Enzyme Research最新文献_第2页

Plackett-Burman Design for rGILCC1 Laccase Activity Enhancement in Pichia pastoris: Concentrated Enzyme Kinetic Characterization. 提高毕赤酵母rGILCC1漆酶活性的Plackett-Burman设计:浓缩酶动力学表征。

Enzyme Research Pub Date : 2017-01-01 Epub Date: 2017-03-21 DOI: 10.1155/2017/5947581

Edwin D Morales-Álvarez, Claudia M Rivera-Hoyos, Ángela M Cardozo-Bernal, Raúl A Poutou-Piñales, Aura M Pedroza-Rodríguez, Dennis J Díaz-Rincón, Alexander Rodríguez-López, Carlos J Alméciga-Díaz, Claudia L Cuervo-Patiño

{"title":"Plackett-Burman Design for rGILCC1 Laccase Activity Enhancement in Pichia pastoris: Concentrated Enzyme Kinetic Characterization.","authors":"Edwin D Morales-Álvarez, Claudia M Rivera-Hoyos, Ángela M Cardozo-Bernal, Raúl A Poutou-Piñales, Aura M Pedroza-Rodríguez, Dennis J Díaz-Rincón, Alexander Rodríguez-López, Carlos J Alméciga-Díaz, Claudia L Cuervo-Patiño","doi":"10.1155/2017/5947581","DOIUrl":"https://doi.org/10.1155/2017/5947581","url":null,"abstract":"Laccases are multicopper oxidases that catalyze aromatic and nonaromatic compounds with concomitant reduction of molecular oxygen to water. They are of great interest due to their potential biotechnological applications. In this work we statistically improved culture media for recombinant GILCC1 (rGILCC1) laccase production at low scale from Ganoderma lucidum containing the construct pGAPZαA-GlucPost-Stop in Pichia pastoris. Temperature, pH stability, and kinetic parameter characterizations were determined by monitoring concentrate enzyme oxidation at different ABTS substrate concentrations. Plackett-Burman Design allowed improving enzyme activity from previous work 36.08-fold, with a laccase activity of 4.69 ± 0.39 UL-1 at 168 h of culture in a 500 mL shake-flask. Concentrated rGILCC1 remained stable between 10 and 50°C and retained a residual enzymatic activity greater than 70% at 60°C and 50% at 70°C. In regard to pH stability, concentrated enzyme was more stable at pH 4.0 ± 0.2 with a residual activity greater than 90%. The lowest residual activity greater than 55% was obtained at pH 10.0 ± 0.2. Furthermore, calculated apparent enzyme kinetic parameters were a Vmax of 6.87 × 10-5 mM s-1, with an apparent Km of 5.36 × 10-2 mM. Collectively, these important stability findings open possibilities for applications involving a wide pH and temperature ranges.","PeriodicalId":11835,"journal":{"name":"Enzyme Research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2017/5947581","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34923533","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: 9

The Importance of Surface-Binding Site towards Starch-Adsorptivity Level in α-Amylase: A Review on Structural Point of View. α-淀粉酶表面结合位点对淀粉吸附水平的重要性:从结构角度综述

Enzyme Research Pub Date : 2017-01-01 Epub Date: 2017-12-05 DOI: 10.1155/2017/4086845

Umi Baroroh, Muhammad Yusuf, Saadah Diana Rachman, Safri Ishmayana, Mas Rizky A A Syamsunarno, Jutti Levita, Toto Subroto

{"title":"The Importance of Surface-Binding Site towards Starch-Adsorptivity Level in α-Amylase: A Review on Structural Point of View.","authors":"Umi Baroroh, Muhammad Yusuf, Saadah Diana Rachman, Safri Ishmayana, Mas Rizky A A Syamsunarno, Jutti Levita, Toto Subroto","doi":"10.1155/2017/4086845","DOIUrl":"https://doi.org/10.1155/2017/4086845","url":null,"abstract":"Starch is a polymeric carbohydrate composed of glucose. As a source of energy, starch can be degraded by various amylolytic enzymes, including α-amylase. In a large-scale industry, starch processing cost is still expensive due to the requirement of high temperature during the gelatinization step. Therefore, α-amylase with raw starch digesting ability could decrease the energy cost by avoiding the high gelatinization temperature. It is known that the carbohydrate-binding module (CBM) and the surface-binding site (SBS) of α-amylase could facilitate the substrate binding to the enzyme's active site to enhance the starch digestion. These sites are a noncatalytic module, which could interact with a lengthy substrate such as insoluble starch. The major interaction between these sites and the substrate is the CH/pi-stacking interaction with the glucose ring. Several mutation studies on the Halothermothrix orenii, SusG Bacteroides thetaiotamicron, Barley, Aspergillus niger, and Saccharomycopsis fibuligera α-amylases have revealed that the stacking interaction through the aromatic residues at the SBS is essential to the starch adsorption. In this review, the SBS in various α-amylases is also presented. Therefore, based on the structural point of view, SBS is suggested as an essential site in α-amylase to increase its catalytic activity, especially towards the insoluble starch.","PeriodicalId":11835,"journal":{"name":"Enzyme Research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2017/4086845","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35758451","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: 21

