Concentração de proteínas de soro por ultrafiltração e estudo das propriedades funcionais

Iltchenco, Sidiane; Preci, Daiane; Bonifacino, Carla; Fraguas, Eugenia Franco; Steffens, Clarice; Panizzolo, Luis Alberto; Colet, Rosicler; Fernandes, Ilizandra Aparecida; Abirached, Cecilia; Valduga, Eunice; Steffens, Juliana

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CLINIC AND SURGERY • Cienc. Rural 48 (5) • 2018 • https://doi.org/10.1590/0103-8478cr20170807 copiar

Concentração de proteínas de soro por ultrafiltração e estudo das propriedades funcionais

Autoria SCIMAGO INSTITUTIONS RANKINGS

RESUMO:

O objetivo do estudo foi avaliar o processo de Ultrafiltração (UF) na recuperação dos constituintes do soro de leite. Planejamentos fatoriais sequenciais foram realizados, variando a temperatura (5 a 40°C), a pressão (1 a 3 bar) e visando maximizar a concentração de proteínas usando membrana de 100kDa em sistema dead end. Baseados nos melhores resultados, foram realizados experimentos com de 50kDa e 10kDa. Em relação a membrana de 50kDa, a retenção de proteínas foi cerca de três vezes maior em relação a membrana de 100kDa. O concentrado obtido por membrana UF de 10kDa, 10°C e 2 bar, em escala laboratorial, mostrou uma retenção média de proteína de 80%, maior solubilidade protéica, estabilidade da emulsão e a identificação das frações β-lactoglobulins (18.3kDa) e α-lactalbumin (14.2kDa). Portanto, o uso de membranas de 100 e 50kDa são alternativas recomendáveis industrialmente à concentração de proteínas de soro de leite, e/ou como etapa anterior ao fracionamento de constituintes do soro usando membrana ≤10kDa, visando aplicações futuras em difentes áreas (alimentícia, farmacêutica, química, etc).

Palavras-chave:
Ultrafiltração; soro de leite; dead end; solubilidade; estabilidade de emulsão

Autoria

Sidiane Iltchenco

Universidade Regional Integrada do Alto Uruguai e das Missões (URI), Erechim, RS, Brasil. Universidade Regional Integrada do Alto Uruguai e das MissõesBrazilErechim, RS, BrazilUniversidade Regional Integrada do Alto Uruguai e das Missões (URI), Erechim, RS, Brasil.

Daiane Preci

Carla Bonifacino

Universidad de la Republica, 11800, Montevideo, Uruguay.Universidad de la RepublicaUruguayMontevideo, UruguayUniversidad de la Republica, 11800, Montevideo, Uruguay.

Eugenia Franco Fraguas

Universidad de la Republica, 11800, Montevideo, Uruguay.Universidad de la RepublicaUruguayMontevideo, UruguayUniversidad de la Republica, 11800, Montevideo, Uruguay.

Clarice Steffens ^*E-mail: claristeffens@uricer.edu.br. *Corresponding author.

http://orcid.org/0000-0003-4394-125X

Luis Alberto Panizzolo

Universidad de la Republica, 11800, Montevideo, Uruguay.Universidad de la RepublicaUruguayMontevideo, UruguayUniversidad de la Republica, 11800, Montevideo, Uruguay.

Rosicler Colet

Ilizandra Aparecida Fernandes

Cecilia Abirached

Universidad de la Republica, 11800, Montevideo, Uruguay.Universidad de la RepublicaUruguayMontevideo, UruguayUniversidad de la Republica, 11800, Montevideo, Uruguay.

Eunice Valduga

Juliana Steffens

http://orcid.org/0000-0001-6607-2283

E-mail: claristeffens@uricer.edu.br. ^*Corresponding author.

The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

SCIMAGO INSTITUTIONS RANKINGS

Universidad de la Republica, 11800, Montevideo, Uruguay.Universidad de la RepublicaUruguayMontevideo, UruguayUniversidad de la Republica, 11800, Montevideo, Uruguay.

Figuras | Tabelas

Figuras (5)
Tabelas (5)

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Figure 1
Contour curves for whey flow (L/m²h), obtained in the Design 1 (a) and 2 (b), respectively

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Figure 2
Permeate flows of water (“a” and “c”) and whey (“b” and “d”) as a function of time using a membrane of 100 kDa in the Design 1 and 2, respectively.

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Figure 3
Pareto chart with the estimated effect (absolute value) of acidity (°Dornic) for the concentrate (a) and permeate (b) obtained after the UF process of factorial design 2² - Design 1, respectivelly.

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Figure 4
Contour curves for acidity (a) and rejection coefficient for total solids (b) of concentrate whey obtained in UF process with a flat membrane of 100 kDa in the Design 2.

