Homologous expression, purification and characterization of a novel high-alkaline and thermal stable lipase from Burkholderia cepacia ATCC 25416

Wang, Xiaoqing; Xiang, Yu; Xu, Yan

doi:10.1016/j.enzmictec.2009.05.004

Cited by 56 publications

(36 citation statements)

References 34 publications

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“…The pH-optimum for BPL activity is the same results as others BL [23,24,25]. BPL was found to be stable at pH at high temperature (70°C and 100°C) (Figure 4d; Figure 5), similar as the lipase from Burkholderia cepacia ATCC 25416 and Bacillus pumilus RK31 appears to be tolerant to temperature, as were reported to be stable in the range temperature between 30°C to 60°C [29,30]. The same findings were described by fairolniza, the lipase from bacillus sp was thermostable in the temperature range of 55 to 75°C; and considerable activity (75%) was retained.…”

Section: Effect Of Ph and Temperature On The Activity And Stability Osupporting

confidence: 71%

Biochemical characterization of a new thermostable lipase from Bacillus pumilus strain

Faouzi

Bergadi

Sayari³

et al. 2015

Turk J Bioch

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Objective: Research and characterization of new thermostable lipases from bacterial strains isolated from tannery waters in the old medina of Fez. Methods: Gene which encodes the 16S ribosomal RNA for a bacterial species was amplified via PCR and sequenced (Bacillus pumilus HF544325). The extracellular lipase from B. pumilus is purified by gel filtration (Sephacryl S-200) and cation exchange chromatography (Mono S sepharose cation exchanger). The N-terminal sequences of purified Bacillus pumilus lipase were determined by automated Edman's degradation, using an Applied Biosystems 470 A protein sequencer equipped with PTH 120A analyser. The activity of lipase was examined within the pH range of 6.0-10.0 and the effect of pH on lipase stability was determined by incubating the lipase fraction in various buffer solutions ranging from 3.0 to 10.0 for 24 h at room temperature. Results:The results showed that Bacillus pumilus is a strain that produce non-inducible lipase. This enzyme has a molecular weight of 27 kDa and presents a maximal activity at pH 8 and 45°C. The 18 N-terminal amino acid residues showed a high degree of homology with other Bacillus lipase sequences. After treatment in 100°C for 5 min, the thermostable enzyme maintains 60% of its activity, which is greater than that those founded in previous works. The enzyme retained 100% of its activity after 30 min incubation at 70°C. Conclusion: This newly isolated lipase is thermostable and it has a significant difference which was observed when the biochemical properties of the Bacillus pumilus lipase were compared to others microbial lipases. The Bacillus pumilus lipase can be considered as a good candidature for industrial and biotechnological applications. Key Words: Bacillus pumilus, lipase, purification, thermostable Conflict of Interest: The authors have no conflict of interest. ÖZETAmaç: Çalışmanın amacı Fez şehrinde bulunan tabakhane sularından izole edilen bakteri suşla-rından, sıcaklığa dayanıklı yeni bir lipaz enzimini saflaştırarak karakterize etmektir. Metod: Bakteri türleri için 16S ribosomal RNA'yı kodlayan gen PCR ile çoğaltılarak sekanslanmıştır (Bacillus pumilus HF544325). Ekstraselüler lipaz jel filtrasyonu (Sefakril S-200) ve katyon değiştirici kromatografi kullanılarak (Mono S Sefaroz katyon değiştirici) B. pumilus'den saflaştırılmıştır. Saflaştırılan Bacillus pumilus lipazının N-terminal dizisi Edman degredasyon yöntemi (Applied Biosystems 470 A protein sekanslama cihazı ile PTH120A analizör) ile saptanmıştır. Lipaz aktivitesi 6.0-10.0 pH aralığında incelenmiş, pH'nın lipaz enziminin dayanık-lılığına olan etkisi lipaz fraksiyonunu pH'sı 3.0 ile 10.0 arasında değişen çeşitli tampon çözel-tilerinde 24 saat oda sıcaklığında inkübe ederek araştırılmıştır. Bulgular: Sonuçlar Bacillus pumilus suşunun indüklenmeyen lipaz enzimini ürettiğini gös-termektedir. Lipaz enzimi 27 kDa molekül ağırlığına sahip olup, en yüksek aktiviteyi pH 8 ve 45°C'de göstermektedir. Amino ucundaki 18 amino asit diğer Bacillus lipaz dizileri ile yüksek oranda ...