Recycle of Immobilized Endocellulases in Different Conditions for Cellulose Hydrolysis. 固定化内纤维素酶在不同水解条件下的循环利用。

Enzyme Research Pub Date : 2017-01-01 Epub Date: 2017-03-29 DOI: 10.1155/2017/4362704

D F Silva, A F A Carvalho, T Y Shinya, G S Mazali, R D Herculano, P Oliva-Neto

引用次数: 9

Production of Recombinant Trichoderma reesei Cellobiohydrolase II in a New Expression System Based on Wickerhamomyces anomalus. 重组里氏木霉纤维生物水解酶ⅱ的新表达体系构建

Enzyme Research Pub Date : 2017-01-01 Epub Date: 2017-08-30 DOI: 10.1155/2017/6980565

Dennis J Díaz-Rincón, Ivonne Duque, Erika Osorio, Alexander Rodríguez-López, Angela Espejo-Mojica, Claudia M Parra-Giraldo, Raúl A Poutou-Piñales, Carlos J Alméciga-Díaz, Balkys Quevedo-Hidalgo

{"title":"Production of Recombinant Trichoderma reesei Cellobiohydrolase II in a New Expression System Based on Wickerhamomyces anomalus.","authors":"Dennis J Díaz-Rincón, Ivonne Duque, Erika Osorio, Alexander Rodríguez-López, Angela Espejo-Mojica, Claudia M Parra-Giraldo, Raúl A Poutou-Piñales, Carlos J Alméciga-Díaz, Balkys Quevedo-Hidalgo","doi":"10.1155/2017/6980565","DOIUrl":"https://doi.org/10.1155/2017/6980565","url":null,"abstract":"Cellulase is a family of at least three groups of enzymes that participate in the sequential hydrolysis of cellulose. Recombinant expression of cellulases might allow reducing their production times and increasing the low proteins concentrations obtained with filamentous fungi. In this study, we describe the production of Trichoderma reesei cellobiohydrolase II (CBHII) in a native strain of Wickerhamomyces anomalus. Recombinant CBHII was expressed in W. anomalus 54-A reaching enzyme activity values of up to 14.5 U L-1. The enzyme extract showed optimum pH and temperature of 5.0-6.0 and 40°C, respectively. Enzyme kinetic parameters (KM of 2.73 mM and Vmax of 23.1 µM min-1) were between the ranges of values reported for other CBHII enzymes. Finally, the results showed that an enzymatic extract of W. anomalus 54-A carrying the recombinant T. reesei CBHII allows production of reducing sugars similar to that of a crude extract from cellulolytic fungi. These results show the first report on the use of W. anomalus as a host to produce recombinant proteins. In addition, recombinant T. reesei CBHII enzyme could potentially be used in the degradation of lignocellulosic residues to produce bioethanol, based on its pH and temperature activity profile.","PeriodicalId":11835,"journal":{"name":"Enzyme Research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2017/6980565","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35390935","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: 7

"In Silico" Characterization of 3-Phytase A and 3-Phytase B from Aspergillus niger. 黑曲霉3-植酸酶A和3-植酸酶B的“计算机”鉴定。

Enzyme Research Pub Date : 2017-01-01 Epub Date: 2017-11-20 DOI: 10.1155/2017/9746191

Doris C Niño-Gómez, Claudia M Rivera-Hoyos, Edwin D Morales-Álvarez, Edgar A Reyes-Montaño, Nury E Vargas-Alejo, Ingrid N Ramírez-Casallas, Kübra Erkan Türkmen, Homero Sáenz-Suárez, José A Sáenz-Moreno, Raúl A Poutou-Piñales, Janneth González-Santos, Azucena Arévalo-Galvis

