Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
• • C—CHEMISTRY; METALLURGY
  • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F1/00—Treatment of water, waste water, or sewage
  • C02F1/68—Treatment of water, waste water, or sewage by addition of specified substances, e.g. trace elements, for ameliorating potable water
  • C02F1/681—Treatment of water, waste water, or sewage by addition of specified substances, e.g. trace elements, for ameliorating potable water by addition of solid materials for removing an oily layer on water
• • C—CHEMISTRY; METALLURGY
  • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F3/00—Biological treatment of water, waste water, or sewage
  • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
  • C02F3/344—Biological treatment of water, waste water, or sewage characterised by the microorganisms used for digestion of mineral oil
• • C—CHEMISTRY; METALLURGY
  • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
  • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
  • C09K3/00—Materials not provided for elsewhere
  • C09K3/32—Materials not provided for elsewhere for absorbing liquids to remove pollution, e.g. oil, gasoline, fat
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
  • C12N1/14—Fungi; Culture media therefor
  • C12N1/145—Fungal isolates
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
  • C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
  • C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
  • C12R2001/00—Microorganisms ; Processes using microorganisms
  • C12R2001/645—Fungi ; Processes using fungi

Definitions

• the invention relates to the degradation of the hydrocarbon constituents of crude petroleum and petroleum products in any environment. It particularly relates to the degradation by emulsification, solubilization and breakdown of said petroleum materials by application of an effective amount of Geotrichu marinum or its enzymatic active principles.
• Milz & Frazer Jour. Petr. Techn. 24:255-262 (1972). Milz and Frazer found that an oil slickof 40 gallons of crude oil would cover a 200 x 30 foot area 10 minutes after being spilled into the open sea, which slick would expand to an area 100 feet wide and one-half mile long after one hour.
• German Patent No. 2547742 discloses a typical system, in which polymers are used to absorb crude petroleum in an oil slick, where the oil must be then removed by skimming of the spill-polluted water en ironment.
• the art totally lacks any effective method, compound or composition capable of degrading crude petroleum and petroleum products in actual, non-laboratory conditions and environments, such as open ocean spills of crude petroleum.
• the art particularly lacks any such degradative method, compound or composition utilizing a microorganism such as a bacterium or fungus having such actual environment effectiveness.
• c products in an environment is such that:
• the method, compound and composition must be easy to effect and manufacture, while safe to personnel applying the composition or carrying out the process at all stages and times of its preparation and use.
• the present invention relates to a method, compound and compositions for effecting the degradation of crude petroleum and petroleum products in any environment.
• the method comprises
• the compound and compositions of the present invention comprise the fungus Geotrichum marinum Thirumalachar sp nov . (which term herein comprises generically its cultural derivatives and mutants) and a biologically pure culture of the fungus Geotrichum marinum Thirumalachar sp no .
• ATCC 20614 which fungus, cultural derivatives, mutants and culture are capable of effecting degradation of crude petroleum and petroleum products in an environment.
• An enzymatic active principle compound which degrades, by emulsification, solubilzation and breakdown, the hydrocarbon constituents of petroleum materials, and which is produced by and isolated from the Geotrichum marinum fungus, is also provided by the present invention, as are compositions comprising the fungus or the enzymatic active principle compound produced by the fungus and a carrier medium.
• the method, compound and compositions of the invention are particularly efficacious when utilized to degrade crude petroleum and petroleum products in a salt water environment, including the open ocean.
• the present invention overcomes the lackings and drawbacks of the prior art by providing a method, compound and composition for effecting degradation of crude petroleum and petroleum products which demonstrates efficacy in actual, non-laboratory environments, including
• OMPI inhospitable climates and various land and open water conditions, yet is non-toxic and non- deleterious, generates no deleterious products or chemicals, is long-acting and rapid in the onset of its initial activity, does not require further support of sustenance after application to the crude petroleum or petroleum product, is, together with the degraded, solubilized crude petroleum or petroleum products, self-dissipating, so as to require no retrieval and disposal of any petroleum-laden component or material, and is easy and safe to manufacture and utilize.
• the preferred method of the invention provides for the degradation of crude petroleum and petroleum products in an environment.
• the method comprises contacting said petroleum material withan effective amount of a new species of fungus, Geotrichum marinum Thirumalachar sp nov., isolated from marinesoil.
• the new fungus, Geotrichum marinum Thirumalachar sp nov., has been deposiited in accordance with the provisions of MPEP ⁇ 608.01 (p) (1981), and may be obtained from the permanent colleciton of the American Type Culture Collection, Rockville, Md., where it has been deposited under an unrestricted deposit as Geotrichum marinum ATCC 20614.
• An "effective amount" of the fungus Geotrichum marinum Thirumalachar sp nov. is an amount sufficient to degrade, by emulsification, solubilization, break down and consumption, the
