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project „Use of integrated bioprocesses in production of lactic acid” (058-0581990-1997, Key Researcher Srđan Novak, Full Professor)
supported by Ministry od Science, Education and Sports of the Republic of Croatia
Advances in biotechnological
processes for lactic acid production
Anita Slavica, Antonija Trontel, Božidar Šantek, Srđan Novak
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
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Industrial bioprocesses for LA production - state of the art
fermentation
chemical synthesis of LA
microorganism
simple
carbohydrates
fermentation
LA
LA - lactic acid
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
2/21
Industrial bioprocesses for LA production - state of the art consolidation of subsidiaries
subsidiary
investment
production of food and
feed products
land holding and real estate
production of sugar
and by-products
engineering and
production of equipment
production of LA
and its derivatives
production of energy
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
3/21
Industrial bioprocesses for LA production - world raw sugar
market price
source: FINASUCRE
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4/21
Use of produced LA
- traditional use (food industry, pharmaceutical industry,...)
- production of other chemicals and biodegradable
polymers (polylactides, PLA) drives global LA demand
O
O-
HO
O
O
O
LA
O
n
PLA unit
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
5/21
Advances in industrial bioprocesses for LA production (1)
- main principle of new bioprocesses
fermentation
microorganism
simple
carbohydrates
fermentation
LA
simultaneous (semi-)solid substrate saccharification and fermentation to LA
microorganism
renewable raw
material
(polysaccharides)
hydrolysis
simple
carbohydrates
fermentation
LA
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
6/21
Advances in industrial bioprocesses for LA production (2)
- microorganism
Lactobacillus amylovorus DSM 20531T - lactic acid bacterium with proteolytic, amylolytic
and fermentative activity
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
7/21
Advances in industrial bioprocesses for LA production (2)
- microorganism
Lactobacillus amylovorus DSM 20531T
cells stained by Gram
colonies
corn grits particle
the total magnification of microscope 1000X
MRS medium / 45C / t = 12 h
cgs100 / 45C / t = 31 h
cgs100 - corn grits suspension in demineralized water
S0 = 100 g L-1
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
8/21
Advances in industrial bioprocesses for LA production (3)
- bioreactors and media
stirred tank bioreactor
horizontal rotating tubular bioreactor
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
9/21
Advances in industrial bioprocesses for LA production (3)
- bioreactors and media
media: MRS-starch100,
MRS-cg100 and
cgs100
medium cgs60
MRS-starch100 and MRS-cg100 - starch and corn grits in MRS medium without glucose
S0 = 100 g L-1
cgs100 and cgs60- corn grits suspension in demineralized water
S0 = 100 and 60 g L-1
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
10/21
Advances in industrial bioprocesses for LA production (4)
- comprehensive analytical toolbox
cgs100 / 45C / pH 5.5
Methods:
carbohydrates, end products of metabolism,
analytical HPLC-IEX
- gravimetric
- spectrophotometric
- chromatographic
- enzymatic
- SDS-PAGE
t=0h
t=2h
t=6h
t = 72 h t = 216 h
activity of phosphoketolase (EC 4.2.1.22)
C6H11O5PO42- + PO43- + H+  C2H3OPO42- + C4H7O3PO42- + H2O
a-amylase (EC 3.2.1.1)
preparative LC-AF SDS-PAGE
2.0
amino acids, GC
A280 (-)
1.6
1.2
0.8
0.4
0.0
0
2
4
6
8 10 12 14 16 18 20 22
V (mL)
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
11/21
Efficiency of the new bioprocesses for LA production
L. amylovorus DSM 20531T SACCHARIFICATION
MRS-starch100
(%)
stirred tank bioreactor
45C / pH 5.5 / 400 min-1
t = 146,0 h
100
MRS-cg100
stirred tank bioreactor
45C / pH 5.5 /400 min-1