{"title":"\"In Silico\" Characterization of 3-Phytase A and 3-Phytase B from Aspergillus niger.","authors":"Doris C Niño-Gómez, Claudia M Rivera-Hoyos, Edwin D Morales-Álvarez, Edgar A Reyes-Montaño, Nury E Vargas-Alejo, Ingrid N Ramírez-Casallas, Kübra Erkan Türkmen, Homero Sáenz-Suárez, José A Sáenz-Moreno, Raúl A Poutou-Piñales, Janneth González-Santos, Azucena Arévalo-Galvis","doi":"10.1155/2017/9746191","DOIUrl":"https://doi.org/10.1155/2017/9746191","url":null,"abstract":"Phytases are used for feeding monogastric animals, because they hydrolyze phytic acid generating inorganic phosphate. Aspergillus niger 3-phytase A (PDB: 3K4Q) and 3-phytase B (PDB: 1QFX) were characterized using bioinformatic tools. Results showed that both enzymes have highly conserved catalytic pockets, supporting their classification as histidine acid phosphatases. 2D structures consist of 43% alpha-helix, 12% beta-sheet, and 45% others and 38% alpha-helix, 12% beta-sheet, and 50% others, respectively, and pI 4.94 and 4.60, aliphatic index 72.25 and 70.26 and average hydrophobicity of -0,304 and -0.330, respectively, suggesting aqueous media interaction. Glycosylation and glycation sites allowed detecting zones that can affect folding and biological activity, suggesting fragmentation. Docking showed that H59 and H63 act as nucleophiles and that D339 and D319 are proton donor residues. MW of 3K4Q (48.84 kDa) and 1QFX (50.78 kDa) is similar; 1QFX forms homodimers which will originate homotetramers with several catalytic center accessible to the ligand. 3K4Q is less stable (instability index 45.41) than 1QFX (instability index 33.66), but the estimated lifespan for 3K4Q is superior. Van der Waals interactions generate hydrogen bonds between the active center and O2 or H of the phytic acid phosphate groups, providing greater stability to these temporal molecular interactions.","PeriodicalId":11835,"journal":{"name":"Enzyme Research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2017/9746191","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35749045","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

Inhibition of α-Amylases by Condensed and Hydrolysable Tannins: Focus on Kinetics and Hypoglycemic Actions. 浓缩和水解单宁对α-淀粉酶的抑制作用:动力学和降糖作用的研究

Enzyme Research Pub Date : 2017-01-01 Epub Date: 2017-05-14 DOI: 10.1155/2017/5724902

Camila Gabriel Kato, Geferson de Almeida Gonçalves, Rosely Aparecida Peralta, Flavio Augusto Vicente Seixas, Anacharis Babeto de Sá-Nakanishi, Lívia Bracht, Jurandir Fernando Comar, Adelar Bracht, Rosane Marina Peralta

{"title":"Inhibition of α-Amylases by Condensed and Hydrolysable Tannins: Focus on Kinetics and Hypoglycemic Actions.","authors":"Camila Gabriel Kato, Geferson de Almeida Gonçalves, Rosely Aparecida Peralta, Flavio Augusto Vicente Seixas, Anacharis Babeto de Sá-Nakanishi, Lívia Bracht, Jurandir Fernando Comar, Adelar Bracht, Rosane Marina Peralta","doi":"10.1155/2017/5724902","DOIUrl":"https://doi.org/10.1155/2017/5724902","url":null,"abstract":"The aim of the present study was to compare the in vitro inhibitory effects on the salivary and pancreatic α-amylases and the in vivo hypoglycemic actions of the hydrolysable tannin from Chinese natural gall and the condensed tannin from Acacia mearnsii. The human salivary α-amylase was more strongly inhibited by the hydrolysable than by the condensed tannin, with the concentrations for 50% inhibition (IC50) being 47.0 and 285.4 μM, respectively. The inhibitory capacities of both tannins on the pancreatic α-amylase were also different, with IC50 values being 141.1 μM for the hydrolysable tannin and 248.1 μM for the condensed tannin. The kinetics of the inhibition presented complex patterns in that for both inhibitors more than one molecule can bind simultaneously to either the free enzyme of the substrate-complexed enzyme (parabolic mixed inhibition). Both tannins were able to inhibit the intestinal starch absorption. Inhibition by the hydrolysable tannin was concentration-dependent, with 53% inhibition at the dose of 58.8 μmol/kg and 88% inhibition at the dose of 294 μmol/kg. For the condensed tannin, inhibition was not substantially different for doses between 124.4 μmol/kg (49%) and 620 μmol/kg (57%). It can be concluded that both tannins, but especially the hydrolysable one, could be useful in controlling the postprandial glycemic levels in diabetes.","PeriodicalId":11835,"journal":{"name":"Enzyme Research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2017/5724902","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35066284","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: 48

