CS244963B1 - Streptomyces aureofaciens ccm 3766 strain producing chlorotetracycline and resistant against phagus infection - Google Patents
Streptomyces aureofaciens ccm 3766 strain producing chlorotetracycline and resistant against phagus infection Download PDFInfo
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- CS244963B1 CS244963B1 CS844327A CS432784A CS244963B1 CS 244963 B1 CS244963 B1 CS 244963B1 CS 844327 A CS844327 A CS 844327A CS 432784 A CS432784 A CS 432784A CS 244963 B1 CS244963 B1 CS 244963B1
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- strain
- infection
- streptomyces aureofaciens
- lysis
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- 241000186984 Kitasatospora aureofaciens Species 0.000 title claims abstract description 18
- 208000015181 infectious disease Diseases 0.000 title claims abstract description 11
- CYDMQBQPVICBEU-UHFFFAOYSA-N chlorotetracycline Natural products C1=CC(Cl)=C2C(O)(C)C3CC4C(N(C)C)C(O)=C(C(N)=O)C(=O)C4(O)C(O)=C3C(=O)C2=C1O CYDMQBQPVICBEU-UHFFFAOYSA-N 0.000 title claims description 8
- CYDMQBQPVICBEU-XRNKAMNCSA-N chlortetracycline Chemical compound C1=CC(Cl)=C2[C@](O)(C)[C@H]3C[C@H]4[C@H](N(C)C)C(O)=C(C(N)=O)C(=O)[C@@]4(O)C(O)=C3C(=O)C2=C1O CYDMQBQPVICBEU-XRNKAMNCSA-N 0.000 title claims description 8
- 238000004519 manufacturing process Methods 0.000 abstract description 19
- 238000000855 fermentation Methods 0.000 abstract description 15
- 230000004151 fermentation Effects 0.000 abstract description 15
- 230000009089 cytolysis Effects 0.000 abstract description 8
- 230000003115 biocidal effect Effects 0.000 abstract description 6
- 238000000034 method Methods 0.000 abstract description 5
- 239000002028 Biomass Substances 0.000 abstract description 2
- 239000003242 anti bacterial agent Substances 0.000 abstract description 2
- 230000002411 adverse Effects 0.000 abstract 1
- 229940088710 antibiotic agent Drugs 0.000 abstract 1
- 230000008033 biological extinction Effects 0.000 abstract 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract 1
- 239000002689 soil Substances 0.000 description 6
- 239000002054 inoculum Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 230000028070 sporulation Effects 0.000 description 3
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 240000008042 Zea mays Species 0.000 description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 235000005822 corn Nutrition 0.000 description 2
- 238000009776 industrial production Methods 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 241000197194 Bulla Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 235000019764 Soybean Meal Nutrition 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 208000002352 blister Diseases 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 150000002540 isothiocyanates Chemical class 0.000 description 1
- 238000005065 mining Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000019612 pigmentation Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 235000002639 sodium chloride Nutrition 0.000 description 1
- 238000009331 sowing Methods 0.000 description 1
