Application of Enzymes in Sunflower Oil Extraction: Antioxidant Capacity and Lipophilic Bioactive Composition

Ribeiro, Suellen A. O.; Nicacio, Antonio E.; Zanqui, Ana B.; Biondo, Polyana B. F.; Abreu-Filho, Benício A. de; Visentainer, Jesui V.; Gomes, Sandra T. M.; Matsushita, Makoto

doi:10.5935/0103-5053.20150335

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Articles • J. Braz. Chem. Soc. 27 (5) • May 2016 • https://doi.org/10.5935/0103-5053.20150335 copy

Application of Enzymes in Sunflower Oil Extraction: Antioxidant Capacity and Lipophilic Bioactive Composition

Authorship SCIMAGO INSTITUTIONS RANKINGS

A 2³ factorial design with center point in triplicate was used to evaluate the application of different enzymes in extraction of sunflower oil, comparing its quality with the oil obtained by conventional methods (solvent and pressing) in relation to antioxidant capacity, phytosterol and tocopherol contents, and fatty acid composition. Three enzymes were used; Pectinex Ultra SPL, Celluclast 1.5 L and Alcalase 2.4 L; and three factors were evaluated: enzyme concentration, sample/water ratio and extraction time. Enzymatic aqueous extraction produced oil with better quality in relation to some of the evaluated parameters: highest antioxidant capacity against peroxyl radicals (371 µmol Trolox g^-1 of oil), total phytosterols (183 mg 100 g^-1 of oil) and omega-3 fatty acid content, as well the lowest content of saturated fatty acids. The application of enzymes in sunflower oil extraction represents an environmentally friendly methodology, free of toxic solvent residues and providing a final product of high quality.

Keywords:
oilseed; clean method; enzymes; lipids; antioxidant capacity

Factors	Symbol	Unit	Type	Level
Factors	Symbol	Unit	Type	−1	0	+1
Enzymes	X₁	%	numeric	6	8	10
Sample/Water	X₂	g mL^-1	numeric	1/6	1/8	1/10
time	X₃	hours	numeric	4	6	8

Run	Coded variable			Actual variable
Run	X₁ ^a a Enzyme concentration;	X₂ ^b b sample/water ratio;	X₃ ^c c extraction time;	X₁ ^a a Enzyme concentration; / %	X₂ ^b b sample/water ratio; / (g mL^-1)	X₃ ^c c extraction time; / h	Y ^d d enzymatic aqueous extraction yield. / %
1	−1	−1	−1	6	1/6	4	27.4
2	+1	−1	−1	10	1/6	4	29.7
3	−1	+1	−1	6	1/10	4	22.3
4	+1	+1	−1	10	1/10	4	23.1
5	−1	−1	+1	6	1/6	8	34.7
6	+1	−1	+1	10	1/6	8	36.6
7	−1	+1	+1	6	1/10	8	29.4
8	+1	+1	+1	10	1/10	8	28.3
9	0	0	0	8	1/8	6	27.6
10	0	0	0	8	1/8	6	27.3
11	0	0	0	8	1/8	6	27.3

Source	Degrees of freedom	Sum of squares	Mean square	F-value	p-value
X₁^a a Enzyme concentration;	1	1.78	1.78	5.60	0.0643
X₂^b b sample/water ratio;	1	79.70	79.70	251.02	< 0.0001
X₃^c c extraction time.	1	87.98	87.98	277.11	< 0.0001
X₁X₂	1	2.57	2.57	8.08	0.0361
Curvature	1	5.15	5.15	16.22	0.0100
Residual	5	1.59	0.32	−	−
Lack of fit	3	1.51	0.50	13.29	0.0708
Pure error	2	0.08	0.04	−	−
Total	10	178.75	−	−	−

Method	Yield / %
Solvent	55.1^a a Maximum extraction yield obtained from the factorial design. Mean (n = 3) ± standard deviation. Different letters in the same column indicate significant difference by Tukey’s test (p < 0.05). ± 0.7
Pressing	36.8^b ± 0.5
Enzymatic^a a Maximum extraction yield obtained from the factorial design. Mean (n = 3) ± standard deviation. Different letters in the same column indicate significant difference by Tukey’s test (p < 0.05).	36.6^b ± 0.6

