared to a less preferred one Formation of form fora in the form of other salts in the same (to be determined by calculation) stuohioaetrisohen Quantities are fed.

Furthermore, nitrogen buss as salt, ammonia, Απωoniurneals or ammonium hydroxide, preferably in the form of an ammonium bond » especially as annnonium hydroxide, in amounts of 0.08 to 0.20, preferably from 0.1 to 0.15 g of nitrogen per g of dried kneader generated cells are supplied.

The temperature of the culture can range from about 20 to 45 ° 0, preferably from 25 to 40 ° 0, to maintain the correct temperature it is usually necessary to warm the Geaisoh initially while bit progressive Waohstua is generated in the culture Warne so that cooling is then necessary, above the desired tea temperature to pause. This can be done in an economical way by circulating part of the goods through a cooler or by using cooling coils in the reaction vessel as described above.

If air is used, the amount of air supplied is

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ten hydrocarbons (including the goods circulated), based on the added water content (including NH ₄ OH) 1 to 10% by weight, preferably 1 to 5% by weight, in particular 0.5 to 2% by weight .

'' The fluid retention time for the preferred bacteria, näslioh Mioroooccua cerifioaris (Arthrobacter), i.e. the liquid volume in the reaction vessels divided by the Amount of output goodea fed in per hour (and that for Keeping the liquid level constant in the reaction vessels withdrawn products), is generally 1 to 10 hours « preferably 1 to 3 hours, in particular 1.5 to 2.5 hours. These preferred residence times do not only improve the selectivity » but also reduce the need for oxygen.

example 1 Influence of various starting materials on the bacterial yield

and the growth of animals

Miorococous oerifioana was in an aqueous salt medium bred which a lirik gas oil fraction as the sole source for carbon. The product cells were removed by centrifugation harvested, washed once with water, centrifuged off again, washed twice with acetone and filtered off. Like that in "Official Method of Analysis", 9th edition, 1960 « Protein assessment procedures described in Section 39 »133 (e) and (f) when rats were fed a diet which contained these bacterial cells as the only protein obtained the following results. With these (plague, the sample to be examined is added to the feed in such an amount,

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because the protein content of the feed is 9.09 * “The had are fed with this putter for 4 weeks. The weight unahae and feed consumption is rewarded every week. A bath of the fourth week is the Gewiohtssunahae, sucked on the used putter length, calculated and the value »it that of a control series carried out at the same time, in which the feed contains casein as protein. For both test series (old casein and bit of the au Sample) 10 rats are used.

Biological nutritional value according to the rat feeding

Paraffin, cell aging method, starting material Qew .-) tau yield (casein »100)

Lirik-Gaeöl 60 59 59 Parafflngateoh 76 97 53

Hexadeoan 100 105 76

The improvement in nutritional value is likely the fact rubbing the ears, that the hydrocarbons not uagesetate «- • toffrüoketand made from dea Gaeöl and paraffin slack from the Waohstua of animals hemet, while pure or relatively pure (i.e. purified as described above) paraffins none disadvantageous effects «intrinsic» This can be seen from the following additional tereuohe values. Di · Aoeton-Waeohfluesigkeit dea Idrik-aaeöl became unaffected by driving off the acetone at Ataoephärendruok and then ia Vakuua conientriiert. laoh dea Xrlnhieren dee Ölruoketandes ait 5 £ lfer Xatronlauie sweoks Intfernunf of organic acids became · the neutral · Iraq ·

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tion analyzed by mass spectrometry. This resulted in the following composition: 31.4 $ paraffins, 19.2 $ naphthenes, 49.4 $ aromatics. This material was then mixed with a standardized casein (standard regulation of the American Nutritional Research Council) in amounts of 1, 3 or 5 GeW ₀ -Jt, and this casein was used in the protein evaluation test according to "Official Methods of Analysis", 9th edition » 1960, Section 99o.135 (o) and (f) added to rat food as the only protein · In the same way, a number of casein samples were prepared which contained 1, 3, 5, 7 and 10 56 purified paraffins (adsorption on 5A molecular sieve and subsequent purification), the paraffins essentially consisting of pure straight-chain paraffins with an aroma t tightness of about 70 ropes per million.

