CS226614B1 - Kmen mikroorganizmu Trichoderma reesei MHC 22 CCF 1855 - Google Patents
Kmen mikroorganizmu Trichoderma reesei MHC 22 CCF 1855 Download PDFInfo
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- CS226614B1 CS226614B1 CS26981A CS26981A CS226614B1 CS 226614 B1 CS226614 B1 CS 226614B1 CS 26981 A CS26981 A CS 26981A CS 26981 A CS26981 A CS 26981A CS 226614 B1 CS226614 B1 CS 226614B1
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- Czechoslovakia
- Prior art keywords
- mhc
- trichoderma reesei
- ccf
- microorganism strain
- beta
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Description
POPIS VYNÁLEZU
<”> K AUTORSKÉMU OSVEDČENIU 226614 (11} (Bl)
(22) Přihlášené 14 01 81(21) (PV 269-81) (51) Int. CL3C 12 N 1/14 (40) Zveřejněné 26 08 83
ÚŘAD PRO VYNÁLEZY (45) Vydané 15 02 86
A OBJEVY (75)
Autor vynálezu
FARKAŠ VLADIMÍR ing. CSc., BRATISLAVA, LABUDOVÁ IVICA ing., KOŠICE,BAUER ŠTEFAN dř. ing. DrSe., BRATISLAVA, FERENCZY LAJOŠ dř., SZEGED (54) Kmen mikroorganizmu Trichoderma reesei MHC 22 CCF 1855 1
Vynález sa týká nového kmene mikroorganizmu Trichóderma reesei MHC 22 CCF 1855, ktorýprodukuje celulázový enzýmový komplex obsahujúci zvýšená hladinu beta-glukozidázy.
Imperfektná mikroskopická huba Trichoderma reesei QM 9414 produkuje při povrchovéj asubmerznej kultivácii na vhodnom substráte celulázový enzýmový komplex, schopný hydrolyzo-vať natívnu celulózu, to znamená komplexnú zmes polysacharidov typu celulózy a hemicelulózna mono- a nižšie oligosacharidy. Pretože hladina beta-glukozidázy v produkovanom celulá-zovom systéme je veTmi nízká (Sternberg D: Appl. Environ. Microbiol. 31, 648 /1976/) v hyd-rolyzáte celulózy převláda celobióza ako hlavný degradačný produkt (Ghose T. K., BisariaV. S. : Biotechnol. Bioeng. 21 , 131 /1979/)· Nakol’ko celobióza je len velmi slabo utilizova-ná kvasinkami používanými na áalšie zhodnocovanie celulózových hydrolyzétov je žiadúce, abyvšetka celobióza v hydrolyzáte bola konvertovaná najprv na glukózu. V praxi sa to doterazriešilo přidáváním exogénnej beta-glukozidázy z iného zdroja ku zmesi enzýmov z Trichodermareesei (Sternberg D., Vijaykumar P,, Reese E. T.: Can. J. Microbiol. 23, 139 /1977/), aleborozkladom celobiózy v kontínuálnom reaktore obsahujúcom imobilizovanú beta-glukozidázu(Sachdev R. K., Ghose T. K.: Proceedings of the Symposium on Bioconversion of CellulosicSubstances into Energy, Chemicals and Microbial Protein (I. K. Ghose, ed.) Indián Instituteof Technology, Delhi, /1977/.
Kmen Trichoderma reesei MHC 22 CCF 1855, ktorý je predmetom tohto vynálezu, odstraňujetieto nevýhody tým, že produkuje za rovnakých podmienok zhruba trojnásobné vyššiu hladinuenzýmu beta-glukozidázy, δο sa prejaví v zvýšení koncentrácie glukózy na úkor celobiózyv hydrolyzáte celulózy. Napr.<po 2 hod. hydrolýze je mólový poměr glukóza : celobióza 0,28s enzýmom Trichoderma reesei QM 9414, zatiaT ěo pri použití enzýmu z kmeňa MHC 22 je tentopoměr 4,3. Kmen MHC 22 CCF 1855 je uložený v zbierke Katedry botaniky University Karlovy 226614
Claims (2)
- 226614 2 v Praze
- 2, Benátská 2. Kmen MHC 22 sa získal ožarovaním konídií kmene Trichoderma reeseiQM 9414 získaného z American Culture Collection of Pungi (QM), University of Massachusetts,Amherst, Massachusetts, USA, UF-svetlom počas 5 minút. Hlavnou výhodou nového kmeňa je zvý-šená produkcia beta-glukozidézového komponente produkovaného celulolytického enzýmovéhosystému. Pri pěstovaní na 1 % mikrokryštalickej celulóze ako zdroji uhlíka dosahuje Speci-fická aktivita beta-glukozidázy 0,50 až 0,70 j/mg extracelulérneho proteinu, zatial’ čos materským kmeňom QM 9414 je Specifická aktivita beta-glukozidézy 0,20 až 0,25 j/mg extra-celulárneho proteinu. Jednotkou aktivity beta-glukozidázy sa rozumie množstvo enzýmu, ktoré katalyzuje hyd-rolýsu 1 /umólu p-nitrofenyl-beta-D-glukopyranozidu za 1 minutu za podmienok stanoveniapodTa (Berghem L. E. R, Pettersson L. G. : Eur. J, Biochem. 