CS277576B6 - Strain of acremonium chrysogenum ccf-2035 micro-organism - Google Patents
Strain of acremonium chrysogenum ccf-2035 micro-organism Download PDFInfo
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- CS277576B6 CS277576B6 CS886340A CS634088A CS277576B6 CS 277576 B6 CS277576 B6 CS 277576B6 CS 886340 A CS886340 A CS 886340A CS 634088 A CS634088 A CS 634088A CS 277576 B6 CS277576 B6 CS 277576B6
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- ccf
- acremonium chrysogenum
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- 241000228431 Acremonium chrysogenum Species 0.000 title claims abstract description 10
- 244000005700 microbiome Species 0.000 title description 4
- HOKIDJSKDBPKTQ-GLXFQSAKSA-N cephalosporin C Chemical compound S1CC(COC(=O)C)=C(C(O)=O)N2C(=O)[C@@H](NC(=O)CCC[C@@H](N)C(O)=O)[C@@H]12 HOKIDJSKDBPKTQ-GLXFQSAKSA-N 0.000 claims abstract description 10
- 238000004519 manufacturing process Methods 0.000 abstract description 8
- 238000009395 breeding Methods 0.000 abstract description 3
- 230000001488 breeding effect Effects 0.000 abstract description 3
- 229920001817 Agar Polymers 0.000 description 5
- 239000008272 agar Substances 0.000 description 5
- 239000002054 inoculum Substances 0.000 description 4
- 229930006000 Sucrose Natural products 0.000 description 3
- 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 3
- 240000008042 Zea mays Species 0.000 description 3
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 3
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 3
- 235000005822 corn Nutrition 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000002689 soil Substances 0.000 description 3
- 239000005720 sucrose Substances 0.000 description 3
- 239000004470 DL Methionine Substances 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- FFEARJCKVFRZRR-UHFFFAOYSA-N methionine Chemical compound CSCCC(N)C(O)=O FFEARJCKVFRZRR-UHFFFAOYSA-N 0.000 description 2
- 229930182817 methionine Natural products 0.000 description 2
- 235000006109 methionine Nutrition 0.000 description 2
- 230000000644 propagated effect Effects 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 101710143086 Paralytic peptide 2 Proteins 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- QYDYPVFESGNLHU-UHFFFAOYSA-N elaidic acid methyl ester Natural products CCCCCCCCC=CCCCCCCCC(=O)OC QYDYPVFESGNLHU-UHFFFAOYSA-N 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 230000002070 germicidal effect Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- QYDYPVFESGNLHU-KHPPLWFESA-N methyl oleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC QYDYPVFESGNLHU-KHPPLWFESA-N 0.000 description 1
- 229940073769 methyl oleate Drugs 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000219 mutagenic Toxicity 0.000 description 1
- 230000003505 mutagenic effect Effects 0.000 description 1
- 238000005580 one pot reaction Methods 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 239000013587 production medium Substances 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 235000012424 soybean oil Nutrition 0.000 description 1
- 239000003549 soybean oil Substances 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Vynález sa týká kmeňa mikroorganizmu Acremonium chrysogenum CCF-2035 produkujúceho cefalosporín C. Tento kmeň bol získaný z kmeňa Acremonium chrysogenum CCM F-750 šlachtením zameraným na zvýšenie produkcie cefalosporínu C.The invention relates to a strain of Acremonium chrysogenum CCF-2035 producing cephalosporin C. This strain was obtained from strain Acremonium chrysogenum CCM F-750 breeding aimed at increasing production cephalosporin C
Description
Vynález sa týká kmeňa mikroorganizmu Acremonium chrysogenum CCF-2035 produkujúceho cefalosporín C /dalej CFS-C/. Tento kmeň bol získaný z kmeňa Acremonium chrysogenum CCM F-750, šlachtením zameraným na zvýšenie produkcie CFS-C.The invention relates to a cephalosporin C-producing strain of Acremonium chrysogenum CCF-2035 (hereinafter CFS-C). This strain was obtained from Acremonium chrysogenum strain CCM F-750, by breeding aimed at increasing CFS-C production.
Východzí kmeň dosahoval produkčnú aktivitu v baničkách 10 200 ug/ml a‘ v tančíku 22 400 ug/ml.The starting strain achieved a production activity in the flasks of 10,200 ug / ml and in the tank of 22,400 ug / ml.