Characterization of Pectinase from Bacillus subtilis Strain Btk 27 and Its Potential Application in Removal of Mucilage from Coffee Beans. 枯草芽孢杆菌Btk 27株果胶酶的特性及其在去除咖啡豆黏液中的潜在应用

Enzyme Research Pub Date : 2017-01-01 Epub Date: 2017-09-11 DOI: 10.1155/2017/7686904

Oliyad Jeilu Oumer, Dawit Abate

{"title":"Characterization of Pectinase from Bacillus subtilis Strain Btk 27 and Its Potential Application in Removal of Mucilage from Coffee Beans.","authors":"Oliyad Jeilu Oumer, Dawit Abate","doi":"10.1155/2017/7686904","DOIUrl":"https://doi.org/10.1155/2017/7686904","url":null,"abstract":"The demand for enzymes in the global market is projected to rise at a fast pace in recent years. There has been a great increase in industrial applications of pectinase owing to their significant biotechnological uses. For applying enzymes at industrial scale primary it is important to know the features of the enzyme. Thus, this study was undertaken with aims of characterizing the pectinase enzyme from Bacillus subtilis strain Btk27 and proving its potential application in demucilisation of coffee. In this study, the maximum pectinase activity was achieved at pH 7.5 and 50°C. Also, the enzyme activity was found stimulated with Mg2+ and Ca2+ metal ions. Moreover, it was stable on EDTA, Trixton-100, Tween 80, and Tween 20. Since Bacillus subtilis strain Btk27 was stable in most surfactants and inhibitors it could be applicable in various industries whenever pectin degradation is needed. The enzyme Km and Vmax values were identified as 1.879 mg/ml and 149.6 U, respectively. The potential application of the enzyme for coffee processing was studied, and it is found that complete removal of mucilage from coffee beans within 24 hours of treatment indicates the potential application in coffee processing.","PeriodicalId":11835,"journal":{"name":"Enzyme Research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2017/7686904","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35556243","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: 53

Molecular Analysis of CYP21A2 Gene Mutations among Iraqi Patients with Congenital Adrenal Hyperplasia 伊拉克先天性肾上腺增生患者CYP21A2基因突变的分子分析

Enzyme Research Pub Date : 2016-09-29 DOI: 10.1155/2016/9040616

Ruqayah G Y Al-Obaidi, Bassam M S Al-Musawi, Munib A. Al-Zubaidi, C. Oberkanins, S. Németh, Yusra G. Y. Al-Obaidi

{"title":"Molecular Analysis of CYP21A2 Gene Mutations among Iraqi Patients with Congenital Adrenal Hyperplasia","authors":"Ruqayah G Y Al-Obaidi, Bassam M S Al-Musawi, Munib A. Al-Zubaidi, C. Oberkanins, S. Németh, Yusra G. Y. Al-Obaidi","doi":"10.1155/2016/9040616","DOIUrl":"https://doi.org/10.1155/2016/9040616","url":null,"abstract":"Congenital adrenal hyperplasia is a group of autosomal recessive disorders. The most frequent one is 21-hydroxylase deficiency. Analyzing CYP21A2 gene mutations was so far not reported in Iraq. This work aims to analyze the spectrum and frequency of CYP21A2 mutations among Iraqi CAH patients. Sixty-two children were recruited from the Pediatric Endocrine Consultation Clinic, Children Welfare Teaching Hospital, Baghdad, Iraq, from September 2014 till June 2015. Their ages ranged between one day and 15 years. They presented with salt wasting, simple virilization, or pseudoprecocious puberty. Cytogenetic study was performed for cases with ambiguous genitalia. Molecular analysis of CYP21A2 gene was done using the CAH StripAssay (ViennaLab Diagnostics) for detection of 11 point mutations and >50% of large gene deletions/conversions. Mutations were found in 42 (67.7%) patients; 31 (50%) patients were homozygotes, 9 (14.5%) were heterozygotes, and 2 (3.2%) were compound heterozygotes with 3 mutations, while 20 (32.3%) patients had none of the tested mutations. The most frequently detected mutations were large gene deletions/conversions found in 12 (19.4%) patients, followed by I2Splice and Q318X in 8 (12.9%) patients each, I172N in 5 (8.1%) patients, and V281L in 4 (6.5%) patients. Del 8 bp, P453S, and R483P were each found in one (1.6%) and complex alleles were found in 2 (3.2%). Four point mutations (P30L, Cluster E6, L307 frameshift, and R356W) were not identified in any patient. In conclusion, gene deletions/conversions and 7 point mutations were recorded in varying proportions, the former being the commonest, generally similar to what was reported in regional countries.","PeriodicalId":11835,"journal":{"name":"Enzyme Research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2016-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86184587","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 10

