• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    A novel culture of the microorganism Streptomyces caligosus DS 14,486 and a novel proteolytic enzyme produced from cultivating the culture are disclosed. The enzyme has utility in the depilation of animal skins.
    公开号:SU1001862A3
    申请号:SU772519807
    申请日:1977-08-24
    公开日:1983-02-28
    发明作者:Беллок Андре；Флоран Жан；Люнель Жан；Палла Жан-Клод；Манси Дениз
    申请人:Рон-Пуленк Эндюстри (Фирма)；
    IPC主号:
    专利说明:

    It also interacts with proteins of the deep skin layers so that the skin obtained after such treatment has a surface damaged by grooves and grooves. The purpose of the invention is to obtain an enzyme with a more selective action, providing more efficient enzymatic epilation, without damaging the skin. The goal is achieved by the fact that, according to the method of obtaining a new proteolytic enzyme from Strepto myces caligosus DS ItjSS strain NRRL 8195, the indicated production is cultivated under aerobic conditions on a nutrient medium containing carbon, nitrogen, mineral salts and, if necessary, growth factors, at initial pH 8 and 23 ° C, followed by the separation of the cultural liquid and the release of the enzyme, whereby the cultivation is usually carried out with aeration of the medium at aeration rate of 0.3- 3 liters of air per 1 liter of nutrient medium per 1 min. The closest to Streptomyces caligosus DS is Streptomyces nobori toens is, since the latter produces melaine pigment on organic media, it develops a more or less thick dark (brown gr However, it doesn’t differ from the known strain, because Streptomesces noboritoensis does not form repetitive spirals of spore chains, it does not liquefy gelatin or does weakly, does not give a soluble pigment, or gives only a slightly soluble brownish pigment only on asparagine-glucose agar, produces a thick red-brown pigment on nutrient agar, then to AK Streptomyces caligosus DS lAi86 forms spore chains that regularly twist into tight helixes, liquefy gelatin, give a black soluble pigment on glucose-aspartic agar and give a grayish-brown soluble pigment on a nourishing agar; Streptomyces noboritoens is also forming on nitrate-containing synthetic agar, on Sakha624 rose an unpainted vegetative mycelium, while Streptomyces caligosus DS on this medium forms a thick, vegetative mycelium with jpOBaTO; to that. however, Streptomyces noboritoensis does not metabolize either rhamnose or sucrose, but it assimilates (and very limitedly) only arabinose, inositol and xylose, whereas Streptomyces caligosus DS absorbs all these carbon sources very well.  The streptomycin strain, which is the producer of a new proteolytic enzyme, has been extracted from a sample of land and has been assigned a DS number. A sample of this strain is stored in the Nortern National Research laboratory Museum US, (Agricultural Department in PEOPIA, W, USA), where it is registered under the number NRRL 8195 This organism, due to its new qualities, which do not allow it to be identified with the species already described, should be considered as a new species with the name Streptomyces caligosus DS I4it86.  The isolation is carried out according to a general method, which consists in suspending a small amount of earth in distilled sterile water, diluting the suspension to various concentrations, spreading a small amount of each dilution on the surface of a Petri dish containing nutritious agaro After incubating at 2b ° C for several days that allows microorganisms to grow, colonies that can be isolated for the study are selected and transplanted onto nutrient agar in order to get more complete Flax dilution of Streptomyces caligosus DS forms cylindrical spores of 0.8-1.0 microns / 0.6-0.8 microns in size “It forms a spore-like cluster; spore chains can have up to several dozen spores, they twist into tight more or less long spirals, forming mainly 2-3 rings, but quite often up to 6-8 or even up to 12 rings. By the form of its spore formation, Streptomyces caligosus DS is located in Section Spira.   according to the classification of Pridham.  Streptomyces caligosus DS develops well at 26 ° C, does not grow well and does not develop at all. It has a spore gray gray mycelium. The color of its ancient mycelium varies from more or less thick brown to dark brown depending on the nutrient medium.  On organic media, especially on special tyrosine agar - Waxman yeast extract (melanin-forming medium), it gives the substance a rather large amount with melanin color, which gives the medium a dark shade; It is capable of producing soluble blackish pigment in more or less significant quantities in many synthetic substances. In nutrient media maintained at 26 ° C, the enzyme has the following biological characteristics: Formation of chalk — Nina occurs.  