2013 ; 23(9):
|Author||Ribka Litantra, Stefani Lobionda, Joung Han Yim, Hyung Kwoun Kim|
|Affiliation||Division of Biotechnology, The Catholic University of Korea, Bucheon 420-743, Republic of Korea,Faculty of Biotechnology, Atma Jaya Catholic University, Jalan Jenderal Sudirman 51, Jakarta 12930, Indonesia|
|Title||Expression and Biochemical Characterization of Cold-Adapted Lipases from Antarctic Bacillus pumilus Strains|
J. Microbiol. Biotechnol.2013 ; 23(9):
|Abstract||Two lipase genes (bpl1 and bpl3) from Antarctic Bacillus pumilus strains were expressed in
Bacillus subtilis. Both recombinant lipases BPL1 and BPL2 were secreted to the culture medium
and their activities reached 3.5 U/ml and 5.0 U/ml, respectively. Their molecular masses
apparent using SDS-PAGE were 23 kDa for BPL1 and 19 kDa for BPL3. Both lipases were
purified to homogeneity using ammonium sulfate precipitation and HiTrap SP FF column and
Superose 12 column chromatographies. The final specific activities were estimated to be
328 U/mg for BPL1 and 310 U/mg for BPL3. Both lipases displayed an optimum temperature
of 35oC, similar to other mesophilic enzymes. However, they maintained as much as 70% and
80% of the maximum activities at 10oC. Accordingly, their calculated activation energy at a
temperature range of 10-35oC was 5.32 kcal/mol for BPL1 and 4.26 kcal/mol for BPL3, typical
of cold-adapted enzymes. The optimum pH of BPL1 and BPL3 was 8.5 and 8.0, respectively,
and they were quite stable at pH 7.0-11.0, showing their strong alkaline tolerance. Both
lipases had a preference toward medium chain length (C6-C10) fatty acid substrates. These
results indicate the potential for the two Antarctic B. pumilus lipases as catalysts in bioorganic
synthesis, food, and detergent industries.|
|Keywords||lipase, Bacillus pumilus, Antarctic, cold-adapted enzyme|
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