Purification and characterization of organic solvent and detergent tolerant lipase from thermotolerant Bacillus sp. RN2

The aim of this study was to characterize the organic solvent and detergent tolerant properties of recombinant lipase isolated from thermotolerant Bacillus sp. RN2 (Lip-SBRN2). The isolation of the lipase-coding gene was achieved by the use of inverse and direct PCR. The complete DNA sequencing of t...

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Published inInternational journal of molecular sciences Vol. 11; no. 10; pp. 3783 - 3792
Main Authors Kanjanavas, Pornpimon, Khuchareontaworn, Sintawee, Khawsak, Paisarn, Pakpitcharoen, Arda, Pothivejkul, Khajeenart, Santiwatanakul, Somchai, Matsui, Kenji, Kajiwara, Tadahiko, Chansiri, Kosum
Format Journal Article
LanguageEnglish
Published Switzerland MDPI AG 29.09.2010
Molecular Diversity Preservation International (MDPI)
Subjects
Amino acids
Bacillus
Bacillus - enzymology
Bacterial Proteins - chemistry
detergent tolerant
Detergents - chemistry
Enzymes
Hot Temperature
lipase
Lipase - chemistry
organic tolerant
Peptides
Proteins
Solvents
Solvents - chemistry
thermotolerant
Thailand
Bangkok Thailand
thermotolerant
detergent tolerant
Bacillus
lipase
organic tolerant
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Summary:The aim of this study was to characterize the organic solvent and detergent tolerant properties of recombinant lipase isolated from thermotolerant Bacillus sp. RN2 (Lip-SBRN2). The isolation of the lipase-coding gene was achieved by the use of inverse and direct PCR. The complete DNA sequencing of the gene revealed that the lip-SBRN2 gene contains 576 nucleotides which corresponded to 192 deduced amino acids. The purified enzyme was homogeneous with the estimated molecular mass of 19 kDa as determined by SDS-PAGE and gel filtration. The Lip-SBRN2 was stable in a pH range of 9-11 and temperature range of 45-60 °C. The enzyme was a non metallo-monomeric protein and was active against pNP-caprylate (C8) and pNP-laurate (C12) and coconut oil. The Lip-SBRN2 exhibited a high level of activity in the presence of 108% benzene, 102.4% diethylether and 112% SDS. It is anticipated that the organic solvent and detergent tolerant enzyme secreted by Bacillus sp. RN2 will be applicable as catalysts for reaction in the presence of organic solvents and detergents.
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ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms11103783