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Bioprocessing of protein-rich raw material towards
improved functionality
Laitila, Arja; Karsma, Anni; Rosa-Sibakov, Natalia;
Juvonen, Riikka; Sozer, Nesli
32nd Nordic Cereal Congress "Future Food
Security - New Opportunities for Nordic Crops",
7 - 9 September 2015 , Espoo, Finland.
Nordic Cereal Association (2015)
2015
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VTT TECHNICAL RESEARCH CENTRE OF FINLAND LTD
Bioprocessing of protein-rich
raw material towards
improved functionality
Nordic Cereal
Congress 2015,
Espoo
Arja Laitila, Anni Karsma, Natalia RosaSibakov, Riikka Juvonen & Nesli Sozer
VTT Technical Research Centre of Finland Ltd
arja.laitila@vtt.fi
Novel multifunctional plant protein ingredients
with bioprocessing (BIOPROT)
1. Improve protein and nutritional functionality of bran and faba
bean ingredients by tailored bioprocessing with microbes and
enzymes
2. Establish technological functionality of modified plant protein
sources in several food categories (pasta, bread and extruded
snacks)
Finnish funding: MMM Makera
BIOPROT-team
 University of Helsinki, Finland
 Elisa Arte, Rossana Coda,
Kati Katina (coordinator)
 University of Bari, Italy
 Marco Gobbetti, Carlo
Rizzello
08/09/2015 Laitila
Project leaders
 VTT Ltd, Finland
 Riikka Juvonen, Anni Karsma,
Arja Laitila, Atte Mikkelson,
Margherita Re, Natalia RosaSibakov, Nesli Sozer, Irina
Tsitko
 Central Research Institute of
Food and Feed Control, Turkey
 Nurcan Aysar Guzelsoy,
Orhan Eren
Faba bean is an excellent source of protein
 High protein content (25-35%)
 Vitamins, minerals, dietary fibre and
bioactive compounds
 Low cost: Half the price of soy bean and
peas (~200 vs 400 €/t)
 Suitable for gluten-free products
 The limitations on using faba beans are
the presence of antinutritional factors, a
bitter taste and poor technological
functionality
08/09/2015 Laitila
Vicia faba major
Vicia faba minor
Bioprocessing as a tool to improve the technofunctional and nutritional profile
Microbial food cultures
 Bacteria (especially lactic acid
bacteria), yeasts and moulds
 Versatile, multifunctional actions
Enzymatic treatments
 Fast, controlled, targeted
actions
Structure
Safety and
shelf life
Colour
Bioprocessing
Process
performance
08/09/2015 Laitila
Taste
Digestability
and
bioavailability
This study aimed to improve:
 protein bioavailability
 technological functionality
1. Enzymatic treatment with
phytase
2. Lactic acid bacteria, including
exopolysaccharide producing
(EPS) bacteria
 Fermented faba bean
ingredients in extrusion
Phytic acid
 Strong binding affinity to important minerals
 Forms complexes with proteins decreasing
solubility





08/09/2015 Laitila
Microbial EPS such as dextrans
Selected lactic acid bacteria
EPS to deliver viscosifying, texturising and
emulsifying agents in to foods and beverages
Effective at low concentrations
(0.5-1%, d.w.)
Health promoting properties
In situ production: Label-free technology for
modifying texture and nutritional properties
Vicia faba L. var major
Dehulling, milling (partcle size ca 600 µm)
Raw material characterisation
Enzyme selection
LAB selection
Food-grade phytase (Ultra Bio-Logics Inc)
Indigenous and from other plant sources
L. plantarum, Lc. lactis, W. confusa
106 cfu/g
Enzyme treatment
Fermentation
2, 10 and 20 U (33-167-333 nkat)
Ratio (s:l) = 1 solid : 2 water
RT and 55°C; 1,2,4 h
0.5 kg and 10-15 kg
33% flour + 62 % water + 5% sucrose*
(*only EPS production)
25°C 24 h
Freeze drying and milling
Pasta
Extrusion
Baking
100 -50 – 25 % mixed with rice flour
08/09/2015 Laitila
LAB = lactic acid bacteria
Phytic acid (mg/kg)
Treatment with food-grade phytase was effective
Native
Incubation time (h)
Phytic acid content in faba beans was reduced up to 89 %
08/09/2015 Laitila
Phytase treatment increased protein release of faba bean
(in vitro digestion model Nordlund et al, 2013, J Cereal Sci, 58, 200-208)
Phytase treatment
 enhanced the solubilisation and release of protein from faba bean in
the gastric stage of in vitro digestion
 increased Ca, Mg, Fe, Zn availability (data not shown)
08/09/2015 Laitila
Bioprocessing of faba beans with LAB
08/09/2015 Laitila
Faba beans are natural source for
exopolysaccharide (EPS) producing LAB
 100-1000 cfu/g → >108 cfu/g
 Weissella and Leuconostoc
bacteria isolated and
characterised
 In situ production of EPS in
protein rich matrix ?
