Strengthening capacity of Institute ofMicrobiology & Biotechnology (IMBT)
forbioindustry development

Duong Van Hop.IMBT.VNU
Tokyo. 23 Feb.2010

1Content• Establishment of IMBT• NITE,Japan & IMBT,VNU, Vietnam
cooperation project is important factor forbuilding up capacity of VTCC, IMBT.
• Current activities of IMBTTokyo. 23 Feb.20102

1

Priority policy for biotechnology• Policy• Budget ( equipment &abroad training)• Fund for R-D

Tokyo. 23 Feb.20103

Some Government’sdirectives• Directive No18/ND-CP: National strategy for
BT to 2010 (13/4/1993)• Directive No. 50-CT/TW : Development andapplication of biotechnology to industrializationand modernization of the country (3/2005)• Directive No 115/CP-Ttg: Management oforganization for Science and Technology(6/2006)• Directive 80/ND-CP : Supporting Spin off
development (5/2008)

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Annual Budget• Before 1993: 200 000 USD• 1994-2003: around 2-4 Millions USD• 2004-up to Date: around 25-50 Millions
USD

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5

The need of NBRC for Bioindustry

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Establishment of IMBT
10/1996: Centre of Biotechnology Establishment .
5/2007 : Institute of Microbiology & Biotechnology( Act 661/QĐ-TTg)

Tokyo. 23 Feb.2010

7Mission• Conducting fundamental research ,application, education and consultant in thefields of biotechnology and microbiology• Development and technology tranfering for
novel bioproducts• International collaboration for conducting
Msc and PhD courses in biotechnology
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84

MANPOWER & INFRASTRUCTURE

Facility & Equipment: 3 millions USD
Lab. Area:
1000 square meters

Research staff: 42
Professors: 04,
Dr. Biology: 08
MSc:
07
BSc:
17Admin. staff: 06
PhD & MSc students: 6

Equipment:
For analytical work
- Conventional chromatography and FPLC,HPLCsystems- Multi-function UV-VIS spectrophotometers
For molecular biology work
- 1D & 2D electrophoresis apparatuses- PCR machines- Automatic sequencing machine (ABI 3.100 Avant)- DNA gel apparatuses- Freeze - centrifuges
For microbiology work
- Incubators, incubation shakers, fermentors- Deep freezers, culture-boxes- High-quality microscopes- Freeze-dryers, Freezing room-etc.
Fermentation pilot lab.

Tokyo. 23 Feb.2010

9NITE,Japan & IMBT,VNU, Vietnam
cooperation projectisan important factor
forbuilding up capacity of VTCC, IMBTTokyo. 23 Feb.2010

105Starting points (2003)

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11Signing of Memorandum of Understanding (MOU)
between the Ministry of Science and Technology (MOST) of Viet Namand the National Institute of Technology and Evaluation of Japanfor the Joint Research Program on Conservation and Sustainable Use of
Biological Resources

Signing for MOU between
Dr. Le Minh Sat, Vice
Director General ofDepartment of Science andTechnology, MOST, andMr. Masahiro Miyazaki,
Director General ofDepartment ofBiotechnology, NITE, on15 March 2004, Kisarazu,Chiba, Japan
The project : “Taxonomic and Ecological Studies of Microorganisms inVietnam and the Utilization” was started in April 2004
Tokyo. 23 Feb.2010126

Agreement on a collaborative
work with Viet Nam
JapanViet Nam
MOUProject
Agreement

NITE

Work Plan¾Collaborative Research¾Exchange of Expertise¾Technology Transfer¾Material Transfer

Ministry of Scienceand Technology(MOST)
Center ofBiotechnology,Vietnam National
University, Hanoi(VNUH)Tokyo. 23 Feb.2010

13Objectives
Overal objective:
Buiding up capacity of VTCC in culture collection
management

Specific objectives:- Technology transfer- Manpower development- VTCC image- Benefit sharing (utilization and publication)- Mutual understanding
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Technology transfer• Sampling• Isolating• Identification• Reservation• Documentation• Antibiotic screening

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15

Work flow of the project
Sampling

Preservation

Isolation
Identification

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16

8

Sampling

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17

Isolation
Microbes

MethodFilamenous fungi(8 methods)

Dilution met.,
UV radiation met,
Aeration met,
Surface sterile met.,
Moisture chamber met.,
Skeman’micromenipulator met.,
Diriec isolation met.,
Washing met.,

Actinomyces(3 methods)

SDS‐YE method for general actinomycetes
RC method for motile actinomycetes
DH method for isolation of Streptomyces

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Tokyo. 23 Feb.201019

Identification• Observation-Normal light microscopy-Scanning Electronic Microscope (SEM)-Fluorecent light microscopy• Molecular techniques-DNA sequencing(18s,16s,ITS,D1D2 ,23s rDNA genes)-DNA hybridization• Chemotaxonomy techniques :-Analysing menaquinone, ubiquinone, cell wall sugars, faty acids• Phylogenetic construction:-Homologous search (BLAST)- Phylogenetic tree construction (Clustal ,Phylip,Treview)

Tokyo. 23 Feb.20102010Observation

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21

New Genus
Tricladiella
Tetraploa
Lateriramulosa

Triramulispora？
Ordus
Scolecobasidium

Diprocladiella

Varicosporium?

