RICE PARBOILING WASTEWATER IN THE MAXIMIZATION OF CAROTENOIDS
               BIOPRODUCTION BY <i>Phaffia rhodozyma</i>

The interest in carotenoid production from natural sources has increased based in their possible use as dyes and their powerful biological antioxidant capacity. This study evaluated the use of rice parboiling wastewater (RPW) as an alternative substrate for the bioproduction of carotenoids using the yeast Phaffia rhodozyma and found it to be promising as the only source of nutrients, reaching a concentration of 0.6 μg mL^-1 (259.1 μg g^-1) in 48 h. To increase the potential use of this industrial effluent, a study of supplementation was carried out to enhance the production of carotenoids using the methodology of experimental design (a 2_IV ^6-2fractional factorial design sequenced by a 2⁴ central composite design). The conditions for maximizing the production of carotenoids were (g L^-1) malt extract (16.25), peptone (8.75), sucrose (15) and rice parboiling wastewater (87.5), with a pH of 5 at 25 °C and 150 rpm for 144 h, which produced a concentration 5.3 μg mL^-1 (628.8 μg g^-1).

Biodye; submerged cultive; response surface methodology; yeast

Authorship

Danielle Alves da Silva Rios

Universidade Federal do Rio Grande/FURG - Rio Grande - RS - Brasil Universidade Federal do Rio Grande/FURGBrazilRio Grande, RS, BrazilUniversidade Federal do Rio Grande/FURG - Rio Grande - RS - Brasil

Thais de Matos de Borba

Susana Juliano Kalil

Janaína Fernandes de Medeiros Burkert ^**Autor para correspondência: Janaína Fernandes de Medeiros Burkert, email: jfmb@vetorial.net

Universidade Federal do Rio Grande/FURG - Cx. P. 474 - 96201-900 - Rio Grande - RS - Brasil - jfmb@vetorial.net Universidade Federal do Rio Grande/FURGBrazilRio Grande, RS, BrazilUniversidade Federal do Rio Grande/FURG - Cx. P. 474 - 96201-900 - Rio Grande - RS - Brasil - jfmb@vetorial.net

*Autor para correspondência: Janaína Fernandes de Medeiros Burkert, email: jfmb@vetorial.net

SCIMAGO INSTITUTIONS RANKINGS

Figures | Tables | Formulas

Figures (4)
Tables (4)
Formulas (2)

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Figure 1:
Cultivation with rice parboiling wastewater (■ biomass, ● pH, ▲ volumetric carotenoids, ♦ specific carotenoids) at 25 ºC and 150 rpm (n = 3).

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Figure 2:
Main effects of carotenoids production for the responses biomass concentration (g L^-1) (A), specific production (μg g^-1) (B) and volumetric production of carotenoids (μg mL^-1) (C). Y.E.: yeast extract, M.E.: malt extract, P: peptone, S: sucrose and W: rice parboiling wastewater (*p < 0.1 **p < 0.15).

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Figure 3:
Contour diagrams for volumetric carotenoids production (μg mL^-1) as a function of malt extract and peptone concentration (A), malt extract (g L^-1) and sucrose (g L^-1) (B), malt extract (g L^-1) and rice parboiling wastewater (g L^-1), peptone (g L^-1) and sucrose (g L^-1) and rice parboiling wastewater (g L^-1) (E), sucrose (g L^-1) and rice parboiling wastewater (g L^-1) (F).

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Table 1:
Levels and concentrations of the variables studied at the factorial designs.

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Table 2:
Partial characterization of substrates of culture media.

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Table 3:
ANOVA and Tukey's test for the different yeast extract concentrations (n = 3).

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Table 4:
2⁴ Central Composite Design with real values of the variables (X_n), C:N ratio and the maximum responses (Y_n) of biomass concentration, specific production of carotenoids, volumetric production of carotenoids and relative desviation (%) and residual sucrose (%).

Thumbnail

Figure 1: Cultivation with rice parboiling wastewater (■ biomass, ● pH, ▲ volumetric carotenoids, ♦ specific carotenoids) at 25 ºC and 150 rpm (n = 3).

Figure 2: Main effects of carotenoids production for the responses biomass concentration (g L^-1) (A), specific production (μg g^-1) (B) and volumetric production of carotenoids (μg mL^-1) (C). Y.E.: yeast extract, M.E.: malt extract, P: peptone, S: sucrose and W: rice parboiling wastewater (*p < 0.1 **p < 0.15).

Figure 3: Contour diagrams for volumetric carotenoids production (μg mL^-1) as a function of malt extract and peptone concentration (A), malt extract (g L^-1) and sucrose (g L^-1) (B), malt extract (g L^-1) and rice parboiling wastewater (g L^-1), peptone (g L^-1) and sucrose (g L^-1) and rice parboiling wastewater (g L^-1) (E), sucrose (g L^-1) and rice parboiling wastewater (g L^-1) (F).

Table 1: Levels and concentrations of the variables studied at the factorial designs.

