Int. J. Mol. Sci. 2010, 11(10), 3783-3792; doi:10.3390/ijms11103783
Article

Purification and Characterization of Organic Solvent and Detergent Tolerant Lipase from Thermotolerant Bacillus sp. RN2

Pornpimon Kanjanavas 1 email, Sintawee Khuchareontaworn 1 email, Paisarn Khawsak 1 email, Arda Pakpitcharoen 1 email, Khajeenart Pothivejkul 2 email, Somchai Santiwatanakul 3 email, Kenji Matsui 4 email, Tadahiko Kajiwara 4  and Kosum Chansiri 1,* email
1 Department of Biochemistry, Faculty of Medicine, Srinakharinwirot University, Bangkok 10110, Thailand 2 Department of Biology, Faculty of Sciences, Srinakharinwirot University, Bangkok 10110, Thailand 3 Department of Pathology, Faculty of Medicine, Srinakharinwirot University, Bangkok 10110, Thailand 4 Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi, Japan
* Author to whom correspondence should be addressed.
Received: 21 August 2010; in revised form: 17 September 2010 / Accepted: 18 September 2010 / Published: 29 September 2010
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
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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.
Keywords: lipase; organic tolerant; detergent tolerant; Bacillus; thermotolerant

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MDPI and ACS Style

Kanjanavas, P.; Khuchareontaworn, S.; Khawsak, P.; Pakpitcharoen, A.; Pothivejkul, K.; Santiwatanakul, S.; Matsui, K.; Kajiwara, T.; Chansiri, K. Purification and Characterization of Organic Solvent and Detergent Tolerant Lipase from Thermotolerant Bacillus sp. RN2. Int. J. Mol. Sci. 2010, 11, 3783-3792.

AMA Style

Kanjanavas P, Khuchareontaworn S, Khawsak P, Pakpitcharoen A, Pothivejkul K, Santiwatanakul S, Matsui K, Kajiwara T, Chansiri K. Purification and Characterization of Organic Solvent and Detergent Tolerant Lipase from Thermotolerant Bacillus sp. RN2. International Journal of Molecular Sciences. 2010; 11(10):3783-3792.

Chicago/Turabian Style

Kanjanavas, Pornpimon; Khuchareontaworn, Sintawee; Khawsak, Paisarn; Pakpitcharoen, Arda; Pothivejkul, Khajeenart; Santiwatanakul, Somchai; Matsui, Kenji; Kajiwara, Tadahiko; Chansiri, Kosum. 2010. "Purification and Characterization of Organic Solvent and Detergent Tolerant Lipase from Thermotolerant Bacillus sp. RN2." Int. J. Mol. Sci. 11, no. 10: 3783-3792.

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