Overexpression of Soluble Recombinant Human Lysyl Oxidase by Using Solubility Tags: Effects on Activity and Solubility 利用溶解度标签过表达可溶性重组人赖氨酸氧化酶:对活性和溶解度的影响

Enzyme Research Pub Date : 2016-01-31 DOI: 10.1155/2016/5098985

Madison A Smith, J. Gonzalez, Anjum Hussain, Rachel N. Oldfield, Kathryn A. Johnston, Karlo Lopez

{"title":"Overexpression of Soluble Recombinant Human Lysyl Oxidase by Using Solubility Tags: Effects on Activity and Solubility","authors":"Madison A Smith, J. Gonzalez, Anjum Hussain, Rachel N. Oldfield, Kathryn A. Johnston, Karlo Lopez","doi":"10.1155/2016/5098985","DOIUrl":"https://doi.org/10.1155/2016/5098985","url":null,"abstract":"Lysyl oxidase is an important extracellular matrix enzyme that has not been fully characterized due to its low solubility. In order to circumvent the low solubility of this enzyme, three solubility tags (Nus-A, Thioredoxin (Trx), and Glutathione-S-Transferase (GST)) were engineered on the N-terminus of mature lysyl oxidase. Total enzyme yields were determined to be 1.5 mg for the Nus-A tagged enzyme (0.75 mg/L of media), 7.84 mg for the Trx tagged enzyme (3.92 mg/L of media), and 9.33 mg for the GST tagged enzyme (4.67 mg/L of media). Enzymatic activity was calculated to be 0.11 U/mg for the Nus-A tagged enzyme and 0.032 U/mg for the Trx tagged enzyme, and no enzymatic activity was detected for the GST tagged enzyme. All three solubility-tagged forms of the enzyme incorporated copper; however, the GST tagged enzyme appears to bind adventitious copper with greater affinity than the other two forms. The catalytic cofactor, lysyl tyrosyl quinone (LTQ), was determined to be 92% for the Nus-A and Trx tagged lysyl oxidase using the previously reported extinction coefficient of 15.4 mM−1 cm−1. No LTQ was detected for the GST tagged lysyl oxidase. Given these data, it appears that Nus-A is the most suitable tag for obtaining soluble and active recombinant lysyl oxidase from E. coli culture.","PeriodicalId":11835,"journal":{"name":"Enzyme Research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2016-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89921758","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 6

Characterization of a Hyperthermostable Alkaline Lipase from Bacillus sonorensis 4R 索诺芽孢杆菌4R超耐热碱性脂肪酶的研究

Enzyme Research Pub Date : 2016-01-21 DOI: 10.1155/2016/4170684

H. Bhosale, Uzma Shaheen, T. Kadam

{"title":"Characterization of a Hyperthermostable Alkaline Lipase from Bacillus sonorensis 4R","authors":"H. Bhosale, Uzma Shaheen, T. Kadam","doi":"10.1155/2016/4170684","DOIUrl":"https://doi.org/10.1155/2016/4170684","url":null,"abstract":"Hyperthermostable alkaline lipase from Bacillus sonorensis 4R was purified and characterized. The enzyme production was carried out at 80°C and 9.0 pH in glucose-tween inorganic salt broth under static conditions for 96 h. Lipase was purified by anion exchange chromatography by 12.15 fold with a yield of 1.98%. The molecular weight of lipase was found to be 21.87 KDa by SDS-PAGE. The enzyme activity was optimal at 80°C with t 1/2 of 150 min and at 90°C, 100°C, 110°C, and 120°C; the respective values were 121.59 min, 90.01 min, 70.01 min, and 50 min. The enzyme was highly activated by Mg and t 1/2 values at 80°C were increased from 150 min to 180 min when magnesium and mannitol were added in combination. The activation energy calculated from Arrhenius plot was 31.102 KJ/mol. At 80–120°C, values of ΔH and ΔG were in the range of 28.16–27.83 KJ/mol and 102.79 KJ/mol to 111.66 KJ/mol, respectively. Lipase activity was highest at 9.0 pH and stable for 2 hours at this pH at 80°C. Pretreatment of lipase with MgSO4 and CaSO4 stimulated enzyme activity by 249.94% and 30.2%, respectively. The enzyme activity was greatly reduced by CoCl2, CdCl2, HgCl2, CuCl2, Pb(NO3)2, PMSF, orlistat, oleic acid, iodine, EDTA, and urea.","PeriodicalId":11835,"journal":{"name":"Enzyme Research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2016-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88006002","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 42