Biochemical properties and three-dimensional structures of two extracellular lipolytic enzymes from Bacillus subtilis

Eggert, Thorsten; Pouderoyen, Gertie van; Pencreach, Gaëlle; Douchet, Isabelle; Verger, Robert; Dijkstra, Bauke W.; Jaeger, Karl‐Erich

doi:10.1016/s0927-7765(02)00033-4

Cited by 49 publications

(35 citation statements)

References 42 publications

(89 reference statements)

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“…The highest lipolytic activity exhibited by isolated B. subtilis was 0.155 U/ml (0.416 U/mg protein). Bacillus subtilis produces several extracellular hydrolytic enzymes and among them a phospholipase, a lipase LipA and an esterase LipB have been identiWed [8]. Eggert et al [7] claimed that LipA and LipB of wild-type B. subtilis 168 were diVerentially expressed depending on the composition of the growth medium: LipA was produced in rich (LB supplemented with glucose) and in minimal medium (SMS supplemented with tryptophan and glucose), whereas LipB was present only in rich medium.…”

Section: Discussionmentioning

confidence: 99%

“…B. subtilis, B. pumilus, B. licheniformis and B. alcalophilus are among common bacterial lipase producers [9]. Lipolytic enzymes produced and secreted by B. subtilis are of substantial biotechnological interest and many have, therefore, been identiWed, cloned, and characterized [6][7][8].…”

Section: Introductionmentioning

confidence: 99%

“…From the industrial point of view, utilization of natural nutrients to form costeVective production media is attractive. Bacillus can eYciently utilize complex nutrient sources [8]. However, the literature reports on the development of a medium composed of natural nutrients such as vegetable oils for high lipolytic activity of isolated Bacillus species are limited [13,15,17]; and, to our knowledge, none of them reports on B. subtilis.…”

Section: Introductionmentioning

confidence: 99%

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Effects of lipidic carbon sources on the extracellular lipolytic activity of a newly isolated strain of Bacillus subtilis

Takaç

Marul

2008

J Ind Microbiol Biotechnol

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An isolate exhibiting high extracellular lipolytic activity was identified as Bacillus subtilis by 16S rRNA gene sequence analysis. The enzyme activity of the isolate was improved by using different concentrations of lipidic carbon sources such as vegetable oils, fatty acids and triglycerides. Lipolytic activity was assayed spectrophotometrically using p-nitrophenyl palmitate. One percent (v/v) of sesame oil provided the highest activity with 80 and 98% enhancements with respect to 1% (v/v) concentrations of linoleic acid and triolein as the favored fatty acid and triglyceride, respectively. Glucose presented a repressive effect on lipase production. Lipase secreted by B. subtilis was partially purified by ultrafiltration and anion exchange chromatography; and the purified enzyme was tested for its residual activity in the presence of EDTA, SDS, Triton X-100, Tween 20, Tween 80 and protease. The present work reports, for the first time, that the lipolytic activity of a B. subtilis strain can be improved by using inexpensive vegetable oils; and also that B. subtilis lipase is suitable for use in detergents.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Effects of lipidic carbon sources on the extracellular lipolytic activity of a newly isolated strain of Bacillus subtilis

Takaç

Marul

2008

J Ind Microbiol Biotechnol

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“…This feature is called interfacial activation. This probable mechanism also hints at the presence of a cap or a lid which when closed covers the catalytic site (the triad of serine, histidine and aspartic acid) making the enzyme inactive but in the presence of the interface, opens to reveal the active site, a hypothesis that has been proven [14]. This type of structure can be imagined to be like the character of the popular arcade game Pacman, and is in fact often depicted in a similar fashion.…”

Section: How the Lipase Workmentioning

confidence: 91%

Lipase Immobilization Techniques for Biodiesel Production: An Overview

Nigam¹,

Mehrotra²,

Vani³

et al. 2014

IJREB

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The growing energy needs and depleting fuel sources compel us to look towards production of biodiesel, an appropriate alternative. The industrially used chemical catalysis process is beset with problems that enzymatic production using lipases could avoid. In this light, the immobilization of lipases plays an important role in the optimization of the production process. This review discusses the various techniques that have been studied for lipase immobilization, namely adsorption, covalent attachment, entrapment, cross-linked enzyme agglomerates and whole-cell biocatalysts, while highlighting their benefits and drawbacks. It also sheds light on the future of enzyme immobilization and its industrial application.

