ACTA BIOLOGICA CRACOVIENSIA

16 TH INTERNATIONAL SYMPOSIUM ON CAROTENOIDS
misleading and cannot be regarded as a monomolecular film. In
the literature, however, β-carotene was applied for Langmuir
monolayer formation [1,2]. In the above mentioned papers, the
film-forming abilities of β-carotene were inferred only from surface
pressure-area isotherm, and none of the other complementary
method was used to probe the structure of layers formed at
the interface. Since our BAM experiments indicated the domain
formation of β-carotene dropped from a wide variety of organic
solvents, we tried to improve its spreading behaviour by adding a
small amount of surface active substance (isopropanol or n-pentanol)
into the solution. This method of adding different additives
to the spreading solution to improve the stability and monolayer
structure has long been known [3] and was further applied by
many authors (see e.g. [4]). Unfortunately, all these experiments
failed in obtaining a homogeneous monolayer from β-carotene,
indicating lack of film-forming properties.
REFERENCES
SAKAKIBARA K, IFUKU S, TSUJII Y, KAMITAKAHARA H, TAKANO T, NAKATSUBO
F. 2006. Biomacromolecules, 7: 1960.
SHIBATA A, KIBA Y, AKATI N, FUKUZAWA K, TERADA H. 2001. Chem. Phys.
Lipids 113: 11-22.
DERVICHIAN DG. 1939. Nature 144: 629.
MINONES J, TRILLO S, GARCIA FERNANDEZ P, SANZ PEDRERO J. 1968.
Colloid Interafce Sci. 26: 518.
3.20.
Degrading of carotenoids by the DyP
peroxidase MsP2 from Marasmius
scorodonius
Kateryna Zelena1 , Nicole Lehnert1 , Ulrich Krings1 ,
Györgyi Horváth2 , Péter Molnár2 , Erika Turcsi3 ,
József Deli3 , Ralf G. Berger1 1 Leibniz University Hannover, Institut for Food Chemistry,
Callinstr. 5, D-30167 Hannover, Germany,
kateryna.zelena@lci.uni-hannover.de,
nicole.lehnert@lci.uni-hannover.de, krings@ lci.uni-hannover.de,
rg.berger@lci.uni-hannover.de
2 Department of Pharmacognosy, University of Pecs, Med. School,
Rokus u. 2, H-7624 Pecs, Hungary, Peter.Molnar@aok.pte.hu,
Gyorgyi.Horvath@aok.pte.hu
3 Department of Biochemistry and Medical Chemistry, University of
Pécs, Medical school, Szigeti út 12, H-7624 Pécs, Hungary,
Erika.Turcsi@aok.pte.hu, Jozsef.Deli@aok.pte.hu
From the perspective of human physiology and nutritional sciences,
the symmetric cleavage of carotenoids yielding retinoids
has attracted much attention. Retinoids act as regulating and signalling
molecules and as visual pigments. The excentric cleavage
of the carotenoids' polyene chain yields a wealth of apocarotenoids,
with the plant hormone abscisic acid being the most
prominent example. Many apocarotenoids, such as α- and
β-ionone, geraniol and β-damascenone act as potent flavour compounds
(Winterhalter and Rouseff, 2002). Apart from the generation
of retinoids, plant hormones or flavour compounds, there is
a strong interest of the detergent and food industries in
carotenoid degradation for bleaching fabrics, whey and other substrates.
Few data are available on microbial carotenoid degradation
pathways and the enzymes involved.