- 239000004455 soybean meal Substances 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 125000000185 sucrose group Chemical group 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Infekcia kultúry kmeňa Streptomyces aureofaciens produkujúceho chlórtetracyklín aktinofágom má za následek lýzu citlivej populácie kmeňa. Lýza nadobúda rozsah od čiastočného rozpadu biomasy mycélia až po jeho úplný zánik. Miere poškodenia produkčného mycélia odpovedajú poruchy fermentačného procesu a nižšia produkcie antibiotika v porovnaní s neinfikovanou kultúrou; pri úplnej lýze je kultúra bez produkcie antibiotika. Kmeň Streptomyces aureofaciens CCM 3 766 je odolný voči aktinofágovi a pri infekcii kultúry tohto kmeňa nedochádza k lýze mycélia a nasledujúcim nežiadúcim změnám vo fermentačnom proceseInfection of Streptomyces aureofaciens strain chlorothetracycline-producing actinophage results in lysis sensitive strain population. Lysis ranges from mycelium biomass partial decay to its complete extinction. Production damage rate mycelia respond to fermentation disorders process and lower antibiotic production compared to uninfected culture; in complete lysis, the culture is non-production antibiotics. Streptomyces aureofaciens CCM strain 3,766 is resistant to actinophage and infection the culture of this strain does not occur mycelial lysis and subsequent adverse events changes in the fermentation process
Description
HOFBAUER HENRICH ing., BANSKÁ BYSTRICA; KRUMPHANZL VLADIMÍR prof. ing. DrSc., PRAHA; KÚDELA OTAKAR RNDr. CSC., BRATISLAVA; ŽIGOVÁ E VELÍ NA ing., BANSKÁ BYSTRICA,· BÁRTA MIROSLAV RNDr., PRAHA; MARTINCEK LUBOMÍR ing., BANSKÁ BYSTRICA/ VYSKOČIL PAVEL ing., PRAHA; BULLA JÁN ing., BANSKÁ BYSTRICA,'ETTLER PETR ing. CSc., PRAHA (54) Kmen Streptomyces aureofaciens CCM 3 766 produkujůci chlórtetracyklín a odolný voči fágovej infekcii 1 2HOFBAUER HENRICH ing., BANSKÁ BYSTRICA; KRUMPHANZL VLADIMÍR prof. ing. DrSc., Prague; KUELA OTAKAR RNDr. CSC., BRATISLAVA; ŽIGOVÁ E MAKES IN ing., BANSKÁ BYSTRICA, · BÁRTA MIROSLAV RNDr., PRAGUE; MARTINCEK LUBOMÍR ing., BANSKÁ BYSTRICA / VYSKOČIL PAVEL ing., PRAGUE; BULLA JAN ing., BANSKÁ BYSTRICA, 'ETTLER PETR ing. CSc., PRAGUE (54) Streptomyces aureofaciens strain CCM 3,766 producing chlorotetracycline and resistant to phage infection 1 2
Infekcia kultúry kmeňa Streptomyces aureofaciens produkujúceho chlórtetracyklín aktinofágom má za následek lýzu citlivej populácie kmeňa. Lýza nadobúda rozsah od čiastočného rozpadu biomasy mycélia až po jeho· úplný zánik. Miere poškodenia produkčného mycélia odpovedajú poruchy fermentačného procesu a nižšia produkcie antibiotika v porovnaní s neinfikovanou kultúrou; pri úplnej lýze je kultura bez produkcie antibiotika.Infection of a culture of Streptomyces aureofaciens producing chlorotetracycline with actinophage results in lysis of a sensitive strain of the strain. Lysis extends from the partial decay of mycelium biomass to its complete disappearance. The rates of damage to the production mycelium correspond to disturbances in the fermentation process and lower antibiotic production compared to the uninfected culture; on complete lysis, the culture is free of antibiotic production.
Kmen Streptomyces aureofaciens CCM 3 766 je odolný voči aktinofágovi a pri infekcii kultúry tohto kmeňa nedochádza k lýze mycélia a nasledujúcim nežiadúcim změnám vo fermentačnom procese.The strain Streptomyces aureofaciens CCM 3,766 is resistant to actinophage, and infection of the culture of this strain does not result in lysis of the mycelium and subsequent undesirable changes in the fermentation process.
Vynález sa týká kmeňa mikroorganizmu Streptomyces aureofaciens CCM 3 766 (intéřrté označenie 5/211} produkujúceho antibiotikum chlórtetracyklín a· vyznačujúceho sa odolnosťou voči fágovej infekcii.The present invention relates to a strain of the microorganism Streptomyces aureofaciens CCM 3,766 (intere designation 5/211) producing the antibiotic chlorotetracycline and characterized by resistance to phage infection.