EPS
producers
Total LAB
10,0
9,0
8,0
7,0
6,0
5,0
4,0
3,0
2,0
1,0
0,0
log 10 CFU/g
 Intensive LAB growth
0h
24h
 EPS-negative: Lactobacillus
plantarum
 Antimicrobial potential
 Reduction of antinutritive factors
and protein modification
08/09/2015 Laitila
Leuconostoc lactis
Weissella confusa
Lactobacillus plantarum E76 (EPS-negative) vs
Weissella confusa E3403 after 24h fermentation
08/09/2015 Laitila
Intensive dextran production during faba bean
fermentation
7,0
6,5
Dextran (g/ 100g dm)
6,0
4,9
5,0
4,0
3,0
2,0
1,4
0,8
1,0
0,2
0,0
L. plantarum
Lc. lactis_Control
Lc. lactis_EPS
W. confusa_Control
W. confusa_EPS
Control = without added sucrose
08/09/2015 Laitila
University of Helsinki: Katina
FESEM micrographs after 24h fermentation
Starch granules
Protein matrix
EPS negative
L. plantarum
In situ produced dextran
Polymeric EPS matrix
Weissella bacteria
VTT: Field emission scanning electron
microscopy (FESEM) / Tsitko & Laitila
08/09/2015 Laitila
EPS = exocellular polysaccharide
Fermentation releases proteins from the cell wall
matrix and significantly increases the amount of
free amino acids (AA)
Total free amino acids
Sample
g/kg
Control
5.7
L. plantarum
7.6
Lc. lactis
10.4
W. confusa
9.8
08/09/2015 Laitila
University of Bari: Rizzello
Fermentation had a
positive impact on
amino acid profiles
→The amount of
essential AAs increased
Bioprocessed faba bean ingredients in extrusion
08/09/2015 Laitila
Fermentation with L. plantarum improved
mechanical properties of extruded snacks
Decreased hardness
Increased crispiness (50 and 25% addition)
18,0
0,45
Native faba
bean flour
16,0
0,40
14,0
L. plantarum
fermented
faba bean
Hardness (N)
12,0
10,0
8,0
6,0
Crispiness Index
0,35
0,30
0,25
0,20
0,15
4,0
0,10
2,0
0,05
0,0
0,00
100%
50%
25%
Level of faba bean raw material in the sample
08/09/2015 Laitila
100%
50%
25%
Level of faba bean raw material in the sample
In protein-rich material, no additional impacts
with EPS producers
100 %
50 %
25 %
Native faba
L. plantarum
fermented
W. confusa
fermented
(EPS)
Challenge: Burning (Maillard reaction), especially with EPS producing LAB
Future: Other applications such as in baking
08/09/2015 Laitila
Benefits of faba bean bioprocessing
Enzymatic treatments
 Phytase treatment as a fast
tool to increase
 protein solubility
 mineral availability
08/09/2015 Laitila
Microbial food cultures
 Release of proteins and
modification of amino acid
profile by fermentations
 Reduction of antinutritional
compounds
 Biocontrol
 Faba beans provides an
excellent base for EPS
production → functionality in
other applications such as
baking, vegetable pastes
Take home messages
 Well-characterised microbes and enzymes as a tool for
improving nutritional profile and functional properties of
protein-rich legume ingredients and products
→ New product innovations
08/09/2015 Laitila
TECHNOLOGY FOR BUSINESS