53, 55 /1973/). Morfologie:mycélium vláknité, husto vetvené, zriedkavo septované, hemisférický apex. Priemer hýf 0,6až 1,5/um. Konídionosiče kratšie a rozvětvené, fialídy zvyíajne jednotlivé, striedavé, ale-bo protistojace. Konídiá hojné, zeleno až zelenohnedo pigmentované, elipticko ovoidné,priemer 2,3 až 3,0 ^um, alebo niekedy elipsoidné, priemer 2,5 až 3,0 x 3,8 až. 4,5 /lm. Přisubmerznej kultivácii občasné chlamydospory, sférické, priemer do 8,0/um. Koniliácia indu-kovatel’ná svetlom. V p8de obsahujúcej dostatok věetkých živin produkuje žitý, vo vodě roz-pustný pigment. Mikroorganizmus rastie na: D-glukóze, D-galaktóze, D-manóze, laktóze, melibióze,D-fruktóze, D-xylóze, L-arabinóze, D-arabinóze, celobióze, eskulíne, trehalóze, škrobe,etanole, glycerole, D-erytritole, D-ribitole, D-manitole, D-sorbitole, duleitole, m-inozi-tolei1 acetáte, sukcináte, fumaráte, maláte, citráte, glukoronáte, celulóze, alebo rastiena: maltóze, rafinoze, L-sorboze, arbutíne, salicíne, inulíne, laktáte, tartráte a maloná-te, nerastie na: sacharóze, alfa-metyl-D-glukozide, melezitóze, L-ramnóze. Příklad Krneň Trichoderma reesei MHC 22 se kultivuje vo fermentori v živnom médiu nasledovnéhozlošenia (v gramoch na liter): močovina 0,3; síran amonný 1,4; dihydrogénfosforečnan dra-selný 0,2; heptahydrát síranu horečnatého 0,3; chlorid vápenatý 0,3; kvasníčný extrakt 0,1;mikrokryštalická celuldtea 10,0; Tween 80 2,0; peptón 1,0; síran železnatý 5.10”^; hydrátsíranu manganatého l,4.10“6; chlorid zinočnatý 1,7.10-6; chlorid kobalnatý 2.10“^. Teplota kultiváoie je 28 °C, prevzduáňovanie 0,15 až 0,30 v/v/min., rýchlosť otéčokmiešadla 300 až 1,500 ot./min v závislosti na konštrukcii fermentora. Ako inokulum sa po-užije baničková kultúra v logaritmickej fáze rastu vypěstovaná submerzne z konídií v rovna-kom médiu aké bolo použité vo fermentorovej kultivácii. Objem inokula je 10 % z objemu fer-mentačnej zmesi. Kultivácia prebieha počas 5 až 7 dní, pričom sa sleduje celková celulázo-vá aktivita (Mandels M., Antreotti R., Roche C.: Biotechnol. Bioeng. Symp. 6, 21 /1976/)a aktivita beta-glukozidázy v médiu. pH kultúry sa počas kultiváoie udržiava v rozmedzí4,0 až 4,5. Keň celulázová aktivita v médiu dosiahne maximélnej hodnoty (okolo 3 až 3,5jednotiek FP) ml médium sa scentrifuguje a číry supernatant sa použije ako zdroj celuloly-tických enzýmov. Gelulolytický enzýmový komplex z Trichoderma reesei MHC 22 sa dá použit pri sacharifi-kácii celulózy a lignocelulózových substrátov a je potřebné pri enzýmovej hydrolýze celu-lózy dosiahnuť maximélnej konverzie na glukózu. Vynález má použitie v potravinárstve, far-mácii, pivovarníctve a vinérstve.'· PREDMET VYNÁLEZU Kmen mikroorganizmu Trichoderma reesei MHC 22 GCF 1855, produkujúci celulolytickýenzýmový komplex, obsahujúci zvýšená hladinu beta-glukozidázového komponentu.
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| Application Number | Priority Date | Filing Date | Title |
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| CS26981A CS226614B1 (cs) | 1981-01-14 | 1981-01-14 | Kmen mikroorganizmu Trichoderma reesei MHC 22 CCF 1855 |
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| CS26981A CS226614B1 (cs) | 1981-01-14 | 1981-01-14 | Kmen mikroorganizmu Trichoderma reesei MHC 22 CCF 1855 |
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| CS226614B1 true CS226614B1 (cs) | 1984-04-16 |
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| CS26981A CS226614B1 (cs) | 1981-01-14 | 1981-01-14 | Kmen mikroorganizmu Trichoderma reesei MHC 22 CCF 1855 |
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1981
- 1981-01-14 CS CS26981A patent/CS226614B1/cs unknown
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