The average growth of the rats in these experiments was compared with the average waohetum of rats fed pure casein (10 rats per sample) of 4 weeks compared. The results can be found in the following table:

Growth of the rats, Jl of the oasein hydrocarbon control sample

amount in casein, Lirik-Gaeöl- Paraffins purified with molecular sieve wt .-} * residue

1 77.5 103

3 75.0 100

5 61.9 104

7-94

10-103

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Further evidence of the inhibitory effect of the old acetone extractable material is the fact that Mioroooooue oerifloans HO-1, when grown on paraffin and not washed old acetone, was practically worthless as a protein additive, since the rats were fed old food containing this material as a single protein, 5 g increased in two weeks, their weight gain so only 14 1 · those was that at the same time old pure "casein was ersielt.

^ Example 2
Importance of lingering

Mioroooocus cerifioans was continuously in dea following Bred Mediua:

g / l

Paraffin oil from 10 to 20

Sorbitan aonostearate
("Span 60 ^M ) -Polyoxyethylated sorbitol fatty acid ester has an average of 60 oxyethylene groups (" Tween 60 ") 0.6 to 1.2

5
1

0.5 0.2 0.2 0.2 0.2

Concentrated Aaaoniuahydroxydlösung was added as needed - among other things to keep the pH value at 7.

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H ₃ PO ₄ o7 H ₂ O KOl • 4 HgO OaOl ₂ • 7 H ₂ O MgSO ₄ MnSOj FeSO ₄ HaOl

The above nutrients were in the desired concentrations dissolved or dispersed in tap water.

The temperature of the reaction mixture was kept at 30 ° C. and the fermentation was carried out in a 7.5 liter New Brunowiok fermentation vessel with a pulp volume of 4. The air flow rate and the rate of agitation were modified so that the aeration efficiencies, determined according to the Sulflt-Ojcydation method of Cooper and Pernet rom (Industrial and Engineering Chemistry, Volume 36, 1944, pages 405-509) »in the range from 3.5 to 7, 0 Hillieol O ₂ per liter per hour.

You dwell time in the fermentation vessel was in the area varies from 1.3 to 4.0 hours, and the effect of these changes results from the following table:

Selectivity, wt% dry cells,

soaked up in cells and 0O ₉ transferred

Dwell time, hours led ParaffingatsolT

4.0 107

4.0 102

2.0 120

2.0 128

2.0 134

2.0 133

2.0 121

1.3 138

This is a significant step forward, isn't it only in view of the increased selectivity, but also because this 30-year improvement in both the oxygen demand and

also reduces the amount of heat to be dissipated by 50 Jt. This Versuoh was made with a pure straight chain

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Paraffinkohlenwaseeretoff, namely n-hexadecane, as a starting material repeated, and analyzes for unreacted hydrocarbons were carried out by steam fractometry. In these" parts could calculate the selectivities as Jt Hettoprodukt which per unit of hydrocarbon consumed was formed. My results are as follows

Lingers, selectivity, weight -φ dried cells, hours based on hydrocarbon consumed

4 105-2

2 116 ± 6

Each of these values represents an average value from 5 to 6 tests «

Example 3

Significance of the air supply speed Hioroooccue oerifloans was searched for, as described in Example 2, using a Faraffingatsohkonsentratlon of 20 g / l. The stirring speed and the air supply speed were varied in alternate experiments, the following results being obtained!

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Ventilation effectiveness (Trash mol //)

Residence in the reaction vessel, Hours.

In the case of delivery delays, VA / min.

^ Hydrocarbon _f based on inorganic salts Ια aqueous Mli

4 * 8 7.0 4th 4th 1 1.5 2 2 1200 1400

Charging speed, X./mn,

Cell yield, 5 &, related

on sugeeetsstee wax 44 «4 9S, 1

Example 4

Influence of the reduced u £ @ n & Verweilzelt AEIE oxygen ausnutzun _r / i and the Wärassentwleklung

Xn the same way as described in dsm Examples 2 and 3, were continuously under © FesraentationsVersuohe

same conditions with the exception of the time in the reaction vessel durohgeftüirto The conditions were the following:

Fermentation medium -Aa- BATH ORIGINAL n-hexadecane 10.0 85% H ₃ PO ₄ 2.5 KOl 1.0 Ha ₂ SO ₄ 0.5 MgSO ₄ 0.25 MnSO ₄ ⁴ HgO 0.04 FeSO ₄ 7Η ₂ Ο 0.04 OaOl ₂ 0.5 The rest of the tap water

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Reaction vessels, Faaeungsveraugen 7.5 1, FLUID VOLUME 4 1

Taffeta feed rate 3.0 l / min.