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Figure 5
Image of electrophoresis gels SDS-PAGE for UF permeate and concentrate samples of membrane of 10 kDa. From left to right: Column 1- molar mass marker; Column 2- In nature whey; Column 3- concentrated whey after UF; Column 4- permeated whey after UF.

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Table 1
Matrix of the factorial design 2² (real and coded values) - Designs 1 and 2 and values of permeate flow (Lm^-2h^-1) for water and whey using membrane of 100 kDa.

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Table 2
Matrix of the factorial design 2² (real and coded values) - Design 1 and response in terms of pH and acidity (°D) for the concentrates and permeates obtained after UF.

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Table 3
Tests the factorial design 2² (real and coded values) - Design 2 and response in terms of pH, acidity (°D), conductivity and rejection coefficient (protein, solids, lactose and minerals) for the concentrates and permeates obtained after UF.

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Table 4
In nature whey constituents, concentrate and permeate fractions and rejection (%) obtained by UF with membrane of 10kDa at 10°C and 2 bar.

Thumbnail

Table 5
Values total protein, soluble protein, K_h and K_s for in nature whey, concentrate and permeate whey by membrane of 50 and 10 kDa.

Figure 1 Contour curves for whey flow (L/m²h), obtained in the Design 1 (a) and 2 (b), respectively

Figure 2 Permeate flows of water (“a” and “c”) and whey (“b” and “d”) as a function of time using a membrane of 100 kDa in the Design 1 and 2, respectively.

Figure 3 Pareto chart with the estimated effect (absolute value) of acidity (°Dornic) for the concentrate (a) and permeate (b) obtained after the UF process of factorial design 2² - Design 1, respectivelly.

Figure 4 Contour curves for acidity (a) and rejection coefficient for total solids (b) of concentrate whey obtained in UF process with a flat membrane of 100 kDa in the Design 2.

Figure 5 Image of electrophoresis gels SDS-PAGE for UF permeate and concentrate samples of membrane of 10 kDa. From left to right: Column 1- molar mass marker; Column 2- In nature whey; Column 3- concentrated whey after UF; Column 4- permeated whey after UF.

Table 1 Matrix of the factorial design 2² (real and coded values) - Designs 1 and 2 and values of permeate flow (Lm^-2h^-1) for water and whey using membrane of 100 kDa.

Tests	------------------Independent variables^*--------------------		--------------------Permeate flow (L m^-2h^-1) ----------------
Tests	X₁	X₂	Water	Whey
----------------------------------------------------------------------------------Design 1-------------------------------------------------------------------------
1	20 (-1)	1.0 (-1)	120.33	15.41
2	40 (1)	1.0 (-1)	165.68	18.17
3	20 (-1)	3.0 (1)	389.05	23.35
4	40 (1)	3.0 (1)	431.92	24.82
5	30 (0)	2.0 (0)	440.39	22.17
6	30 (0)	2.0 (0)	433.51	22. 29
7	30 (0)	2.0 (0)	443.57	22.53
---------------------------------------------------------------------------------Design 2--------------------------------------------------------------------------
1	5 (-1)	1.0 (-1)	114.86	14.17
2	15 (1)	1.0 (-1)	170.97	15.53
3	5 (-1)	3.0 (1)	392.22	17.53
4	15 (1)	3.0 (1)	434.04	18.73
5	10 (0)	2.0 (0)	226.02	17.70
6	10 (0)	2.0 (0)	230.78	17.73
7	10 (0)	2.0 (0)	222.84	17.76

^*X₁= Temperature (°C); X₂= Pressure (bar). Fixed dependent variables: feed volume (200mL).

Table 2 Matrix of the factorial design 2² (real and coded values) - Design 1 and response in terms of pH and acidity (°D) for the concentrates and permeates obtained after UF.

Tests	---Independent variables^*----		-------------------------pH-----------------------			------------------Acidity^** (°D)---------------
Tests	X₁	X₂	Concentrate	Permeate	Concentrate	Permeate
1	20 (-1)	1.0 (-1)	5.00^b (±0.01)	5.12^a (±0.03)	27.10^a (±0.85)	22.83^b (±0.29)
2	40 (1)	1.0 (-1)	4.37^b (±0.03)	4.49^a (±0.01)	31.73^a (±0.64)	28.50^b (±0.50)
3	20 (-1)	3.0 (1)	5.14^b (±0.03)	5.20^a (±0.03)	26.30^a (±0.20)	23.03^b (±0.06)
4	40 (1)	3.0 (1)	4.40^b (±0.01)	4.51^a (±0.01)	30.33^a (±0.29)	27.67^b (±0.58)
5	30 (0)	2.0 (0)	4.66^b (±0.01)	5.01^a (±0.03)	26.43^a (±0.51)	23.17^b (±0.29)
6	30 (0)	2.0 (0)	4.65^b (±0.01)	4.98^a (±0.01)	26.50^a (±0.50)	23.50^b (±0.50)
7	30 (0)	2.0 (0)	4.64^b (±0.01)	5.0^a (±0.02)	26.33^a (±0.58)	23.33^b (±0.58)

^*X₁= Temperature (°C); X₂= Pressure (bar); ^**mean (± standard deviations) followed by same letters on lines represents no significant difference at 5% level (t student test). Fixed independent variables: feed volume (200mL), UF time of 90min.

