Studies on lipolytic isoenzymes from a thermophilic Bacillus sp.: Production, purification and biochemical characterization

Nawani, Neerupma; Kaur, Jagdeep

doi:10.1016/j.enzmictec.2006.07.006

Cited by 41 publications

(20 citation statements)

References 25 publications

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“…Many enzymes were produced by bacteria and yeast showed maximum activities at high temperatures, such as Pseudomonas aeruginosa (70°C), a thermophilic Bacillus sp. (60 to 70°C) [26], and the yeast Kurtzmanomyces sp.…”

Section: Resultsmentioning

confidence: 99%

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: Resultsmentioning

confidence: 99%

Biochemical characterization of a new thermostable lipase from Bacillus pumilus strain

Faouzi

Bergadi

Sayari³

et al. 2015

Turk J Bioch

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“…The structural features of thermophilic and mesophilic lipases are compared with that of CLPs in order to arrive for stabilizing mutations (Suhre & Claverie 2003). But it was very hard to correlate thermostability with specific amino acids interactions (Nawani & Kaur 2007). Therefore directed evolution through random mutagenesis and tedious and time-consuming screening for maximum thermostability had been more effective (Eijsink et al 2004;Jaeger & Eggert 2004;Reyes-Duarte et al 2005).…”

Section: Thermal Stabilitymentioning

confidence: 99%

Cold active lipases – an update

Kavitha

2016

Frontiers in Life Science

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Cold active lipases (CLPs) are gaining importance nowadays as they are increasingly used in fine chemical synthesis, bioremediation, food processing and as detergent additive. These enzymes exhibit high catalytic activity at low temperatures and flexibility to act at low water medium. Since they are active at low temperatures consume less energy and also stabilize fragile compounds in the reaction medium. CLPs are commonly obtained from psychrophilic microorganisms which thrive in cold habitats. Compared to mesophilic and thermophilic lipases, only a few CLPs were studied and industrially exploited so far. CLPs (C. antarctica lipase-A and C. antarctica lipase-B) from Candida antarctica isolated from Antarctic region are the well studied and industrially employed, and many are being followed up. This review updates the CLPs reported recently and the industrial applications of CLPs. ARTICLE HISTORY

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“…Comparing these values to other kinetic analyses using the same substrate, Nawani, Kaur [42] analyzed lipases from thermophilic Bacillus sp, finding a V max = 0.032 µmoles/mL and Km = 0.19 mM. Brabcová et al [47] found a higher Km for the same substrate for an extracellular lipase from the mesophilic fungus Geotrichum candidum 4013: 0.406 mM.…”

Section: Kinetics Parameters Of the Enzymementioning

confidence: 99%

“…PTI-001, Nawani, Kaur. [42] found pH 8 and temperature 60˚C as optimum conditions. Figure 3(c) shows that the lipases partially purified of R. pusillus incubated at different pH were less stable at more alkaline pH values (8.5 and 9.5) after 150 minutes, with steeper decline than for the samples kept at pH 6.5 and 7.5.…”

Section: Open Accessmentioning

confidence: 99%

Partial purification, immobilization and preliminary biochemical characterization of lipases from Rhizomucor pusillus

Ferrarezi¹,

Pivetta²,

Bonilla-Rodriguez³

et al. 2013

AER

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Lipases have important applications in biotechnological processes, motivating us to produce, purify, immobilize and perform a biochemical characterization of the lipase from Rhizomucor pusillus. The fungus was cultivated by solid state fermentation producing lipolytic activity of about 0.5 U/mL(4U/g). A partial purification by gel filtration chromatography in Sephacryl S-100 allowed obtaining a yield of about 85% and a purification factor of 5.7. Our results revealed that the purified enzyme is very stable with some significant differences in its properties when compared to crude extract. The crude enzyme extract has an optimum pH and temperature of 7.5˚C and 40˚C, respectively. After purification, a shift of the optimum pH from 7 to 8 was observed, as well as a rise in optimumtemperature to 60˚C and an increase in stability. The enzyme was immobilized on CNBr-Agarose and Octyl-Agarose supports, having the highest immobilization yield of 94% in the second resin. The major advantage of immobilization in hydrophobic media such as Octyl is in its hyper activation, which in this case was over 200%, a very interesting finding. Another advantage of this type of immobilization is the possibility of using the derivatives in biotechnological applications, such as in oil enriched with omega-3 as the results obtained in this study display the hydrolysis of 40% EPA and 7% DHA from sardine oil, promising results compared to the literature.

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Studies on lipolytic isoenzymes from a thermophilic Bacillus sp.: Production, purification and biochemical characterization

Cited by 41 publications

References 25 publications

Biochemical characterization of a new thermostable lipase from Bacillus pumilus strain

Biochemical characterization of a new thermostable lipase from Bacillus pumilus strain

Cold active lipases – an update

Partial purification, immobilization and preliminary biochemical characterization of lipases from Rhizomucor pusillus

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