Protein production

World-class competence in the development of protein production systems in microbes, plant cells and plants

VTT offers world-class competence in the development of protein production systems in microbes, plant cells and plants. We provide custom recombinant protein production in our Expression service. We improve protein production hosts and carry out research to understand the molecular background and physiology of protein production. Our customers benefit by getting proteins produced efficiently or by advancing their own protein production technology. Partner with VTT to fulfil your protein production needs.

The major global enzyme companies Genencor and AB Enzymes, have been our long-term research partners. In a protein expression commission for a client, we produced a recombinant enzyme at a level of 28 g/l.

More than 30 years of experience in protein production technology in different host systems

VTT is a leading research institute in the development of the filamentous fungus Trichoderma reesei as a production host. Our work in T. reesei has included native enzyme and recombinant protein production, the development of molecular and systems biology tools, the enhancement of productivity by strain engineering and the systems-wide investigation of the host physiology and optimisation of the fermentation process. We express recombinant proteins and improve their productivity also in plants and plant cells, yeasts and bacteria. We develop efficient high throughput screening procedures in different protein production hosts for enzyme discovery and engineering.

Our strong competence and state of the art technology

Our most important resources are our skilful and experienced personnel, excellent gene technology and systems biology tools and proprietary technology for protein expression and recovery and production system development. We can also provide technology transfer of protein production systems and technology to our customers. Our protein production research is well connected with VTT’s fermentation technology up to the pilot scale and with our enzyme development, protein engineering and metabolic modelling competences. We have worked mainly with industrial enzyme, chemical and therapeutic companies.

Our protein production expertise can be also applied to liquid biofuel and platform chemicals production in biorefineries and in the food, feed and pulp and paper industries.

​VTT Expression Service - From gene to protein

VTT Expression Service can help researchers and companies in production of recombinant proteins. VTT has more than 30 years of experience in protein production technology research in different hosts systems such as:

  • The bacterium Escherichia coli
  • The yeasts Saccharomyces cerevisiae, Pichia pastoris and Kluyveromyces lactis
  • The filamentous fungus Trichoderma reesei
  • Baculovirus/insect cells
  • Plants and plant cell cultures


This knowledge of many expression hosts enables the best case-specific choice for each protein. The expression vector and host strain are selected from our versatile collection, and purification tags can be added to expression constructs when required. We have the facilities and know-how to cultivate these production hosts in bioreactors up to pilot scale (1.2 m2), and expertise on downstream processing and protein purification.

The Expression Service offers a customer-oriented solution to help researchers and companies to successfully produce their recombinant proteins. We can provide the customer with complete tailor-made service for producing a protein of interest, even including cloning of the target gene. Alternatively, we can help the customer with any individual step along this route from gene isolation to purification and analysis of the final product.

The Expression Service offers flexible co-operation with its partners along the entire gene-to-product axis, or within a selected segment of the project. In that case the suitable parts of protein production can be chosen to complement the partner's own expertise.

 

 

 

The Expression Service offers flexible co-operation with its partners along the entire gene-to-product axis, or within a selected segment of the project. In that case the suitable parts of protein production can be chosen to complement the partner's own expertise.

2015

Enabling Low Cost Biopharmaceuticals: A Systematic Approach to Delete Proteases from a Well-Known Protein Production Host Trichoderma reesei. Landowski CP, Huuskonen A, Wahl R, Westerholm-Parvinen A, Kanerva A, Hänninen AL, Salovuori N, Penttilä M, Natunen J, Ostermeier C, Helk B, Saarinen J, Saloheimo M.
PLoS One. 2015 Aug 26;10(8):e0134723. http://www.ncbi.nlm.nih.gov/pubmed/26309247

Swollenin from Trichoderma reesei exhibits hydrolytic activity against cellulosic substrates with features of both endoglucanases and cellobiohydrolases. Andberg, M., Penttilä, M. and Saloheimo, M., Bioresource Technol. 2015, 181:105-113. http://www.ncbi.nlm.nih.gov/pubmed/25643956

Protein body formation in leaves of Nicotiana benthamiana: a concentration-dependent mechanism influenced by the presence of fusion tags. Saberianfar, R., Joensuu, J.J., Conley, A. and Menassa, R. Plant Biotechnol. J. 2015, in press. http://www.ncbi.nlm.nih.gov/pubmed/25640969