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“…Among esterases, carboxyl ester hydrolases (EC 3.1.1.1) hydrolyze esters of short chain carboxylic acids (≤12) and triacylglycerol hydrolases or lipases (EC 3.1.1.3) display maximum activity towards insoluble long chain (≥12) acylglycerides [1]. In organic media, they catalyze reactions such as esterification, inter-esterification and trans-esterification [2].…”

Section: Introductionmentioning

confidence: 99%

Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425T

Akatin

2015

Turk J Bioch

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Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425 T [Thermus sp. NCCB 100425 T ' den yeni bir ısıya dayanıklı esterazın saflaştırılması ve karakterizasyonu] ABSTRACT Objective: The purpose of the present study was to purify and characterize an esterase from a thermophilic bacterium Thermus sp. NCCB 100425 T and to check its suitability for industrial applications. Methods: Thermus sp. NCCB 100425 T esterase was purified by using ammonium sulphate precipitation and hydrophobic interaction chromatography and then characterized biochemically. Results: The purity of the enzyme was observed as a single band on native-and SDS-PAGE. In the presence of p-nitrophenyl acetate (pNPA) as a substrate, the optimum pH and temperature of the enzyme were found to be 7.5 and 60°C, respectively. K m and V max values are calculated as 18.32 mM and 96.15 U/mg protein, respectively, with pNPA. pH stability was investigated in the range of pH 4.0-9.0 at 4°C and 60°C. After 7 days incubation, activity of pure enzyme was retained 85-90% for all pH at 4°C and 59±5.2% for pH 8.0 at 60°C. It was determined that approximately 80% of enzyme activity was retained between 30-60°C after 7 days incubation. In the presence of 10% ethanol and DMSO, the enzyme activity was retained 96±2.7% and 78±2.5%, respectively. Additionally, it was detected that some metal ions affect the enzyme activity at different ratios. T esterazı amonyum sülfat çöktürmesi ve hidrofobik etkileşim kromatografisi kullanılarak saflaştırıldı ve biyokimyasal özellikleri incelendi. Bulgular: Doğal-ve SDS-PAGE'de tek bant şeklinde görülen saf enzimin, p-nitrofenil asetat (pNPA) varlığında optimum pH ve sıcaklık değerleri sırasıyla 7,5 ve 60°C olarak belirlendi. Yine pNPA bir substrat olarak kullanıldığında K m ve V max değerlerinin 18,32 mM ve 96,15 U/ mg protein olduğu tespit edildi. pH kararlılığı pH 4,0-9,0 aralığında 4°C ve 60°C'de incelendi. 7 günlük bir inkübasyondan sonra saf enzimin aktivitesinin 4°C'de bütün pH'larda yaklaşık %85-90 oranında korunduğu, bu oranın 60°C'de pH 8,0'de ise %59±5,2 olduğu görüldü. 30-60°C'ler arasında 7 gün inkübe edilen enzim, orijinal aktivitesini %80 oranında korudu. %10 nihai konsantrasyondaki etanol ve DMSO varlığında enzim aktivitesi sırasıyla %96±2,7 ve %78±2,5 oranlarında korundu. Ayrıca, bazı metal iyonlarının enzim aktivitesini farklı oranlarda etkilediği tespit edildi. Sonuç: Bütün bu sonuçlar değerlendirildiğinde Thermus sp. NCCB 100425 T esterazının özel-likle yüksek sıcaklık-ve pH-kararlılığı nedeniyle endüstriyel ve/veya klinik uygulamalar için avantajlara sahip olabileceği açıkça görülmektedir. Thus, this study can be considered as a step to obtain an industrially suitable esterase having excellent features. ) and solvents were purchased from Merck A.G. (Darmstadt, Germany). All chemicals were analytical and microbiological grade. Conclusion Materials and Methods Chemicals Bacterial strain and crude enzyme extractionThermus sp. NCCB 100425 T bacterial strain isolated from a geotherm...

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Biochemical properties and three-dimensional structures of two extracellular lipolytic enzymes from Bacillus subtilis

Cited by 49 publications

References 42 publications

Effects of lipidic carbon sources on the extracellular lipolytic activity of a newly isolated strain of Bacillus subtilis

Effects of lipidic carbon sources on the extracellular lipolytic activity of a newly isolated strain of Bacillus subtilis

Lipase Immobilization Techniques for Biodiesel Production: An Overview

Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425T

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