scholarly journals Characterization of a novel laccase purified from the fungus Hohenbuehelia serotina and its decolourisation of dyes.

2015 ◽  
Vol 63 (2) ◽  
Author(s):  
Yingyin Xu ◽  
Yuxiao Lu ◽  
Rui Zhang ◽  
Hexiang Wang ◽  
Qinghong Liu
Keyword(s):  
Molecular Weight ◽  
Enzyme Activity ◽  
Enzymatic Activity ◽  
Inhibitory Effect ◽  
Single Band ◽  
White Rot ◽  
White Rot Fungus ◽  
Sds Page

A novel laccase was purified from the white rot fungus, Hohenbuehelia serotina, to investigate the applications of this laccase in the decoloration of various dyes. SDS-PAGE revealed a single band of this laccase corresponding to a molecular weight of approximately 57.8 kDa. The enzyme showed activity towards several substrates, the most sensitive of which was 2,2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS). The highest enzymatic activity using ABTS as a substrate was observed at pH 6.8 and 30°C. The enzyme activity was found to be significantly enhanced in the presence of Zn(2+) ions and inhibited by Fe(2+) ions. Moreover, SDS and β-mercaptoethanol were inhibitory, and inhibition by L-cysteine was observed while EDTA and DMSO had almost no inhibitory effect. The laccase could effectively decolorize seven different dyes within 30 minutes at 40°C.

scholarly journals PURIFICATION AND CHARACTERIZATION OF LACCASE INVOLVED IN THE DECOLOURIZATION OF SYNTHETIC DYES AND 2,3,7,8-TCDD CONGENER DEGRADATION BY THE WHITE ROT FUNGUS ISOLATED FROM BAVI FOREST OF VIETNAM

2018 ◽  
Vol 55 (4C) ◽  
pp. 180 ◽  
Author(s):  
Phung Khac Huy Chu
Keyword(s):  
Enzymatic Activity ◽  
Metal Ions ◽  
Textile Industry ◽  
White Rot ◽  
White Rot Fungus ◽  
Synthetic Dyes ◽  
Sds Page ◽  
Extracellular Laccase ◽  
Optimum Ph

The fungal strain FBV40 was isolated from soil containing decayed wood in Ba Vi National Forest and capable of producing an extracellular laccase in the TSH1 medium. Two isozyme such as Lac1 and Lac2 were purified were estimated to be 55 and 60 kDa by SDS-PAGE. The optimum pH and temperature for the enzymatic activity of Lac1 were 3.0 and 60°C with ABTS using as the substrate. Kinetic constants Km and Vmax of Lac 1 were 0.3 µM and 200,000 µM/mins with ABTS as substrate. Cl-, SDS, and EDTA at any concentration (2 mM; 5 mM and 10 mM) strongly inhibited the activity of laccase. The enzyme was stable in the presence of several metal ions including Ni2+ (1 mM), Cu2+ (1 mM and 3 mM), Ca2+ (3 mM and 4 mM); in the presence of Cu2+ (2 mM) and Ca2+ (0.5 mM, 1.0 mM and 2.0 mM), laccase even showed the increase in the activity. The presence of metal ions Mn2+, Mg2+, Fe2+ completely inhibited the enzymatic activity at any examined concentration. The crude enzyme, as well as Lac 1, was able to decolourization MN.FBN dye from the textile industry from Ministry of Defence. This strain was able to degrade 2,3,7,8-TCDD isotop with initial concentration 2,000 ng-TEQ/L at rates over 46.8 % after ten days cultivation in the TSH1 medium. In the presence of three strains FBV40, FBVLa1 and FBD154 with the ratio 1:1:1, the degradation of this congener was achieved more than 95 % at the same time cultivation.

scholarly journals Characterization of cellulase from E. coli BPPTCC-EGRK2

2018 ◽  
Vol 52 ◽  
pp. 00024
Author(s):  
Mas Gunawan Haryanto ◽  
Siswa Setyahadi ◽  
Muhammad Sahlan ◽  
Masafumi Yohda ◽  
Yosuke Fukutani ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Enzyme Activity ◽  
Animal Feed ◽  
Maximum Velocity ◽  
Cellulase Production ◽  
Luria Bertani ◽  
E Coli ◽  
Sds Page ◽  
Weight Analysis