2014

Cloning and characterisation of a novel acidic cutinase from Sirococcus conigenus. Nyyssölä, A., Pihlajaniemi, V., Häkkinen, M., Kontkanen, H., Saloheimo, M. and Nakari-Setälä, T. Appl. Microbiol. Biotechnol. 2014, 98: 3639-3650. http://www.ncbi.nlm.nih.gov/pubmed/24121867

Screening of candidate regulators for cellulase and hemicellulase production in Trichoderma reesei and identification of a factor essential for cellulase production. Häkkinen, M., Valkonen, M., Westerholm-Parvinen, A., Aro, N., Arvas, M., Vitikainen, M., Penttilä, M., Saloheimo M. and Pakula, T.. Biotechnology for Biofuels 2015, 7:14. http://www.ncbi.nlm.nih.gov/pubmed/24472375

Comparison of intracellular and secretion-based strategies for production of human α-galactosidase A in the filamentous fungus Trichoderma reesei. Smith, W., Jäntti, J., Oja, M. and Saloheimo, M. BMC Biotechnol. 2014, 14: 91. http://www.ncbi.nlm.nih.gov/pubmed/25344685

Intracellular pH responses in the industrially important fungus Trichoderma reesei. Valkonen, M., Penttilä, M. and Bencina, M.. Fung. Genet. Biol. 2014, 70: 83-90. http://www.ncbi.nlm.nih.gov/pubmed/25046860
Scale-up of hydrophobin-assisted recombinant protein production in tobacco BY-2 suspension cells. Reuter, L., Bailey, M., Joensuu, J., Ritala, A.. Plant Biotechnol. J. 2014, 12: 402-410. http://www.ncbi.nlm.nih.gov/pubmed/24341724

Influence of elastin-like polypeptide and hydrophobin on recombinant hemagglutinin accumulations in transgenic tobacco plants. Phan, H., Hause, B., Hause, G., Arcalis, E., Stoger, E., Maresch, D., Altmann, F., Joensuu, J., and Conrad, U. PLoS ONE 2014, 9:e99347. http://www.ncbi.nlm.nih.gov/pubmed/24914995

Plant-based solutions for veterinary immunotherapeutics and prophylactics. Kolotilin, I., Topp, E., Cox, E., Devriendt, B., Conrad, U., Joensuu, J., Stäger, E., Warzecha, H.,McAllister, T., Potter, A., McLean, M., Hall, C. and Menassa, R..Veterinary Research 2014, 45:117. http://www.ncbi.nlm.nih.gov/pubmed/25559098

Protein production – ER quality control and secretion stress responses in Trichoderma reesei. Saloheimo, M., Pakula, T., Aro, N. and Joensuu, J.. In: Gupta, V.J. et al. (Eds.) Biology and Biotechnology of Trichoderma. Elsevier 2014, Waltham, MA, USA.

2013

Directing enzymatic cross-linking activity to the air-water interface by a fusion protein approach. Paananen, A., Ercili-Cura, D., Saloheimo, M., Lantto, R. and Linder, M..
Soft matter 2013, 9: 1612.

Intracellular protein production in Trichoderma reesei (Hypocrea jecorina) with hydrophobin fusion technology. Mustalahti, E., Saloheimo, M. and Joensuu, J.. New Biotechnol. 2013, 30: 262-268. http://www.ncbi.nlm.nih.gov/pubmed/21971507

Sturucture-function relationships in hydrophobins: probing the role of charged side chains. Lienemann M, Gandier J.-A., Joensuu J.J., Iwanaga A., Takatsuji Y., Haruyama T., Master E., Tenkanen M. and Linder M.B.. Applied and Environmental Microbiology 2013, 79:5533-5538. http://www.ncbi.nlm.nih.gov/pubmed/23835172

Unconventional microbial systems for the cost-efficient production of high-quality protein therapeutics. Corchero, J., Gasser, B., Resina, D. Smith, W., Parrilli, E., Vázquez, F., Abasolo, I., Giuliani, M., Jäntti, J., Ferrer, P., Saloheimo, M., Mattanovich, D., Schwartz, S., Tutino, L. and Villaverde, A.. Biotechnology Advances 2013, 31: 140-153. http://www.ncbi.nlm.nih.gov/pubmed/22985698