Popisovaný, kmeň mikroorganizmu Acremonium chrysogenum CCF-2035 bol získaný tak, že materský kmeň Acremonium chrysogenum CCM-740, bol vystavený mutagénnemu šlachteniu UV-svetlom.The described strain of the microorganism Acremonium chrysogenum CCF-2035 was obtained by subjecting the parent strain of Acremonium chrysogenum CCM-740 to mutagenic UV light.
Šlachtenie bolo robené tak, že vysporulovaná kultúra rodičovského kmeňa na šikmom agare SP-S o zložení: peptón 1 %, maltóza 3,8 %, tekutá rafináda /ťažká šťava/ o obsahu 60 %, sacharózy 4 %, agar 2,4 % sa zmyla 10 ml sterilněj vody, přefiltrovala cez vatovú zátku umiestnenú na pipete, za účelom. oddelenia hýf a artrospór od konídií. Z takejto suspenzie konídií, vhodné nariedenej sa očkovali Petriho misky s pódou SP-S.Breeding was performed by sporulating the culture of the parent strain on SP-S slant agar composed of: peptone 1%, maltose 3.8%, liquid raffinate / heavy juice / containing 60%, sucrose 4%, agar 2.4% with washed with 10 ml of sterile water, filtered through a cotton plug placed on a pipette, in order to. separation of hyphae and arthrospores from conidia. Petri dishes with SP-S pod were inoculated from such a conidia suspension, suitably diluted.
Mutagenný zásah sa robil pomocou UV světla germicídnou lampou Philips TUV 30 W s rozdělením spektrálnéj energie - vlnová dížka 253,7 nm až 93,5 %, ostatně vlnové dížky 6,5 %. Vzdialenosť misky od zdroja žiarenia bola 30 cm. Po ožiarení sa misky okamžité preniesli do inkubátora na teplotu 28 °C, kde sporulovali 11 dní. Vysporulované kolonie boli za aseptických podmienok izolované a přenesené na šikmý agar pódy SP-S. Ako pomnožovacia submerzná póda sa použila póda IP-V o zložení: sacharóza 1,5 %, C2H7O2N 0,5 %, DL-metionín 0,1 %, KH2PO4 0,027 %, kukuřičný výluh /100 %-ný/ 0,8 %, metyloleát 0,08 %. Po 72 hodinovéj kultivácii na rotačnom trepacom stroji 4 Hz pri teplote 27 °C, sa získalo vegetativně inokulum /1.vegetatívny stupeň/.Mutagenesis was performed using UV light with a Philips TUV 30 W germicidal lamp with a spectral energy distribution - wavelength of 253.7 nm to 93.5%, and wavelengths of 6.5%. The distance of the dish from the radiation source was 30 cm. After irradiation, the plates were immediately transferred to an incubator at 28 ° C, where they sporulated for 11 days. The recommended colonies were isolated under aseptic conditions and transferred to SP-S slant agar. IP-V soil was used as the propagation submerged medium with the following composition: sucrose 1.5%, C 2 H 7 O 2 N 0.5%, DL-methionine 0.1%, KH 2 PO 4 0.027%, corn leachate / 100 % / 0.8%, methyl oleate 0.08%. After culturing for 72 hours on a 4 Hz rotary shaker at 27 ° C, a vegetative inoculum (1st vegetative stage) was obtained.
ml tohoto inokula sa očkovali 500 ml varné banky s obsahom 60 ml pódy IP-V a kultivovali pri teplote 27 °C na rotačnom trepacom stroji 48 hodin /II. vegetatívny stupeň/.ml of this inoculum were inoculated into 500 ml boiling flasks containing 60 ml of IP-V pod and cultured at 27 ° C on a rotary shaker for 48 hours / II. vegetative degree.
Z nařasteného inokula sa 4 ml očkovali 500 ml varné banky plněné 40 ml produkčněj pódy PP-II/as o zložení: škrob kukuřičný 3,2 %, dextrin 6 %, sacharóza 1,5 %, glukóza 0,5 %, CaCO3 1,1 %, /NH4/2SO4 1,5 %, KH2PO4 0,5 %, MgSO4 0,3 %, kukuřičný výluh 100 %-ný 8,7 %, sojový olej 1,9 %. DL-metionín do pódy nepřidáváme, pretože nároky produkčného mikroorganizmu na túto aminokyselinu sú dostatočne zabezpečené inými zložkami pódy napr. kukuřičným výluhom.From the grown inoculum, 4 ml were inoculated into a 500 ml boiling flask filled with 40 ml of PP-II / as production medium consisting of: corn starch 3.2%, dextrin 6%, sucrose 1.5%, glucose 0.5%, CaCO 3 1 , 1%, / NH 4 / 2SO 4 1.5%, KH 2 PO 4 0.5%, MgSO 4 0.3%, corn extract 100% 8.7%, soybean oil 1.9%. We do not add DL-methionine to the soil, because the demands of the production microorganism on this amino acid are sufficiently ensured by other components of the soil, eg corn steep liquor.
