Effects of enzymatic hydrolysis (Flavourzyme®) assisted by ultrasound in the structural and functional properties of hydrolyzates from different bovine collagens

VIDAL, Alessandra Roseline; FERREIRA, Thaísa Egiéli; MELLO, Renius de Oliveira; SCHMIDT, Michele Mantelli; KUBOTA, Ernesto Hashime; DEMIATE, Ivo Mottin; ZIELINSKI, Acácio Antonio Ferreira; DORNELLES, Rosa Cristina Prestes

doi:10.1590/fst.16717

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Research Paper • Food Sci. Technol 38 (Suppl. 1) • Dec 2018 • https://doi.org/10.1590/fst.16717 linkcopy

Effects of enzymatic hydrolysis (Flavourzyme®) assisted by ultrasound in the structural and functional properties of hydrolyzates from different bovine collagens

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

This study aimed at evaluating the functional (antioxidant and antimicrobial activity) and structural properties of different bovine collagen hydrolysates obtained through ultrasound assisted enzymatic hydrolysis (Flavourzyme®). The use of previous or simultaneous ultrasound strengthened the enzymatic hydrolysis of bovine collagen samples, generating higher number of bands in wave number (cm^-1). The best treatment was the enzymatic hydrolysis, while for the powder fiber and hydrolysate, it was the ultrasound hydrolysis. The sample that showed the highest inhibiting action to the gram-negative bacteria Salmonella choleraesuis growth and gram-positive bacteria Staphylococcus aureus hydrolyzed with enzyme and simultaneous ultrasound. Therefore, the enzymatic hydrolysis (Flavourzyme®) was concluded to enable the hydrolysis of samples, providing structural rupture and better functionality to the different bovine collagen hydrolysates.

Keywords:
antioxidant activity; antimicrobial activity; FTIR; hydrolysis degree; ultrasound

Types of collagen	Natural fiber	Powder fiber	Gelatin 1	Gelatin 2	Hydrolysate 1	Hydrolysate 2
Name/abbrev.	Fibra/FB	Fibra pó/FP	Gelita®/GEL	Rousselot®/ROU	Peptiplus®/PTL	Peptan B®/PEP
Manufacturer	Novaprom Food Ingredients Ltda (Lins – SP - Brazil)	Novaprom Food Ingredients Ltda (Lins – SP- Brazil)	Gelita do Brasil Ltda (Cotia – SP- Brazil)	Rousselot Gelatinas do Brasil Ltda (Amparo – SP- Brazil)	Gelita do Brasil Ltda (Cotia – SP- Brazil)	Rousselot Gelatinas do Brasil Ltda (Amparo – SP- Brazil)
Description	Particle size between 1.80 and 1.92 mm, raw material: bovine skin, alkaline treatment, pH between 7 and 9.5	Particle size between 0.45 and 0.57 mm, raw material: bovine skin, alkaline treatment, pH between 7 and 9.5	Raw material: leather or other sources, partial alkaline hydrolysis (severe treatment), 236 g Bloom; 40 mP viscosity; 11% humidity; pH 5.6; <= 2% ash	Raw material: leather or other sources, partial alkaline hydrolysis (severe treatment), 250g Bloom; 3.8 mP viscosity; 12.4% humidity; pH 5.5; <=2% ash	Raw material: bovine leather/ swine skin, chemical + enzymatic hydrolysis, 39 mP viscosity; 5.9% humidity; 92.1% protein; pH 5.7; <=2% ash	Raw material: bovine leather/ swine skin, chemical + enzymatic hydrolysis, 4 mP viscosity; 7.2% humidity; 92.15% protein; pH 6; 0.65% ash

Abbr.	Description of treatments	Enzyme	Ultrasound	T °C	Time (h)	pH
CC	Crude collagens	No	No	50	2	7.0
HU	Ultrasound hydrolysis	No	Yes	50	2	7.0
EH	Enzymatic hydrolysis	Yes	No	50	2	7.0
EHU	Enzymatic Hydrolysis with simultaneous ultrasound	Yes	Yes	50	2	7.0
UEH	Previous ultrasound followed by enzymatic hydrolysis	Yes	Yes	50	4	7.0