Variables	Level -1 (g L^-1)	Level 0 (g L^-1)	Level +1 (g L^-1)
	2_IV ^6-2fractional factorial
Yeast Extract	1	5.5	10
Malt Extract	1	5.5	10
Peptone	1	5.5	10
Sucrose	5	12.5	20
RPW*	0	90	180
pH	4	5	6
	CCD 2⁴
Malt Extract	8.75	12.5	16.25
Peptone	8.75	12.5	16.25
Sucrose	15	25	35
RPW*	87.5	125	162.5

*RPW (rice parboiling wastewater).

Table 2: Partial characterization of substrates of culture media.

Substrates	Carbon (%)	Hydrogen (%)	Nitrogen (%)	Total sugars (mg L^-1)	Total soluble solids (g L^-1)	Phosphorus (mg L^-1)
Yeast extract	38.40	5.65	10.67	ND	ND	ND
Malt extract	39.24	6.54	10.93	ND	ND	ND
Peptone	43.47	6.71	14.47	ND	ND	ND
RPW*	0.16	2.25	3.02	67.0	3.97	30.7

*RPW (rice parboiling wastewater); ND (not determined).

Table 3: ANOVA and Tukey's test for the different yeast extract concentrations (n = 3).

YE (g/L)	Y₁ (g L^-1)	Y₂ (μg g^-1)	Y₃ (μg mL^-1)
0	9.3 ± 0.1^a	359.5 ± 23.1^a	3.0 ± 0.1^a
0.5	8.2 ± 0.3^b	385.9 ± 5.6^a	3.0 ± 0.1^a
1	8.8 ± 0.2^ab	345.4 ± 31.4^a	2.8 ± 0.2^a

Means with different superscript letters within the same column are significantly different (p < 0.05). YE &#091;yeast extract (g L^-1)&#093;; Y₁ &#091;biomass (g L^-1)&#093;; Y₂ &#091;specific carotenoids (μg g^-1)&#093;; Y₃ &#091;volumetric carotenoids (μg mL^-1)&#093;.

Table 4: 2⁴ Central Composite Design with real values of the variables (X_n), C:N ratio and the maximum responses (Y_n) of biomass concentration, specific production of carotenoids, volumetric production of carotenoids and relative desviation (%) and residual sucrose (%).

Trial	X₁	X₂	X₃	X₄	C:N	Y₁	Y₂	Y₃	Y₄	Y₅
1	8.75	8.75	15	87.5	8.2	7.6	504.0	3.4	2.9	6.2
2	16.25	8.75	15	87.5	7.6	9.7	628.8	5.3	-1.9	7.8
3	8.75	16.25	15	87.5	6.0	7.1	370.5	2.4	-4.2	6.4
4	16.25	16.25	15	87.5	5.8	8.6	576.5	5.0	2.0	17.3
5	8.75	8.75	35	87.5	15	10.1	336.0	3.1	0.0	4.4
6	16.25	8.75	35	87.5	13.9	10.0	502.4	5.0	13.6	18.4
7	8.75	16.25	35	87.5	9.8	9.3	312.2	2.7	7.4	11.9
8	16.25	16.25	35	87.5	9.5	9.4	530.1	3.9	0.0	20.1
9	8.75	8.75	15	162.5	8.3	6.7	593.6	4.0	7.5	5.0
10	16.25	8.75	15	162.5	7.8	7.2	540.8	3.9	0.0	4.6
11	8.75	16.25	15	162.5	6.1	7.3	420.0	3.0	16.7	12.9
12	16.25	16.25	15	162.5	5.9	8.4	491.6	3.9	15.4	6.3
13	8.75	8.75	35	162.5	15.1	10.4	379.5	3.6	2.8	4.0
14	16.25	8.75	35	162.5	14.1	10.9	355.3	2.9	0.0	11.9
15	8.75	16.25	35	162.5	9.9	9.0	485.3	3.5	11.4	8.2
16	16.25	16.25	35	162.5	9.5	7.9	399.6	2.3	0.0	9.2
17	12.50	12.50	25	125	9.1	8.6	402.6	2.6	-20.7	3.1
18	12.50	12.50	25	125	9.1	8.0	424.2	2.7	-29.6	5.4
19	12.50	12.50	25	125	9.1	9.6	351.9	2.8	-25.0	3.2

X₁ &#091;malt extract (g L^-1)&#093;; X₂ &#091;peptone (g L^-1)&#093;; X₃ &#091;sucrose (g/L)&#093;; X₄ &#091;RPW (g L^-1)&#093;; Y₁ &#091;biomass (g L^-1)&#093;; Y₂ &#091;specific carotenoids (μg g^-1)&#093;; Y₃ &#091;volumetric carotenoids (μg mL^-1)&#093;; Y₄ &#091;Relative desviation (%)&#093;; Y₅ &#091;Residual Sucrose (%)&#093;.

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RICE PARBOILING WASTEWATER IN THE MAXIMIZATION OF CAROTENOIDS BIOPRODUCTION BY <i>Phaffia rhodozyma</i>

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[1] *Autor para correspondência: Janaína Fernandes de Medeiros Burkert, email: jfmb@vetorial.net