Figure 1. At top, aspect of the different wheat bran fractions (from left to right): coarse, fine and middlings.

At bottom, percentage of the different fractions obtained in the citrus juice extraction process (courtesy of

Grupo Leche Pascual).

Figure 2. Wheat Bran Fractionation Protocol to derive fibre concentrates, oligosaccharides and ferulic acid.

Bran (10 - 40% suspension: 90 – 100 oC)

De-starch (Termamyl) (>100U/g :15 - 20 min)

Oligosaccharides

Protease (Alcalase) (20ml/g; 60oC : 3.5h)

‘Xylanase’ (Pentopan Mono) (7.5U/g ; 5h : 60oC)

Supernatant

Residue:‘Insoluble’ fibre

Residue: FIBRE CONCENTRATE

maltose / peptide mix

Growth medium etc

Further Enzyme

treatment / fractionation

Esterase

Ferulic Acid

Figure 3. Untreated bran, bran fibre concentrate and oligosaccharide extract.

Figure 4. NSP content and composition of fibre prepared from wheat milling fractions. ara = arabinose, xyl

= xylose, man = mannose, gal = galactose, glc = glucose, uronic = uronic acid. AB2 (Germ). Names on the

x-axis identify the location of the wheat fraction sampling point in the flour milling process.

Untreated fibre concentrate oligosaccharide

Figure 5. Yield of Pentopan-soluble material from milling fractions and corresponding Prebiotic Index (PI).

PI positive control (FOS) = 3.7 and negative control = -1.8.

Table 1. Summary of technological advantages and sensory screening for oligosaccharides in food products.

Food product Intended

application

Advantages Sensory screening

Bread roll fibre enrichment The dough rises quicker. Acceptable

Rye loaf fibre enrichment The dough rises quicker. Acceptable

Rye bar fibre enrichment The dough rises quicker. Acceptable

Natural yoghurt fibre enrichment Thicker texture of the yoghurt Not acceptable

Flavoured yoghurt fibre enrichment Thicker texture of the yoghurt Acceptablility flavour

dependant

Figure 6. TAC of ethanolic and aqueous extracts of wheat samples. TAC = Total antioxidant capacity. TE =

Trolox equivalents.

Figure 7. TAC of hydrolysed/fermented wheat bran. TAC = Total antioxidant capacity. TE = Trolox

equivalents

A B

2 (G

erm

)A F

‘G’ C

oarse

‘G’ F

ine

FBF

CBF

BF3

BF4

4MD/B

2BF

BK

Bulk

0

5

10

15

20

25

TA

C (m

mo

lTE

/gD

W)

Milling Fraction

aqueous

ethanolic

Control (0h) Control (24h) Driselase Viscozyme L0

10

20

30

40

50

TA

C (mm

olT

E/g

DW

)

Treatment

aqueous

Figure 8. Bioconversion with Pseudomonas fluorescens BF13-1p of (A) food-grade ferulic acid in buffer

(pH 7) and (B) ferulic acid recovered from wheat bran after removal of reducing sugars and its re-suspension

in buffer. : Ferulic acid; : Vanillin; : Vanillic acid.

0,0

0,2

0,4

0,6

0,8

1,0

1,2

0 1 2 3 4 5 6 7 8

B Vanillin molar yield: 55.1%

Selectivity: 68%

hours

mm

ol/

L

0,0

0,2

0,4

0,6

0,8

1,0

1,2

0 1 2 3 4 5 6 7 8

A

Selectivity: 71%

hours

mm

ol/

L

Vanillin molar yield: 63.2%

Figure 9. Protocol for the integrated processing of citrus pomace. MW: Microwave drying, DIC:

Instantaneous Controlled Pressure Drop, SE: Solvent Extraction, USE: Ultrasound Assisted Solvent

Extraction, CO2: Supercritical CO2 extraction.

Citrus

fiber

Texturizing

Fruit

paste

Formulation

with other

ingredients

Snack barBreakfast

cereals

Filler for

bakery

Functional

drink

Baking Extrusion Texturizing

Cloud

Concentration

Debittered

cloud

Debittering

(resin

adsorption)

Polyphenols

extract

Elution

Homogenization

Citrus peels

Blanching Enzyme

Enzyme

Decanting

Pasteurization

Extraction/Debittering

SE

USE

CO2

Polyphenols

extract

Drying

Milling

MW

DIC

Extraction/Debittering

Route 1 Route 2 Route 3

Centrifugation

To 1

or 2

From

3

From

3

Figure 10. Colour of the non-debittered and debittered orange fibre obtained (right) compared with the

commercial reference fibre (left); commercial is the whiter.

Figure 11. Visual comparison of the NAMASTE cloud obtained from the acid water (right) with commercial

orange cloud (centre) and the commercial cloud analogue lipid emulsion (left)

Table 2. Summary of the technological advantages and sensory screening for citrus fibres

Citrus fibre

addition

Intended technology

application

Advantages Sensory

screening

frozen potato

croquette Less fat absorption, fibre

enrichment

More stable dough, (easier to handle,

less sticky, less loss during production)