Formation - and Tyrosinase Forms Dilution of Desire - Tina Occurs Acquisition of Cellulose - Vines Formation of nit- None on REEDs from the nitrifuge bottle containing nitrates, occurs on synthetic media.  Amidone is hydrolyzed. Dispersing Nutrient medium without coagulation on skimmed milk. The results of growing Streptomyces caligosus DS mtSS and various nutrient media are shown in Table. Cultivation was carried out for about 3 weeks with, besides those indicated.  These characteristics are observed on nutrient agars and broths commonly used to determine the morphological characteristics of Streptomyces strains, and they are grown on agar media on oblique agars.  The variety of growth media used is prepared according to known formulas, indicated, for example, in case 3, the composition of the medium is indicated by the letter WH by the number assigned to it.  The components of other nutrient media are also known. LJ Streptomyces caligosus DS 8 peculiarity () to assimilate various carbon or nitrogen sources to ensure its development and the degree of development observed on the host medium with glucose substitution by different, respectively tested carbon sources, or different, respectively The tested sources of nitrogen ();) are shown in table.  2, To obtain a new proteolytic enzyme, the cultivation of Streptomyces caligosus DS IlifSS strain NRRL 8195 can be carried out by aerobic growth on the surface or in depth, the latter method () is particularly convenient. . For this purpose, sowing and fermentation techniques are used, as well as various types of apparatus that are commonly used in the fermentation industry. The fermentation medium is known in principle 7 and contains sources of assimilable carbon and nitrogen, mineral elements and, if necessary, growth factors, all these elements can to be introduced in the form of individual substances or in the form of complex mixtures that are found in biological products of different passageways. As a source of digestible carbon, you can use carbohydrates such as glucose, sucrose, maltose, dextrins, amidone, or other carbonaceous substances, such as glycerin or some other organic acids (lactic and citric acids) o Some animal or vegetable oils (lard and soybean oil) can be beneficial to replace these different carbon sources or may be added to them. Suitable sources of digestible nitrogen are extremely diverse.  They can be very simple chemicals, for example, inorganic or organic ammonium salts, urea, certain amino-containing acids.  They can also be made with complex substances containing nitrogen in the protein form: casein, lactalbumin, Thloten and their hydrolyzates, soybean, peanut, fish meal, meat extracts, yeast, soluble distillers and soaked grains. Among the added elements, some may have buffering or neutralizing effects, such as alkaline or alkaline-earth phosphates71.  nmx of elements or calcium carbonate and magnesium o Others bring the ionic equilibrium necessary for the development of Streptomyces caligosus DS and for the processing of the enzyme, such as alkali and alkaline-earth chlorides and sulfates of o. Finally, some act more specifically as activators of metabolic reactions Streptomyces cnl igosus DS are salts of zinc, cobalt,; (iron, copper, and manganese measures of growth factors are substances such as vitamins, such as riboflavin, folic acid, pantothenic acid, o.  The pH of the fermentation medium at the start of the dilution should be in the range of, 8, preferably 6.2-7.0. The optimum fermentation temperature is in the range of 25-30 ° C, but satisfactory growth is obtained at 23-33 ° C.  Aeration of fermentation can vary widely.  However, it was found that aeration of 0.3–3 l of air per liter of broth per 1 min is the most optimal.  The maximum yield of enzyme 2 199 RP is obtained after 2-8 days of development, this time significantly depends on the nutrient therapy used.  The enzyme can be isolated from the fermentation wort in the following manner. The fermentation wort can be filtered, if necessary, in the presence of a filtering agent at a pH that is created in the medium at the end of the preparation process.  The filtrate is concentrated to 1/5 of the original volume, then the enzyme is precipitated by the addition of a solvent, for example acetone.  