Download Research group: MariZyme

Faculty: Food Science and Nutrition and Physical Sciences
Research group: MariZyme, Science Institute, Department of Chemistry
Physical location: Dunhagi 5, 107 Reykjavik, Iceland
MariZyme
www.marizyme.is
Leader: Prof. Ágústa Guðmundsdóttir ([email protected])
General description: MariZyme is an internationally recognized research group in marine enzyme science and
technology1,2 focusing on basic research and practical applications of trypsin and other serine proteases from Atlantic cod
(Gadus morhua).
Senior scientists: Prof. Ágústa Guðmundsdóttir, Prof. Jón Bragi Bjarnason, Dr. Bjarki Stefánsson and Dr. Hilmar
Hilmarsson.
Graduate students: Guðrún Birna Jakobsdóttir and Karen Ósk Pétursdóttir. Recently graduated students (last 5 years):
Hólmfríður Sveinsdóttir (Ph.D) 2009, Helga M. Pálsdóttir (Ph.D) 2006, Una Björk Jóhannsdóttir (MS) 2009, Guðrún
Jónsdóttir (MS) 2006 and Linda Helgadóttir, (MS) 2005.
Funding: The Technology Development Fund , The AVS Research Fund, The University of Iceland Research
Fund, The University Research Fund of Eimskipafélag Íslands, The Icelandic Graduate Research Fund, The
Icelandic Science Fund, Nordic Industrial Fund, EU COST928 Research Program.
Research outputs: Cold adapted proteolytic enzymes, like Atlantic cod trypsin, are generally better suited for
enzymatic processes at lower temperatures than their mesophilic counterparts. Practical applications of cod trypsin and
chymotrypsin include their use for therapeutic purposes and in food processing. These enzymes have higher catalytic
efficiencies and sensitivity to inactivation by heat, low pH and autolysis than their mesophilic analogues 1-2. Native
proteins also appear to be more readily hydrolyzed by the cold-adapted fish proteases3. Several concepts for treatment of
infections and dermatological disorders originate from our research on cod trypsin (Penzyme®). Penzyme was tested for
its efficacy to reduce the adhesion of three strains of human pathogens to cell monolayers. The human pathogens tested
were Staphylococcus aureus (SA), its drug resistant derative Methicillin-resistant Sthaphylococcus aureus (MRSA) and
Streptcoccus pyogenes bacteria4. The bacteria were preincubated with 10 and 20 U/ml concentrations of Penzyme (Fig.1)
followed by incubation on cell monolayers. The results demonstrated that Penzyme reduces cell adhesion of all the
bacteria tested. Penzyme showed high efficacy against MRSA adhesion. However, the highest efficacy of Penzyme was
observed against Staphylococcus aureus (SA) (Fig. 1). The direct cell adhesion was lower for the group A streptococci
strains available in our cell line repertoire. New applications of cod trypsin have been developed and the results evaluated
for pharmaceutical or medical technology products. The genes for most of the serine proteases have been isolated and
expressed in yeast and bacteria5-8. The main purpose of the expression studies is to make derivatives of the enzymes with
increased stability and to produce the enzymes and its derivatives on a large-scale.
The trypsin profile in cod embryos and larvae was studied as this proteolytic enzyme has been shown to be a suitable
short-term indicator for the nutritional status of early cod larvae 9. Trypsin and chymotrypsin activities were shown to be
involved in the embryogenesis of Atlantic cod 10. Surprisingly, these proteolytic digestive enzymes seem to be produced
in relatively low amounts in the larvae at the beginning of first feeding (Fig 2). MALDI-TOF analysis of the proteome of
Atlantic cod larvae gave an important insight into changes in their global protein expression pattern in response to
exposure to fish protein hydrolysates and probiotic bacteria11-12. Treatment of cod larvae with probiotic bacteria resulted
in down-regulation of several proteins related to immune responses whereas the upregulated proteins can be linked to
growth and development11. Feeding of cod larvae with a protein hydrolysate affected the abundance of several proteins
known to be involved in energy metabolism indicating enhanced development in the treatment group compared to the
control bacteria12. Despite visible morphological and functional changes with age, the pattern of abundant proteins was
largely conserved in the cod larvae from days 6 to 24-post hatch9. However, many of the proteins were present in
different amounts in the two age groups but keratins showed the most pronounced developmental stage specific pattern.
