STATISTICAL OPTIMIZATION OF MINERAL SALT AND UREA CONCENTRATION FOR CELLULASE AND XYLANASE PRODUCTION BY <i>Penicillium echinulatum</i> IN SUBMERGED FERMENTATION

Reis, L. dos; Ritter, C. E. T.; Fontana, R. C.; Camassola, M.; Dillon, A. J. P.

doi:10.1590/0104-6632.20150321s00003099

Acessibilidade / Reportar erro

Brasil

Brazilian Journal of Chemical Engineering

Español English

Brasil

Español English

sumário « anterior atual seguinte »

Sumário

BIOPROCESS ENGINEERING • Braz. J. Chem. Eng. 32 (1) • Jan-Mar 2015 • https://doi.org/10.1590/0104-6632.20150321s00003099 copy

STATISTICAL OPTIMIZATION OF MINERAL SALT AND UREA CONCENTRATION FOR CELLULASE AND XYLANASE PRODUCTION BY Penicillium echinulatum IN SUBMERGED FERMENTATION

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

Penicillium echinulatum S1M29 is a mutant with cellulase and xylanase production comparable to the most studied microorganisms in the literature. However, its potential to produce these enzymes has not been fully investigated. This study aimed at optimizing salt and urea concentrations in the mineral solution, employing the response surface methodology. A 2^5-1 Fractional Factorial Design and a 2³ Central Composite Design were applied to elucidate the effect of salts and urea in enzyme production. Lower concentrations of KH₂PO₄ (2.0 g.L^-1), (NH₄)₂SO₄ (1.4 g.L^-1), MgSO₄.₇H₂O (0.375 g.L^-1) and CaCl₂ (0.375 g.L^-1) were most suitable for the production of all enzymes evaluated. Nevertheless, higher concentrations of urea (0.525 g.L^-1) gave the best results for cellulase and xylanase production. The maximum FPase (1,5 U.m.L^-1), endoglucanase (7,2 U.m.L^-1), xylanase (30,5 U.m.L^-1) and β-glucosidase (4,0 U.m.L^-1) activities obtained with the planned medium were, respectively, 87, 16, 17 and 21% higher when compared to standard medium. The experimental design contributed to adjust the concentrations of minerals and urea of the culture media for cellulase and xylanase production by P. echinulatum, avoiding waste of components in the medium.

Cellulolytic enzyme; Experimental design; Medium composition; Shake flask

Run	Experimental factors					Cellulases (U.mL^-1)			Xylanases (U.mL^-1)
Run	X1	X2	X3	X4	X5	FPase	β-glucosidades	Endoglucanases	Xylanases (U.mL^-1)
1	(-1) 1.4	(-1) 0.3	(-1) 2.0	(-1) 0.3	(+1) 0.6	0.89	2.46	5.52	34.35
2	(+1) 2.8	(-1) 0.3	(-1) 2.0	(-1) 0.3	(-1) 0.3	0.73	1.08	5.21	31.06
3	(-1) 1.4	(+1) 0.6	(-1)2.0	(-1) 0.3	(-1) 0.3	0.7	1.38	4.63	30.6
4	(+1) 2.8	(+1) 0.6	(-1) 2.0	(-1) 0.3	(+1) 0.6	0.77	2.5	6.35	32.33
5	(-1) 1.4	(-1) 0.3	(+1) 4.0	(-1) 0.3	(-1) 0.3	0.8	1.89	5.1	30.3
6	(+1) 2.8	(-1) 0.3	(+1) 4.0	(-1) 0.3	(+1) 0.6	0.81	2.06	5.29	33.04
7	(-1) 1.4	(+1) 0.6	(+1) 4.0	(-1) 0.3	(+1) 0.6	0.69	2.04	4.71	30.17
8	(+1) 2.8	(+1) 0.6	(+1) 4.0	(-1) 0.3	(-1) 0.3	0.78	1.35	4.69	26.85
9	(-1) 1.4	(-1) 0.3	(-1) 2.0	(+1) 0.6	(-1) 0.3	0.67	1.74	4.74	32.19
10	(+1) 2.8	(-1) 0.3	(-1)2.0	(+1) 0.6	(+1) 0.6	0.71	2.52	4.95	36.16
11	(-1) 1.4	(+1) 0.6	(-1) 2.0	(+1) 0.6	(+1) 0.6	0.98	2.51	4.37	34.64
12	(+1) 2.8	(+1) 0.6	(+1) 4.0	(+1) 0.6	(-1) 0.3	0.6	2.15	3.95	28.89
13	(-1) 1.4	(-1) 0.3	(+1) 4.0	(+1) 0.6	(+1) 0.6	0.66	2.15	4.86	30.98
14	(+1) 2.8	(-1) 0.3	(+1) 4.0	(+1) 0.6	(-1) 0.3	0.69	1.74	4.41	29.26
15	(-1) 1.4	(+1) 0.6	(+1) 4.0	(+1) 0.6	(-1) 0.3	0.68	1.73	4.54	27.76
16	(+1) 2.8	(+1) 0.6	(+1) 4.0	(+1) 0.6	(+1) 0.6	0.68	1.55	3.79	28.91
17	(0) 2.1	(0) 0.45	(0) 3.0	(0) 0.45	(0) 0.45	0.73	1.49	5.77	31.82
18	(0) 2.1	(0) 0.45	(0) 3.0	(0) 0.45	(0) 0.45	0.76	1.84	5.1	33.59
19	(0) 2.1	(0) 0.45	(0) 3.0	(0) 0.45	(0) 0.45	0.76	1.71	5.5	31.01

