Možnosti produkce pigmentů kvasinkami
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Authors
Gonová, Dominika
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A
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Vysoké učení technické v Brně. Fakulta chemická
Abstract
Karotenoidy sú prirodzene sa vyskytujúce pigmenty syntetizované kvasinkami, baktériami, vláknitými hubami i rastlinami. V posledných rokoch vzrastá záujem o štúdium týchto pigmentov a ich mikrobiologickú produkciu najmä vďaka významným biologickým účinkom pripisovaným práve karotenoidom. Táto bakalárska práca je koncipovaná ako porovnávacia štúdia šiestich karotenogenných kvasiniek, a to dvoch kmeňov druhu Rhodotorula glutinis, Rhodotorula mucilaginosa, Sporobolomyces roseus, Sporobolomyces metaroseus a Sporobolomyces pararoseus. Ich kultivácia prebiehala na rôznych živných médiách, kde bol aplikovaný oxidačný stres v podobe peroxidu vodíka a tiež bol použitý odpadný materiál – vaječné cestoviny ako nutričný zdroj. Všetky študované kmene boli schopné využívať odpadný substrát ako zdroj živín, pričom u rodu Sporobolomyces boli na takomto médiu zaznamenané zvýšené produkcie sledovaných metabolitov. Pôsobením oxidačného stresu sa u väčšiny študovaných kvasiniek prejavili nadprodukcie karotenoidov i sprievodných lipidických látok. Kvasinka Sporobolomyces roseus CCY 19-6-4 sa pri kultivácii na cestovinovom médiu síce vyznačovala najvyššou produkciou karotenoidov (1923,78 µg/g -karoténu), ale na druhej strane sa u nej prejavil výrazný pokles biomasy. Ako potencionálne najlepší producent biomasy (9,85 g/l) i karotenoidov (780,24 µg/g -karoténu) sa ukázala kvasinka Rhodotorula glutinis CCY 20-2-26. Medzi prírodné pigmenty patrí aj pulcherimín syntetizovaný najmä kvasinkami, ale aj niektorými sporotvornými baktériami. V poslednej dobe sa intenzívne študuje jeho antagonistický efekt, ktorý vykazuje voči viacerým mikroorganizmom, čo sa javí ako veľmi významné z hľadiska biologickej kontroly. Druhá časť práce je venovaná práve štúdiu regulácie produkcie pulcherimínu a jeho antimikrobiálnej aktivite. Ako kvasinky produkujúce tento pigment boli použité kmene Metschnikowia pulcherrima 145, Metschnikowia pulcherrima 147, Mestchnikowia pulcherrima 149 a Metschnikowia andauensis 129, ktoré boli kultivované na rozličných živných médiách obsahujúcich rôzne koncentrácie iónov kovov a tiež peroxid vodíka ako exogénny stres. V rámci štúdia antagonistického efektu pulcherimínu boli ako testované mikroorganizmy využité grampozitívna baktéria Lactobacillus sakei, gramnegatívna baktéria Escherichia coli, ďalej kvasinka Candida glabrata a vláknitá pleseň Phanrochaete chrysosporium. Všetky kvasinky rodu Metschnikowia boli schopné produkcie pulcherimínu, a teda aj antagonistického efektu, iba v médiách obohatených o železité ióny, pričom intenzita sfarbenia bola priamo úmerná práve koncentrácii železa. Antimikrobiálna aktivita pulcherimínu sa neprejavila iba voči baktérií Escherichia coli.
Carotenoids are naturally occurring pigments synthesized by yeast, bacteria, filamentous fungi and plants. In recent years, the interest in the study of these pigments and their microbiological production is increasing mainly due to significant biological effects attributed to carotenoids. This work is conceived as comparative study of six carotenogenic yeasts: two strains of species Rhodotorula glutinis, Rhodotorula mucilaginosa, Sporobolomyces roseus, Sporobolomyces metaroseus a Sporobolomyces pararoseus. Their cultivation was carried out in various media where the oxidative stress was applied in the form of hydrogen peroxide and also waste material – egg pasta – was used as a nutrition source. All studied strains were able to use waste substrate as a source of nutrients, in case of the genus Sporobolomyces increasing production of metabolites was observed. Under the oxidative stress, the majority of the studied yeast showed overproduction of carotenoids and accompanying lipid substances. Sporobolomyces roseus CCY 19-6-4 was characterized by the highest production of carotenoids (1923.78 µg/g -carotene) at the egg pasta media, but on the other hand, it showed a significant decline in biomass. As potentially the best producer of biomass (9.85 g/l) and carotenoids (780.24 µg/g of -carotene), the yeast Rhodotorula glutinis CCY 20-2-26 was observed. Pulcherrimin, synthesized mainly by yeast but also by some spore-forming bacteria, also belongs among natural pigments. Recently, its antagonistic effect that was shown against several microorganisms has been intensively studied, as it seems to be very relevant in regard to biological control. The second part of the work deals with the regulation of pulcherrimin production and its antimicrobial activity. As the pigment-producing yeast strains were used Metschnikowia pulcherrima 145, Metschnikowia pulcherrima 147, Mestchnikowia pulcherrima 149, Metschnikowia andauensis 129, which were cultured on various media containing different concentrations of metal ions and hydrogen peroxide as exogenous stress. In the study of pulcherrimin antagonistic effect, gram-positive bacteria Lactobacillus sakei, gram-negative bacteria Escherichia coli, yeast Candida glabrata and filamentous fungi Phanrochaete chrysosporium were used as tested microorganisms. All studied yeasts were able to produce pulcherrimin, and therefore to show antagonist effect, only in media supplemented with iron, while the color intensity was proportional to the concentration of iron. Antimicrobial activity of pulcherrimin was not observed only against bacteria Escherichia coli.