Our previous research has also involved studies of the native form of serine proteases from Antarctic krill (Euphausia
superba) as well as production of the recombinant form of Euphauserase, an important proteolytic enzyme intended for
medical use13. Stabilized derivatives of the enzyme were created by genetic engineering and expressed in Pichia
pastoris14-15. Other research projects focused on the isolation, purification and characterization of a bacteriocin, termed
carnocin UI49, from Carnobacterium piscicola with the purpose of using the bacteriocin as a food preservative16.
Box 2. Trypsin and chymotrypsin
Box 1. Antibacterial activity of cod trypsin4.
activities in Atlantic cod eggs and larvae10.
SA adhesion reduction by Penzyme in Clone cells
4,5
4,216
Penzyme 20 U/ml
3,5
100
2,5
2
1,775
1,5
1
Milliunits per mg of protein
3,23
3
CFU / Cell
120
Control
4
80
60
40
0,601
0,5
20
I
II
III
IV
V
0
SA 213
SA MRSA
0
-1
0
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
Days past fertilization
Fig. 1. Left columns: CFU colony forming units of Staphylococcus
aureus (SA, blue) strain after Penzyme treatment compared to PBS
buffer control (red). Right columns: CFU colony forming units of
Streptococcus pyogenes (SP, blue) strain after Penzyme treatment
compared to PBS buffer control (red). About 7 fold reduction in
adhesion of the bacterial strains is seen against SA while about 2 fold
reduction is seen SP.
Fig. 2. Trypsin and chymotrypsin activities
in Atlantic cod eggs and yolk-sac larvae
(black diamonds: trypsin activity, white
diamonds: chymotrypsin activity).
Surprisingly, these proteolytic digestive
enzymes seem to be produced in relatively
low amounts in the larvae at the beginning of
first feeding.
Select publications (involving one or more group members): 1) Guðmundsdóttir and Pálsdóttir (2005) Marine
Biotechnol. 7, 77-88; 2) Gudmundsdóttir and Bjarnason (2007). In Bob Rastall (Ed.) Novel enzyme technology for food
applications pp. 205-214. Woodhead Publ. Ltd., Cambridge, UK; 3) Stefansson et al. (2010). Comp. Biochem. and
Physiol. Part B, 186-194; 4) Hilmarsson et al. J. Bact. (2010) (in prep.) . 5) Jónsdóttir, et al., (2004). Prot. Expr. and Purif.
33, 110-122; 6) Pálsdóttir and Gudmundsdóttir (2007) Prot. Expr. and Purif. 51, 243-252; 7) Pálsdóttir and
Gudmundsdóttir (2007) Comp. Biochem and Biophys. 146, 26-34; 8) Pálsdóttir and Gudmundsdóttir (2008) Food
Chemistry 111, 408-414; 9) Sveinsdóttir et al. (2008) Comp. Biochem. and Physiol. Genomics and Proteomics. Part D 3,
243-250; 10) Sveinsdóttir et al. (2006) Aquaculture 260, 307-314; 11) Sveinsdóttir et al. (2009) Comp. Biochem. and
Physiol. Part D4, 249–254; 12) Sveinsdóttir and Gudmundsdóttir (2010) Aquaculture Nutrition (in press). 13)
Gudmundsdóttir (2002). Biol. Chem. 383, 1125-1131; 14) Kristjánsdóttir and Gudmundsdóttir (2000) Eur. J. Biochem.
267, 2632-2639; 15) Benjamin et al., (2001). Eur. J. Biochem. 268, 127-131: 16) Stoffels et al. (1994) Microbiology,
140:1443-1450;
Collaboration: Dr. Charles S, Craik (UCSF, USA); Med. Dr. Aftab Jasir and Med. Dr. Claes Schalén (Lund University,
Sweden); Aake Larsson (Enzymatica, Lund, Sweden); Dr. Maria Luisa Tutino (University of Naples, Federico II, Italy);
Dr. Phillip Cash (Aberdeen University, UK); Dr. David L. Brautigan (University of Virginia, USA); Guðjón
Thorkelsson, Dr. Sjöfn Sigurgísladóttir, Sigurjón Arason, Dr. Hörður G. Kristinsson (Matís and University of Iceland);
Dr. John LaCava (The Rockefeller University, USA).