Isthmolongispora
Ceratosporella

New Genus
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New Genus

2211

Molecular techniques

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23Reservation• Slant tranfering• Deep freezer• Lyophilization ( freezed drying)&liquid drying• Liquid nitrogen

Liquid nitrogen

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Lyophilization

Deep freezer

24

Slant tranfering

12Documentation

Culture’data

VTCC’website
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Antibiotic screening

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26

13

Working together=Technology transfer

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27

Manpower development• 12 visits (Scientists and researchers) from VTCC toNITE (1-2 months )• 6 visits of scientists to attended annual ACM
meeting (I.II, III,IV,V)• Conducting 14 Technical training workshops at IMBT
for a number of 120 Vietnam participants• Annual project workshops (2004-2009)

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28

14

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A lecture on entomopathogenic fungi and the genus Kickxellales wasTokyo. 23 Feb.201030
held on 10 September, 2004 by
Dr. Yuko Kurihara.

15

ACM-6. Hanoi 29-31 Nov.2009Tokyo. 23 Feb.2010

31

Image of VTCC
• Up-grade quality:
Number /isolates/documented• Daily access visiting VTCC
homepage:30-50• Domestic Recognised and requested
for servicing,deposit,distribution• International recorgnized:
visit,deposit, distribution• Member of WFCC:
http://wdcm.nig.ac.jp/hpcc.htm

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32

VTCC/ WDCM933/ Vietnam Type Culture Collection16Benefit sharing (utilization and publication)• Number of 1200/2000 sequences of isolates• 2 paper published,1 submitted and3 manuscripts• Starting biomass projects (2009-2011)

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Mutual understanding

Life in Vietnam• 12 visits of VTCC membersto Japan (1-2 monthsperiods)• 70 visits of NITE to Vietnam(3-7days)• 6 ACM visits to Asian
countries (Thailand,Indonesia, China, Korea)

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17• Briefly introduction to current activities ofInstitute of microbiology and biotechnology(IMBT)

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VIETNAM NATIONAL UNIVERSITY, HANOIInstitute of Microbiology & Biotechnology

Establishment :May , 2007
Address:
E2, 144 Xuan Thuy
Cau Giay,Hanoi,Vietnam
Tel: +84 43 768 0907
Fax: +84 43 768 0907
Website: http://www.biotechvnu.edu.vn
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Main activities• Microbial Diversity• Enzyme & protein works• Bioactive compounds from microbes• Applications of Molecular biology• Bio-product Development• Graduate Training (M.sc, PhD)
• International Colaboration

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4 Main divisions

VTCC(Microbialgene pool)
1

R-D
Activities
International
graduatecourses32
Bioproductdevelopment(Spin off)

4

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VTCC
• Enrichment (isolation, identification)• Maintaining• Distribution

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Database management of VTCC
Bacteria

Fungi

Internet

Database(Microsoft Access)YeastFile.html
ExportUploadPC server & client

Actinomycetes

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20

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Database in Microsoft Access

Enrichment of the cultures
2005

2006

2007

500

552

612

700

900

950

-----

50

Actinomyces

425

500

1023

1634

2108

3500

Yeast

381

470

523

643

780

850

Filamentous fungi

700

1314

1723

2123

2681

3000

Cell lines, plasmids

18

20

21

21

24

28

2024

2856

6469

8378

MicrobeAerobic bacteria
Anaerobic bacteria

Total

2004

3902 5121

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20094221Documented cultures
2000
1800
1600
1400
1200
1000
800
600
400
200
0number

2004 2005 2006 2007 2008 2009

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43

Some bioproduct and applications
BioproductApplication

Customer

Taq ADN Polymerase

for PCR and DNA finger
printing

5 Molecular labs. In Hanoi, Ho
Chi Minh cities

T4-ADN ligaza

for cloning and ADN
modification

3 Molecular labs. In Hanoi, Ho
Chi Minh cities

Digestive multienzymes

for animal feed

National Institute for animalhusbandry

probiotic

For animal feed

National Institute for animalhusbandry

Microbial preparationfor environmental
pollutiopn treatment

composting and producing
microbial fertilizers from urban
celulosic waste

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Hue and Ho Chi Minh Cities

4422

VTCC-Y- 0826

Cells of strain No. VTCC-Y-0826

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Colony of strain No. VTCC-Y-0826

45

VTCC-A- 1723 (VN07-A-0345)-Strain No. VN07-A-0345

Conidiophore and spores ofstrain No. VN07-A-0345

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VTCC-B-557

Colony of strain No. VTCC-B-557

Cells, endospores and parasporal crystalsof Strain No. VTCC-B-557

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Deposit and distribution
350300250200domestic Dep.
foreign Dep.
Dom.Dist.