• the novel fungus, Geotrichum marinum Thirumalachar sp nov . was isolated from marine soil and bred to a pure form of superior efficacy with respect to hydrocarbon degradation by strain selection techniques, known to the art, which extended over several generations . This involved the utilization of standard techniques in growing daughter generations and selecting single cell colonies, which were then grown in known fermentation media to measure the capacity of those cultures to degrade petroleum incorporated into said media. This also provided superior, enhanced mixing properties in the Geotrichum marinum with respect to crude petroleum or petroleum products, allowing ready mixing with the petroleum hydrocarbon and emulsification and metabolization thereof in the process of further growth of the fungus.
• the fungus Geotrichum marinum Thirumalachar sp nov. has many of the general characteristics of Oncocladium Wallroth (Sensu Hughes, Canadian Jour. Bot. 46:941 (1968)), in having only the side branches fragmenting into arthroconidia.
• the fungus including its biologically pure form, is a white filamentous fungus with septate branching hyphae, which fragment the branches into said arthrospores with growth. In an old culture, the arthrospores appear as a group of yeast cells.
• a filamentous fungus is involved, its habitat en masse is like that of a yeast cell: it remains as a mass of rectangular free cells in the early stages of growth, and thereafter rounds off into ovate to rhomboidal cells in a mature condition.
• the fungus when cultured on both agar surface cultures and submerged liquid cultures, produces a strong apple fruit flavor characteristic of the strain.
• Other species of Geotrichum are known, including the common Geotrichum gracitis isolated from cow's milk. None of these species, however, possess the unique hydrocarbon degradation properties of .Geotrichum marinum in either its isolated or biologically purified form.
• This rapid growth and biomass-generating capacity of the novel Geotrichum marinum fungus provides it with capabilities to maintain itself and its degradation of hydrocarbon constituents of petroleum materials in all types of environments, including open ocean, over time, and to continue its growth activity and rapid metabolization of said hydrocarbon constituents.
• the novel Geotrichum marinum fungus grows readily on most of the known media used for culturing fungi under laboratory conditions, including sold agar media such as potato dextrose agar, Sabaouraud's agar, the mycelium is white, thin and crusty, becoming rough and chalky white with the formation of a mass of ⁇ onidia; it is never creamy and yeast-like as in the other known Geotrichum species.
• the fungus may be grown in submerged culture on a variety of known media, and is capable of utilizing various carbon sources, such as alcohols, fatty acids from oils, sugars and polysaccharides, and nitrogen sources, both organic and inorganic in nature.
• various carbon sources such as alcohols, fatty acids from oils, sugars and polysaccharides, and nitrogen sources, both organic and inorganic in nature.
• the addition of various amino acids and vitamins has an effect on the growth of Geotrichum marinum and its production of enzymatic active principle compounds, which comprises its hydrocarbon emulsifying and breakdown constituent agent.
• Boosting of the pseudosolubilization properties of the fungus to achieve breakdown of the petroleum material maybe provided by addition of these known amino acids, vitamins and growth-promoting substsances.
• the preferred method of the invention involves utilization of the novel Geotrichum marinum fungus in any form and level of biological strength or purity for degradation of petroleum materials.
• a particularly preferred method of the invention utilizes the biologically pure form, produced by the aforementioned strain selection techniques, of the Geotrichum marinum Thirumalachar sp nov. for contacting with the crude petroleum or petroleum product in an environment to cause degradation.
• the environment in which the method of the invention demonstrates efficacy comprises any land or water area, enclosed or open, including salt water bodies and open ocean locations. No loss of efficacy results from petroleum presence in
• These environments also include man-made localities and objects, such as the inside of closed or open storage tanks and vessels, the interior of reaction vessels, chemical processing apparatus and piping, the interior portions of internal combustion engines and related fuel storage tanks or pumps, and the interior of pipelines and related pumping equipment.
• the environments in which the method of the invention is efficacious includes virtually any locality in which the Geotrichum marinum fungus or its enzymatic active principle compounds can be introduced. This would also comprise underground oil formations or other naturally occurring oil formations.
• the method of the invention maybe effected in parallel or concert with, or otherwise concurrently or as a part of other known industrial and chemical processes compatible therewith. These processes would include those not destructive of the Geotrichum marinum fungus, such as secondary degradation of undesired petroleum products after an initial separatory or extractive procedure.
• Another preferred embodiment of the method of the invention for effecting degradation of crude petroleum and petroleum products in an environment comprises the application to said petroleum materialof an effective amount of a composition comprising the novel Geotrichum marinum Thirumalachar sp nov. fungus and a carrier medium.
• the carrier medium comprises any of the standard commercially available carrier media.
• a particularly preferred carrier medium is particulate wheat bran, which also functions as a potential nutrient source (fungi culture medium)
• OMPI for the f ngus.
• the presence of the wheat bran is not, however, in any manner necessary to the practice of the invention.
• Other particularly preferred carrier media include micronized cereal, micronized corn cob, vegetable waste products (such as cereal husks), hay and seaweed powder. Mixtures of the particularly preferred carriers may also be utilized.
• a further preferred embodiment of the method of the invention comprises the application of an effective amount of Geotrichum marinum and a carrier medium to crude petroleum or petroleum products in an environment, followed by further addition of a fungi culture medium to said petroleum material after application of the fungus.
