Biorefjnery development for the production of succinic acid and - - PowerPoint PPT Presentation

Biorefjnery development for the production of succinic acid and value- added coproducts from sunfmower meal

M.N. Efthymiou, C. Pateraki, H. Papapostolou, A. Koutinas mneuth@gmail.com Heraklion 06.2019

Sunfmower Meal (SFM) Production

Food Crude Oil Crude Oil De- hulling Flaking Cooking SFM Presser / Expelle r

Solvent Extracti

• n

Animal Feed Defatted SFM Biofuel s Sunfmow er Seeds

Source:https://apps.fas.usda.gov/psdo nline/app/index.html#/app/advQuery

Sunflower Oil Sunflower Meal 5 10 15 20 25

3,7 4,8 19,3 20,7

European Union Global E stim ated production (M T )

Succinic acid

2

Solvents Food & beverages Detergents & Surfactans Polymer es e.g.

PBS

Adipic acid 1,4 Butanediol Maleic anhydrade Succinic acid Maleic acid Formic acid

• C4-dicarboxylic acid
• Conventional production via

hydrogenation of maleic anhydride

• Bio-based production is

50% of the total

• Glucose syrup and glycerol

are the main carbon sources used in industry

• Basfja succiniciproducens,

engineered Escherichia coli and yeast are the main strains used

• Market size expected to

reach 48.7 Million US $ by 2024

Composition of sunfmower meal

Composition (g/g % db) This Study Literature Moisturea 7.6 ± 0.03 3.7-10.4 T

• tal Solids

92.4 ± 0.03 89.6-96.3 Protein 33.7 ± 0.50 26.6-41.8 T

• tal phenolics (CAE)

3.0 ± 0.04 2.4-4.7 Oil 0.1 ± 0.06 0.9-2.0 Ash 7.4 ± 0.11 4.3-9.0 Pectins 5.7 ± 0.20 4.9 Lignin 12.2 ± 0.03 5.9-11.4 Cellulose 13.8 ± 1.83 14.9-23.0 Hemicellulose 10.8 ± 1.92 7.5-12.9 T

• tal Free Sugars

5.3±0.03

• Sucrose

4.6 ± 0.02

• Glucose

0.5 ± 0.04

• Xylose

0.1 ± 0.01

• Galactose

0.2 ± 0.10

• IPb

63.6 ± 2.33

• FANb

28.8 ± 0.12

• a wet basis

b mg/100g db

3

Moisture 7,67% Protein 33,84% Free Sugars 5,34% Oil 0,17% Ash 7,44% Pectins 5,70% Lignin 12,20% Cellulose 13,83% Hemicellul

• se

10,83% T

• tal

phenolic compound s 2,99%

SFM 100 g Ultrasound Phenolic Extraction

EtOH: H2O 1:1 1/10 w/v

Protein rich Solid Residue 91.8 g Protein Extraction & Precipitation Protein Concentrate Solid Residue 72.7 g Pretreatmen t

Η2SO4 1.25% w/v NaOH 2% w/v

Cellulose rich Solid Residue 19.6 g Sugar rich Hydrolysate Crude Phenolic Extract 8.2 g Hydrolysis Total Phenolic content 33% (CAE)

Crude enzymes Aspergillus awamori

Biorefjnery approach

Succinic acid Fermentation

Actinobacilus succinogenes

Recovery 17% Purity 80% Total Sugar concentration 15.7 g/L Concentration 40 g/L Productivity 0.48 g/L/h

Solid State Fermentation - Enzyme Activity

Cellulass es Xylanas es Phytase s

Aspergillus awamori

Conditions

• 5 g SFM
• 1.0 × 106 spores/g

solid

• 30 oC
• Initial moisture

content 50, 55, 60 & 65 % Enzymes

• Cellulases – CMC 1 % w/v, pH 5, 50 oC, 30

min

• Xylanases – Xylane 0.25 % w/v, pH 5, 50
• C, 60 min
• Proteases - Casein 0.75% w/v, pH 7, 50
• C, 30 min
• Phytases - Phytic acid 0.2 % w/v, pH 5.

50 oC, 15 min Defjnition of enzyme activity units

• Proteases & Phytases:

amount of enzyme needed for the production

• f 1 μg Free Amino Nintrogen (FAN) or Inorganic

Phosphorus (IP) per minute

• Cellulases & Xylanases

amount of enzyme needed for the production

• f 1 mg Glucose or Xylose per minute

Solid State Fermentation - Enzyme Activity

25 50 75 100 125 150 20 40 60 80 100 120 Fermentation Time (h) E nzy m e a ctiv ity U / g SF M

 Proteases  107.2 U/g after 94 h  Moisture content 60%

25 50 75 100 125 150 100 200 300 400 500 600 Fermentation Time (h) Enzym e activity U / g SFM

Phytases 512.7 U/g after 94 h Moisture content 60%

25 50 75 100 125 150 0,5 1 1,5 2 2,5 3 3,5 4

50 % 55 %

Fermentation Time (h) E nzy m e a ctiv ity U/g SFM

 Cellulases  3.7 U/g after 94 h  Moisture content 65%

25 50 75 100 125 150 0,5 1 1,5 2 2,5 3 Fermentation Time (h) E nzy m e a ctiv ity U/g SFM

 Xylanases  2.7 U/g after 94 h  Moisture content 60%

Enzymatic Hydrolysis - optimization

Efgect of T emperature

 Hydrolysis yield: 40.5% Cellulose; 61.1% Hemicellulose  Temperature 50 °C

• tal sugars

45 50 55

10 20 30 40 50 60 70

Cellulose Hemicellulose

T emperature (oC) H y d ro ly sis y ie ld % (g /g )

Enzymatic Hydrolysis - optimization

Efgect of initial SFM concentration

Increasing concentrations of SFM led to decreased hydrolysis yields :

20 40 60 80 100 0,00 0,05 0,10 0,15 0,20 0,25 0,30 0,35

50 g/L 75 g/L 100g /L

Hydrolysis Time (h) To ta l S u g a rs p ro d u c e d (g /g d b )

 Cellulose: 17.4 – 3.9 %  Hemicellulose: 27.7 – 5.6 %

50 75 100 10 20 30 40 50 60 70 Cellulose Hemicellulose

SFM Concentration (g/L) H ydrolysis yield % (g/g)

Fed-batch fermentation for succinic acid production

Conditions

 Actinobacillus succinogenes  Temperature: 37 oC  Initial sugar concentration : 60 g/L  Feeding up to 30 g/L

 Succinic acid concentration: 40 g/L  Yield: 0.53 g succinic acid/g consumed sugars  Productivity: 0.48 g/L/h

Conclusions

Sunfmower meal was efgectively valorized through the development of an advanced biorefjnery concept for the production of proteins, phenolic compounds and succinic acid:  Phenolic rich extract (33% CAE)  Protein Concentrate (80 % purity)  Succinic acid production

• 40 g/L
• Yield: 0.53 g succinic acid/g consumed

sugars

• Productivity: 0.48 g/L/h

Acknowledgements

Thank you for your attention!!