	Ultrasound
	Previous	Simultaneous
Protein, mg/mL	7.89^a	4.33^b
Hydrolysis Degree, %	0.00^b	40.80^a
Antioxidant activity, %	33.63^a	38.01^a

Effects	Ptn	DH	AA
Collagen	0.0001	0.0001	0.0001
Enzyme	0.0001	0.0001	0.0001
Ultrasound	0.0001	0.0001	0.4639
Enzyme × Ultrasound	0.0001	0.0001	0.0014
Collagen × Enzyme	0.0001	0.0001	0.0001
Collagen × Ultrasound	0.0120	0.0007	0.0063
Collagen × Enzyme × Ultrasound	0.0001	0.0001	0.0670
Average	6.19	16.43	32.68
VC	2.14	1.32	22.94

Collagen **	Enzyme, %	Sim.US*		Enzyme average	Collagen average	Sim.US^*		Enzyme average	Collagen average	Sim.US^*		Enzyme average	Collagen average
		without	with	Enzyme average	Collagen average	without	with	Enzyme average	Collagen average	without	with	Enzyme average	Collagen average
		Protein, mg/mL				Hydrolysis degree, %				Antioxidant activity, %
FB	0	7.49^aA	7.54^aA	7.52	6.22^B	0.00^aB	0.00^aB	0.00	17.3^A	5.32^bB	20.0^aB	12.7	28.6^B
FB	8	5.66^aB	4.17^bB	4.92	6.22^B	24.6^bA	44.5^aA	34.5	17.3^A	43.9^aA	45.3^aA	44.6	28.6^B
Ultrasound average		6.58	5.86			12.3	22.2			24.6	32.7
FP	0	8.64^aA	8.22^bA	8.43	7.37^A	0.00^bB	5.68^aB	2.84	17.0^AB	15.8^bA	28.7^aA	22.2	20.9^B
FP	8	6.75^aB	5.88^bB	6.32	7.37^A	25.4^bA	37.0^aA	31.2	17.0^AB	24.7^aA	14.3^aB	19.5	20.9^B
Ultrasound average		7.69	7.05			12.7	21.3			20.2	21.5
PEP	0	6.84^aA	6.72^aA	6.78	5.65^C	0.00^aB	1.98^aB	0.99	16.0^B	35.8^aA	45.2^aA	40.5	42.0^A
PEP	8	5.05^aB	4.00^bB	4.53	5.65^C	23.9^bA	38.0^aA	30.9	16.0^B	40.2^aA	46.8^aA	43.5	42.0^A
Ultrasound average		5.95	5.36			11.9	20.0			38.0	46.0
PTL	0	6.41^aA	6.44^aA	6.42	5.39^D	0.00^aB	0.00^aB	0.03	13.7^C	43.1^aA	30.0^bB	36.5	43.6^A
PTL	8	4.80^aB	3.93^bB	4.36	5.39^D	21.6^bA	33.3^aA	27.4	13.7^C	52.2^aA	47.8^aA	50.0	43.6^A
Ultrasound average		5.61	5.18			10.8	16.7			47.6	38.9
GEL	0	7.46^aA	7.60^aA	7.53	6.16^B	0.00^aB	0.00^aB	0.00	17.9^A	21.4^aB	19.8^aA	20.6	29.4^B
GEL	8	5.70^aB	3.89^bB	4.79	6.16^B	23.6^bA	47.9^aA	35.8	17.9^A	47.2^aA	29.2^bA	38.2	29.4^B
Ultrasound average		6.58	5.75			11.8	23.9			34.3	24.5
ROU	0	7.54^aA	7.49^aA	7.52	6.24^B	0.00^aB	0.61^aB	0.31	17.4^A	2.8^bB	25.6^aB	14.2	29.0^B
ROU	8	5.69^aB	4.25^bB	4.97	6.24^B	24.8^bA	44.1^aA	34.5	17.4^A	46.0^aA	41.6^aA	43.8	29.0^B
Ultrasound average		6.61	5.87			12.4	22.4			24.4	33.6
Ultrasound general average		6.50	5.84			12.0	21.1			31.5	32.9

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[1] Source: Rousselot (2014) ; Gelita (2014) .