By adding τοη HH * OH retained pH value «7.0 Charge speed 1500 rev / Hin. Temperature 35 ° C. Results* Dilapidated, hours, 4 2

Selectivity, wt .- * dry ropes, sucked up
hexadeoane consumed 1.13 1.30

Oxygen consumption,
kg / kg dried cells 1.27 0.89

kg / kg dried cells 0.97 0.62

Other · Ε ^ ηΊ " ¹ « * »* experiments have always given the same results. Although the selectivity and the acidic brewing vary somewhat depending on other conditions, the dependence on the time is always the same ·

Example 5 Child development it

Using Miorocoooue cerifioane and suitable salaedlan, as described in Examples 2 and 6, the "logarithmic" growth was observed in 7.5 l reaction vessels at a buoy speed of 1500 rpm and an adequate supply of air, including the partial pressure Keep ace of oxygen in the exhaust gas above 0.007 kg / oe ² su. Bas usual

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Uses for the logarithmic © growth was the concentration dee Carbon dioxide in the exhaust gas, which doubled every 45-10 minutes ο Xn some cases, the cell concentration was also increased monitored, and the results confirmed that The minimum development time for Miorooooous is therefore 45 ± 10 minutes.

Example 6
Significance of the inorganic bellows containing Beaktionaefliuas

Miorooocous oerifieane was cultured batchwise in Sohüttell flasks using τοη 50 ml or 500 ml Erlensteyer flasks in a rotating saddle.lmasohine at 250 rpm, and for a period of W hours on n-hexadsoane using the following inorganic salt medium.

2.0

₄ 1.0

ifeS0 ₄ »7 H ₂ O 0.5

Fe ₂ (SO ₄ J ₅ 0.01

MnCl -4 _g H ₂ O 0.008

H ₂ O 0.002

The pH was adjusted to 7.2 in hydrochloric acid. Bs was get a cell concentration of 2.0 mg / nl, which is below under these conditions a yield of 12.9% was obtained Amount of the «! Led Hexadeoane, means · If the consentration of sodium nitrate was increased to 12 g / l, the line concentration increased to 4.5 »g /» l, doh. the yield rose

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29 yrs. Under the same conditions with the in Example 2 described mediua cell concentrations of 9.4 ng / al and Achieved yields of £ 60.6.

Example 7

Under the conditions described in Example 2 north with a residence time of 2 hours, Mioroooccus cerifloane xlt n-hexadeoane as a substance using hexadeoan concentrations of about 1 to 2 wt. If the hydrocarbon concentration 1 i »· oetrug yielded verifiable only a small influence of the use -of pure oxygen. At a carbon concentration of 2 £, however, the pure oxygen considerably improved the utilization of the hydrocarbon, as the following values are proper

Concentration of n-hexadeoane, g / l

Source of oxygen Selectivity,

g cells / g converted O _i6 carbon dioxide

Degree of conversion dee hexadecans

9.3 10.9 19.1 19.1 air ° 2 O ₂ üuft 1.14 1.20 *
1.13 1.1 96 91 85 73

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Claims (9)

