PRODUCTION AND CHARACTERIZATION OF THERMOPHILIC CARBOXYMETHYL CELLULASE SYNTHESIZED BY <i>Bacillus</i> sp. GROWING ON SUGARCANE BAGASSE IN SUBMERGED FERMENTATION

Padilha, I. Q. M.; Carvalho, L. C. T.; Dias, P. V. S.; Grisi, T. C. S. L.; Silva, F. L. Honorato da; Santos, S. F. M.; Araújo, D. A. M.

doi:10.1590/0104-6632.20150321s00003298

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Brazilian Journal of Chemical Engineering

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BIOPROCESS ENGINEERING • Braz. J. Chem. Eng. 32 (1) • Jan-Mar 2015 • https://doi.org/10.1590/0104-6632.20150321s00003298 copy

PRODUCTION AND CHARACTERIZATION OF THERMOPHILIC CARBOXYMETHYL CELLULASE SYNTHESIZED BY Bacillus sp. GROWING ON SUGARCANE BAGASSE IN SUBMERGED FERMENTATION

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

The production and characterization of cellulase from thermophilic strain Bacillus sp. C1AC5507 was studied. For enzyme production, sugarcane bagasse was used as carbon source. The produced carboxymethyl cellulase (CMCase) had a molecular weight around 55 kDa and its activity varied between 0.14 and 0.37 IU mL^-1 in conditions predicted by Response Surface Methodology. The optimum temperature and pH for the CMCase production were 70 °C and 7.0, respectively. The enzyme activity was inhibited mostly by Cu⁺² and activated mostly by Co⁺², Mn²⁺, Ca⁺² and Fe⁺³. Our findings provide a contribution to the use of natural wastes such as sugarcane bagasse as substrate for growth and production of thermophilic CMCase. Further optimization to increase the production of cellulase enables the use in industrial applications.

Bacillus sp.; CMCase; Sugarcane; Enzyme production; Thermophilic cellulase

Independent Variables	Code Level of Variables
Independent Variables	-α	-1	0	1	+α
Temperature (°C) pH	30 4.2	36 5.0	50 7.0	64 9.0	70 9.8

	Independent Variables		CMCase activity (IU ml^-1)
Run	pH	T (°C)	Experimental	Predicted
1	5.0	36	0.16	0.11
2	5.0	64	0.36	0.33
3	9.0	36	0.15	0.17
4	9.0	64	0.18	0.22
5	7.0	50	0.29	0.29
6	7.0	50	0.31	0.29
7	7.0	50	0.29	0.29
8	7.0	30	0.14	0.15
9	7.0	70	0.37	0.35
10	4.2	50	0.14	0.19
11	9.8	50	0.21	0.15

Source of Variation	Sum of Squares	Degrees of Freedom	Mean Square	F Ratio (Model SIgnificance)	p-value
pH (Quadratic)	0.021330	1	0.021330	159.9724	0.006193^* * Significant factors (p< 0.05); R2 (coefficient of determination) = 0.84; Adjusted R2=0.7316.
Temperature (Quadratic)	0.002600	1	0.002600	19.5018	0.047643^* * Significant factors (p< 0.05); R2 (coefficient of determination) = 0.84; Adjusted R2=0.7316.
Temperature (Linear)	0.038540	1	0.038540	289.0536	0.003442^* * Significant factors (p< 0.05); R2 (coefficient of determination) = 0.84; Adjusted R2=0.7316.
1L x 2L	0.007225	1	0.007225	54.1875	0.017959^* * Significant factors (p< 0.05); R2 (coefficient of determination) = 0.84; Adjusted R2=0.7316.
Lack of fit	0.012622	4	0.003156	23.6663	0.040952^* * Significant factors (p< 0.05); R2 (coefficient of determination) = 0.84; Adjusted R2=0.7316.
Pure error	0.000267	2	0.000133	-	-

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