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Section: Effect Of Ph and Temperature On The Activity And Stability Osupporting

confidence: 71%

Biochemical characterization of a new thermostable lipase from Bacillus pumilus strain

Faouzi

Bergadi

Sayari³

et al. 2015

Turk J Bioch

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“…Interestingly, none of the solvents inactivated the lipase completely, possibly due to low penetration of the solvent in the enzyme structure compared with hydrated solvents. Results reported by Wang et al 1 using lipase from Burkholderia cepacia ATCC 25416 in the presence of 30% (v/v) acetone, chloroform, n-hexane, cyclohexane and n-heptane in the reaction mixture for 2 h at 37 °C showed activity increases of 1.7, 1.8, 1.7, 1.7 and 2.0-fold, respectively.…”

Section: Effect Of Metal Ions and Solvents On Lipasementioning

confidence: 96%

“…1,2 Lipases are important biocatalysts because they act under mild conditions, are highly stable in organic solvents, exhibit broad substrate specificity, and typically present high region and/or stereoselectivity in catalysis. 3,4 Consequently, lipases have gained importance and been applied widely in various fields such as foods, pharmaceuticals, detergents, biosensors, biodiesel, cosmetics, fine chemicals and agrochemicals.…”

Section: Introductionmentioning

confidence: 99%

Biochemical characterisation of lipase from a new strain of Bacillus sp. ITP-001

et al. 2012

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Recebido em 26/9/11; aceito em 24/1/12; publicado na web em 15/5/12Lipases are characterised mainly by catalytic versatility and application in different industrial segments. The aim of this study was to biochemically characterise a lipase from a new strain of Bacillus sp. ITP-001. The isoelectric point and molecular mass were 3.12 and 54 kDa, respectively. The optima lipase activity was 276 U g -1 at pH 7.0 and a temperature of 80 °C, showing greater stability at pH 5.0 and 37 °C. , respectively determined by the Eadie-Scatchard method.

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“…3) and significantly different with that of the ECL from B. cepacia ATCC 25416 (the optimal pH 11.0), B. cepacia DSM 50181 (the optimal pH 5.0), etc. [5,32]. The WCL from Burkholderia sp.…”

Section: Resultsmentioning

confidence: 99%

Production and Characteristics of the Whole-Cell Lipase from Organic Solvent Tolerant Burkholderia sp. ZYB002

Shu

Cheng

et al. 2011

Appl Biochem Biotechnol

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The thermostable and organic solvent tolerant whole-cell lipase (WCL) was produced by Burkholderia sp. ZYB002 with broad spectrum organic solvent tolerance. The production medium of the WCL was primarily optimized, which resulted in the maximum activity of 22.8 U/mL and the 5.1-fold increase of the WCL yield. The optimized culture medium was as follows (% w/v or v/v): soybean meal 2, soybean oil 0.5, manganese sulfate 0.1, K(2)HPO(4) 0.1, olive oil 0.5, initial pH 6.0, inoculum density 2, liquid volume 35 mL in 250-mL Erlenmeyer flask, and incubation time 24 h. The biochemical characterization of the WCL from Burkholderia sp. ZYB002 was determined, and the results showed that the optimal pH and temperature for lipolytic activity of the WCL was 8.0 and 65°C, respectively. The WCL was stable at temperature up to 70°C for 1 h and retained 79.2% of its original activity. The WCL was highly stable in the pH range from 3.0 to 8.5 for 6 h. Ca(2+), K(+), Na(+), NO (3) (-) , etc. ions stimulated its lipolytic activity, whereas Zn(2+) ion caused inhibition effect. The WCL was also relatively stable in n-butanol at a final concentration of 50% (v/v) for 24 h. However, the WCL was strongly inhibited in Triton X-100 at a final concentration of 10% (v/v). The WCL with thermal resistance and organic solvent tolerance showed its great potential in various green industrial chemical processes.

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Homologous expression, purification and characterization of a novel high-alkaline and thermal stable lipase from Burkholderia cepacia ATCC 25416

Cited by 56 publications

References 34 publications

Biochemical characterization of a new thermostable lipase from Bacillus pumilus strain

Biochemical characterization of a new thermostable lipase from Bacillus pumilus strain

Biochemical characterisation of lipase from a new strain of Bacillus sp. ITP-001

Production and Characteristics of the Whole-Cell Lipase from Organic Solvent Tolerant Burkholderia sp. ZYB002

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