Carotenoids are naturally occurring pigments synthesized by yeast, bacteria, filamentous fungi and plants. In recent years, the interest in the study of these pigments and their microbiological production is increasing mainly due to significant biological effects attributed to carotenoids. This work is conceived as comparative study of six carotenogenic yeasts: two strains of species Rhodotorula glutinis, Rhodotorula mucilaginosa, Sporobolomyces roseus, Sporobolomyces metaroseus a Sporobolomyces pararoseus. Their cultivation was carried out in various media where the oxidative stress was applied in the form of hydrogen peroxide and also waste material – egg pasta – was used as a nutrition source. All studied strains were able to use waste substrate as a source of nutrients, in case of the genus Sporobolomyces increasing production of metabolites was observed. Under the oxidative stress, the majority of the studied yeast showed overproduction of carotenoids and accompanying lipid substances. Sporobolomyces roseus CCY 19-6-4 was characterized by the highest production of carotenoids (1923.78 µg/g -carotene) at the egg pasta media, but on the other hand, it showed a significant decline in biomass. As potentially the best producer of biomass (9.85 g/l) and carotenoids (780.24 µg/g of -carotene), the yeast Rhodotorula glutinis CCY 20-2-26 was observed. Pulcherrimin, synthesized mainly by yeast but also by some spore-forming bacteria, also belongs among natural pigments. Recently, its antagonistic effect that was shown against several microorganisms has been intensively studied, as it seems to be very relevant in regard to biological control. The second part of the work deals with the regulation of pulcherrimin production and its antimicrobial activity. As the pigment-producing yeast strains were used Metschnikowia pulcherrima 145, Metschnikowia pulcherrima 147, Mestchnikowia pulcherrima 149, Metschnikowia andauensis 129, which were cultured on various media containing different concentrations of metal ions and hydrogen peroxide as exogenous stress. In the study of pulcherrimin antagonistic effect, gram-positive bacteria Lactobacillus sakei, gram-negative bacteria Escherichia coli, yeast Candida glabrata and filamentous fungi Phanrochaete chrysosporium were used as tested microorganisms. All studied yeasts were able to produce pulcherrimin, and therefore to show antagonist effect, only in media supplemented with iron, while the color intensity was proportional to the concentration of iron. Antimicrobial activity of pulcherrimin was not observed only against bacteria Escherichia coli.
Description
Citation
GONOVÁ, D. Možnosti produkce pigmentů kvasinkami [online]. Brno: Vysoké učení technické v Brně. Fakulta chemická. 2016.
Document type
Document version
Date of access to the full text
Language of document
cs
Study field
Biotechnologie
Comittee
prof. RNDr. Ivana Márová, CSc. (předseda)
doc. RNDr. Alena Španová, CSc. (místopředseda)
doc. Ing. Bohuslav Rittich, CSc. (člen)
doc. Ing. Pavel Diviš, Ph.D. (člen)
prof. Ing. Stanislav Obruča, Ph.D. (člen)
doc. RNDr. Renata Mikulíková, Ph.D. (člen)
Date of acceptance
2016-06-15
Defence
1. Studentka seznámila členy komise s náplní a cílem bakalářské práce.
2. Byly přečteny posudky na bakalářskou práci.
3. Studentka akceptovala všechny připomínky oponenta a na všechny otázky odpověděla v plné šíři.
Diskuse
prof. Márová:
Příprava media ?
Lze použít pulcherimin jako antimikrobiální činidlo ?
doc. Obruča:
Pulcherimin je extra nebo intracelulární ?
Úprava těstovin do media ?
Result of defence
práce byla úspěšně obhájena
Document licence
Standardní licenční smlouva - přístup k plnému textu bez omezení