• Fungi culture media useful in this embodiment are again well known in the antoibiotic and drug arts, and may include those used for culturing fungi in the laboratory, such as potato dextrose agar or Sabouraud's agar.
• a preferred media for such post-fungus application addition is particulate wheat bran.
• Other media including micronized cereal, micronized corn cob, vegetable waste products (such as cereal husks), hay, seaweed powder and mixtures thereof, may also be used.
• Another preferred embodiment of the method of the invention comprises a variant on direct contact of the Geotrichum marinum fungus per se with the crude petroleum or petroleum product.
• Degradation of the petroleum materials in this additional embodiment is effected by growing Geotrichum marinum Thirumalachar sp nov . in a liquid fungi culture medium, harvesting the resulting broth, and thereafter applying an effective amount of said broth to said petroleum
• the growth may be carried out by those procedures of fermentation and growth well known to the art for the production ofantibiotics and enzymes, such as the procedure described in Unitd States Letters Patent No. 4,082,613, issued April 4, 1978, at column 6, line 18 et seq. , which description is specifically incorporated herein and made a part hereof.
• the growth maybe continued until the nutrition provided- by the fungi culture medium is substantially- exhausted, or may be terminated prior to that stage, so as to provide a broth which will contain sufficient remaining nutrition to attenuate the growth and initiation of degradation of the hydrocarbon constituents of the petroleum material.
• Geotrichum marinum in a nutrient media in aerated, agitated fermentors.
• Particularly preferred nutrients include corn starch, glucose, particulate wheat bran, cotton seed hydrolysate, such as PROFLOTM, manufactured by Trader's Oil Mill Co., P.O. Box 1837, Fort Worth, Texas 76101, mineral salts, and other carbon-nitrogen source materials.
• the broth containing the novel Geotrichum marinum Thirumalachar sp nov. fungus may be applid to the petroleum materials in the environment in
• OMPI any effective manner.
• a preferred method of applying the fungus-containing broth is to apply the broth to the petroleum materials in the form of a finely divided spray, by utilizing well-known spray devices.
• An alternate method of application comprises the further step of air drying the broth under vacuum so as to recover a dry, powdery material. The dry material may then be applied in an effective amount to the petroleum material, such as by dusting from an airplane or helicopter or by broadcasting from apparatus or vehicle, such as a boat.
• the invention also provides a preferred method for preparing a compound for effecting degradation of petroleum materials in an environment.
• the method comprises growing Geotrichum marinum Thirumalachar sp nov. in a liquid fungi culture medium to produce a broth, harvesting the resulting broth, and isolating from said broth an enzymatic active principle compound or compounds. It is particularly preferred to utilize the biologically pure Geotrichum marinum culture having the characteristics of ATCC 20614.
• the active principle compound can be separated from the broth by techniques well known in the antibiotic, drug and enzyme arts, including absorption, elution and precipitation techniques, and may thereafter be concentrated or reduced to a dry form by well-known methods familiar to those arts.
• the enzymatic active principle compound may be isolated by air drying said broth under vacuum, for example, which results in the recovery of a dry active principle material.
• Geotrichum marinum in aerated and agitated fermentors in a media which contains certain nutrients.
• Particularly preferred nutrients include glucose, particulate wheat bran, cottonseed hydrolysate, such as PROFLO 1 ", manufactured by
• fungi culture medium for use in this method contains glucose, cotton seed hydrolysate and salt water, and an optimum culture further contains particulate wheat bran in addition to said glucose, cottonseed hydrolysate and salt water.
• the preferred compounds and compositions of the invention for effecting degradation of crude petroleum and petroleum products comprise the enzymatic active principle compound or compounds produced by the novel Geotrichum marinum Thirumalachar sp nov. fungus, the fungus itself, the biologically pure culture of the fungus, and a composition comprising any of said compounds and a carrier medium.
• the enzymatic active principle compound of the invention is produced by the method of the invention hereinbefore described.
• OMPI which may be reccovered as a dry material, constitutes the enzyme(s) produced by Geotrichum marinum Thirumalachar sp nov. which effects the degradation of the hydrocarbon constituents of the crude petroleum and petroleum products by emulsification, solubilization or pseudosolubilization and break-down. It is non- toxic and non-deleterious to the environment, and leads to no formation of toxic or deleterious products during or after hydrocarbon degradaiton.
• the biologically pure culture of the invention is that culture of the novel Geotrichum marinum Thirumalachar sp nov.
• compositions of the invention comprise the combintion of the enzymatic active principle compound produced by the Geotrichum marinum fungus, the Geotrichum marinum fungus, or the biologically pure culture of the fungus with a carrier medium.
• the carrier media heretofore set forth are suitable for combination with said fungi or compounds to produce the composition.
• compositions comprise the enzymatic active principle compound, either in liquid or dry form, and particulate wheat bran, or the biologically pure culture of the fungus Geotrichum marinum Thirumalachar sp nov. and particulate wheat bran.
• Micronized cereal, micronized corncob, vegetable waste products (such as cereal husks), hay, and seaweed powder may be used in place of or in further combination with said particulate wheat bran.
• compositions of the invention comprise the broth resulting from growing Geotrichum marinum Thirumalachar sp nov. in a liquid fungi culture medium and a carrier medium.