t = 153,5 h
100
cgs100
stirred tank bioreactor
45C / pH 5.5 / 400 min-1
t = 216,0 h
100
cgs60
horizontal rotating tubular bioreactor
20-35C / pH 4.0-6.2 / 10 min-1
t = 118,0 h
(+)
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
12/21
Efficiency of the new bioprocesses for LA production
L. amylovorus DSM 20531T SACCHARIFICATION FERMENTATION
MRS-starch100
(%)
(%)
stirred tank bioreactor
45C / pH 5.5 / 400 min-1
t = 146,0 h
100
96.3
YLA/S
(g g-1)
0.91
MRS-cg100
stirred tank bioreactor
45C / pH 5.5 /400 min-1
t = 153,5 h
100
90.7
0.85
cgs100
stirred tank bioreactor
45C / pH 5.5 / 400 min-1
t = 216,0 h
100
44.7
0.52
cgs60
horizontal rotating tubular bioreactor
20-35C / pH 4.0-6.2 / 10 min-1
t = 118,0 h
(+)
(+)
(max = 8.90 g L-1)
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
13/21
Efficiency of the new bioprocesses for LA production
L. amylovorus DSM 20531T SACCHARIFICATION FERMENTATION
MRS-starch100
(%)
(%)
stirred tank bioreactor
45C / pH 5.5 / 400 min-1
t = 146,0 h
100
96.3
YLA/S
(g g-1)
0.91
MRS-cg100
stirred tank bioreactor
45C / pH 5.5 /400 min-1
t = 153,5 h
100
90.7
0.85
cgs100**
stirred tank bioreactor
45C / pH 5.5 / 400 min-1
t = 216,0 h
100
44.7
0.52
cgs60**
horizontal rotating tubular bioreactor
20-35C / pH 4.0-6.2 / 10 min-1
t = 118,0 h
(+)
(+)
(max = 8.90 g L-1)
**(Ala, Gly, Val, Leu, Ile, Pro, Asn, Asp, Met, Glu, Phe, Lys, Trp, Tyr)
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
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Youth on the move
project „Use of integrated bioprocesses in production of lactic acid” (058-0581990-1997, Key Researcher Srđan Novak, Full Professor)
supported by Ministry od Science, Education and Sports of the Republic of Croatia
Bachelor of Biotechnology Master of Bioprocess Engineering or
Master of Molecular Biotechnology
Martina Andlar
Ana Batušić
Tea Blaško
Vanda Baršić
†
Filip Cingesar
Vedrana Buljan
Ines Ivanuša
Ivana Dusper
Dijana Petrić
Tomislav Grzunov
Ivana Gusić
Nuša Jelovac
Željka Kosovec
Nikolina Mataga
Iva Marković
Morana Peklić
Jadranka Sučević
Vedrana Tadić
PhD
Antonija Trontel
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
15/21
Where the new bioprocess for LA production is now (1)
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
16/21
Where the new bioprocess for LA production is now (1)
Innovative bioprocess was developed
√
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
17/21
Where the new bioprocess for LA production is now (2)
Innovative bioprocess was developed
√
More resource efficient, greener and competitive
bioprocess was developed
√
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
18/21
Where the new bioprocess for LA production is now (3)
Innovative bioprocess was developed
√
More resource efficient, greener and competitive
bioprocess was developed
√
The performance of education system was enhanced
and the entry of young people to the labour market was
facilitated (Youth on the move)
√
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
19/21
Where the new bioprocess for LA production is now (4)
project „Use of integrated bioprocesses in production of lactic acid” (058-0581990-1997, Key Researcher Srđan Novak, Full Professor)
supported by Ministry od Science, Education and Sports of the Republic of Croatia
The project 058-0581990-1997 is following
EUROPE2020
A European strategy for smart, sustainable and inclusive growth.
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
20/21
Thank you for your attention!
Contact
Anita Slavica, Associate Professor
University of Zagreb, Faculty of Food Technology and Biotechnology (http://www.pbf.unizg.hr)
Department of Biochemical Engineering
Laboratory of Biochemical Engineering, Industrial Microbiology, Malting and Brewing Technology
Pierottijeva 6/IV HR-10000 Zagreb
Tel.: +385-(0)1-4605-142
Fax.: +385-(0)1-4836-424
e-mail: [email protected]
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020
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