ITACONIC ACID PRODUCTION FROM SORGHUM BRAN – A BIOREFINING APPROACH AMINA AHMED EL-IMAM; C. DU; P. S. DYER 27 TH AUGUST, 2015
Jan 19, 2018
ITACONIC ACID PRODUCTION FROM SORGHUM BRAN – A BIOREFINING APPROACH
AMINA AHMED EL-IMAM; C. DU; P. S. DYER
27TH AUGUST, 2015
OUTLINEIntroduction
a. Itaconic acid fermentation and potential mitigantsb. Sorghum branc. Biorefineries d. Motivation & Objectives
Methods & Findings a. Compositional analysesb. Hydrolysate preparationc. Determination of potential inhibitorsd. Hydrolysate enrichment and detoxificatione. Fermentation
Summary
INTRODUCTION • ITACONIC ACID (IA):
• A versatile drop-in organic acid monomer• Produced by fermentation of sugars by Aspergillus terreus• High cost limits application – need for cheaper substrates
• SORGHUM BRAN (SB): • Sorghum (Sorghum bicolor) is an important cereal • Used in making various foods, feed, ethanol etc• Grain processing generates bran as waste
• BIOREFINERY• Facility that integrates biomass conversion with fuels, power and
chemical production• Most promising route to creation of biobased industries
INTRODUCTION
Sorghum grains
Sorghum bran
Pap/Ogi
Itaconic acid Fermentation
Figure 1. Reference: El-Imam, A. A., & Du, C. (2014). Fermentative Itaconic Acid Production. J Biodivers Biopros Dev, 1(119),
Pretreatment• Physical• Biological• Chemical
Uses/Applications
PROPOSED FLOWCHART FOR ITACONIC ACID PRODUCTION
MOTIVATION & OBJECTIVESMOTIVATION:
• Sorghum waste is relatively underutilised though it accumulates in large amounts
• Renewable Itaconic acid is beneficial to the environment
OBJECTIVES:A. Conduct compositional analyses of sorghum branB. Screen several isolates for desirable traitsC. Determine potential inhibitors of itaconic acid
fermentationD. Detoxification of the mediumE. Produce itaconic acid in economical amounts
A: COMPOSITIONAL ANALYSES
• a
Proximate composition of sorghum brans. Values are means ± SD of triplicate measurements
COMPONENT WHITE BRAN
RED BRAN
MOISTURE 11.6 ± 0.3 6.3 ± 0.1
PROTEIN 19.2 ± 0.6 21.4 ± 0.1
STARCH 52.9 ± 0.1 57.0 ± 0.2
LIPID 3.2 ± 0.8 3.01 ± 0.7
LIGNIN 6.7 ± 0.01 10.5 ± 0.04
ASH 1.8± 0.04 1.4± 0.05
SUM (%) 95.4 99.6
High Starch, protein and lipid contents
Low ash content:
OR
PROCESSFEEDSTOCK PRODUCTS APPLICATIONS
SORGHUM GRAINS
MILLING
DILUTE ACID HYDROLYSIS
SUGARS
STARCH
ENZYMATIC HYDROLYSIS
BRAN
A. terreus FERMENTATION
HYDROLYSISRESIDUE
S. cerevisiae/YEAST FERMENTATION
ETHANOL
VARIOUS INDUSTRIAL USES
FOOD
FEED
INDUSTRIAL PRODUCTS
HEAT
PYROLYSIS PRODUCTS
ITACONIC ACID
FUEL/ CHEMICAL
PROPOSED BIOREFINING ROUTE FOR THE PRODUCTION OF ITACONIC ACID AND HIGH VALUE CO-PRODUCTS
PHENOLICS
B: HYDROLYSATE / MEDIA PREPARATION DILUTE ACID HYDROLYSATE
+
10% Bran slurry 1% H2SO4 Bioreactor
DEFINED MEDIA/SOLUTIONS • Fermentation medium and all solutions prepared according to standard
methods
Glucose conc. in
hydrolysate (g/L) 22.08 ± .011
C: FERMENTATIONMICROORGANISMS• 44 strains of Aspergillus terreus - BDUN collection • 1 strain - DSMZ
SUBMERGED FERMENTATION• 1 x 106 spores/ml• Incubated at 37oC and 200 rpm for 7 days
ITACONIC ACID ESTIMATION• Jasco® HPLC with a Hi-Plex column and R.I. detector
HYDROLYSATE INHIBITOR ANALYSES• Determined using a Waters® Techsphere ODS-2 column and a Photodiode
Array detector.
C: FERMENTATION
Figure 2: Screening strains of Aspergillus terreus for itaconic acid production on glucose medium.
49-1
49-2
49-3
49-4
49-5
49-6
49-7
49-8
49-9
49-1
049
-11
49-1
249
-13
49-1
449
-15
49-1
649
-17
49-1
849
-19
49-2
049
-21
49-2
249
-23
49-2
449
-25
49-2
649
-27
49-2
849
-29
49-3
049
-31
49-3
249
-33
49-3
449
-35
49-3
649
-37
49-3
849
-39
49-4
049
-41
49-4
249
-43
49-4
449
-45
0
5
10
15
20
25
30
35
40
45
50
YIELDS ON GLUCOSE MEDIUM
ITACONIC ACID
Aspergillus terreus BDUN Strain ID
Itaco
nic
acid
yie
ld (g
/L)
D: DETERMINATION OF POTENTIAL INHIBITORS• Tannins:-
• Phosphorus:- Determined by ICPMS-CCT, along with other elements
• Sulphates:- The effect of neutralization salt Na2SO4 on defined medium was investigated
• Common fermentation Inhibitors:- estimated by HPLC
SAMPLE C.E. (g/100 g)
RED BRAN 0.18 ± 12.7
INHIBITOR NAME
CONCENTRATION (mg/L)
HMF FURFURALVANILLIC ACID
FERULIC ACID
ACETIC ACID SYR
Sorghum Hydrolysate 59.0 30.1 5.6 19.6 ND ND
COMMON FERMENTATION INHIBITORS
TIME COURSE EXPERIMENTSOrdinary Hydrolysate (55g/L) Supplemented Hydrolysate (120g/L)
2 3 4 5 7 90.00
1.00
2.00
3.00
4.00
5.00
6.00RBDAH
49-549-2249-45
TIME (DAYS)
ITA
CO
NIC
AC
ID Y
IELD
(g/L
)
2 3 4 5 7 90.00
5.00
10.00
15.00
20.00
25.00
30.00GLUCOSE MEDIUM
49-5
49-22
49-45
TIME (DAYS)
ITA
CO
NIC
AC
ID Y
IELD
(g/L
)
SUMMARYSUBSTRATE
• Sorghum bran is nutritionally suitable for bioconversions• Inhibitors: tannins and phosphates present
SCREENING • Three strains show promise on the hydrolysate• Glucose augmentation improved yields by ~87%
FERMENTATION• About a quarter of the yield of the glucose medium obtained by 49-22• Peak IA yield was obtained on Day 4
FUTURE WORK• More detoxification attempts are currently being run and the results
appear promising with up to 200% increase in yield
ACKNOWLEDGEMENTSThe Federal Government of NigeriaBioenergy and Fungal Biology groups
THANK YOU FOR LISTENING