Lipase from marine Aspergillus awamori BTMFW032: Production, partial purification and application in oil effluent treatment

被引:66
作者
Basheer, Soorej M. [1 ]
Chellappan, Sreeja [1 ]
Beena, P. S. [1 ]
Sukumaran, Rajeev K. [2 ]
Elyas, K. K. [1 ]
Chandrasekaran, M. [1 ]
机构
[1] Cochin Univ Sci & Technol, Dept Biotechnol, Microbial Technol Lab, Cochin 682022, Kerala, India
[2] NIIST, Div Biotechnol, Thiruvananthapuram, Kerala, India
关键词
STATISTICAL MEDIUM OPTIMIZATION; RESPONSE-SURFACE METHODOLOGY; BIOCHEMICAL-PROPERTIES; EXTRACELLULAR LIPASES; MEDIUM COMPONENTS; WASTE-WATER; ALKALINE; ESTERASE; STRAIN; NIGER;
D O I
10.1016/j.nbt.2011.04.007
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Marine fungus BTMFW032, isolated from seawater and identified as Aspergillus awamori, was observed to produce an extracellular lipase, which could reduce 92% fat and oil content in the effluent laden with oil. In this study, medium for lipase production under submerged fermentation was optimized statistically employing response surface method toward maximal enzyme production. Medium with soyabean meal-0.77% (w/v); (NH4)(2)SO4-0.1 M; KH2PO4-0.05 M; rice bran oil-2% (v/v); CaCl2-0.05 M; PEG 6000-0.05% (w/v); NaCl-1% (w/v); inoculum-1% (v/v); pH 3.0; incubation temperature 35 degrees C and incubation period-five days were identified as optimal conditions for maximal lipase production. The time course experiment under optimized condition, after statistical modeling, indicated that enzyme production commenced after 36 hours of incubation and reached a maximum after 96 hours (495.0 U/ml), whereas maximal specific activity of enzyme was recorded at 108 hours (1164.63 U/mg protein). After optimization an overall 4.6-fold increase in lipase production was achieved. Partial purification by (NH4)(2)SO4 precipitation and ion exchange chromatography resulted in 33.7% final yield. The lipase was noted to have a molecular mass of 90 kDa and optimal activity at pH 7 and 40 degrees C. Results indicated the scope for potential application of this marine fungal lipase in bioremediation.
引用
收藏
页码:627 / 638
页数:12
相关论文
共 61 条
[1]   Optimization of critical medium components using response surface methodology for lipase production by Rhizopus delemar [J].
Acikel, Ue ;
Ersan, M. ;
Acikel, Y. Sag .
FOOD AND BIOPRODUCTS PROCESSING, 2010, 88 (C1) :31-39
[2]   BASIC LOCAL ALIGNMENT SEARCH TOOL [J].
ALTSCHUL, SF ;
GISH, W ;
MILLER, W ;
MYERS, EW ;
LIPMAN, DJ .
JOURNAL OF MOLECULAR BIOLOGY, 1990, 215 (03) :403-410
[3]  
[Anonymous], 1946, Biometrika, DOI [DOI 10.2307/2332195, DOI 10.1093/BIOMET/33.4.305]
[4]  
*AOAC INT, 2006, OFF METH 989 05 FAT
[5]   Statistical media optimization and alkaline protease production from Bacillus mojavensis in a bioreactor [J].
Beg, QK ;
Sahai, V ;
Gupta, R .
PROCESS BIOCHEMISTRY, 2003, 39 (02) :203-209
[6]   Isolation and characterization of three distinct forms of lipases from Candida rugosa produced in solid state fermentation [J].
Benjamin, S ;
Pandey, A .
BRAZILIAN ARCHIVES OF BIOLOGY AND TECHNOLOGY, 2000, 43 (05) :453-460
[7]   Characterization and distribution of esterase activity in activated sludge [J].
Boczar, BA ;
Forney, LJ ;
Begley, WM ;
Larson, RJ ;
Federle, TW .
WATER RESEARCH, 2001, 35 (17) :4208-4216
[8]  
Box GEP., 1960, Technometrics, V2, P455, DOI [10.1080/00401706.1960.10489912, DOI 10.1080/00401706.1960.10489912, 10.2307/1266454]
[9]   Microwave-assisted rapid characterization of lipase selectivities [J].
Bradoo, S ;
Rathi, P ;
Saxena, RK ;
Gupta, R .
JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS, 2002, 51 (02) :115-120
[10]   Optimization of extracellular lipase production by Geotrichum sp using factorial design [J].
Burkert, JFM ;
Maugeri, F ;
Rodrigues, MI .
BIORESOURCE TECHNOLOGY, 2004, 91 (01) :77-84