The crude product can be purified by fractional precipitation with solid inorganic salts or aqueous concentrated solutions, such as ammonium sulfate and / or enzyme solvents, such as acetone. The product can also be purified by dialysis through a membrane, preferably through a regenerated cellulose membrane. Dilution of Streptomyces caligosus NRPL 8195 in the described conditions allows to obtain the enzyme 2 199 RP proteinaceous substance, having the appearance of a powder of dark brown color, quite soluble in water, very poorly soluble in a concentrated aqueous solution of neutral salts 628 (for example, ammonium sulfate and in water-alcohol or water-acetone mixtures, practically insoluble with alcohols and anhydrous ketones; its mass is 27,000 and its isoelectric point lies at 3j7 Proteolytic activity of a new proteolytic enzyme appears on a large number of protein substrates such as casein, hemoglobin, fibrin (lysis of blood clots) J / l or milk (coagulation The activity on casein is determined in a manner analogous to Kunits "Peptides soluble in trichloroacetic acid, which are released during hydrolysis, are determined either spectrometrically at 280 nm (then expressed in Kunitz units), or colorimetrically. Activity is expressed in mg of tyrosine, which is formed per minute under the conditions of the determination.  Coagulating activity on milk is determined according to the method proposed by Berridge, and can be expressed in units of starter culture (UP), on-, the definition of which is this: one unit of starter is the amount of enzyme that can cause clotting of 10 cm of milk per 100 s.  Fibrinolytic activity is determined on clots of standard fibrin obtained by the action of thrombin on fibrinogen, and can be expressed in lytic units, the definition of which is the following: a solution of 100 lytic units per cm (100 UL / cM) is a solution that lyses the standard fibrin clot in 30 4INO But the following examples show how the invention can be used in practice.  Product activity is expressed in units of KUNIIZ (UK), as defined above.  This activity is expressed in K / cm when it comes to the solution, and UK / r when it comes to solid matter. Example 1 of a 170 l fermenter is placed: Peptone, g 1200 Yeast extract, g 600 Agar-agar, Water q , Srp, l105 pH of the medium 6 „55.  Sterilize by blrobiru for 40 min at 120 C.  After cooling, due to condensation of the vapor during sterilization by the volume of the broth, it becomes 115. l, it is supplemented to 120 l by adding 5 l of sterile aqueous solution containing 1200 g of monohydrate glucose.  pH 6.80.  Sow 205 cm stirred cultures of Streptomyces caligosus DS} kk86 strain NRRL 8195.  Culture develops during 23 hours with mixing and aeration with sterile air; then it becomes suitable for sowing for productive razdodki, productive wiring is carried out in a fermentation tank on the ZOO l, loaded with the following substances: Distilled solvent, kg Sucrose, kg Soybean oil, l 0.08 Sulfate | Arganese, kg 370 Water, l pH is adjusted to 7.3 by adding 850 cm of soda, then it is sterilized by bubbling steam at a temperature of 122 ° C for 40 I. After cooling due to condensation of vapors during sterilization, the broth volume becomes 400 l, pH 6.60.  Sow the CO of the grafted culture 1 into the 170 l fermentation tank.  The culture is developed under stirring with a rotating agitator (205 rpm) for 9 hours and with aeration of sterile air (20 then a culture pH of 7.30 and a wort volume of 400 liters are obtained.  The proteolytic activity of the wort at pH 7 and 37 C is equal to 3.6 IR / cm ".  Filter 11 l of the wort, prepared as described above, then wash the precipitate with k l of distilled water. The filtrate and washing are combined, then concentrated to 2 l under reduced pressure (5 mm Hg. Art. ), the temperature is not lower than 30 ° C. To the cooled concentrate, 2, 1 of acetone, preliminarily cooled to a mixture, are quickly added while stirring. Continue to mix for 2 minutes, then the insoluble part is separated by centrifugation at and 5000 (for 10 minutes o The insoluble part obtained is successively extracted with 5Q cm, then 150 cm of water at -4C for 2 hours, a section for hours, a section of each undissolved part of the centrifugation at 5,000% for 10 minutes 10 2 800 cm of an aqueous extract are obtained, pH of which is 7 and 7.5.  This extract is maintained at n- + C and g of crystalline ammonium sulfate is added with stirring, stirring. continue for another 15 minutes, after completing the addition, leave it at rest for 1 hour, then the insoluble part is separated by centrifugation at 10,000% for 10 minutes and dissolved in 500 cm by stirring for 1 hour in water, adjusting the pH to 6 -7.5. , and the resulting solution is dialyzed at + 4 ° C for 17 h in distilled water.   Get 920 cm dialyzed extract, which is quickly poured and mixed with 1.1 liters of acetone, cooled to -10 C.  The insoluble enzyme is separated by centrifugation at 5000 cj.  for 10 min at, then dried at low pressure at.  in the presence of a dehydrating agent (RuO, 9.5 g of enzyme are finally obtained, the enzymatic activity of which is as follows: Protease activity, IR / g Coagulating activity, (J P / g Fibrinolytic 920 activity, UL / r Example 2, ZbO wort, prepared according to example 1 is filtered on a pressed filter in the presence of 25 kg of a filtering aid. To 200 liters of the obtained filtrate add an AOOL of methanol cooled to -10 Co. After cooling the mixture to -10 ° C, the resulting precipitate is separated by cold centrifugation and dried n and at low pressure, was separated with 229 g of enzyme activity, and 290 K / g of Table.  