150
100
5002006

2007

2008

2009

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R-D activities• Screening bioactive compounds frommicrobes ( enzymes, antifungi, antibacteria
and protease inhibitor)• Improvement of industrial microbes ( enzymes,antimicrobes :phytase, protease, ningnamycin,natamycin)• Valuable recombinant proteins (T4 ligase, Taq
polymerase, RT) for diagnostics
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49

Screening for enzym producers
Chitinase

Proteinase

MT5.1
B4.2

DCTK14.2.1
DCTK8A.2
B7.1

B4.2
B5.2

B5.2

TK18.3.2

TK8A.1

B2.3

B5.2
TK18.3.2

B7.1

Cellulaza

Amylaza
B4.2

B7.1

TK13.4

DCTK18.3

B7.1

TK14.6

TK18.3.2

DCB4.2

MT5.1

TK8A.2
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25

Screening bioactive compound against fungi andbacteria

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Screening for probiotic bacteria

Enzyme activity

Screening criteria:- Tolerant to intestinal condition (Low pH, bile

Anti harmful bacteria(Salmonella, Shigella, E.coli)salt, temperature)-Anti harmful bacteria (lactic bacteria)-Enzyme producers-Co-existing

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IMPROVING ACTIVITIES OF ENZYME

Mutant UL12-1514

Mutant UL12-4623

Mutant UL12-6910

Mutant UL12-3368

Negative control

Original strain UL12

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Some projects on going• Development process for enzymatic
production for animal feed.
• Enzymatic process for producing chitosan
oligomers• Development of animal probiotic• Production of L-lactic from cassava• Bioremediation of dioxin contaminated soils
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5427

FISH for studying diversity of bacteria from dioxin
contaminated soil in Danang Airport
DAPI

GAM42aFISH probes:ALP968
BET42aGAM42aα- Proteobateriaβ- Proteobacteria
γ - Proteobacteria
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Diversity of bacteria from dioxin contaminated soil

ALP 7%
BET 27%

ND 30%GAM 36%FISH probes:• ALP968• BET42a• GAM42a

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α- Proteobateriaβ- Proteobacteria
γ - Proteobacteria

56

28

Diversity of 14 isolates obtained from dioxin contaminatedsoils by ARDRA analysing
MspIM

B11

B45 B50 R02 R03 R04 R05 R06 R07

R08

R09 R10 R11 R12HaeIII

Using MspI and HaeIII RE for ARDRA analysing

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DGGE analysis of carbazol-degrading bacteria

Enrichment of carbazol-degrading
bacteria from sample B52-Car and D78car
B52-car

D78-3

D78-carControl

D78-2 D78-1 B52-3 B52-2 B52-1

PCR-DGGE analysing of rADN 16s

Archromobacter sp.
Alcaligenes sp.

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L-lactic fermentation from cassava
Kinetic of lactic ferm entation by L.rhamnosus
from hydrolysed cassava in 50 liters volum e60

The result showed that when using
strain L.rhamnosus for L-fermentationfrom the hydrolysed cassava powder (equally to 80 g/l reducing sugars in 50
liter fermenter ), the amount of L-lactic

5040

acid obtained is about 53 g/l equally tothe yield of 66.3 %.

30
20100135duration of fermentation (day)pH

Total acid (g/l)

Lactic acid (g/l)
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Some results from field test
• On piglets• On chicken

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30

On piglets
Negative *Positive*Exp

Compare to
Negative

396

454

462

Increase by
16.7%

Yield (gam feed/gam of 1.53weight)

1.39

1.42

Reduced by
7.2%

Diarrhea rate (case)

135

175

Reduced by 31%

Indicators
Growth rate(gam/pc/day)

254Piglets: 21 days age/weight: 7-8 kg. 24 pcs/case;Exp.Duration: 50 days
• Negative: feed without probiotic or antibiotic• Positive :feed with antibiotic• Source: Dr. Viet. Natl.Inst.Hus.Man.
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On chicken
Negative *Positive*Exp

Compare to
Negative

2318

2509

2500

Increased by
7.9%

Yield (gam feed/gam of 2.10weight)

1.92

1.96

Reduced by 6.6%

Indicators
Growth rate
(gam/pc/49 days)

Chicken:from 1 day age; 25 pcs/case;Exp.Duration: 49 days
• Negative: feed without probiotic or antibiotic• Positive :feed with antibiotic• Source: Dr. Viet. Natl.Inst.Hus.Man.
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Spin off
• Probiotic in production• Microbial preparation for composting• Bioproduct for biocontrol

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Post graduate training

Opening ceremony international Master course in biotechnology
The cooperation between VNU, Hanoi and Liege Univ.Belgium(Feb.2008)
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