Experimental and bioinformatic investigation of the proteolytic degradation of the C-terminal domain of a fungal tyrosinase. Faccio, G., Arvas, M., Thöny-Meyer, L. and Saloheimo, M.. J. Inorg. Biochem. 2013, 121: 37–45. http://www.ncbi.nlm.nih.gov/pubmed/23333757

Swollenin aids in the amorphogenesis step during the enzymatic hydrolysis of biomass. Gourlay, K., Hu, J., Arantes, V., Andberg, M., Saloheimo, M., Penttilä, M. and Saddler, J.. Bioresource Technol. 2013, 142: 498-503. http://www.ncbi.nlm.nih.gov/pubmed/23759433

2012

The cargo and the transport system: secreted proteins and protein secretion in Trichoderma reesei (Hypocrea jecorina). Saloheimo, M. and Pakula, T.M.. Microbiology 2012, 158: 46 – 57. http://www.ncbi.nlm.nih.gov/pubmed/22053009

Bacterial tyrosinases and their applications. Faccio, G., Kruus, K., Saloheimo, M. and Thöny-Meyer, L.. Process Biochemistry 2012, 47: 1749 - 1760.

Re-annotation of the CAZy genes of Trichoderma reesei and transcription in the presence of lignocellulosic substrates. Häkkinen, M., Arvas, M., Oja, M., Aro, N., Penttilä, M., Saloheimo M. and Pakula, T.M.. Microbial Cell Factories 2012, 11:134. http://www.ncbi.nlm.nih.gov/pubmed/23035824

Xylanase XYN IV from Trichoderma reesei showing exo- and endo-xylanase activity. Biely, P., Tenkanen, M., Vršanská, M., Puchart, V., Wong, Penttilä, M., Saloheimo, M. and Siika-aho, M.. FEBS J. 2012, 280: 285-301. http://www.ncbi.nlm.nih.gov/pubmed/23167779

Bioseparation of recombinant proteins from plant extract with hydrophobin fusion technology. Joensuu, J.J., Conley, A., Linder, M. and Menassa, R.. In: Methods in Molecular Biology. Volume 824: Recombinant Gene Expression: Reviews and Protocols. 3rd ed. Springer 2012, 527-534. 

Transient expression using agroinfiltration and its applications in molecular farming. Menassa, R., Ahmad, A. and Joensuu, J.J.. In: Molecular Farming in Plants: Recent Advances and Future Prospects. Wang, A. & Ma, S. (eds). Springer2012, 183-198.

2011

Correlation of gene expression and protein production rate - a system wide study. Arvas M., Pakula T., Smit B., Rautio J., Koivistoinen H., Jouhten P., Lindfors E., Wiebe M., Penttilä M., Saloheimo M.. BMC Genomics. 2011, 12:616. http://www.ncbi.nlm.nih.gov/pubmed/22185473

Sulfhydryl oxidases: sources, properties, production and applications. Faccio G., Nivala O., Kruus K, Buchert J., Saloheimo M.. Appl. Microbiol. Biotechnol. 2011, 91:957-966. http://www.ncbi.nlm.nih.gov/pubmed/21732243

The effects of disruption of phosphoglucose isomerase gene on carbon utilisation and cellulase production in Trichoderma reesei Rut-C30. Limón M.C., Pakula T., Saloheimo M., Penttilä M..
Microb. Cell Fact. 2011 10:40. http://www.ncbi.nlm.nih.gov/pubmed/21609467

Production and characterisation of AoSOX2 from Aspergillus oryzae, a novel flavin-dependent sulfhydryl oxidase with good pH and temperature stability. Faccio G., Kruus K., Buchert J., Saloheimo M.. Appl. Microbiol. Biotechnol. 2011, 90:941-949. http://www.ncbi.nlm.nih.gov/pubmed/21327412

Influence of growth temperature on the production of antibody Fab fragments in different microbes: a host comparative analysis. Dragosits M., Frascotti G., Bernard-Granger L., Vázquez F., Giuliani M., Baumann K., Rodríguez-Carmona E., Tokkanen J., Parrilli E., Wiebe M.G., Kunert R., Maurer M., Gasser B., Sauer M., Branduardi P., Pakula T., Saloheimo M., Penttilä M., Ferrer P., Luisa Tutino M., Villaverde A., Porro D., Mattanovich D.. Biotechnol. Prog. 2011, 27:38-46. http://www.ncbi.nlm.nih.gov/pubmed/21312353