Run	Experimental factors			Cellulases (U.mL^-1)						Xylanases (U.mL^-1)
	X₁	X₂	X₃	FPase		β-glucosidades		Endoglucanases		Xylanases (U.mL^-1)
	X₁	X₂	X₃	Observed	Predicted	Observed	Predicted	Observed	Predicted	Observed	Predicted
1	(-1) 0.375	(-1) 0.375	(-1) 0.375	1.08	1.03	1.42	1.28	8.44	7.36	36.38	34.3
2	(-1) 0.375	(-1) 0.375	(+1) 0.525	1.09	0.97	1.51	1.9	9.06	8.35	36.62	33.15
3	(+1) 0.525	(-1) 0.375	(-1) 0.375	1.07	0.91	1.3	1.53	7.37	6.36	35.68	34.2
4	(+1) 0.525	(-1) 0.375	(+1) 0.525	1.01	0.87	1.57	1.23	6.59	6.64	34.67	33.28
5	(-1) 0.375	(+1) 0.525	(-1) 0.375	0.9	0.8	0.93	1.45	7.49	6.33	34.2	32.4
6	(-1) 0.375	(+1) 0.525	(+1) 0.525	1.11	1.04	2.6	1.75	6.87	6.89	37.63	35.92
7	(+1) 0.525	(+1) 0.525	(-1) 0.375	0.85	0.73	1.25	1.71	6.52	6.23	27.92	28.22
8	(+1) 0.525	(+1) 0.525	(+1) 0.525	1.17	0.99	0.74	1.06	6.13	6.19	33.02	31.93
9	(0) 0.45	(0) 0.45	(-2) 0.30	0.66	0.75	1.41	1.16	4.87	5.71	30.02	30.94
10	(0) 0.45	(0) 0.45	(+2) 0.60	0.83	0.96	1.05	1.11	6.93	6.65	31.27	33.51
11	(-2) 0.30	(0) 0.45	(0) 0.45	1.07	1.12	2.05	1.91	7.76	8.71	32.32	35.25
12	(+2) 0.60	(0) 0.45	(0) 0.45	0.77	0.94	1.57	1.52	6.98	7.01	30.95	31.18
13	(0) 0.45	(-2) 0.30	(0) 0.45	0.92	1.03	1.51	1.38	6.74	7.55	32.72	35.33
14	(0) 0.45	(+2) 0.60	(0) 0.45	0.79	0.91	1.45	1.38	5.79	5.96	31.57	32.11
15	(0) 0.45	(0) 0.45	(0) 0.45	0.74	0.82	1.82	1.72	6.88	6.42	30.43	32.56
16	(0) 0.45	(0) 0.45	(0) 0.45	0.89	0.82	1.52	1.72	5.45	6.43	33.18	32.56
17	(0) 0.45	(0) 0.45	(0) 0.45	0.75	0.82	1.86	1.72	6.48	6.42	33.04	32.56
18	(0) 0.45	(0) 0.45	(0) 0.45	0.76	0.82	1.81	1.72	6.8	6.43	30.83	32.56
19	(0) 0.45	(0) 0.45	(0) 0.45	0.86	0.82	1.62	1.72	5.35	6.43	33.11	32.56
20	(0) 0.45	(0) 0.45	(0) 0.45	0.79	0.9	1.92	1.72	5.85	6.75	32.19	32.87

