Effect of Transglutaminase on the Film Properties Obtained by Blending Nigella Sativa Protein Concentrate and Pectin

Laialy Ali, Mohammed Sabbah1*, Jehad Abbadi2

1Department of Nutrition and Food Technology, An-Najah National University, Nablus P.O. Box 7, Palestine. 2Department of Biology, Faculty of Science and Technology, Al-Quds University, P.O. Box 20002, Jerusalem, Palestine.

* Corresponding author: m.sabbah@najah.edu

Problem statement(plastic wastes)

Plastic Facts

Degradation of plastic

wastes require more than 200

years

Producing plastic about 350 million tons/year

Greatest quantity of

plastic wastesis derived from food packages

wastes

3Reference: Galloway et al., (2020). One Earth, 2, 5-7.

Plastic Pollution

3

4

Incineration

Plastic degradation

Landfilling

Bio-plastic

Suggested solutions to solve the problem of plastic

wastes disposal

Recycling

Reference: North and Halden (2013). Reviews on environmental

health, 28(1), 1–8. 5

6

Bioplastics application

Edible films applications

6

Edible films : are a primary packaging made from biopolymer materials.

Reference: Aguirre-Joya et al., (2018). In Food packaging and preservation (pp. 1-61).

Main component of edible films

provide good barrier

for oxygen and carbon

dioxide gases

produces edible films

with distinctive

mechanical properties

blocks water

transmission

7Reference: Pavlath, A. E., & Orts, W. (2009). Edible films and coatings for food applications (pp. 1-23).

Strategic improvement of edible films

8

CrosslinkingBlending protein/polysaccharide

Reference : Benbettaïeb et al., (2016). Comprehensive Reviews in Food Science and Food Safety, 15(4), 739-752.

Objectives of study

11

Blending

protein + polysaccharide

Cross linkage by TGase

Physical properties

Water content/uptake

Biodegradability

Materials-NSPC

12

Materials-Pectin (PEC)

13

Pectin

Safe

Bioactive components

carrier

Gases barrier

Polymeric

matrix

Reference: Espitia et al., (2014). Food Hydrocolloids, 35, 287-296.

Materials-TGase

14Reference: Giosafatto et al., (2020). International Journal of Molecular Sciences, 21(10), 3656.

Materials-Glycerol (GLY)

Glycerol

Cheap

Available

safe

Elasticity

non volatile

Plasticizer

15Reference : Vieira et al., (2011). European Polymer Journal, 47(3), 254-263.

Methodology

Methodology

Evaluations of films properties

Thickness and Mechanical properties

Water content

Water uptake

Biodegradable test

Preparation of films

Extraction of protein

1 2 3

NSDS Grinding for 5 min Dissolving in (DW) and stirring for

2h

pH 12 by

Centrifugation

3800 rpm for 20 minCollection of

supernatant

pH 5.4Centrifugation

3800 rpm for 20 min

Collection the pellet Drying Nigella Sativa protein

Concentrate (NSPC) 16

1. Protein extraction

2. Preparation NSPC /PEC / GLY with/without TGase

NSPC PECpH 7.5 and Stirring for

30min

Adding

GLY (30%)

Stirring for 30min

Without TGaseIncubation at 37℃ for 2 h in water

bath

Pouring on polystyrene

Petri dishes Drying Peeling of film 17

With TGase

3. Evaluation of films properties

19

Micrometer

Texture AnalyzerOven Desiccator

Water Bath

ProteaseBalance

Results and Discussion

Next

19

Blended NSPC Films in the presence or absence of different concentration of TGase

20

Thickness

According to pectin concentration, TGase concentration and both PEC and TGase; the values significantly different were respectively

reported by a b c at p<0.05.

PEC Thickness but significantly increases at concentration of PEC (60, 100 mg).

Presence of (10U TGase / g protein) significantly the thickness values.

Synergistic effect of the 3◦ blends of (40:6, 40:10 w/w) with (20U TGase/g protein) significantly in thickness.