3 collected the results obtained on various substrates with the proposed enzyme.  The enzyme activity is manifested in a wide pH range: the optimum pH for hydrolysis of casein is about 7.5, and the enzyme retains at least 60 of its maximum activity at pH 5-8.5.  The maximum enzyme activity occurs at a temperature of about 1110, and the activity drops very quickly when the temperature exceeds 50 ° C. The specificity of the enzyme in relation to hair and skin keratin and to the hair bulb glycoproteins and animal hair cannot be deduced from the activity obtained in normal the substrates presented in the examples given, it can only be detected by full-scale hair removal tests as described below. Enzyme epilation using the resulting enzyme is in that the skin being processed is brought into contact with the enzyme in the epilation tank, in which the percentage of water relative to the skin of the frame is preferably 20-25 ° C. The operation lasts for hours, the bath temperature is between 24-30 ° C and pH corresponds to the maximum proteolytic susceptibility of the enzyme, i.e. is between 7 and 8.5; The pH is kept constant during the operation by using buffer solutions, usually created by the addition of trisodium phosphate or borax.  In addition, we can assume that 0, weight, h, skins is that area. ratios in which the efficiency and cost of processing are the best. In practice, the method of processing the skins consists either in immersing them in a solution containing 10-30 g / l of an enzyme whose activity indicator is about 290 g, or in applying a spray from a gun with a solution containing 1-10 g / l of enzyme, whose index is about 290 U c / g (in the case of sheepskin) and in the case of sheepskin, and in the case of cattle hides, the enzyme activity used is optical when its concentration and its index are in the above elations x capacitance for epilation tion is any tank or drum, which are used in known manner to ensure that the removal of hair from the skin was complete, the enzyme must be accompanied by or preceded by mechanical pullout.  It can simply be the friction of one skin over another during mixing, supplemented if necessary by shrinking the hair during a short period of more intense mixing. This can also be done by passing the treated leather through a wool blending machine that allows you to get undamaged and unpaved wool The proposed method may be performed and / or follow a large number of additional operations. It is desirable to clean the hides before epilio It may consist of washing in water in a drum or a barge, mainly intended for washing the skin of salts, which it impregnates during preservation. Washing can be followed by degreasing with ordinary aromatic or chlorinated solvents or detergent water solutions. This operation removes a large number of organic impurities, such as fats. and inorganic substances that stain the skin and clog the hair sacs. Degreasing would be beneficial to accompany the new washing with water in such a way as to obtain a clean skin ( This may be possible, free from contamination and cleaning agents; After epilation, the skins are washed and the water is allowed to drain. The washing water contains intact wool and enzyme, which can be separated by known methods — decantation, filtration. The skins are then introduced into a clean lime ash pan, which hydrolyzes them.  This ashpit is used many times, because the skins that are placed there are practically clean and only slightly contaminate. It is noted that after treatment in the ashpit, the face layer of the coke does not deteriorate and is firmly attached to the dermis, while the skin epilated by alcalase enzymes , has corrupted facial layers, gelatinous when touched, with a torn dermis. During the epilating operation, the discharge of contaminating products is significantly reduced. This is especially noticeable in relation to sulfides and wool toryh now completely no wastewater, whereas they are present in abundance in the debris produced during epilation tion by dissolving fur skins after processing can osusches gvit all normal operations tanning
    production: deaeration, pickling, chrome tanning, splitting and trimming, re-tanning, emulsifying, drying, drying and finishing leather,
    The skins thus obtained have a very fine grain, high qualities in comparison with those obtained by the lime-sulphide method, to them there are no streaks and they have very good adhesion of the face surface, even on the elongated parts of the animal's sides.