Transformation system for Hypocrea jecorina (Trichoderma reesei) that favors homologous integration and employs reusable bidirectionally selectable markers. Steiger M.G., Vitikainen M., Uskonen P., Brunner K., Adam G., Pakula T., Penttilä M., Saloheimo M., Mach R.L., Mach-Aigner A.R.. Appl. Environ. Microbiol. 2011, 77:114-21. http://www.ncbi.nlm.nih.gov/pubmed/21075888

Protein body-inducing fusions for high-level production and purification of recombinant proteins in plants. Conley A.J., Joensuu J.J., Richman A., Menassa R.. Plant Biotechnol. J. 2011, 9:419-433.
http://www.ncbi.nlm.nih.gov/pubmed/21338467

2010

Exploring laccase-like multicopper oxidase genes from the ascomycete Trichoderma reesei: a functional, phylogenetic and evolutionary study. Levasseur A., Saloheimo M., Navarro D., Andberg M., Pontarotti P., Kruus K., Record E.. BMC Biochem. 2010, 11:32. http://www.ncbi.nlm.nih.gov/pubmed/20735824

Secreted fungal sulfhydryl oxidases: sequence analysis and characterisation of a representative flavin-dependent enzyme from Aspergillus oryzae. Faccio G., Kruus K., Buchert J., Saloheimo M..
BMC Biochem. 2010, 11:31. http://www.ncbi.nlm.nih.gov/pubmed/20727152

Detecting novel genes with sparse arrays. Arvas M., Haiminen N., Smit B., Rautio J., Vitikainen M., Wiebe M., Martinez D., Chee C., Kunkel J., Sanchez C., Nelson M.A., Pakula T., Saloheimo M., Penttilä M., Kivioja T. Gene. 2010, 467:41-51. http://www.ncbi.nlm.nih.gov/pubmed/20691772

Array comparative genomic hybridization analysis of Trichoderma reesei strains with enhanced cellulase production properties. Vitikainen M., Arvas M., Pakula T., Oja M., Penttilä M., Saloheimo M.. BMC Genomics. 2010, 11:441. http://www.ncbi.nlm.nih.gov/pubmed/20642838

Discovery of a new tyrosinase-like enzyme family lacking a C-terminally processed domain: production and characterization of an Aspergillus oryzae catechol oxidase. Gasparetti C., Faccio G., Arvas M., Buchert J., Saloheimo M., Kruus K.. Appl. Microbiol. Biotechnol. 2010, 86:213-26. http://www.ncbi.nlm.nih.gov/pubmed/19798497

Hydrophobin fusions for high-level transient protein expression and purification in Nicotiana benthamiana. Joensuu J.J., Conley A.J., Lienemann M., Brandle J.E., Linder M.B., Menassa R..
Plant Physiol. 2010, 52:622-633. http://www.ncbi.nlm.nih.gov/pubmed/20018596

 

 

Biorevolutionists – HYBER centre of excellence

The world population is growing exponentially while, on the other hand, the natural resources are decreasing. Therefore, sustainable material development at low cost and minimum environmental impact will be the only way to warrant human wellbeing.

HYBER - a Centre of Excellence of the Academy of Finland - studies nature's miracles on a molecular level to create sustainable materials of the future.

VTT’s role in HYBER is to develop designer proteins needed in producing novel biomaterials. Check out this video that summarizes this amazing project.

 

 

Christopher LandowskiChristopher Landowski

I have received my B.S. with Honors in Biochemistry at the University of Wisconsin in 1994. My Ph.D. I received at the University of Michigan, College of Pharmacy, Department of Pharmaceutical Sciences in 2005. My thesis project was “Designing anticancer prodrugs for targeting drug transporters and activation enzymes”.

I began my postdoctoral training in Harvard Medical School in the Brigham and Women’s Hospital, Renal Division. As fellow my research focused on the nutrient transporters and their role in cancer progression. I continued this work in Switzerland at University of Bern, Medical School, Institute for Biochemistry and Molecular Medicine and was principal investigator of the Bernese Cancer League grant “Therapeutic Significance of the TRPV6 Calcium Channel in Prostate Cancer”.

