Enzyme Research最新文献_第6页

Molecular Cloning and Optimization for High Level Expression of Cold-Adapted Serine Protease from Antarctic Yeast Glaciozyma antarctica PI12. 南极酵母Glaciozyma antarctica PI12冷适应丝氨酸蛋白酶的分子克隆及高效表达优化

Enzyme Research Pub Date : 2014-01-01 Epub Date: 2014-06-30 DOI: 10.1155/2014/197938

Norsyuhada Alias, Mu'adz Ahmad Mazian, Abu Bakar Salleh, Mahiran Basri, Raja Noor Zaliha Raja Abd Rahman

{"title":"Molecular Cloning and Optimization for High Level Expression of Cold-Adapted Serine Protease from Antarctic Yeast Glaciozyma antarctica PI12.","authors":"Norsyuhada Alias, Mu'adz Ahmad Mazian, Abu Bakar Salleh, Mahiran Basri, Raja Noor Zaliha Raja Abd Rahman","doi":"10.1155/2014/197938","DOIUrl":"https://doi.org/10.1155/2014/197938","url":null,"abstract":"Psychrophilic basidiomycete yeast, Glaciozyma antarctica strain PI12, was shown to be a protease-producer. Isolation of the PI12 protease gene from genomic and mRNA sequences allowed determination of 19 exons and 18 introns. Full-length cDNA of PI12 protease gene was amplified by rapid amplification of cDNA ends (RACE) strategy with an open reading frame (ORF) of 2892 bp, coded for 963 amino acids. PI12 protease showed low homology with the subtilisin-like protease from fungus Rhodosporidium toruloides (42% identity) and no homology to other psychrophilic proteases. The gene encoding mature PI12 protease was cloned into Pichia pastoris expression vector, pPIC9, and positioned under the induction of methanol-alcohol oxidase (AOX) promoter. The recombinant PI12 protease was efficiently secreted into the culture medium driven by the Saccharomyces cerevisiae α-factor signal sequence. The highest protease production (28.3 U/ml) was obtained from P. pastoris GS115 host (GpPro2) at 20°C after 72 hours of postinduction time with 0.5% (v/v) of methanol inducer. The expressed protein was detected by SDS-PAGE and activity staining with a molecular weight of 99 kDa. ","PeriodicalId":11835,"journal":{"name":"Enzyme Research","volume":"2014 ","pages":"197938"},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2014/197938","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32561390","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 24

Mode of action of lactoperoxidase as related to its antimicrobial activity: a review. 乳过氧化物酶的作用模式与其抗菌活性的关系：综述。

Enzyme Research Pub Date : 2014-01-01 Epub Date: 2014-09-16 DOI: 10.1155/2014/517164

F Bafort, O Parisi, J-P Perraudin, M H Jijakli

引用次数: 0

A Fractional Factorial Design to Study the Effect of Process Variables on the Preparation of Hyaluronidase Loaded PLGA Nanoparticles. 用分数析因设计研究工艺变量对制备透明质酸酶负载PLGA纳米颗粒的影响。

Enzyme Research Pub Date : 2014-01-01 Epub Date: 2014-12-10 DOI: 10.1155/2014/162962

K Narayanan, V M Subrahmanyam, J Venkata Rao

引用次数: 48

Partial Purification and Characterization of a Heat Stable α-Amylase from a Thermophilic Actinobacteria, Streptomyces sp. MSC702. 嗜热放线菌链霉菌 MSC702 中热稳定的 α 淀粉酶的部分纯化和特性。

Enzyme Research Pub Date : 2014-01-01 Epub Date: 2014-10-08 DOI: 10.1155/2014/106363

Renu Singh, Vijay Kumar, Vishal Kapoor

{"title":"Partial Purification and Characterization of a Heat Stable α-Amylase from a Thermophilic Actinobacteria, Streptomyces sp. MSC702.","authors":"Renu Singh, Vijay Kumar, Vishal Kapoor","doi":"10.1155/2014/106363","DOIUrl":"10.1155/2014/106363","url":null,"abstract":"A partial purification and biochemical characterization of the α-amylase from Streptomyces sp. MSC702 were carried out in this study. The optimum operational conditions for enzyme substrate reaction for amylolytic enzyme activity from the strain were evaluated. The optimum pH, temperature, and incubation period for assaying the enzyme were observed to be 5.0, 55°C, and 30 min, respectively. The extracellular extract was concentrated using ammonium sulfate precipitation. It was stable in the presence of metal ions (5 mM) such as K(+), Co(2+), and Mo(2+), whereas Pb(2+), Mn(2+), Mg(2+), Cu(2+), Zn(2+), Ba(2+), Ca(2+), Hg(2+), Sn(2+), Cr(3+), Al(3+), Ag(+), and Fe(2+) were found to have inhibitory effects. The enzyme activity was also unstable in the presence of 1% Triton X-100, 1% Tween 80, 5 mM sodium lauryl sulphate, 1% glycerol, 5 mM EDTA, and 5 mM denaturant urea. At temperature 60°C and pH 5.0, the enzyme stability was maximum. α-amylase retained 100% and 34.18% stability for 1 h and 4 h, respectively, at 60°C (pH 7.0). The enzyme exhibited a half-life of 195 min at 60°C temperature. The analysis of kinetic showed that the enzyme has K m of 2.4 mg/mL and V max of 21853.0 μmol/min/mg for soluble potato starch. The results indicate that the enzyme reflects their potentiality towards industrial utilization. ","PeriodicalId":11835,"journal":{"name":"Enzyme Research","volume":"2014 ","pages":"106363"},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4220580/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32817162","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A Method for Fast Assessment of OP/CB Exposure in the Japanese Quail (Coturnix coturnix japonica) Using Combined Esterases Enzyme Activity as Biomarkers. 用联合酯酶活性作为生物标志物快速评价日本鹌鹑(Coturnix Coturnix japonica)暴露于OP/CB的方法