22

Tensile Strength

22

According to pectin concentration, TGase concentration and both PEC and TGase; the values significantly different were respectively

reported by c

TS significantly to double TS of NSPC films at concentration of PEC 100mg.

TS of films significantly in presence of 10U/g protein.

TS of films significantly to 7 times of TS of NSPC films at 400mg of (PEC) in presence of 10U/g protein.

23

Elongation at Break

According to pectin concentration, TGase concentration and both PEC and TGase; the values significantly different were respectively

reported by c

EB significantly at concentration of PEC (60, 100 mg).

EB of films significantly in presence of 10U/g protein.

EB of films significantly at (60, 100mg) of (PEC) in presence of 20U/g protein compared with NSPC fims with 10U/g protein.

24

Young’s Modulus

According to pectin concentration, TGase concentration and both PEC and TGase; the values significantly different were respectively

reported by c

No significant differences in YM values with increasing PEC and TGase concentrations.

YM value significantly at concentration of PEC 400mg with low concentration of TGase.

25

Water content

Values inducated by (*) was significabtly different compared to the same film in the absence of TGase (p ≤ 0.05). 26

Water uptake

Values inducated by (*) was significabtly different compared to the same film in the absence of TGase (p ≤ 0.05). 27

Film biodegradation

Values inducated by (*) was significabtly different compared to the same film in the absence of TGase (p ≤ 0.05). 28

Conclusion

• Concentration of pectin and concentration of TGase or both have significantly role on mechanical properties of NSPC based films.

• Crosslinked NSPC/PEC(40:40 w/w) with low TGase concentration generates films with high tensile strength values.

• Crosslinked NSPC/PEC with high TGase concentration forms films with high elongation at break values except high concentration of pectin (400 mg).

• Low concentration of enzyme increases water content and uptake of films. Also, it decreases biodegradability rate that means film more resistance.

28

References

Galloway, T., Haward, M., Mason, S., Babayemi, J., Hardesty, B., Krause, S., . . . Horton, A. (2020). Science-

based solutions to plastic pollution. One Earth, 2, 5-7.

North, E. J., and Halden, R. U. (2013). Plastics and environmental health: the road ahead. Reviews on

environmental health, 28(1), 1–8.

Aguirre-Joya, J. A., De Leon-Zapata, M. A., Alvarez-Perez, O. B., Torres-León, C., Nieto-Oropeza, D. E., Ventura-Sobrevilla, J. M., . . . Ramos-Aguiñaga, M. E. (2018). Basic and applied concepts of edible packaging for foods. In Food packaging and preservation (pp. 1-61): Elsevier.

Pavlath, A. E., & Orts, W. (2009). Edible films and coatings: why, what, and how? In Edible films and coatings

for food applications (pp. 1-23): Springer.

Sabbah, M., Altamimi, M., Di Pierro, P., Schiraldi, C., Cammarota, M., & Porta, R. (2020). Black edible films from protein-containing defatted cake of Nigella sativa seeds. International Journal of Molecular Sciences, 21(3), 832.

Benbettaïeb, N., Gay, J. P., Karbowiak, T., & Debeaufort, F. (2016). Tuning the functional properties of polysaccharide–protein bio‐based edible films by chemical, enzymatic, and physical cross‐linking.

Comprehensive Reviews in Food Science and Food Safety, 15(4), 739-752.

Giosafatto, C. V. L., Fusco, A., Al-Asmar, A., & Mariniello, L. (2020). Microbial transglutaminase as a tool to improve the features of hydrocolloid-based bioplastics. International Journal of Molecular Sciences, 21(10),

3656.

Espitia, P. J. P., Du, W.-X., de Jesús Avena-Bustillos, R., Soares, N. d. F. F., & McHugh, T. H. (2014). Edible films from pectin: Physical-mechanical and antimicrobial properties-A review. Food Hydrocolloids, 35, 287-296. 30