• Particularly preferred compositions are those comprising the broth and particulate wheat bran, micronized cereal, micronized corn cob, vegetable waste products (such as cereal husks), hay, seaweed powder, alone or in combination, in admixture.
• compositions of the invention effect degradationof crude petroleum and petroleum products.
• the unique enzyme compound or compounds produced by the novel Geotrichum marinum Thirumalachar sp nov. fungus causes an emulsification and solubilization or pseudosolubilization, due to its surface active properties, of the hydrocarbon constituents of the petroleum materials. Once so solubilized, the fungus then breaks down the hydrocarbon constituents and utilizes the breakdown products as a growth media or nutrient, resulting in the metabolization of the petroleum material.
• the result is a biomass of non-toxic, non-deleterious fungus or fungus plus carrier medium left in the environment, either on the surface of a land area, on the surface of a body of water, or located in an artificial environment such as the interior of an oil storage tank. If exposed to the elements and general weather cycles, the fungus biomass, once deprived of a further source
• OMPI of sustenance on completion of the hydrocarbon constituent breakdown and degradation of the petroleum material is itself rapidly degraded and absorbed into the environment, with no toxic or deleterious effects. If the biomass is in an enclosed artificial environment, it need only be removed and placed in a settling tank, dump area or otherwise exposed to the elements and general weather cycle to effect its degradation in turn. Degradation of the crude petroleum and petroleum products is therefore effected by the methods, compounds and compositions of this invention, without concomitant production of further toxic or deleterious substances serving to further foul the environment which is sought to be cleansed.
• the flask was incubated on a reciprocating shaker at a temperature of 24-28 degrees C Very rapid growth was observed, and, within 48 hours, a mass of septate hyphae with arthroconidia was formed. Growth was complete at the end of 72 hours, with the depletion of nutrition.
• the broth was semi-viscous with a strong apple flavor.
• the resulting broth was then placed dropwise into sea water to determine its hydrophilic-hydrophobic characteristics.
• the broth demonstrated hydrophobic characteristics: it did not mix readily with water (unless titurated) , and remained floating on the surface, forming a filmy layer which microscopically showed filaments of fungi and conidia.
• a peroleum hydrocarbon (Shell X-100 Grade motor oil) was then placed on the sea water/broth.
• the fungus broth mixed readily with the oil film, and gradually hydrolyzed, emulsified and consumed the petroleum hydrocarbon.
• the broth produced as described was also incubated for a 15- to 20-day period in a flask containing a mixture consisting of sea water and a layer of petroleum hydrocarbon (Shell X100 Grade motor oil).
• the fungus first pseudosolubilized the oil, and gradually grew as a thin film covering the upper surface of the water-oil layer of the flask.
• the fungus growth though slow, was continuous in the petroleum oil-sea water mixture, despite the lack of any addition or presence of any added nitrogen source in the flask.
• Incubation/fermentation of the fungus was effected by placing the seeded flask on a reciprocating shaker at 24 to 28 degrees C. Forty-eight hours later, there was a profuse growth of the fungus, showing development of mycelia with conidia. By 72 to 80 hours, all of the added petroleum hydrocarbon had been emulsified and consumed into a uniform mass of fungus. There was no separation out of oil or an oil layer on standing, and when a 10% dilution of the broth was made in water, solubilization resulted.
• a liquid fungi culture nutrient medium comprising 10 grams of particulate, powdered wheat bran and lOOcc of sea water was prepared.
• the medium was sterilized by autoclaving for 30 minutes at 15 lbs. steam pressure, cooled and was then inoculated with 5cc of an inoculum of Geotrichum marinum Thirumalachar sp nov. , prepared as described in Example 1.
• the flask was then placed on a reciprocating shaker at 24-28 degrees C for incubation/fermentaiton.
• the fungus grew very rapidly on the wheat bran, forming mycelia and conidia, demonstrating the growth-supporting properties of the bran for Geotrichum marinum.
• a fungi culture medium comprising 50 grams of particulate, powdered wheat bran and 50cc of sea water. The medium was heated by steaming for 20 minutes at 100 degrees C. The medium was cooled and then inoculatd with lOcc of a spore suspension of Geotrichum marinum Thirumalachar sp nov. The flask was well mixed and incubated for 10 days at 20-28 degrees C. The growth of the fungus was profuse, with the hyphae and coniidia forming a pellicle on the wheat bran particles, particulalrly the dried bran husks.
• a fungus broth grown in the medium described in Example 1 was prepared.
• the broth was seeded onto sea water in which petroleum oil (Shell X-100 Grade motor oil) had been added, by atomizing the broth and spraying it on the oil/water combination.
• the oil/water combination was thereafter sprinkled with a small quantity of particulate, powdered wheat bran.
• the oil and enmeshed fungus layer immediately formed a thick pellicle around the floating particulate wheat bran, the bran serving to hold the petroleum oil and fungus together.
• the oil was emulsified and broken down in 24 to 48 hours.
• the bran/fungus layer continued to demonstrate viability, however, since the fungus was provided with a growth substrate in the bran.
• the dry sprayable composition is prepared by growing Geotrichum on a powdered or particulate carrier, such as wheat bran or corn cob powder, in a Kohi rotary drum fermentor, followed by air-drying, or by admixing Geotrichum broth with a carrier, which combination is thereafter air- dried.
• the Geotrichum composition is dusted on the spill by known means; on open ocean, application with use of Shell Pipeline Corporation Oil Herder may be particularly effective, helping to prevent the spill's spread on the water, as well as breaking down the spill.

