PREPARATION AND CHARACTERIZATION OF IMMOBILIZED SPORES WITH LACCASE ACTIVITY FROM <i>Bacillus pumilus</i> W3 ON DEAE-CELLULOSE AND THEIR APPLICATION IN DYE DECOLORIZATION

Zhou, Wen; Guan, Zheng-Bing; Cai, Yu-Jie; Chen, Yu; Zhang, Ning; Liao, Xiang-Ru

doi:10.1590/0104-6632.20170341s20150492

Wen Zhou

The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China. Phone: + 86 510 8532 7725; Fax: + 86 510 8532 7725. E-mail: guanzb525@163.com; liaoxiangru@163.com

Zheng-Bing Guan ^**To whom correspondence should be addressed

The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China. Phone: + 86 510 8532 7725; Fax: + 86 510 8532 7725. E-mail: guanzb525@163.com; liaoxiangru@163.com

Yu-Jie Cai

The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China. Phone: + 86 510 8532 7725; Fax: + 86 510 8532 7725. E-mail: guanzb525@163.com; liaoxiangru@163.com

Yu Chen

The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China. Phone: + 86 510 8532 7725; Fax: + 86 510 8532 7725. E-mail: guanzb525@163.com; liaoxiangru@163.com

Ning Zhang

The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China. Phone: + 86 510 8532 7725; Fax: + 86 510 8532 7725. E-mail: guanzb525@163.com; liaoxiangru@163.com

Xiang-Ru Liao ^**To whom correspondence should be addressed

The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China. Phone: + 86 510 8532 7725; Fax: + 86 510 8532 7725. E-mail: guanzb525@163.com; liaoxiangru@163.com

*To whom correspondence should be addressed

Orgnaniccompounds	Concentration(%, v/v)	Residual activity (%)
Orgnaniccompounds	Concentration(%, v/v)	Freespores	Immobilizedspores
Control		100.00±0.73	100.00±1.14
methanol	10	110.74±4.29	99.53±3.38
	50	16.68±1.98	82.54±2.11
ethanol	10	104.2±3.22	86.2±5.88
	50	40.79±2.56	51.24±2.23
acetone	10	106.12±4.23	193.87±2.18
	50	1.8±0.23	21.34±1.08
acetonitrile	10	126.09±4.48	180.78±6.56
	50	3.42±0.33	55.23±1.27
Dimethylsulfoxide	10	84.64±1.19	108.02±4.59
	50	6.78±0.56	33.55±1.76
ethyl acetate^a a The products generated from laccase oxidizing substrate were in the water phase. It could not be measured in the presence of 50% (v/v) ethyl acetate because the reaction mixture was an emulsion. "−" represents no reaction and "+" represents weak reaction.	10	100.84±5.23	149.41±6.78
	50	-	+
Dichloromethane^b b The products generated from laccase oxidizing substrate were in the organic phase. The sample containing dichloromethane was measured with the water phase. The products extracted into the organic phase were not measured (only the sample containning 10% of diclorornethane has the situation that a small amount of products extracted into the organic phase). In addition, half of its absorbance value was taken as the final value when the samples containing 50% dichloromethane were measured.	10	36.47±3.23	22.05±1.22
	50	48.62±3.12	103.24±5.59
Dimethylformamide	10	79.42±4.56	128.89±3.45
	50	3.54±0.75	2.33±0.54

Metal ions (10 mM)	Residual activity (%)
Metal ions (10 mM)	Free spores	Immobilized spores
Control	100.00±1.42	100.00±1.10
Na⁺	97.22±2.67	102.67±3.03
Mg²⁺	93.09±4.18	98.78±6.44
Al³⁺	91.55±3.50	111.22±2.16
K⁺	96.52±2.51	122.78±6.02
Ca²⁺	91.32±1.34	113.56±5.14
Mn²⁺	17.48±1.50	28.44±2.26
Fe³⁺	2.07±0.78	5.00±1.62
Co²⁺	30.36±4.17	92.78±2.31
Ni²⁺	75.25±2.84	96.11±4.67
Cu²⁺	79.21±2.59	95.11±4.30
Zn²⁺	78.32±1.50	110±6.35

Inhibitors	Concentration (mM)	Residual activity (%)
Inhibitors	Concentration (mM)	Free spores	Immobilized spores
Control		100.00±1.22	100.00±2.87
EDTA	10	78.18±4.23	64.69±4.68
	25	75.63±6.14	66.67±3.74
	50	59.25±1.75	51.38±1.39
Cysteine	0.01	73.93±2.35	131.49±2.87
	0.1	5.77±1.22	89.33±2.01
	1	0.36±0.15	71.01±1.13
NaN3	0.0005	68.89±1.33	37.68±0.66
	0.005	79.01±2.26	85.38±1.28
	0.05	51.35±1.39	98.81±3.35
NaCl	100	97.49±2.22	82.87±5.59
	500	99.37±1.38	89.33±1.19
	1000	12.35±0.51	1.58±0.33
SDS	0.1	89.61±6.88	92.23±5.29
	1	50.2±4.75	81.29±3.01
	10	30.25±3.35	10.14±1.15
PMSF	0.1	96.91±3.33	91.44±5.43
	1	101.25±2.34	91.88±4.56
	10	26.44±0.69	65.22±1.76

[1] *To whom correspondence should be addressed

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PREPARATION AND CHARACTERIZATION OF IMMOBILIZED SPORES WITH LACCASE ACTIVITY FROM Bacillus pumilus W3 ON DEAE-CELLULOSE AND THEIR APPLICATION IN DYE DECOLORIZATION

Abstract