Basidiomycete fungi are a potent source of unique lignin
decomposing enzymes. In a screening more than 100 basidiomycetes
in an agar plate based assay, 37 strains also showed
a strong carotenoid degrading activity. Some β-carotene cleaving
enzyme activities were described previously (Zorn et al., 2003). In
particular, the DyP peroxidase MsP2 of the "garlic-mushroom"
Marasmius scorodonius was characterized on the biochemical
SESSION 3
and molecular level (Scheibner et al., 2008). In the current work,
the gene encoding MsP2 was cloned from the cDNA and functionally
expressed in different Escherichia coli strains using pCold (a
cold shock induced expression) system. Fusing the MsP2
sequence with the N-terminal His tag was effective for the purification
of the target protein. Biologically active peroxidase was
obtained as proven by β-carotene destaining. Aside from degrading
β-carotene, the stable MsP2 enzyme was able to effectively
decolorize lycopene, lutein and capsanthin as well.
This study, on the part of the Hungarian authors was supported by the
Grants OTKA K76176, K 83898 (Hungarian Scientific Research
Foundation) and PTE ÁOK KA-34039-35/2009.
REFERENCES
WINTERHALTER P, ROUSEFF RL. 2002. Carotenoid-derived aroma compounds:
an introduction. In: Winterhalter P, Rouseff RL (eds)
Carotenoid-derived aroma compounds. American Chemical
Society, Washington, p 1.
ZORN H, LANGHOFF S, SCHEIBNER M, BERGER RG. 2003. Cleavage of
beta,beta-carotene to flavor compounds by fungi. Applied
Microbiology and Biotechnology 62: 331-336.
SCHEIBNER M, HÜLSDAU B, ZELENA K, NIMTZ M, DE BOER L, BERGER RG,
ZORN H. 2008. Novel peroxidases of Marasmius scorodonius
degrade β-carotene. Applied Microbiology and Biotechnology 77:
1241-1250.
3.21.
Isolation and characterization of two novel
capsorubin like carotenoids in the red mamey
(Pouteria sapota)
Enrique Murillo1 , Yesury Mosquera1 , Tibor Kurtán2 ,
Gergely Gulyás-Fekete3 , József Deli3 1Department of Biochemistry, Faculty of Exact Natural Sciences
and Technology, University of Panama, Panama,
emurillo29@hotmail.com
2University of Debrecen, Faculty of Sciences, Department of
Organic Chemistry, Egyetem tér 1, 4032, Debrecen, Hungary,
kurtan.tibor@science.unideb.hu
3University of Pécs, Medical School, Department of Biochemistry
and Medical Chemistry, Szigeti út 12, 7624, Pécs, Hungary,
jozsef.deli@aok.pte.hu; gergely.gulyas@aok.pte.hu
The most common C 40 carotenoids have six-membered rings as
end-groups (β or ε rings), carotenoids with five-membered rings (κrings)
are rare. Capsorubin and capsanthin isolated from paprika
are the most well-known carotenoids with κ-end group. The κ-end
group of these carotenoids always bear a hydroxyl group. Mamey is
a fruit native in Panama, Central America and Mexico, much appreciated
for its pleasant taste and attractive red-orange colour of
flesh. Recently, we reported the complete isolation and characterization
of the sapotexanthin (β,κ-caroten-6'-one) – which bears nonhydroxylated
κ-end group- from red mamey pulp. In our poster we
present evidence showing the existence of two new and unknown
di-keto-di-κ carotenoids which are structurally related to capsorubin.
The carotenoids were isolated and characterized by combining
the techniques of open column chromatography, TLC and HPLC-
DAD, HPLC-MS and qualitative tests of reduction and acetylation.
Three carotenoids (A, B and C) were isolated from the pulp of red
mamey that have two keto groups conjugated to the polyene chain.
UV/Vis spectra of the three carotenoids and their reduction products
are the same as those of capsorubin which suggests that A, B
and C have the same chromophore as capsorubin. Carotenoid A
was identified as capsorubin (3,3'-dihydroxy-κ,κ-caroten-6,6'dione)
by co-chromatography (TLC and HPLC), MS and qualitative
tests. The carotenoid B has one hydroxyl group and its molar mass
is 584, while C has no hydroxyl group and its molar mass is 568.
60 ACTA BIOLOGICA CRACOVIENSIA Series Botanica