    Comparison data on the epilation of cattle hides using the following three methods are given in te: epile using the proposed enzyme (A); epilci using a proteolytic enzyme from you, subttlfs (B) and epilci using a known method of dissolving a hair in a bath containing lime and sodium sulfide (c) o
    Positively
    D-Manitoll Positive L-Tyrosine
    Positive but slow
    Positively
     II
     II
    Positively
    Negative Positive ... D-Sorbi Inosito Salicin Indicators
    Type of wool:
    hair bags
    surface
    To the touch
    Flexibility
    Finishing by car
    Tension resistance
    kg / mm
    Elongation at break,%
    Deflection arrow in the formation of folds, mm
    Very clean Few clean Okrug Very smooth
    Slightly dense - Slightly dense on
    Good average
    1.5 66
    7.5 G11 1L .... L.J ... 1.:. - DL-Proline and - and L-Histidine, - Limited L-Tryptophan - and Substrate I Reactions Activity Casein Proteolysis Ik / g Milk Coagulation Y500 UP / r Fibriogen Lysis fibrin - 4920 Oi / mg of fibrin 1LO Reactant at pH 7.5 and at 37 ° C, the substrate concentration is 5 g / l. Reaction at pH 6.35 and when reconstituted with milk. Reaction pH 7.5 and at. .... .- / .L.i ... l ....... L.L ....... Continued table. 2 Betaine Table 3 - Table A Epilation method
    27 100186228
    invention formula Sources of information
     1. (Jnoco6 obtained by proteolyticism 1 "USSR author's certificate
    Who enzyme from Streptonyces caligo- (f 66002, class A 22 B 5/08,.
    SUS DS strain NRRL 8195, about t l-52 "Gracheva IM Enzyme technologists by the fact that the realization of drugs M, Food industry cultivation of the specified production, 1975, s, k7-,
    under aerobic nutritional conditions3. Waksman S, A The Actinomycede containing carbon sources, tes The Willfams and Wilkins Comzo, mineral salts and in the case of non-Jurap, Baltimore, 19b1, v.v J, 2-,
    obkhodimosti growth factor, at the beginning-C, Pridham T.G. et col. Hickey and
    nom pH 5.8-7.8 and a temperature of 23-33C g Tresners Agar. - Antibiotics Annuc followed by separating culture al., 1956-950.
    fluid and enzyme release. Waksman S.A. The Actinomyce2. The method according to p. 1, about tl and h and y- 15tes Sennets Agar, 19b1, v. 2, p.
    ti and with the fact that cultivation6. H.D. Tresner et F. Yanda conducted under aeration of the medium with sc.-G. of Bacteriology, 1958, 76 239-2.
    the growth rate of aeration of 0.3–3 l of air per liter, J, UK Patent No. 1377105,
    nutrient medium in 1 min; cl, C 12T 13/10, published „197
    taken into account in the examination
    权利要求:
    Claims (2)
    [1]
    Claim
    1. A method of obtaining a proteolytic enzyme from Streptornyces caligosus DS 14486 strain NRRL 8195, t l and _ 5, characterized in that they carry out the cultivation of the specified producer in aerobic conditions on a nutrient medium containing sources of carbon, nitrogen, mineral salts and, if necessary growth factor, at an initial pH of 5.8-7.8 and a temperature of 23-33 ° C g, followed by separation of the culture fluid and isolation of the enzyme.