Landscapes

• Chemical & Material Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Engineering & Computer Science (AREA)
• Organic Chemistry (AREA)
• Health & Medical Sciences (AREA)
• Wood Science & Technology (AREA)
• Zoology (AREA)
• Genetics & Genomics (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Microbiology (AREA)
• Biotechnology (AREA)
• General Health & Medical Sciences (AREA)
• General Engineering & Computer Science (AREA)
• Biochemistry (AREA)
• Mycology (AREA)
• Environmental & Geological Engineering (AREA)
• Biomedical Technology (AREA)
• Medicinal Chemistry (AREA)
• Hydrology & Water Resources (AREA)
• Oil, Petroleum & Natural Gas (AREA)
• Water Supply & Treatment (AREA)
• General Chemical & Material Sciences (AREA)
• Molecular Biology (AREA)
• Materials Engineering (AREA)
• Botany (AREA)
• Tropical Medicine & Parasitology (AREA)
• Virology (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Public Health (AREA)
• Biodiversity & Conservation Biology (AREA)
• Micro-Organisms Or Cultivation Processes Thereof (AREA)

Applications Claiming Priority (1)

Publications (1)

Family

ID=22175579

Family Applications (1)

Country Status (2)

Families Citing this family (2)

Family Cites Families (7)

• 1983
  • 1983-11-14 WO PCT/US1983/001795 patent/WO1985002196A1/en not_active Ceased
  • 1983-11-14 EP EP84900168A patent/EP0162840A1/de not_active Withdrawn

Non-Patent Citations (1)

Also Published As

Similar Documents

Legal Events