H42089 Eeso Research and Eng «CSo. Claims Claims 1 ·) A process for the production of protein-rich foods, characterized in that Cj to Q-xq hydrocarbons, a liquid »aqueous» inorganic salts containing enrichment medium and an oxygen-containing gas are continuously fed with stirring to a substance vessel which contains the hydrocarbons »the aqueous medium and contains bacteria capable of growing by reacting the hydrocarbons, continuously removing a mixture of bacteria, aqueous medium and unreacted hydrocarbons and separating the bacteria. 2. A process for the production of protein-rich foods, characterized in that O ₁ - to O · »- hydrocarbons», an aqueous, inorganic salt-containing, growth-restricting, aqueous ZüohtungsBedium and an excess oxygen-containing gas are continuously fed with vigorous stirring to a reaction vessel, which the Hydrocarbons, the aqueous medium and bacteria containing »which are able to grow under conversion of the hydrocarbons» »loh in the logarithmic growth phase (the exponential 909810/0380 "'*" ■ bad original increase) and are present in a concentration, Al is equal to or greater than that »that is to say, the exponential · Bacterial increase is equal to the continuously sucked-off amount, while nan is continuously a mixture Bacteria »waives aqueous medium and unreacted hydrocarbons and separates the bacteria. 3 · Terfahren according to claim 1 or 2, characterized by » that the material separated from the bacteria is circulated into the reaction vessel surüok. 4. Terfahren according to claim 1 or 2 »characterized in that that the liquid β dwell time is equal to 1.5 to 4 times the minimum development time of the bacteria concerned measured. 5. The method according to claim 1 or 2 »characterized in that that the oxygen-containing gas dea reaction vessel in Mangen Clay 0.5 to 4 components air / rough part reaction fluidity today per Jt dried bacterial concentration in reaction flasks / liquid residence time is supplied in hours. 6 · Terfahren according to claim 1 or 2 »characterized in that a normally gaseous hydrocarbon is used will. 7. Terfahren according to claim 1 or 2 »thereby known se is worthwhile, that light petrol is used as the hydrocarbon. - 33 909810/0380 8. The method according to claim 1 or 2, characterized in that βίβ hydrocarbons boiling between 190 and 400 ° C · straight-chain «paraffins are used, which _{contain less all O 9} I wt.-Jt aromatics. 9 * A process for the production of protein-rich hand-made ingredients, characterized * in that a mixture of 0 »1 or 10 wt. A mixture of bacteria, aqueous medium and unconverted paraffin hydrocarbons is continuously drawn off into an aqueous Züohtungsmedium giving off inorganic salts and an oxygen-containing gas with stirring to a reaction vessel seeded with Microooccue oerifloans. Bacteria with a liquid content of 80 to 95 C-ew.- £ _t based on the Trookenzellen, refrains and the bacteria dries. 10. Process according to claim 9, characterized in that one with a residence time in the reaction vessel of 1.5 to 2.5 Hours worked 11. The method according to claim 9, characterized in that add up to 5% by weight of straight-chain paraffinic hydrocarbons to the reaction vessel containing mixture and an oxygen-enriched air with an oxygen content of more every 90 yrs. - 34 - 909810/0380 UA2089 12. The method according to claim 91, characterized in that the aqueous Züohtungeaediu containing the inorganic salts is supplied in the following amounts, the weight percentages for the salt refer to the dried bacteria obtained per unit of time, O ₉ 05 to 0.25 wt. -5t H ₅ PO ₊ , 0.025 to 0.05 wt. J & Ha ₂ SO ₄ , 0.05 to 0.10 GeWo * ECl, 0.02 to 0.05 wt. ^ HgSO ₄ , 0.02 to 0.05% by weight of OaOl ₂ , 0.005 to 0.02% by weight of FeSO ₄ and 0.005 to 0.02 13. The method according to claim 9 »characterized in that ■ works at a Terwell time in the reaction vessel of 1 to 2 hours, a 4 to 8 wt .- ^ straight chain ^ paraffins containing Geeisch and oxygen-enriched air with an oxygen content of more than 90 Supplied in such an amount that the amount of oxygen supplied is 1.5 to 2.5 kg per kg of dried bacteria obtained as product per side unit, and that the aqueous ZÜhungsnediuiB containing the inorganic salts is supplied in the following amounts, whereby the Percentages by weight for the amount of salt refer to the dried bacteria obtained per unit of time: 0.05 to 0.25% by weight -5 & H ₅ PO ₄ , 0.025 to 0.05 % by weight 4 Ha ₂ SO ₄ , 0.05 to 0.10 6EW <> - £ XCl, 0.02 to 0.05 MgSO _4, 0.02 to 0.05 wt £ CaCl _2, 0.005 to 0.02 ßew. _jt-FeSO4 and 0.005 to 0.02 wt. - £ MnSO ₄ . - 35 909810/0380 U42089 Η «Bacterial, protein-string, non-toxic foodstuff with biological nutritional value and an aromatic content τοη less than ale O ₁ OS Gewo- #, consisting of a protein-Xährkoaponente and a light protein-Hearing component, whereby the protein component non-viable cells of Microooccus oerifloans ale predominate Bacterial protein contains "which are essentially free of mononuclear and polynuclear aroaatic hydrocarbons and substances harmful to growth. 15. Protein-rich, non-toxic cattle feed additive alt biological nutritional value and an aromatic content of less than O # 05 wt. 5t, whose protein component consists essentially of consists of non-viable cells of Microcoocua oerifleans, which are practically free from mononuclear and polynuclear aroaatisohen Are hydrocarbons and, based on Trookenbasls, only the following substances, which are harmless to the diet, in changing, but not exceeding the following maximum values Consentrations included! a) Beter, 0.5 wt .- #, b) straight chain O ₁₁ - to C ₅₀ -Paräff ^ hydrocarbons, 2.0 OeWoH ^ and o) inorganic salts, 9.0 wt. 16. Method of sun feeding animals with non-toxic animal protein, characterized in that the animal · old non-viable cells fed by Mioroooooue oerifioans - 36 - 909810/0380 ₉ which are practically free of mononuclear and polynuclear aromatics eowle τοη for the growth of harmful substances. - 37 909810/0380