An amylopullulanase (ApuNP1) from Geobacillus thermoleovorans NP1: biochemical characterization and its potential industrial applications.

An amylopullulanase was produced by Geobacillus thermoleovorans NP1. The optimum enzyme production occurred at 45°C and pH 7.0 (12 hr). NP1 amylopullulanase (ApuNP1) exhibited the maximal activity at 50°C and pH 6.0 and was stable between 30-50°C, and pH 3.0-12.0 for 24 hr. The enzyme showed two bands with molecular weights of 112 and 107 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The amylopullulanase retained 100% of its activity in the presence of 10 mM of Ca2+, Ba2+, Zn2+, Mg2+, Cu2+, EDTA, and PMSF. While the enzyme showed resistance to 5% of TritonX-100, Tween 20, and Tween 80, the activity was inhibited by 5% ß-mercaptoethanol and H2O2. While the hydrolysis products of pullulan were maltose, maltotriose, and maltodextrin, the starch was hydrolyzed to maltose, maltotriose, and maltodextrin units. This shows that NP1 pullulanase is a type II pullulanase (amylopullulanase). After the liquefaction assay, 12% glucose content was measured with a refractometer in the presence of 20% starch. According to the wash performance tests, the mixture of ApuNP1 and 1% detergent removed almost all of the stains. This novel thermo-acidic amylopullulanase has a potency to be used in detergent, starch, food, baking, textile, and cosmetic industries.

Isolation and characterization of a cold-active, alkaline, detergent stable α-amylase from a novel bacterium Bacillus subtilis N8.

A cold-active alkaline amylase producer Bacillus subtilis N8 was isolated from soil samples. Amylase synthesis optimally occurred at 15°C and pH 10.0 on agar plates containing starch. The molecular weight of the enzyme was found to be 205 kDa by performing SDS-PAGE. While the enzyme exhibited the highest activity at 25°C and pH 8.0, it was highly stable in alkaline media (pH 8.0-12.0) and retained 96% of its original activity at low temperatures (10-40°C) for 24 hr. While the amylase activity increased in the presence of ß-mercaptoethanol (103%); Ba2+, Ca2+, Na+, Zn2+, Mn2+, H2O2, and Triton X-100 slightly inhibited the activity. The enzyme showed resistance to some denaturants: such as SDS, EDTA, and urea (52, 65, and 42%, respectively). N8 α-amylase displayed the maximum remaining activity of 56% with 3% NaCl. The major final products of starch were glucose, maltose, and maltose-derived oligosaccharides. This novel cold-active α-amylase has the potential to be used in the industries of detergent and food, bioremediation process and production of prebiotics.

Associations of polymorphisms in HPC2/ELAC2 and SRD5A2 genes with benign prostate hyperplasia in Turkish men.

Benign prostate hyperplasia (BPH) is the most common benign tumor in elderly men for which the HPC2/ELAC2 and SRD5A2 genes are known genetic factors. The HPC2/ELAC2 gene features Ser217Leu and Ala541Thr polymorphisms and the SRD5A2 gene Ala49Thr and Val89Leu polymorphisms. The aim of this study was to examine relationships between these polymorphisms and BPH in Turkish men using amplification by the polymerase chain reaction (PCR) method. Polymorphisms were determined by using restriction fragment length polymorphism (RFLP) with suitable restriction: TaqI?, Fnu4HI, Mwo I and Rsa I. We found statistically significant relationship between the SRD5A2 gene Ala49Thr (OR=2.3; CI 95%, 1.04-5.1; p=0.01<0.05) , but not the other polymorphisms, and BPH. For the first time, our data demonstrate that the correlation between SRD5A2 gene Ala49Thr and polymorphisms is statistically significant in Turkish men with BPH.

Highly thermostable and alkaline alpha-amylase from a halotolerant-alkaliphilic Bacillus sp. AB68 / Alfa-amilase alcalina termoestável de Bacillus sp AB68 halotolerante-alcalifílico

An alkaliphilic and highly thermostable alpha-amylase producing Bacillus sp. was isolated from Van soda lake. Enzyme synthesis occurred at temperatures between 25ºC and 40ºC. Analysis of the enzyme by SDS-PAGE revealed a single band which was estimated to be 66 kDa. The enzyme was active in a broad temperature range, between 20ºC and 90ºC, with an optimum at 50ºC; and maximum activity was at pH 10.5. The enzyme was almost completely stable up to 80ºC with a remaining activity over 90 percent after 30 min pre-incubation. Thermostability was not increased in the presence of Ca2+. An average of 75 percent and 60ºC of remaining activity was observed when the enzyme was incubated between pH 5 and 9 for 1 h and for 2 h, respectively. The activity of the enzyme was inhibited by SDS and EDTA by 38 percent and 34 percent, respectively.