Enzyme Research Pub Date : 2014-01-01 Epub Date: 2014-01-09 DOI: 10.1155/2014/812302

Kasim Sakran Abass

{"title":"A Method for Fast Assessment of OP/CB Exposure in the Japanese Quail (Coturnix coturnix japonica) Using Combined Esterases Enzyme Activity as Biomarkers.","authors":"Kasim Sakran Abass","doi":"10.1155/2014/812302","DOIUrl":"https://doi.org/10.1155/2014/812302","url":null,"abstract":"The aims of this study were to investigate the presence of different esterase activities in plasma and liver for Japanese quail and to combine determination of both carboxylesterase and cholinesterase as biochemical biomarker in order to identify the effects of carbamate and organophosphate compounds exposure. Carboxylesterase exhibits larger sensitivity to carbamate and organophosphate compounds than to cholinesterase and is present at higher levels. This permitted nature and distribution of carboxylesterase or cholinesterase to be measured. One predominant toxicological form of enzyme level constant in its patterns of motivation and inhibition with cholinesterase was identified in plasma with an apparent Michaelis constant for butyrylthiocholine iodide of 0.394 mM. Carboxylesterase activity in liver was considered by its preferential hydrolysis of the S-phenyl thioacetate. A concentration dependent decrease of carboxylesterase and cholinesterase has demonstrated during in vitro incubation of malathion, parathion, and trichlorfon in the range 0.125-2 mM, while with methomyl was in the range 0.25-4 mM. When quail (n = 15) was exposed orally for 48 h to concentrations of carbamate or organophosphate compounds of 3-200 mg/kg, the percentage inhibition of cholinesterase was in each case larger than that of carboxylesterase and reached statistical significance (P < 0.05) at lower concentrations. ","PeriodicalId":11835,"journal":{"name":"Enzyme Research","volume":"2014 ","pages":"812302"},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2014/812302","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32115380","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 11

Esterase Active in Polar Organic Solvents from the Yeast Pseudozyma sp. NII 08165. 酵母假酶在极性有机溶剂中活性的酯酶。

Enzyme Research Pub Date : 2014-01-01 Epub Date: 2014-04-03 DOI: 10.1155/2014/494682

Deepthy Alex, Anju Shainu, Ashok Pandey, Rajeev K Sukumaran

{"title":"Esterase Active in Polar Organic Solvents from the Yeast Pseudozyma sp. NII 08165.","authors":"Deepthy Alex, Anju Shainu, Ashok Pandey, Rajeev K Sukumaran","doi":"10.1155/2014/494682","DOIUrl":"https://doi.org/10.1155/2014/494682","url":null,"abstract":"Esterases/lipases active in water miscible solvents are highly desired in biocatalysis where substrate solubility is limited and also when the solvent is desired as an acyl acceptor in transesterification reactions, as with the case of biodiesel production. We have isolated an esterase from the glycolipid producing yeast-Pseudozyma sp. NII 08165 which in its crude form was alkali active, thermo stable, halo tolerant and also capable of acting in presence of high methanol concentration. The crude enzyme which maintained 90% of its original activity after being treated at 70°C was purified and the properties were characterized. The partially purified esterase preparation had temperature and pH optima of 60°C and 8.0 respectively. The enzyme retained almost complete activity in presence of 25% methanol and 80% activity in the same strength of ethanol. Conditions of enzyme production were optimized, which lead to 9 fold increase in the esterase yield. One of the isoforms of the enzyme LIP1 was purified to homogeneity and characterized. Purified LIP1 had a K m and V max of 0.01 and 1.12, respectively. The purified esterase lost its thermo and halo tolerance but interestingly, retained 97% activity in methanol. ","PeriodicalId":11835,"journal":{"name":"Enzyme Research","volume":"2014 ","pages":"494682"},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2014/494682","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32318193","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 16

