Extraction of proteins from Palmaria palmata Rsa Jnsdttir - - PowerPoint PPT Presentation

Extraction of proteins from Palmaria palmata Rósa Jónsdóttir

Background

2

• Increasing demand for high quality protein both for food and feed
• The access to high quality protein is becoming more challenging
• Important to find alternative food sources including protein alternatives

that contain all the essential amino acids fulfilling human requirements

Utilisation of Palmaria palmata

3

• Palmaria palmata - dulse
• Good nutritional value
• Suitable for human food and animal feed application
• Utilized as food in Iceland for centuries

Background

✓ High protein content (up to 35%) – Comparable to high protein vegetables e.g. soybeans ✓ Main polysaccharide is xylan (34-35%) ✓ P. palmata has a rigid cell wall consisting mainly of β- (1→4)/β-(1→3)-D-xylans along with some fractions of cellulose and β-(1→4)-xylans ✓ Due to the strength of the cell wall, traditional protein extraction methods might give limited results

Objectives

✓ Develop fractionation processes for preparation

• f protein-rich products and extracts to be used

as food, nutraceutical or feed ingredients ✓ Explore the use of enzymes (xylanse and protease) as processing tool ✓ Study protein yield, quality and the bioactivity

• f the different fractions

• P. palmata from Norway (NIBIO)
• Preprocessing: wet-milling
• Enzyme hydrolyzation
• Filtration
• Chemical analysis (water, fat, salt, ash, protein)
• Protein

– SDS-PAGE; Nitrogen; – Amino acids (SINTEF)

• Bioactivity (TPC, ORAC, DPPH, MC, ACE)

Material and methods

Enzyme Producer Optimum conditions Activity pH Temperature Umamizyme Amano enzymes Inc. 7.0 50 Endo- and exo-peptidase complex ProteAX Amano enzymes Inc. 6.0 60 Endo- and exo-peptidase complex Xylanase Matis ltd 6.5 60 Endo-1,4-β-xylanase Xylanase is derived from a bacterium belonging to the genus Planifilum, of the family Thermoactinomycetaceae according to 16S rRNA sequence analysis and blasting to the NCBI nr database (Matís, unpublished results).

Enzymes Optimum conditions and activity

Flow chart of the extraction process

Freeze drying Freeze drying

100 µm sieve

Extraction yields of dry matter of both fractions liquid extract and solid phase

10 20 30 40 50 60 70 Control Umamizyme Xylanase Xylanase and Umamizyme Umamizyme Extraction yield (%dw)

Yield

Liquid extract Solid phase

Amino acid composition and protein content of P. palmata

10

Values for untreated seaweed (n=1) are presented as % of total amino acid content. Values for control, xylanase, xylanase + Umamizyme and Umamizyme (n=2) are presented as % of total amino acid content. Values for nitrogen content are presented as % dry weight ± SD (n=4).

Essential amino acids, % of total amino acid content

Amino acid composition and protein content of P. palmata

11

• All EAA were present in the samples
• EAA ratio higher in all solid phase samples compared to liquid extract
• EAA ration higher in liquid extract when treated with protease

― Protease break down proteins into peptides and amino acids passing the 100 µm sieve after the hydrolysis => resulting in higher amino acid content in liquid extract. Essential amino acids (EAA)

Protein content of P. palmata, untreated and enzymatically treated, using different N conversion factor

54.9 33.4 5 10 15 20 25 30 35 40 45 50 55 60 Untreated Liquid extract Solid phase Liquid extract Solid phase Liquid extract Solid phase Liquid extract Solid phase Control Xylanase Xylanase + Umamizyme Umamizyme Protein content (% dw)

N = 6.25 Calculated conversion factor

Values are presented as % dry weight ± SD (n=4).

Conversion factor 4.7 3.6 4.1 2.5 3.8 2.9 3.5 3.2 3.4

Bioactivity of different fractions of P. palmata antioxidant activity

13

0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 Control ProteAX ProteAX and Umamizyme Umamizyme Xylanase Xylanase and Umamizyme Umamizyme

DPPH, IC50 values

Liquid extract Solid phase

10 20 30 40 50 60 70 80 90 100 Control ProteAX ProteAX + Umamizyme 75 Umamizyme 75 Xylanase Xylanse + Umamizyme 200 Umamizyme 200

Metal chelating ability (%)

Liquid extact Solid phase

DPPH antioxidant activity, IC50 values High antioxidant activity in liquid extracts Metal chelating ability (%) High antioxidant activity in some solid phase and liquid extract

ACE (angiotensin converting enzyme) inhibition activity of different fractions of P. palmata

14

IC50 (mg/mL) Liquid extract Solid phase Control >1.00 >1.00 ProteAX 0.45 ± 0.11 >1.00 ProteAX + Umamizyme 75 0.38 ± 0.02 >1.00 Umamizyme 75 0.69 ± 0.05 >1.00 Xylanase >1.00 >1.00 Xylanase + Umamizyme 200 1.14 ± 0.08 >1.00 Umamizyme 200 >1.00 >1.00

Blood pressure

Conclusion

15

• P. palmata contains high protein content of good quality that can be

extracted with good results using enzymatic treatment with xylanse

• Xylanase disrupted the rigid cell wall, resulting in higher protein yield in

the solid phase

• Protein extract from P. palmata is high in essential amino acids and

would therefore be suitable as food and nutraceutical ingredient

• The results indicate that hydrolysis with protease can be a beneficial

method to extract bioactive hydrolysates from P. palmata

Publications

Acknowledgement

17

Málfríður Bjarnadóttir, Matís Björn Viðar Aðalbjörnsson, Matís Anna Nilsson, Matís Guðmundur Óli Hreggviðsson, Matís Ólafur H. Friðjónsson, Matís Rasa Slizyte, SINTEF Ocean Michael Y. Roleda, NIBIO

rosa.jonsdottir@matis.is