Bacillus sp AB68 alcalif¨ªlico produtor de alfa-amilase alcalina termoest¨¢vel foi isolado do lago Van soda. A s¨ªntese da enzima ocorreu entre 25ºC e 40ºC. A an¨¢lise da enzima por SDS-PAGE revelou uma ¨²nica banda estimada em 66 kDa. A enzima foi ativa em uma ampla faixa de temperatura, entre 20ºC e 90ºC, com um ¨®timo a 50ºC. A atividade m¨¢xima foi em pH 10,5. A enzima foi est¨¢vel at¨¦ 80ºC, mantendo 90 por cento de atividade ap¨®s 30 min de pr¨¦-incubação. A termoestabilidade não aumentou na presença de Ca2+. Quando incubada em pH entre 5 e 9 por 1h e por 2h, a enzima manteve 75 por cento e 60 por cento de atividade, respectivamente. SDS e EDTA causaram redução de 38 por cento e 34 por cento na atividade da enzima, respectivamente.

Highly thermostable and alkaline α-amylase from a halotolerant-alkaliphilic Bacillus sp. AB68.

An alkaliphilic and highly thermostable α-amylase producing Bacillus sp. was isolated from Van soda lake. Enzyme synthesis occurred at temperatures between 25°C and 40°C. Analysis of the enzyme by SDS-PAGE revealed a single band which was estimated to be 66 kDa. The enzyme was active in a broad temperature range, between 20°C and 90°C, with an optimum at 50°C; and maximum activity was at pH 10.5. The enzyme was almost completely stable up to 80°C with a remaining activity over 90% after 30 min pre-incubation. Thermostability was not increased in the presence of Ca(2+). An average of 75% and 60°C of remaining activity was observed when the enzyme was incubated between pH 5 and 9 for 1 h and for 2 h, respectively. The activity of the enzyme was inhibited by SDS and EDTA by 38% and 34%, respectively.

Highly thermostable, thermophilic, alkaline, SDS and chelator resistant amylase from a thermophilic Bacillus sp. isolate A3-15.

A thermostable alkaline alpha-amylase producing Bacillus sp. A3-15 was isolated from compost samples. There was a slight variation in amylase synthesis within the pH range 6.0 and 12.0 with an optimum pH of 8.5 (8mm zone diameter in agar medium) on starch agar medium. Analyses of the enzyme for molecular mass and amylolytic activity were carried out by starch SDS-PAGE electrophoresis, which revealed two independent bands (86,000 and 60,500 Da). Enzyme synthesis occurred at temperatures between 25 and 65 degrees C with an optimum of 60 degrees C on petri dishes. The partial purification enzyme showed optimum activity at pH 11.0 and 70 degrees C. The enzyme was highly active (95%) in alkaline range of pH (10.0-11.5), and it was almost completely active up to 100 degrees C with 96% of the original activity remaining after heat treatment at 100 degrees C for 30 min. Enzyme activity was enhanced in the presence of 5mM CaCl2 (130%) and inhibition with 5mM by ZnCl2, NaCl, Na-sulphide, EDTA, PMSF (3mM), Urea (8M) and SDS (1%) was obtained 18%, 20%, 36%, 5%, 10%, 80% and 18%, respectively. The enzyme was stable approximately 70% at pH 10.0-11.0 and 60 degrees C for 24h. So our result showed that the enzyme was both, highly thermostable-alkaline, thermophile and chelator resistant. The A3-15 amylase enzyme may be suitable in liquefaction of starch in high temperature, in detergent and textile industries and in other industrial applications.

Sorption of basic dyes from aqueous solution by activated sludge.

The adsorption of dyes in the solutions using activated sludge might be a promising approach in wastewater treatment units. The adsorption of Basic Red 18 and Basic Blue 9 from aqueous solution by dried activated sludge was investigated with in a batch system. The activated sludge had the highest dye uptake capacity, having the monolayer adsorption capacity 285.71 and 256.41 mg g(-1) for Basic Red 18 and Basic Blue 9, respectively, at pH value of 7.0 and 20 degrees C. Langmuir and Freundlich adsorption models were used for the mathematical description of the adsorption equilibrium and the equilibrium data fixed very well with both the Langmuir and Freundlich models. The R(L) values showed that, activated sludge was favorable for the adsorption of basic dyes. The suitability of the kinetic models for the adsorption of dyes on the activated sludge was also discussed. It was clear that the adsorption kinetics of dyes to dried activated sludge obeyed pseudo second-order adsorption kinetics.

Isolation of alcohol tolerant, osmotolerant and thermotolerant yeast strains and improvement of their alcohol tolerance by UV mutagenesis.

In this study, the yeast strains were isolated from grapes by serial dilution technique to determine their alcohol-, sugar- and thermotolerance. 34 wild type yeast strains were isolated and alcohol-, sugar- and thermotolerance of these strains were determined. The maximum alcohol tolerance was found to be 9% (v/v) in yeast strain which is named Y2. Thermotolerance behavior of 6 strains were investigated. The strains were treated with UV light with intervals of 20, 30, 40 and 50 seconds. Selected resistant colonies were investigated for alcohol tolerance. It was found that alcohol tolerance increased from 9% (v/v) to 12% (v/v) on Y2 strain.