Table 3 Tests the factorial design 2² (real and coded values) - Design 2 and response in terms of pH, acidity (°D), conductivity and rejection coefficient (protein, solids, lactose and minerals) for the concentrates and permeates obtained after UF.

Test	---------------pH^*--------------		--------Acidity^* (°D)--------			-Conductivity (mS cm^-1)^*--			----Rejection Coefficient (%)^*-----
	Concentrate	Permeate	Concentrate	Permeate	Concentrate	Permeate	P.	S.	L.	M.
1	6.41^b (±0.01)	6.50^a (±0.02)	12.33^a (±0.58)	12.17^a (±0.76)	5.12^b (±0.02)	5.22^a (±0.03)	70.33	21.79	4.72	20.00
2	6.33^b (±0.01)	6.45^a (±0.03)	13.33^a (±1.15)	12.67^a (±0.50)	5.14^b (±0.03)	5.25^a (±0.05)	64.84	19.64	5.15	18.33
3	6.42^b (±0.03)	6.50^a (±0.04)	12.33^a (±0.29)	12.33^a (±0.58)	5.13^b (±0.05)	5.23^a (±0.03)	67.03	18.04	3.43	21.67
4	6.36^b (±0.03)	6.47^a (±0.01)	13.67^a (±0.58)	13.17^a (±0.29)	5.15^b (±0.04)	5.24^a (±0.04)	64.84	17.50	3.86	20.00
5	6.38^b (±0.01)	6.46^a (±0.03)	13.00^a (±1.00)	12.33^a (±0.58)	5.14^b (±0.04)	5.26^a (±0.03)	72.53	18.93	4.72	21.67
6	6.39^b (±0.02)	6.46^a (±0.02)	12.67^a (±0.51)	12.33^a (±0.58)	5.15^b (±0.03)	5.25^a (±0.05)	70.33	18.75	4.51	23.33
7	6.36^b (±0.02)	6.44^a (±0.03)	12.67^a (±0.58)	12.00^a (±1.00)	5.13^b (±0.04)	5.24^a (±0.04)	71.43	18.75	4.08	23.33

^*mean (± standard deviations) followed by same letters on lines represents no significant difference at 5% level (t student test). Fixed independent variables: feed volume (200mL), UF time of 90min. P= protein; S=solids; lactose; M= minerals.

Table 4 In nature whey constituents, concentrate and permeate fractions and rejection (%) obtained by UF with membrane of 10kDa at 10°C and 2 bar.

Constituents	In nature	Concentrate	Permeate	Rejection (%)
Protein (g 100 mL^-1)	0.82^b±0.03	1.46^a±0.03	0.16^c ±0.02	80.48
Solids (g 100 mL^-1)	5.81^b± 0.03	6.78^a± 0.02	4.15^c ± 0.09	28.57
Lactose (g 100 mL^-1)	4.96^a± 0.01	4.41^b±0.08	4.16^c± 0.0	16.12
Minerals (g 100 mL^-1)	0.57^a± 0.04	0.48^b±0.03	0.49^b±0.02	14.04
Conductivity (mS cm^-1)	5.68^a± 0.04	5.61^a± 0.06	5.77^a ± 0.09	-
Acidity (°Dornic)	12.67^a± 0.57	13.33^a± 0.57	13.00^a ± 0.50	-
pH	6.51^c± 0.01	6.55^b± 0.01	6.58^a± 0.02	-

^*mean (± standard deviations) followed by same letters on lines represents no significant difference at 5% level (Tukey´s test).

Table 5 Values total protein, soluble protein, K_h and K_s for in nature whey, concentrate and permeate whey by membrane of 50 and 10 kDa.

Sample	Total protein (%)¹	Soluble protein (%)¹	K_h (min^-1)	K_s (min^-1)
In nature whey	12.39^b	41.39^b	(6.7±2.0).10^{-4 b}	(7.1±0.3).10^-3a
Concentrate 50 kDa	12.69^b	57.82^a	(1.1±0.1).10^-3a	(5.8±0.6).10^-3b
Concentrate 10 kDa	22.66^a	54.40^a	(7.2±0.1).10^-8c	(1.2±0.2). 10^-3c

^*means (± standard deviations) followed by same letters on lines/columns represents no significant difference at 5% level (Tukey´s test). ¹Expressed in dry basis.

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Concentração de proteínas de soro por ultrafiltração e estudo das propriedades funcionais

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[1] E-mail: claristeffens@uricer.edu.br. ^*Corresponding author.