Purification and Characterization of Organic Solvent and Detergent Tolerant Lipase from Thermotolerant Bacillus sp. RN2 |
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| Authors: | Kanjanavas Pornpimon Khuchareontaworn Sintawee Khawsak Paisarn Pakpitcharoen Arda Pothivejkul Khajeenart Santiwatanakul Somchai Matsui Kenji Kajiwara Tadahiko Chansiri Kosum |
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| Affiliation: | Department of Biochemistry, Faculty of Medicine, Srinakharinwirot University, Bangkok 10110, Thailand; E-Mails: kanjanavas@hotmail.com (P.K.); wevansi@hotmail.com (S.K.); pkhla@hotmail.com (P.K.); oom_arda@hotmail.com (A.P.). |
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| Abstract: | 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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| Keywords: | lipase organic tolerant detergent tolerant Bacillus thermotolerant |
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