Příklad 1Example 1
Konídie materského kmeňa Acremonium chrysogenum CCM F-750 asepticky zmyté z Endovej skúmavky 10 ml sterilnej vody a nariedené lOOOkrát sa s 0,5 ml naočkovali na Petriho misky s pódou SP-S. Misky se vystavili 1 minútovému pósobeniu UV světla v aseptickom boxe. Po ožiarení sa misky okamžité preniesli do inkubátora a po 11 dňoch sa 55 samostatných kolonii preočkovalo na šikmé agary s pódou SP-S. Po narastení za 11 dní sa takto získaní izo láty testovali najprv jednobaničkovým testom na produkciu. 5 najlepších izolátov z tohto súboru sa podrobilo druhému stupňu výběru a to v 3 baničkovom teste. Z nich sa získal izolát, ktorý mal o cca 29 % vyššiu produkční! aktivitu, ako východzí rodičovský kmen.Conidia of the parent strain Acremonium chrysogenum CCM F-750 aseptically washed from the End tube with 10 ml of sterile water and diluted 1000-fold were inoculated with 0.5 ml onto SP-S-coated petri dishes. The plates were exposed to UV light for 1 minute in an aseptic box. After irradiation, the plates were immediately transferred to an incubator, and after 11 days, 55 separate colonies were inoculated onto SP-S slant agar slant. After growing for 11 days, the isolates thus obtained were first tested in a one-pot production test. The 5 best isolates from this set were subjected to the second stage of selection in a 3-pot test. An isolate was obtained from them, which had about 29% higher production! activity as the initial parent strain.
Tento kmen bol zaslaný do zbierky mikroorganizmov na Katedru botaniky nižších rastlín, Prírodovedeckej fakulty UK, Praha 2, Benátská 2, kde mu bolo přidělené číslo CCF-2035.This strain was sent to the collection of microorganisms to the Department of Botany of Lower Plants, Faculty of Science, Charles University, Prague 2, Benátská 2, where it was assigned the number CCF-2035.
Příklad 2Example 2
Produkčný kmen Acremonium chrysogenum CCF-2035 bol z lyofilizovanej konzervy postupné pomnožený na šikmý agar SP-S v Roux-flaši. Kmen tvoří kolonie krémovej farby s vrásčitým povrchom, v střede s kráterom róznej velkosti.The production strain of Acremonium chrysogenum CCF-2035 was gradually propagated from a lyophilized can onto SP-S slant agar in a Roux bottle. The stem forms cream-colored colonies with a wrinkled surface, with a crater of various sizes in the middle.
Konídie z Roux-flaši boli v dvojstupňovom inokulačnom postupe pomnožené do vegetatívnej formy. S 1500 ml narasteného inokula bol naočkovaný laboratórny tančík Chemap 0030. Po 136 hodinách kultivácie sa dosiahla produkčná aktivita 35 800 ug/ml cefalosporínu C, vypuštěný objem činil 26 000 ml.The conidia from the Roux bottle were propagated into a vegetative form in a two-step inoculation procedure. The Chemap 0030 laboratory tank was inoculated with 1500 ml of grown inoculum. After 136 hours of cultivation, a production activity of 35,800 ug / ml of cephalosporin C was reached, the volume discharged being 26,000 ml.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS886340A CS277576B6 (en) | 1988-09-26 | 1988-09-26 | Strain of acremonium chrysogenum ccf-2035 micro-organism |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS886340A CS277576B6 (en) | 1988-09-26 | 1988-09-26 | Strain of acremonium chrysogenum ccf-2035 micro-organism |
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| Publication Number | Publication Date |
|---|---|
| CS634088A3 CS634088A3 (en) | 1992-11-18 |
| CS277576B6 true CS277576B6 (en) | 1993-03-17 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CS886340A CS277576B6 (en) | 1988-09-26 | 1988-09-26 | Strain of acremonium chrysogenum ccf-2035 micro-organism |
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| CS (1) | CS277576B6 (en) |
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1988
- 1988-09-26 CS CS886340A patent/CS277576B6/en unknown
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| Publication number | Publication date |
|---|---|
| CS634088A3 (en) | 1992-11-18 |
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