Letno poročilo 2005

Laboratorij za biotehnologijo
Laboratory of Biotechnology
prepre~ijo njegovo vezavo na receptorje. Dolo~ili
smo 3D strukturo ve~ kot {tirim peptidom,
bodisi v okolju membranskih mimetikov ali LPS.
Analizirali smo tretjo generacijo peptidov, ki so
imeli bistveno izbolj{ano antimikrobno delovanje
in smo jih zasnovali na osnovi 3D strukture
peptidov v kompleksu z LPS in v micelah kot
membranskih mimetikih. Ugotavljali smo vpliv
acilacije peptida na spremembo aktivnosti in
interakcijo z membranami ter njegovo 3D
strukturo v zwitterionskih micelah. Na osnovi
rezultatov antimikrobnega delovanja in
nevtralizacije endotoksina skupaj s partnerji EU
projekta pripravljamo prijavo mednarodnega
patenta in nato objavo rezultatov.
Na raziskavah prionskih proteinov smo v
lanskem letu napredovali tudi na podro~ju
vezave spojin, ki se ve`ejo na amiloide, kjer smo
ugotovili tudi delovanje in na~in vezave spojin
naravnega izvora, kot je kurkumin, kar lahko
vodi k novim zdravilom in/ali diagnostiki. Na
tem podro~ju smo intenzivno vklju~eni v
mednarodno sodelovanje in smo za~eli z
izvajanjem EU projekta TSEUR.
Na podro~ju na~rtovanja primarnega metabolizma
smo v preteklem letu zaklju~ili s
koordinatorstvom evropskega projekta ANTICO,
ki ga je tri leta sofinancirala Evropska komisija
pod okriljem 5. Okvirnega projekta. Namen
projekta je bilo dokazati, da lahko s prenosom
dveh klju~nih genov, izoliranih iz glive Aspergillus
niger, ki kodirajo encime primarnega
metabolizma, v druge komercialne mikroorganizme
dvignemo produktivnost slednjih.
Prakti~no pri vseh recipientskih mikroorganizmih,
tako filamentoznih glivah, kvasovkah
in bakterijah, smo uspeli, vsaj v enem primeru,
dokazati pozitiven u~inek vne{enih genov na
pove~ano produkcijo. [e najbolj{e rezultate pa
smo dobili z vnosom skraj{anega gena za kratek
fragment 6-fosfofrukto-1-kinaze (PFK1). Pred
kratkim smo namre~ ugotovili, da pri glivi Aspergillus
niger pride do posttranslacijske
modifikacije nativnega encima, pri tem pa
nastane kraj{i fragment enicma s spremenjeno
kinetiko. Bistvena prednost novo nastalega
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pounds that directly bind to LPS and prevent
its binding to receptors. We determined 3D
structure of more than four peptides either in
the presence of membrane mimetics or LPS. We
analyzed the third generation of peptides showing
essentially improved antimicrobial activity
and were prepared on the basis of the 3D structure
of peptides in the complex with LPS and in
micelles as membrane mimetics. We also investigated
the effect of acylation of the peptide
on the activity and interaction with membranes
as well as its 3D structure in zwitterion
micelles. Based on the results of antimicrobial
activity and of endotoxin neutralization we are
preparing, together with our partners in the
EU project, an international patent application
and subsequent publication of the results.
In the research of prion proteins a progress was
made in the field of binding compounds that
bind to amyloids. We elucidated the action and
the mode of binding of natural compounds
such as curcumin, which could lead to discovery
of new drugs and/or diagnostic tools. In
this research field we are intensively involved in
the international cooperation and we just
started the work on EU project TSEUR.
In the field of designing primary metabolism in
2005 we brought to the end a coordination of
the EU project ANTICO which was financed by
the EC in the 5 FW program for three years.
The aim of the project was to prove that it was
possible to improve the productivity of commercial
microorganisms by transferring two key
genes from the fungus Aspergillus niger coding
for enzymes of primary metabolism. Almost
in all recipient microorganisms, filamentous
fungi, yeasts and bacteria, we succeeded to
prove, at least in one case, a positive effect of
the introduced genes on increased production.
The best results were obtained by the introduction
of a shortened gene for a short fragment
of 6-phosphofructo-1-kinase (PFK1). Recently,
we found that with the fungus A. niger a
posttranslational modification of the native
enzyme occurred, which resulted in a shortened