1. Advances in biotechnological processes for lactic acid production
project „Use of integrated bioprocesses in production of lactic acid” ( , 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 1

2. chemical synthesis of LA
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

3. 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
production of LA
and its derivatives
engineering and
production of equipment
production of energy
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020 3

4. Industrial bioprocesses for LA production - world raw sugar
market price
source: FINASUCRE
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020 4

5. - traditional use (food industry, pharmaceutical industry,...)
Use of produced LA
- traditional use (food industry, pharmaceutical industry,...)
- production of other chemicals and biodegradable
polymers (polylactides, PLA) drives global LA demand
LA PLA unit
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020 5

6. renewable raw material
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)
simple
carbohydrates
hydrolysis
fermentation LA
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020 6

7. 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

8. 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

9. 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

10. 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
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11. Advances in industrial bioprocesses for LA production (4)
- comprehensive analytical toolbox
Methods:
- gravimetric
- spectrophotometric
- chromatographic
- enzymatic
- SDS-PAGE
cgs100 / 45C / pH 5.5
carbohydrates, end products of metabolism, analytical HPLC-IEX
t = 0 h t = 2 h t = 6 h t = 72 h t = 216 h
activity of phosphoketolase (EC )
C6H11O5PO42- + PO43- + H+  C2H3OPO42- + C4H7O3PO42- + H2O
a-amylase (EC )
preparative LC-AF SDS-PAGE
amino acids, GC
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020 11

12. Efficiency of the new bioprocesses for LA production
L. amylovorus DSM 20531T SACCHARIFICATION
MRS-starch (%)
stirred tank bioreactor
45C / pH 5.5 / 400 min-1
t = 146,0 h
MRS-cg100
45C / pH 5.5 /400 min-1
t = 153,5 h
cgs100
t = 216,0 h
cgs60
horizontal rotating tubular bioreactor
20-35C / pH / 10 min-1
t = 118,0 h (+)
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020 12

13. Efficiency of the new bioprocesses for LA production
L. amylovorus DSM 20531T SACCHARIFICATION FERMENTATION YLA/S
MRS-starch (%) (%) (g g-1)
stirred tank bioreactor
45C / pH 5.5 / 400 min-1
t = 146,0 h
MRS-cg100
45C / pH 5.5 /400 min-1
t = 153,5 h
cgs100
t = 216,0 h
cgs60
horizontal rotating tubular bioreactor
20-35C / pH / 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

14. Efficiency of the new bioprocesses for LA production
L. amylovorus DSM 20531T SACCHARIFICATION FERMENTATION YLA/S
MRS-starch (%) (%) (g g-1)
stirred tank bioreactor
45C / pH 5.5 / 400 min-1
t = 146,0 h
MRS-cg100
45C / pH 5.5 /400 min-1
t = 153,5 h
cgs100**
t = 216,0 h
cgs60**
horizontal rotating tubular bioreactor
20-35C / pH / 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 14

15. Bachelor of Biotechnology Master of Bioprocess Engineering or PhD
Youth on the move
project „Use of integrated bioprocesses in production of lactic acid” ( , 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 PhD
Master of Molecular Biotechnology
Martina Andlar Ana Batušić Antonija Trontel
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
The 2nd International Symposium „VERA JOHANIDES“ - BIOTECHNOLOGY IN CROATIA BY 2020 15

16. Where the new bioprocess for LA production is now (1)
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17. Innovative bioprocess was developed √
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

18. Innovative bioprocess was developed √
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

19. Innovative bioprocess was developed √
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

20. The project 058-0581990-1997 is following E U R O P E 2 0 2 0
Where the new bioprocess for LA production is now (4)
project „Use of integrated bioprocesses in production of lactic acid” ( , Key Researcher Srđan Novak, Full Professor)
supported by Ministry od Science, Education and Sports of the Republic of Croatia
The project is following
E U R O P E
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 (
Department of Biochemical Engineering
Laboratory of Biochemical Engineering, Industrial Microbiology, Malting and Brewing Technology
Pierottijeva 6/IV HR Zagreb
Tel.: +385-(0)
Fax.: +385-(0)
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