V priebehu priemyselnej výroby chlórtetracyklínu sa jeho produkcia neustále zvyšuje. Vyššia produkcia sa dosahuje buď rozšiřováním výrbbných fermentačných prevádzok alebo zvyšováním výtažnosti fermentácie. Druhý sposob nevyžaduje, na rozdiel od prvého, významné investičně náklady. Je založený na lepšej produkčnej schopnosti ikmeňa používaného pre biosyntézu, alebo· na lepších fermentačných podmienkach vyplývajúcich z dokonalejšieho riadenia fermentácie. Pre mikrobiálnu priemyselnú výrobu představuje velké nebezpečie možnost fágovej infekcie, ktorej dosledkom je zastavenie výroby na vetmi dlhú dobu. Závédenie mikrobiálnych kmeňov, odolných voči spósobeniu tága, umožňuje výrobu antibiotika i v přítomnosti fagových partikulí, ktoré sa běžnými spósobmi sterilizácie nedajú z výrobného zariadenia odstrániť.During the industrial production of chlorotetracycline, its production is steadily increasing. Higher production is achieved either by extending the mining fermentation plants or by increasing the fermentation yield. The second approach, unlike the first, does not require significant investment costs. It is based on the better production capacity of the stem used for biosynthesis, or on the better fermentation conditions resulting from improved fermentation control. For microbial industrial production, there is a great risk of phage infection, the consequence of which is to stop production for a long time. The irradiation of cue resistant microbial strains allows the production of an antibiotic even in the presence of phage particles, which cannot be removed from the production equipment by conventional sterilization methods.
Kmeň Streptomyces aureofaciens CCM 3 766 bol získaný z kmeňa Streptomyces aureofaciens CM 3 493 nevoděním spontánnej rezistencie .k fágovi, t. j. pěstováním kmeňa. Streptomyces aureofaciens CCM 3 493 v přítomnosti fága a nasledujúcou selekciou odolného mikroorganizmu z fermentačnej půdy obsahujúcej prevažne zlyzované mycélium. Spory získaného kmeňa bolí vysiate na Petriho misku so sporulačnou půdou obsahujúcou fága a, na Petriho misku so sporulačnou půdou bez fága. Zhodný nárast a sporulácia dokázali odolnost kmeňa voči fágovej infekcii.Streptomyces aureofaciens strain CCM 3,766 was obtained from Streptomyces aureofaciens strain CM 3,493 by not conferring spontaneous phage resistance, i. j. by growing the strain. Streptomyces aureofaciens CCM 3,493 in the presence of phage and subsequent selection of a resistant microorganism from a fermentation broth containing predominantly lysed mycelium. The spores of the obtained strain were sown on a phage-containing Petri dish with phage-containing sporula soil and, on a phage-free petri dish. Consistent growth and sporulation demonstrated strain resistance to phage infection.
Týmto sposobom bol získaný kmeň, na ktorý je uplatňovaný patentový nárok.In this way, the strain to which the claim is claimed was obtained.
Hlavným znakom kmeňa Streptomyces aureofaciens CCM je jeho odolnost voči fágovej infekcii. Tvar kolonií pri monospórovom rozseve, ich pigmentácia a farba spór sú rovnaké ako u východzieho kmeňa Streptomyces aureofaciens CCM 3 493. Rast kmeňa Streptomyces aureofaciens CCM 3 766 je na stuzených i kvapalných půdách pomatší ako u východzieho kmeňa. Tak isto sporulácia nastupuje pozdejšie a kmeň Streptomyces aureofaciens CCM 3 766 je citlivější na dodržanie podmienok sporulácie. Produkčná schopnost kmeňa Streptomyces aureofaciens CCM 3 766 je o 10 až 15 °/o nižšia v porovnaní s kmeňom Streptomyces aureofaciens CCM 3 493.The main feature of the strain Streptomyces aureofaciens CCM is its resistance to phage infection. The shape of the colonies in monospore sowing, their pigmentation and the color of the spores are the same as that of the parent strain of Streptomyces aureofaciens CCM 3,493. Also, sporulation occurs later and the Streptomyces aureofaciens CCM 3,766 strain is more sensitive to compliance with sporulation conditions. The production capacity of Streptomyces aureofaciens CCM 3,766 is 10-15% lower than that of Streptomyces aureofaciens CCM 3,493.
