Abstract：
:To improve enzymatic activity of Bacillus pumilus lipases, DNA shuffling was applied to two lipase genes from local B. pumilus isolates. Using a high-throughput activity assay, the mutant with highest activity was selected. This chimeric mutant (L3-3), carrying two crossover positions and three point mutations, has a specific activity 6.4 and 8.2 times higher than the two parent enzymes. The mutant also is more tolerant to various detergents and organic solvents, and has a 9 times longer half-life at 50 °C. Homology modeling of mutant L3-3, based on the highly homologous B. subtilis lipase A, shows that the increased thermostability is likely due to structural rigidification and reduced surface hydrophobicity. Increased specific activity may result from the location of mutations close to the active site. Together, our results show that it is possible to evolve, by DNA shuffling, B. pumilus lipase variants with improved applicability as biocatalysts, even if the two parent enzymes are highly similar.
journal_name
J Biotechnoljournal_title
Journal of biotechnologyauthors
Akbulut N,Tuzlakoğlu Öztürk M,Pijning T,İşsever Öztürk S,Gümüşel Fdoi
10.1016/j.jbiotec.2012.12.016subject
Has Abstractpub_date
2013-03-10 00:00:00pages
123-9issue
1eissn
0168-1656issn
1873-4863pii
S0168-1656(13)00007-2journal_volume
164pub_type
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