I joined VTT in 2008 and I am the research team leader in the protein production team. My own research is aimed at understanding various aspects of protein secretion and improving the main protein production host, Trichoderma reesei. I am the project manager for the Academy of Finland’s Centre of Excellence Project in Molecular Engineering of Biosynthetic Hybrid Materials. 

https://www.researchgate.net/profile/Christopher_Landowski
https://www.linkedin.com/in/christopher-landowski-b6929356
https://scholar.google.fi/citations?user=GqHQUBQAAAAJ&hl=en


Nina AroNina Aro

I am an experienced project manager in the field of biotechnology. I received my PhD in Biochemistry from Department of Biosciences, University of Helsinki in 2003. I have strong technical background in genetic manipulation of filamentous fungi and yeasts, molecular biology, and protein production. At VTT I have mainly worked in customer projects focusing on protein production in the filamentous fungi T. reesei. I have worked with regulation of gene expression in, cell polarity regulation in yeast S. cerevisiae and development of high throughput screening methods. Currently my research focuses on development of novel protein production platforms and improving heterologous protein production in T. reesei and generation T. reesei strains for production of enzyme mixtures for biomass hydrolysis.

I am teamwork-oriented, committed and focused in my work and outgoing as a person. Given my passion for my research field, I believe I am able to deliver results and create long-term value to our customers.


Anne Huuskonen Anne Huuskonen

I received my M.Sc. degree from University of Helsinki in biochemistry. Over the years at VTT (employed at 2003) I have been involved with projects related to protein production in filamentous fungus Trichoderma reesei (improvement of production by e.g. decreasing protease load), physiology of brewer’s yeast under high gravity fermentation and also with metabolic engineering of other yeasts. Currently my focus is on heterologous protein production in T. reesei.


(https://fi.linkedin.com/in/anne-huuskonen-a278aa2) (https://www.researchgate.net/profile/Anne_Huuskonen)


Joensuu JussiJoensuu Jussi

I am a molecular biologist with a passion for recombinant protein production, purification and analytics. With 17 years of working experience in the field I have had a pleasure working with multitude of different expression systems and organisms including bacteria, virus, algae, fungi, yeast, plants and animals.

I obtained my MSc (2000), PhD (2006) and carry a tittle of docent (2010) for molecular genetics at the University of Helsinki. After two post-doctoral periods between 2006 and 2009 (University of Brussels, Belgium and Agriculture and Agri-Food Canada) I was recruited to VTT. I am an IPMA Certified Project Manager and have headed several research projects funded by TEKES and Academy of Finland as well as development projects for pharma and biotech industry.

www.researchgate.net/profile/Jussi_Joensuu
https://www.linkedin.com/in/jussi-j-joensuu-3073b2112

 


   Markku SaloheimoMarkku Saloheimo

I am project manager and senior researcher in the protein production team. I received my PhD Degree in 1991 on Molecular Genetics at the University of Helsinki and worked as a post-doctoral scientist in the University of Hannover, Germany 1992-1993. I received Adjunct Professor (Docent) title at Helsinki University in 1996.

While doing my PhD work and after the post-doc period I have worked at VTT as a Research scientist, Senior scientist and Principal scientist and as a Research team leader since 2000. My scientific work has focused on development of protein production host systems, in particular the fungus Trichoderma reesei, and in discovery and characterisation of novel industrial enzymes with various activities and application areas. In Trichoderma I have worked, for example, with gene regulation mechanisms, the protein secretion system, systems biology and different aspects of recombinant protein production. I have been the project manager for a number of research and development projects, funded both from public sources and by companies. For example, I was the manager of the 5-year company collaboration project aimed at therapeutic protein production in T. reesei. I am an author of over 90 peer-reviewed publications and an inventor in 23 patent families. My tasks as the Research team leader include personnel management, acquiring funding and marketing of our services to companies.

My areas of expertise include molecular and systems biology, recombinant protein production and development of microbial production hosts and processes.