Method	Assay / (μmol Trolox g^-1 of oil)
Method	DPPH	L-ORAC
Solvent	86.71^b ± 3.54	225.71^b ± 2.68
Pressing	106.18^a ± 3.89	181.94^c ± 11.11
Enzymatic	87.98^b ± 3.89	371.53^a ± 12.65

Method	α-Tocopherol / (mg 100 g⁻¹)	Phytosterol / (mg 100 g⁻¹)			Total phytosterol / (mg 100 g⁻¹)
Method	α-Tocopherol / (mg 100 g⁻¹)	Campesterol	Stigmasterol	Sitosterol	Total phytosterol / (mg 100 g⁻¹)
Solvent	48.46^a ± 2.38	17.17^a ± 2.16	19.75^a ± 1.27	130.03^b ± 3.75	166.95^b ± 3.65
Pressing	48.69^a ± 5.43	17.29^a ± 3.31	17.16^a ± 2.06	121.06^b ± 1.29	155.52^b ± 4.01
Enzymatic	40.84^b ± 0.17	19.85^a ± 0.99	19.34^a ± 1.30	143.81^a ± 6.70	183.00^a ± 6.90

Fatty acid	Method / (mg g⁻¹ total lipids)
Fatty acid	Solvent	Pressing	Enzymatic
14:0	0.667^a ± 0.005	0.666^a ± 0.016	0.632^a ± 0.028
15:0	0.118^b ± 0.002	0.133^a ± 0.003	0.107^c ± 0.009
16:0	52.836^a ± 0.246	49.999^c ± 0.427	51.070^b ± 0.382
16:1n-7	1.207^b ± 0.037	1.347^a ± 0.030	1.240^b ± 0.016
17:0	0.387^b ± 0.002	0.453^a ± 0.033	0.444^a ± 0.015
17:1n-7	0.384^a ± 0.014	0.411^a ± 0.037	0.387^a ± 0.022
18:0	31.612^a ± 0.600	30.338^a ± 0.255	30.628^a ± 0.645
18:1n-9	561.817^a ± 4.316	560.623^a ± 1.572	559.393^a ± 1.948
18:1n-7	3.611^b ± 0.121	4.322^a ± 0.165	4.293^a ± 0.160
18:2n-6	294.466^a ± 6.095	299.332^a ± 2.012	301.753^a ± 3.353
18:3n-3	0.246^a ± 0.009	0.259^a ± 0.009	0.226^b ± 0.007
20:0	1.811^b ± 0.064	1.892^b ± 0.029	2.102^a ± 0.029
20:1n-9	1.362^a ± 0.039	1.383^a ± 0.047	1.477^a ± 0.064
20:4n-6	6.755^a ± 0.099	6.206^b ± 0.051	6.181^b ± 0.143
20:5n-3	0.194^c ± 0.008	0.296^b ± 0.015	0.343^a ± 0.011
24:0	2.150^a ± 0.082	1.944^ab ± 0.055	1.988^b ± 0.074
	Summations and ratios / (mg g^-1 total lipids)
SFA	89.582^a ± 0.655	85.425^b ± 0.500	86.971^b ± 0.755
MUFA	568.382^a ± 4.317	568.087^a ± 1.577	566.789^a ± 1.955
PUFA	301.661^a ± 6.096	306.093^a ± 2.013	308.502^a ± 3.356
n-6	301.222^a ± 6.096	305.538^a ± 2.013	307.934^a ± 3.356
n-3	0.440^b ± 0.010	0.555^a ± 0.016	0.568^a ± 0.009
n-6/n-3	685.312^a ± 21.287	551.134^b ± 16.674	542.011^b ± 10.495
PUFA/SFA	3.368^b ± 0.072	3.583^a ± 0.032	3.548^a ± 0.049

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