    [2]
    2. The method according to π. 1, with the fact that cultivation is carried out by aerating the medium with an aeration rate of 0.3 “3 liters of air per 1 liter of nutrient medium for 1 min;
    862 28
    类似技术:
    公开号 | 公开日 | 专利标题
    Arvidson et al.1971|Influence of cultivation conditions on the production of extracellular proteins by Staphylococcus aureus
    McCARTY1952|THE LYSIS OF GROUP A HEMOLYTIC STREPTOCOCCI BY EXTRACELLULAR ENZYMES OF STREPTOMYCES ALBUS: I. Production and Fractionation of the Lytic Enzymes
    US3930954A|1976-01-06|Neutral protease useful for animal tissue and cell culture
    CS211400B2|1982-02-26|Method of microbiological enzymatic production of d-aminoacids
    Foissy1974|Examination of Brevibacterium linens by an electrophoretic zymogram technique
    SU1001862A3|1983-02-28|Process for producing proteolytic enzyme from streptomyces galigous ds 14486 strain no.rrl 8195
    US3649455A|1972-03-14|Production of lipase
    JPH05252949A|1993-10-05|Alkaline protease, its production, its usage and microorganism producing the same protease
    Jönsson et al.1964|Protease production by Aspergillus fumigatus
    Grzywnowicz et al.1989|Comparative characterization of proteolytic enzymes from Trichophyton gallinae and Trichophyton verrucosum
    DE2614114B2|1978-08-31|Process for the production of creatinine amidohydrolase
    CN103391997B|2016-08-03|Enzyme from boolean's Fred ear mould | and preparation method thereof
    US3840433A|1974-10-08|Dehairing of leather
    CA1166986A|1984-05-08|Process for producing the enzyme cholesterase and for hydrolysing cholesterol esters of fatty acids byusing the enzyme itself
    US7186546B2|2007-03-06|Process for the preparation of alkaline protease
    US4064010A|1977-12-20|Purification of uricase
    US3616233A|1971-10-26|Removing esterase from microbial rennin
    US3330738A|1967-07-11|Production of enzyme complex by cytophaga | ncib 9497
    Naganuma et al.1993|Characterization of agarolytic Microscilla isolates and their extracellular agarases
    CN105199983A|2015-12-30|Bacillus amyloliquefaciens and application of protease produced by same in tanning depilation
    NAKAGAWA et al.1984|Terferol, an inhibitor of cyclic adenosine 3', 5'-monophosphate phosphodiesterase I. Isolation and characterization
    JP2882652B2|1999-04-12|Alkaline protease and its producing microorganism
    Femi-Ola et al.2015|Isolation and characterization of feather degrading bacteria from poultry soil
    JPH06261691A|1994-09-20|Production of low allergenic milk protein
    JP3483604B2|2004-01-06|Method for producing amino acids
    同族专利:
    公开号 | 公开日
    JPS5326381A|1978-03-11|
    AU513565B2|1980-12-11|
    DK145827C|1983-08-29|
    NL7709336A|1978-02-28|
    DK373477A|1978-02-25|
    GB1560448A|1980-02-06|
    DE2737711A1|1978-03-09|
    IT1085381B|1985-05-28|
    ES461817A1|1978-11-01|
    FR2362862B1|1979-05-25|
    US4288556A|1981-09-08|
    AU2806677A|1979-02-22|
    FR2362862A1|1978-03-24|
    DK145827B|1983-03-14|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    BE786788A|1971-07-27|1973-01-26|Rhone Poulenc Sa|NEW THROMBOLYTIC ENZYME AND ITS PREPARATION BY FERMENTATION OF A STREPTOMYCES|
    DE2404789C3|1974-02-01|1979-02-15|Roehm Gmbh, 6100 Darmstadt|Process for the production of ready-to-tan pelts from animal hides and skins|
    US4066503A|1976-03-12|1978-01-03|Alexandr Pavlovich Bashkovich|Enzyme of hydrolytic action, enzymatic preparation, method for preparing both and their uses|US4416640A|1981-03-16|1983-11-22|Dennis J. Romano|Life ring|
    EP0455818A4|1989-11-24|1993-05-12|Banyu Pharmaceutical Co., Ltd.|Physiologically active substance be-16627|
    US5609668A|1994-09-30|1997-03-11|Gill; Paul E.|Spill clean-up process|
    US5525139A|1993-07-12|1996-06-11|Gill; Paul E.|Process for bioremediation of soils|
    JP2001164300A|1999-12-06|2001-06-19|Daiwa Kasei Kk|Enzymic depilatory in hide tanning and method for enzymic dehairing|
    US6777219B2|2002-03-13|2004-08-17|Council Of Scientific And Industrial Research|Process for the preparation of alkaline protease|
    US8739734B2|2011-12-02|2014-06-03|Paul E. Gill|Spill clean up material and pet litter, and methods of making and using same|
    法律状态:
    优先权:
    申请号 | 申请日 | 专利标题
    FR7625631A|FR2362862B1|1976-08-24|1976-08-24|
    [返回顶部]