	Interception	(NH₄)₂SO₄	CaCl₂	KH₂PO₄	MgSO₄	Urea	p-level
FPase Xylanases Endoglucanases β-glucosidases	0.858 34.2 6.50 1.13	-0.002 -0.0138 0.004 -0.007	-0.004 -0.59 -0.11 -0.0114	-0.002 -0.147 -0.0188 -0.0049	-0.019 0.127 -0.231 0.059	0.021 0.863 0.091 0.193	0.2690 0.0004^* * Significant factors, P<0.05 0.0358^* * Significant factors, P<0.05 0.0814

	Xylanases		Endoglucanases		FPase		β-glucosidases
Factor	Coefficient	p-level	Coefficient	p-level	Coefficient	p-level	Coefficient	p-level
Mean	33.13	0.0006^* * Significant factors, P<0.05	6.75	1.4.10^-9 ^* * Significant factors, P<0.05	0.82	2.10^-8 ^* * Significant factors, P<0.05	1.52	3.2.10^-10 ^* * Significant factors, P<0.05
X₁ (L)	-1.02	0.042^* * Significant factors, P<0.05	-0.426	0.05^* * Significant factors, P<0.05	-0.043	0.034^* * Significant factors, P<0.05	-0.097	0.021^* * Significant factors, P<0.05
X₁ (Q)	-1.02	0.03^* * Significant factors, P<0.05	-0.43	0.05^* * Significant factors, P<0.05	-0.04	0.035^* * Significant factors, P<0.05	-0.1	0.022^* * Significant factors, P<0.05
X₂ (L)	-0.8	0.0913	-0.396	0.07	-0.03	0.107	-3.10^-7	1
X₂ (Q)	-0.8	0.069	-0.4	0.23	-0.03	0.108	0	1
X₃ (L)	0.641	0.163	0.235	0.24	0.051	0.016^* * Significant factors, P<0.05	-0.013	0.716
X₃ (Q)	0.641	0.132	0.235	0.237	0.051	0.016^* * Significant factors, P<0.05	-0.01	0.717
X₁X₂	-1.03	0.094	0.228	0.406	0.013	0.603	0.008	0.877
X₁X₃	0.053	0.924	-0.15	0.577	0.005	0.834	-0.23	0.002^* * Significant factors, P<0.05
X₂X₃	1.163	0.065	-0.11	0.688	0.0073	0.016^* * Significant factors, P<0.05	-0.09	0.089

	MS	MS	MS	MS
	Standard	2.0	1.5	Planned
KH₂PO₄ (g.L^-1)	2.0	4.0	3.0	2.0
(NH₄) ₂SO₄ (g.L^-1)	1.4	2.8	2.1	1.4
MgSO₄.7H₂O (g.L^-1)	0.3	0.6	0.45	0.375
CaCl₂ (g.L^-1)	0.3	0.6	0.45	0.375
Urea (g.L^-1)	0.3	0.6	0.45	0.525
FPase (U.mL^-1)	0.83^b	1.28^ab	1.24^ab	1.45^a
Xylanase (U.mL^-1)	25.4^b	29^a	26.2^b	30.5^a
Endoglucanase (U.mL^-1)	5.8^b	7.3^a	7.3^a	6.7^ab
β-glucosidase (U.mL^-1)	2.95^a	3.4^a	3.1^a	3.2^a

Brazilian Society of Chemical Engineering Rua Líbero Badaró, 152 , 11. and., 01008-903 São Paulo SP Brazil, Tel.: +55 11 3107-8747, Fax.: +55 11 3104-4649, Fax: +55 11 3104-4649 - São Paulo - SP - Brazil
E-mail: rgiudici@usp.br

Acompanhe os números deste periódico no seu leitor de RSS

[1] *To whom correspondence should be addressed