By Brigitta Kónya („Brigi”) Budapest University of Technology and

Economics

Department of Applied Biotechnology and Food Science

Laboratory of Industrial Microbiology

Gellert ter 4, 1111 Budapest, Hungary

http://www.tub-collection.com/

TUB MICROBIAL CULTURE COLLECTION (= DACT) FOR

EDUCATION AND RESEARCH IN INDUSTRIAL

MICROBIOLOGY

Trichoderma harzianum

欢迎 Welcome!

•Thank you for the invitation, nice to come here!

•Who are we?

•What is the purpose of our laboratory?

•What have we achieved in scientific research in the last 20 years? ( I participate in the lab work in the last 4 years)

Who are we in the world?

•Hungary is in the center of Europe.

•part of the European Union since 2004.

Who are we in Europe?

Hungary

•population: 10 million

•benefit is the central position

•around us there are seven other countries

• Fermentation experiments

• enzyme activity measurements

• Making evaluations, reports

• teaching microbiology lab practice for Hungarian and Brazilian students Research topics:

• enzymatic degradation of cellulose

• solid state fermentation (SSF)

• production of nanosilver particles by fungi

• isolation of microorganisms from soil

• filamentous fungi capable of inhibiting multiresistant Escherichia coli isolate

My main tasks

4600 microorganisms (main groups are below)

Filamentous fungi Yeasts

Actinomycetes Bacteria

Isolation worldwide

• Samples from extreme environmental conditions such as salty soils, alkaline soils, acidic soils, or sludge from the Adriatic sea, 40 meters deep

Soil collection

• Soil samples are collected worldwide for isolation and screening programs

• 2.) Soil, camping, Fusina (near Venice), Italy, July 25, 1982

• 41.) Soil near Mt.Aso vulcano ( 60 km from Kumamoto), Japan, Dec.20, 1984

• 50.) Meadow soil, near Leipzig, Germany, June 15, 1986

• 61.) Soil, Queensland, Australia, Jan.1988

• 65.) Desert sand, Babel el Khashab, Giza, Egypt, Feb.28, 1988

• 146.) Soil near seashore, Coral Island (Koh Lann), near Pattaya, Thailand, Feb. 1994

• 280.) Soil from park, Beijing, China, Dec.11, 1998

• 287.) Soil, Sun City, South Africa, Apr.11, 1997

• 549.) Soil, ruins of the Inca village, 2500 m elevation, Machu Picchu, Peru, August, 2013

Isolation from the

environment

Mucor sp. TUB F-1230 SEM picture (Scanning Electron Microscope)

Different fungal colonies

Storage • Subculture: storage on agar slants, perpetual transfer, serial transfer (for

a few weeks or for a few months)

• We sent 140 strains to ATCC (= American Type Culture Collection)

• Cryopreservation: freezing, storage in liquid nitrogen

• We prefer freeze-drying: storage by lyophilization (viability: 8-50 years)

Biological Experiments: Antagonism, inhibition of

other microbes

Mycoparasitism • Use of Trichoderma against

plant pathogenic fungi Sclerotinia , "biocontrol„

• Sclerotinia, which infects sunflower fields

sclerotium

sclerotia (black balls)

sclerotium

Sclerotinia

Trichoderma harzianum

• sclerotiums, especially strong and resistant forms of the fungus, it survives many years in soil

Analytical measurements

• Ultraviolet–visible spectroscopy (UV-VIS)

• Polyacrylamide gel electrophoresis (PAGE)

• High-performance liquid chromatography (HPLC)

• Mass spectrometry (MS)

Research topics for our lab in the last 20 years

• Production and application of different microbial enzymes (cellulase, hemicellulase, phytase, protease, amylase, chitinase, pectinase, etc.)

• Production and/or separation of R and S enantiomeric compounds from racemic mixtures by microorganisms and/or microbial enzymes

• Biotransformation of steroid and non-steroid compounds

• Basic materials for drugs (pharmaceuticals) as microbial secondary metabolites (lovastatin, cyclosporine, echinocandin)

Achievements • Discovery of novel cellulases and hemicellulases and their reaction

mechanisms for hydrolysis of lignocellulosic biomass, to produce bioethanol. http://www.disco-project.eu

• The aims of the DISCO project are to develop more efficient and cost-effective enzyme tools to produce bioethanol from lignocellulosic biomass, and understand how these enzymes work.

• Lignocellulose is a complex of carbohydrate polymers (cellulose and hemicellolose) tightly bound to lignin and is a major constituent of a wide variety of materials including waste materials from agriculture, forestry, wood-based industries, and municipal solid waste.

EU research project

Solid state fermentation (SSF)

Growth of Trichoderma asperellum on wheat straw in solid state fermentation (SSF)

Growth of Trichoderma virens on wheat straw in solid state fermentation (SSF)

SSF is used for the production of many oriental food. Tempeh, Koji, Sufu, one of the steps in soy sauce making .

Silver nanoparticles production • Silver nanoparticles produced from silver nitrate by Chaetomium

globosum OKI 270 by nitrate reductase enzyme

Livestock breeding Struggle for life in the microenvironment (microscopic scale)

• Inhibition of Escherichia coli in cultivated cattle manure

• E.coli, Salmonella cause illness for cows for example mastitis

• hypothesized that microorganisms which are antagonistic to E. coli develop in the cultivated manure are responsible for this phenomenon.

• elucidate the mechanism responsible for the shorter survival of E. coli

Agricultural Research

• Antibacteril activity of lactic acid bacteria as components in inoculants for silage

• 1. hypothesis is that specific LAB strains interact with rumen microorganisms to enhance rumen functionality and animal performance (live-weight gain, milk production).

• 2. LAB, which are used as inoculants for silage , inhibit detrimental microorganisms in the silage. LAB produce a variety of antimicrobial substances such as bacteriocins.

• The aim of the study was to determine antibacterial activity in LAB silage inoculants cultures.

Thank you for your attention!

Picture: Coremia of Penicillium claviforme on agar media