Immobilization of Aspergillus niger F7-02 Lipase in Polysaccharide Hydrogel Beads of Irvingia gabonensis Matrix. 黑曲霉F7-02脂肪酶在加蓬树基质多糖水凝胶珠中的固定化。

Enzyme Research Pub Date : 2014-01-01 Epub Date: 2014-12-31 DOI: 10.1155/2014/967056

Safaradeen Olateju Kareem, Olayinka Quadri Adio, Michael Bamitale Osho

{"title":"Immobilization of Aspergillus niger F7-02 Lipase in Polysaccharide Hydrogel Beads of Irvingia gabonensis Matrix.","authors":"Safaradeen Olateju Kareem, Olayinka Quadri Adio, Michael Bamitale Osho","doi":"10.1155/2014/967056","DOIUrl":"https://doi.org/10.1155/2014/967056","url":null,"abstract":"The potential of polysaccharide Irvingia gabonensis matrix as enzyme immobilization support was investigated. Lipase of Aspergillus niger F7-02 was immobilized by entrapment using glutaraldehyde as the cross-linking agent and stabilized in ethanolic-formaldehyde solution. The pH and temperature stability and activity yield of the immobilized enzyme were determined. Such parameters as enzyme load, bead size, number of beads, and bead reusability were also optimized. Adequate gel strength to form stabilized beads was achieved at 15.52% (w/v) Irvingia gabonensis powder, 15% (v/v) partially purified lipase, 2.5% (v/v) glutaraldehyde, and 3 : 1 (v/v) ethanolic-formaldehyde solution. There was 3.93-fold purification when the crude enzyme was partially purified in two-step purification using Imarsil and activated charcoal. Optimum lipase activity 75.3 Ug(-1) was achieved in 50 mL test solution containing 15 beads of 7 mm bead size. Relative activity 80% was retained at eight repeated cycles. The immobilization process gave activity yield of 59.1% with specific activity of 12.3 Umg(-1) and stabilized at optimum pH 4.5 and temperature 55°C. Thus the effectiveness and cost-efficiency of I. gabonensis as a polymer matrix for lipase immobilization have been established. ","PeriodicalId":11835,"journal":{"name":"Enzyme Research","volume":"2014 ","pages":"967056"},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2014/967056","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32997448","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 5

Enhanced thermostability of a fungal alkaline protease by different additives. 不同添加剂对真菌碱性蛋白酶热稳定性的影响。

Enzyme Research Pub Date : 2014-01-01 Epub Date: 2014-07-03 DOI: 10.1155/2014/109303

Nilesh P Nirmal, R Seeta Laxman

引用次数: 16

Effect of Chromium(VI) Toxicity on Enzymes of Nitrogen Metabolism in Clusterbean (Cyamopsis tetragonoloba L.). 铬(VI)毒性对四季豆氮代谢酶的影响

Enzyme Research Pub Date : 2014-01-01 Epub Date: 2014-03-18 DOI: 10.1155/2014/784036

Punesh Sangwan, Vinod Kumar, U N Joshi

引用次数: 0

Molecular Dynamics and Metadynamics Simulations of the Cellulase Cel48F. 纤维素酶Cel48F的分子动力学和元动力学模拟。

Enzyme Research Pub Date : 2014-01-01 Epub Date: 2014-05-21 DOI: 10.1155/2014/692738

Osmair Vital de Oliveira

{"title":"Molecular Dynamics and Metadynamics Simulations of the Cellulase Cel48F.","authors":"Osmair Vital de Oliveira","doi":"10.1155/2014/692738","DOIUrl":"https://doi.org/10.1155/2014/692738","url":null,"abstract":"Molecular dynamics (MD) and metadynamics techniques were used to study the cellulase Cel48F-sugar. Cellulase is enzyme that breaks cellulose fibers into small sugar units and is potentially useful in second generation alcohol production. In MD simulations, the overall structure of equilibrated Cel48F did not significantly change along the trajectory, retaining root mean square deviation below 0.15 nm. A set of 15 residues interacting with the sugar chains via hydrogen bonding throughout the simulation was observed. The free energy of dissociation (ΔGdiss.) of the chains in the catalytic tunnel of Cel48F was determined by metadynamics. The ΔGdiss. values of the chains entering and leaving the wild-type Cel48F cavity were 13.9 and 62.1 kcal/mol, respectively. We also mutated the E542 and Q543 to alanine residue and obtained ΔGdiss. of 41.8 and 45.9 kcal/mol, respectively. These mutations were found to facilitate smooth dissociation of the sugar chain across the Cel48F tunnel. At the entry of the Cel48F tunnel, three residues were mutated to alanine: T110, T213, and L274. Contrary to the T110A-Cel48F, the mutants T213-Cel48F and L274-Cel48F prevented the sugar chain from passing across the leaving site. The present results can be a guideline in mutagenesis studies to improve processing by Cel48F. ","PeriodicalId":11835,"journal":{"name":"Enzyme Research","volume":"2014 ","pages":"692738"},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2014/692738","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32453134","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 5