Acceptable

frozen potato

dumpling Less fat absorption, fibre

enrichment

More stable and less sticky dough, more

convenient to prepare at home

Acceptable

frozen filled

potato

dumpling

Fibre enrichment More stable and less sticky dough, more

convenient to prepare at home

Acceptable

cottage cheese

dumpling Fibre enrichment More stable dough, more convenient to

handle at home

Acceptable

meat roll Meat replacer, moisture

control, lower fat content /

fibre enrichment

Replacement of 16-20% meat can be

achieved

Acceptable

Doughnut Fat, oil, moisture control,

fibre enrichment

To avoid fat absorption during frying Improvement

needed

bread roll moisture control, flour

replacer, fibre enrichment

Replacement of 15-20% flour can be

achieved

Acceptable

sweet roll filled

with jam Moisture management Thickening agent (filling),

Flour replacer, fibre

enrichment

Replacement of 15-20% flour can be

achieved in the dough

Improvement

needed

Thicker filling can be achieved Not

Acceptable

Citrus fibre

addition

Intended technology

application

Advantages Sensory

screening

salad dressing Thickening agent Thicker dressing, even distribution of

herbs, more even surface distribution

Not

Acceptable

yoghurt based

salad dressing Thickening agent Light yoghurt (1.5%) becomes thicker;

improved distribution of herbs on

vegetable surface

Not

Acceptable

red fruit sauce

(yoghurt sauce) Thickening agent Light yoghurt (1,5%) becomes thicker;

improved surface distribution on fruits

Not

Acceptable

batter mix Egg replacer, fibre

enrichment

Replacement of eggs Acceptable

Figure 12. Citrus ingredients based beverages: samples elaborated with experimental clouds vs reference

sample (left), samples of fibre enriched orange juice sent for consumer’s testing (center) and polyphenol

enriched nectar (right).

Figure 13. Example of the plot of the Principal Component Analysis of the results of the sensory evaluation

of the juice samples enriched with the NAMASTE fibres (debittered and non-debittered) vs three commercial

juices that claim content in pulp or whole orange.

-4 -3 -2 -1 0 1 2 3

-1.0

-0.5

0.0

0.5

1.0

1.5

2.0

CA factor map

Dim 1 (43.7%)

Dim

2 (

23

.84

%)

1

2

3

4

5

6

7

8

9

10

1112

1314

15

16

17

18

1920 21

2223

24

25

26

27

28

29

30

3132

33

34

35

36

37

38

39

40

41

42

43

44

45

46

4748

49

50

51 52

53

54

55

56

57

58

5960minute.maid

don.simon

graniniazti.des

azti.sin

Figure 14. Food products produced by using wheat bran and citrus by-products.

Figure 15. Effect of HPH treatments on color and appearance of the HPH-citrus-based formulations.

Figure 16. Scavenging activity of doughs added with raw or pre-fermented bran (before (t0) and after (tf)

sourdough fermentation. From left: 1=Sourdough; 2=no bran added; 3=Raw (untreated) bran; 4=bran pre-

fermented with L. casei; 5=bran pre-fermented with L. sakei; 6=bran treated with Driselase® and pre-

fermented with L. casei; 7= bran treated with Driselase® and pre-fermented with L. sakei; 8=ascorbic acid 1

mg/ml).

Figure 17. Fibre content increase (g/100 g) of the NAMASTE bread due to the enrichment with the different

milling fractions and NAMASTE bran (white bars) and total fibre content of conventional bread and

wholegrain bread (grey bars).

A M

ill B

2 (G

erm

)

2A M

ill F

A M

ill ‘G

’ Coar

se

A M

ill ‘G

’ Fin

e

A M

ill F

BF

A M

ill C

BF

B M

ill B

F3

B M

ill B

F4

B M

ill 4

MD/B

2

B M

ill F

B M

ill K

NAM

ASTE

bra

n

Conve

ntional

bre

ad

Conve

ntional

whole

gra

in b

read

0

5

10

15

Fib

re c

on

ten

t (g

/100g

bre

ad

)

Figure 18. Theoretical TAC increase (µmol TE/100 g bread) of the NAMASTE bread due to the enrichment

with the different milling fractions and NAMASTE bran. TE = trolox Equivalent.

A M

ill B

2 (G

erm

)

2A M

ill F

A M

ill ‘G

’ Coar

se

A M

ill ‘G

’ Fin

e

A M

ill F

BF

A M

ill C

BF

B M

ill B

F3

B M

ill B

F4

B M

ill 4

MD/B

2

B M

ill F

B M

ill K

NAM

ASTE

bra

n

0

100

200

300

400

500

TA

C (m

mo

lTE

/100 g

)

Figure 19. Visual differences observed in feeding trial in orange molly using citrus by-products fractions.

Table 3. Comparison of the different NAMASTE products for their selection for assessment in WP5.

Product

(ingredients)/process

Note Exploitability and Feasibility

Pre-fermented Bran Interesting results were collected also on rice bran. It has

antioxidant capacity and probiotic effects. The same

enzymes were tested on rice and wheat with similar

effects.

It is of sure industrial interest, but not within

the consortium. Not enough quantities are

obtained in the laboratory trials due to low

yields.

High Pressure

Homogenization

(HPH) applied on

citrus fibre

It is an innovative application of HPH. It is very

interesting for citrus fibres and for fibres in general. The

main applications of the products obtained from the

application of HPH on Citrus could be in the preparation

of snack filler or beverages (to be evaluated)

It is patentable if possible applications are

described. GLP and JRS (limited to the

powder) are interested for the exploitation.

SELECTED for WP5

Citrus Fibre A screening of possible applications is needed for

debittered and non-debittered formulation

The patentability has to be evaluated in

association with JRS.

SELECTED for WP5

Cloudy Agent To be evaluated also in association with the polyphenol

crude extract

Not patentable but GLP is interested.

SELECTED for WP5

Concentrated bran

fibre

GLP is available to test the product GLP is not interest in the product. IP member

interest should be evaluated. Actual facilities

are not food grade.

Oligosaccaride

fraction

Same as above Same as above

Natural Vanillin The complete production process starting from bran is

not available. RTD addressed to improve ferulic acid

release from bran will be carried out at the laboratory

scale in close cooperation with IFR

Difficult to evaluate; very probably, given the

low rate of ferulic acid release and in turn of

vanillin production achieved, the results can

be of none industrial interest. They will be

mostly published on international journals.