Tiina PakulaTiina Pakula

My research activities at VTT (since 1994) are focussed on molecular and systems biology of the fungus Trichoderma reesei.  Especially I have been working on regulation of cellulase and hemicellulase production, protein secretion and stress responses related to protein production and physiology under protein production conditions. Application of systems biology approaches in order to understand the cellular processes related to enzyme production and to improve the properties of the organism has been among the main topics of interest. My earlier research activities at the University of Helsinki include e.g. molecular biology and replication of bacteriophages (Ph.D. thesis on the bacteriohage PRD1).    


Career at glance: 
Experience and positions held at VTT: 
2014, Certified project manager, IPMA C level 
2011-current, Principal scientist, VTT Technical Research Centre of Finland 
2000-current; Deputy team leader, VTT Technical Research Centre of Finland 
1999-2011, Senior research scientist, VTT Technical Research Centre of Finland 
1994-1999, Research scientist, VTT Technical Research Centre of Finland 
2000-2006 Deputy Team leader of the Academy of Finland Centre of Excellence Program 'Industrial Biotechnology
2009-2012 Leader of the consortium "Signalling in fungi" funded by the Academy of Finland 
Management of a number of other projects with public or company funding  
 
Previous experience: 

M.Sc. Department of Genetics, University of Helsinki, 1988 
Ph.D. Department of Genetics, University of Helsinki, 1992 
Scientific visits to Universidad Autonoma de Madrid, Spain 1990-1991 (EMBO short term/FEBS fellowship) 
1987-1988, Assistant, the University of Helsinki
1988-1989, Research assistant, the University of Helsinki
1990-1991, 1993 Research Scientist, the University of Helsinki


Ann Westerholm-ParvinenAnn Westerholm-Parvinen

My research focuses on improving heterologous protein production in the filamentous fungus Trichoderma reesei . Before joining VTT in 2001 I had received my M.Sc in Biochemistry from Åbo Akademi University and a PhD from the Heidelberg University in Germany. The PhD work was carried out at EMBL in Heidelberg and CSIS in Madrid, studying the function and protein 3D structure of a kinesin-like protein.

At VTT I have mainly worked in customer projects, both for national and international companies. The companies have been startup companies, SMEs, as well as large companies. The projects have been everything between short expression service projects and large projects lasting up to five years. In the larger projects we have for instance been developing better microbial strains for protein expression and secretion, improving strain generation procedures and optimizing cultivation parameters for higher protein expression. I have worked closely together with fermentation experts in improving fermentation conditions for T. reesei. For most of the projects I have been working as the project manager, and I am an IPMA C certified project manager.

My areas of expertise include amongst other molecular biology, protein sequence analyses, heterologous protein production in microbes and physicochemical characterization of proteins.

 


Mari Valkonen Mari Valkonen

I have worked in projects related to protein production in filamentous fungi and other microbes. I joined the Protein Production group at VTT in 1996 and my research activities have included studying the secretory pathway of Trichoderma reesei and protein expression. I did my PhD at VTT with the topic of “Functional studies of the secretory pathway of filamentous fungi - The effect of unfolded protein response on protein production”. I have made two Post Doctoral research visits to the University of Edinburgh, where the research project was about live-cell imaging of the secretory pathway of filamentous fungus Trichoderma reesei using FLIM/FRET state-of-the-art imaging techniques (FLIM/FRET, TIRF). Since then I have worked in both customer and jointly funded projects. The customer projects have been mainly about optimising the expression of heterologous proteins in Trichoderma reesei, but we have also had projects with other production hosts. For most of the projects I have been working as the project manager.

My areas of expertise are molecular biology, cell biology (microscopy & imaging techniques) and heterologous protein production in microbes.


Marika VitikainenMarika Vitikainen

I am a molecular biologist with strong experience in protein production in various microbial hosts. Before joining VTT I worked a decade at National Public Health Instute (currently National Institute for Health and Welfare) in projects involving protein production in Bacillus and pathogenicity of Staphylococcus. During that time I received my PhD in microbiology in 2004 at the University of Helsinki. I came to VTT in 2007 as an Academy of Finland Postdoctoral fellow for the development of genomic tools to characterize fungal strains. After the fellowship I have worked in various projects on protein production and gene regulation in fungal host and also managed expression service projects.

https://fi.linkedin.com/in/marika-vitikainen-40116157
https://www.researchgate.net/profile/Marika_Vitikainen
http://scholar.google.fi/citations?user=yCu5oZAAAAAJ&hl=fi

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