Příklad 1Example 1
Podlá doterajšieho spůsobu fermentácie kmeňa Streptomyces aureofaciens CCM 3 493 sa vegetativně inokulum připravuje submerzne v 500 ml varných baňkách so 60 ml inokulačnej půdy, ktorá obsahuje základné živiny, a to zdroj uhlíka sacharózu, zdroje dusíka kukuřičný extrakt, síran amonný a sójovú muku a soli. Půda na přípravu vegetatívneho inoikula sa naočkuje sporami asi z jednej třetiny kultúry na šikmom agare a inkubuje sa na rotačnom trepacom stroji 22 hodin. Takto připraveným vegetatívnym inokulom naočkujú sa fermentačné baňky objemu 500 ml so 60 ml produkčnej živnej půdy. Zdrojom uhlíka je sacharóza, zdrojom dusíka je kukuřičný extrakt, sójová mučka a amónna sol’. Půda ďalej obsahuje uhličitan vápenatý a dalšie soli. Na začiatku fermentácie sa do půdy přidává sterilný roztok izotiokyanátu. Příprava vegetatívneho inokula a vlastná fermentácia prebiehajú na rotačnom trepacom stroji pri 230 o/min“1, výstřednosti 25 mm a teplote 29 °C. Fermentácia trvá 4 dni, kedy sa dosahuje maximálna produkcia antibiotika. Vyššie uvedeným postupom sa dosahujú produkcie 5 500 až 6 000 (Wg/ml s priemernou hodnotou 5 737 fig chlórtetracyklínu/ml půdy odpovedajúcou indexu 100 °/o.According to the previous method of fermentation of the strain Streptomyces aureofaciens CCM 3,493, a vegetatively inoculum is prepared submerged in 500 ml beakers with 60 ml seed broth containing basic nutrients, namely a carbon source of sucrose, a nitrogen source of corn extract, ammonium sulfate and soybean and salts. The soil for preparing the vegetative inoculum is inoculated with spores from about one third of the slant agar culture and incubated on a rotary shaker for 22 hours. The vegetative inoculum thus prepared is inoculated with 500 ml fermentation flasks with 60 ml of production broth. The carbon source is sucrose, the nitrogen source is corn extract, soybean meal and ammonium salt. The soil further contains calcium carbonate and other salts. At the start of the fermentation, a sterile isothiocyanate solution is added to the soil. Preparation of vegetative inoculum and the fermentation is run net on a rotary shaker at 230 o / min "1, the eccentricity of 25 mm and a temperature of 29 ° C. Fermentation lasts 4 days when maximum antibiotic production is achieved. The above procedure produces 5,500-6,000 (Wg / ml with an average value of 5,737 figs of chlorotetracycline / ml of soil corresponding to an index of 100%).
Příklad 2Example 2
Podta doterajšieho spůsobu fermentácie kmeňa Streptomyces aureofaciens CCM 3 493 v baňkách v příklade 1 sú vegetativně inokulum a z něho očkovaná produkčná půda infikované aktinofágom. Fág lyžuje buňky mycélia citlivej populácie produkčného kmeňa. Za týchto podmienok sú dosahované produkcie 0 až 4 000 ^ug/ml s priemernou produkciou produkujúcich Ikultúr 3 626 ,ug chlórtetracyklínu/ml půdy odpovedajúcou indexu 63 °/o.According to the previous method of fermentation of the strain Streptomyces aureofaciens CCM 3,493 in the flasks of Example 1, the vegetatively inoculum and the actinophage-infected production broth are seeded from it. The phage lyses the mycelial cells of a susceptible population of the production strain. Under these conditions, production of 0-4000 µg / ml is achieved with an average production producing 3,626 µg chlorotetracycline / ml soil corresponding to an index of 63 ° / o.
Příklad 3Example 3
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Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS844327A CS244963B1 (en) | 1984-06-07 | 1984-06-07 | Streptomyces aureofaciens ccm 3766 strain producing chlorotetracycline and resistant against phagus infection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS844327A CS244963B1 (en) | 1984-06-07 | 1984-06-07 | Streptomyces aureofaciens ccm 3766 strain producing chlorotetracycline and resistant against phagus infection |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CS432784A1 CS432784A1 (en) | 1985-11-13 |
| CS244963B1 true CS244963B1 (en) | 1986-08-14 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CS844327A CS244963B1 (en) | 1984-06-07 | 1984-06-07 | Streptomyces aureofaciens ccm 3766 strain producing chlorotetracycline and resistant against phagus infection |
Country Status (1)
| Country | Link |
|---|---|
| CS (1) | CS244963B1 (en) |
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1984
- 1984-06-07 CS CS844327A patent/CS244963B1/en unknown
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| Publication number | Publication date |
|---|---|
| CS432784A1 (en) | 1985-11-13 |
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