Characterization of a thermoalkalophilic esterase from a novel thermophilic bacterium, G2

Çolak, Ahmėt; Şişik, Deniz; Saglam, Nagihan; Güner, Saadettin; Çanakçı, Sabriye; Beldüz, Ali Osman

doi:10.1016/j.biortech.2004.06.003

Cited by 50 publications

(18 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The enzyme can be classified as cold-active and slightly alkaliphilic (Colak et al, 2005), as highest activity was observed in the range of 4-20 • C and at pH 8.0 (Figures 6, 7). Metal ions were found to have a marked effect on the activity of the enzyme (Figure 8).…”

Section: Discussionmentioning

confidence: 99%

A Novel Cold Active Esterase from a Deep Sea Sponge Stelletta normani Metagenomic Library

et al. 2017

View full text Add to dashboard Cite

Esterases catalyze the hydrolysis of ester bonds in fatty acid esters with short-chain acyl groups. Due to the widespread applications of lipolytic enzymes in various industrial applications, there continues to be an interest in novel esterases with unique properties. Marine ecosystems have long been acknowledged as a significant reservoir of microbial biodiversity and in particular of bacterial enzymes with desirable characteristics for industrial use, such as for example cold adaptation and activity in the alkaline pH range. We employed a functional metagenomic approach to exploit the enzymatic potential of one particular marine ecosystem, namely the microbiome of the deep sea sponge Stelletta normani. Screening of a metagenomics library from this sponge resulted in the identification of a number of lipolytic active clones. One of these encoded a highly, cold-active esterase 7N9, and the recombinant esterase was subsequently heterologously expressed in Escherichia coli. The esterase was classified as a type IV lipolytic enzyme, belonging to the GDSAG subfamily of hormone sensitive lipases. Furthermore, the recombinant 7N9 esterase was biochemically characterized and was found to be most active at alkaline pH (8.0) and displays salt tolerance over a wide range of concentrations. In silico docking studies confirmed the enzyme's activity toward short-chain fatty acids while also highlighting the specificity toward certain inhibitors. Furthermore, structural differences to a closely related mesophilic E40 esterase isolated from a marine sediment metagenomics library are discussed.

show abstract

Section: Discussionmentioning

confidence: 99%

A Novel Cold Active Esterase from a Deep Sea Sponge Stelletta normani Metagenomic Library

et al. 2017

View full text Add to dashboard Cite

show abstract

“…The reduction of the ATPase activity in the presence of EDTA can be attributed to its metal chelating effect (Fig. 7) (Colak et al, 2005). Sodium azide and ouabain, common inhibitors of most ATPases and sodium pumps, also inhibit the G2ALT enzyme (Schneider et al, 1998).…”

Section: (A) (B)mentioning

confidence: 99%

“…It is known that metal ions have an important role in maintaining enzyme activity and stabilizating the structure (DiTusa et al, 2001;Colak et al, 2005). The influence of some metal ions on the A. gonensis G2 ATPase activity was examined by using various metal chloride salts at various concentrations.…”

Section: (A) (B)mentioning

confidence: 99%

The ATPase activity of the G2alt gene encoding an aluminium tolerance protein from Anoxybacillus gonensis G2

et al. 2011

View full text Add to dashboard Cite

The G2ALT gene was cloned and sequenced from the thermophilic bacterium Anoxybacillus gonensis G2. The gene is 666 bp long and encodes a protein 221 amino acids in length. The gene was overexpressed in E. coli and purified to homogeneity and biochemically characterized. The enzyme has a molecular mass of 24.5 kDa and it could be classified as a member of the family of bacterial aluminium resistance proteins based on homology searches. When this fragment was expressed in E. coli, it endowed E. coli with Al tolerance to 500 μM. The purified G2ALT protein is active at a broad pH range (pH 4.0-10.0) and temperature range (25°C-80°C) with optima of 6.0 and the apparent optimal temperature of 73°C respectively. Under optimal conditions, G2ALT exhibited a low ATPase activity with K (m) (-) and V (max) (-) values of 10±0.55 μM and 26.81±0.13 mg Pi released/min/mg enzyme, respectively. The ATPase activity of G2ALT requires Mg(2+) and Na(+) ions, while Zn(2+) and Al(3+) stimulate the activity. Cd(2+) and Ag(+) reduced the activity and Li(+), Cu(2+), and Co(2+) inhibited the activity. Known inhibitors of most ATPases, like such as β-mercaptoethanol and ouabain, also inhibited the activity of the G2ALT. These biochemical characterizations suggested that G2ALT belongs to the PP-loop ATPase superfamily and it can be responsible for aluminium tolerance in A. gonensis G2.

show abstract

“…This finding is contrary to those previously reported on lipolytic enzymes, Universitas Scientiarum Vol. 22 (1): http://ciencias.javeriana.edu.co/investigacion/universitas-scientiarum which showed that metallic ions can play an important role in maintaining enzymatic structural stability and integrity [50], for example the presence of Fe 3+ and Mn 2+ at 1 mM increased the relative enzymatic activity of the 499EST esterase of Acidicaldus USBA-GBX-499 [23]. Moreover, under particular conditions, the presence of metallic cofactors facilitated the correct folding of an enzyme [51] and stabilized the structure of the catalytic site of Geobacillus zalihae lipase [52].…”

Section: Effect Of Metallic Ions On the Activity Of The Enzyme 505 Lipmentioning

confidence: 99%

Response Surface Methodology (RSM) for analysing culture conditions of Acidocella facilis strain USBA-GBX-505 and Partial Purification and Biochemical Characterization of Lipase 505 LIP

et al. 2017

View full text Add to dashboard Cite

Using Response Surface Methodology (RSM) we evaluated the culture conditions (nitrogen source, carbon source, pH and agitation rate) that increase the biomass of Acidocella facilis strain USBA-GBX-505 and therefore enhance the production of its lipolytic enzyme, 505 LIP. RSM results revealed that yeast extract and agitation were key culture factors that increased the growth-associated lipolytic activity by 4.5-fold (from 0.13 U.mg -1 to 0.6 U.mg -1). The 505 LIP lipase was partially purified using size-exclusion chromatography and ion-exchange chromatography. Its molecular weight was >77 kDa. The enzyme shows its optimum catalytic activity at 55 °C and pH 7.5. EDTA, PMSF, 1-butanol and DMSO inhibited enzymatic activity, whereas Tween 20, acetone, glycerol and methanol increased it. Metallic ions are not required for the activity of 505 LIP, and even have an inhibitory effect on the enzyme. This study shows the potential use of A. facilis strain USBA-GBX-505 for the production of a newly identified lipolytic enzyme, 505 LIP, which is stable at moderate temperatures and in the presence of organic solvents. These are important characteristics for the synthesis of many useful products.

show abstract

Characterization of a thermoalkalophilic esterase from a novel thermophilic bacterium, G2

Cited by 50 publications

References 34 publications

A Novel Cold Active Esterase from a Deep Sea Sponge Stelletta normani Metagenomic Library

A Novel Cold Active Esterase from a Deep Sea Sponge Stelletta normani Metagenomic Library

The ATPase activity of the G2alt gene encoding an aluminium tolerance protein from Anoxybacillus gonensis G2

Response Surface Methodology (RSM) for analysing culture conditions of Acidocella facilis strain USBA-GBX-505 and Partial Purification and Biochemical Characterization of Lipase 505 LIP

Contact Info

Product

Resources

About