Cellulase enzymes are widely used in various industries such as detergent industry, bioethanol, animal feed, textile and paper. This research focused on characterization of cellulase produced from Eschericia coli BPPT-CC EgRK2, which is a recombinant that can produce protein enzymes endo- β-1,4-glucanase. Eschericia coli BPPT-CC EgRK2 was cultured in 1 litre liquid medium Luria Bertani. Because the bacteria is intracellular, sonication is needed for cell disruption to get the cellulase enzyme. The enzyme activity was then analyzed by CMC substrate at different concentration. The protein content analysis was carried out using Bradford method; the molecular weight analysis was done using SDS-PAGE; while the enzyme kinetics was plotted using Michaelis-Menten model. Our results showed the highest enzyme activity was 2.403 U/ml and the protein concentration was 5.352 mg/ml. The Michaelis-Menten constant (Km) and maximum velocity (Vmax) for CMC substrate hydrolysis were 0.07 μmol/ml and 2.49 μmol/ml/sec, respectively. The cellulase molecular weight was 58 kDa using SDS-PAGE with 7.5% stacking gel. The results indicated that Eschericia coli BPPT-CC EgRK2 is a promising renewable source for cellulase production for industrial application.

Molecular characterization of an endo-type chitosanase from the fish pathogenRenibacteriumsp. QD1

2014 ◽  
Vol 94 (4) ◽  
pp. 681-686 ◽  
Author(s):  
Peichuan Xing ◽  
Dan Liu ◽  
Wen-Gong Yu ◽  
Xinzhi Lu
Keyword(s):  
Molecular Weight ◽  
Enzyme Activity ◽  
Specific Activity ◽  
Fermentation Broth ◽  
Maximal Activity ◽  
Optimum Temperature ◽  
Sds Page ◽  
Ph Range ◽  
Tlc Analysis

Renibacteriumsp. QD1, a bacteria strain capable of hydrolysing chitosan, was isolated from the homogenate of small crabs. An extracellular chitosanase, Csn-A, was purified from the QD1 fermentation broth. The enzyme was purified to homogeneity, with a yield of eight-fold, 67% recovery and a specific activity of 1575 U/mg proteins. The molecular weight of Csn-A was estimated to be 26.1 kDa by SDS-PAGE. Unlike other chitosanases, the purified Csn-A displayed maximal activity at a pH range of 5.3–6.5, and it was stable in a broad pH range of 5.0–10.0. The optimum temperature for chitosanlytic activity was 55°C. The enzyme activity was strongly stimulated by Mn2+but inhibited by Fe3+, Cu2+, Al3+, Zn2+and SDS. TLC analysis demonstrated that Csn-A hydrolysed N-deacetylated polymeric glucosamines into chito-biose and -triose in an endo-type manner. The amino acid seuquence of Csn-A showed close identity with an uncharacterized chitosanase of strain ATCC33209.

Enzymological Characterization of Secreted Proteinases from Candida parapsilosis and Candida lusitaniae

2001 ◽  
Vol 66 (11) ◽  
pp. 1707-1719 ◽  
Author(s):  
Olga Hrušková-Heidingsfeldová ◽  
Jiří Dostál ◽  
Petr Hamal ◽  
Jarmila Pazlarová ◽  
Tomáš Ruml ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Enzyme Activity ◽  
Candida Parapsilosis ◽  
Yeast Culture ◽  
Opportunistic Pathogens ◽  
Novel Drugs ◽  
Sds Page ◽  
Candida Lusitaniae ◽  
Culture Supernatants

Opportunistic pathogens of the genusCandidaproduce secreted aspartic proteinases (Saps) that are considered as one of the virulence factors. While Saps ofC. albicansandC. tropicalishave been studied extensively, information concerning proteinases secreted by other pathogenicCandidaspecies is scarce. To our knowledge, enzymologic characterizations of Sap ofC. parapsilosis(Sapp1p) andC. lusitaniae(Saplp) are limited. We have purified Saps ofC. parapsilosisandC. lusitaniaefrom yeast culture supernatants and detected only one gene product of both Sapp1p and Saplp using isoelectric focusing and N-terminal sequencing. Molecular weight of Saplp determined by SDS-PAGE is approximately 42 000. For the highest enzyme activity, both Sapp1p and Saplp require pH ranging from 3 to 4 and temperature between 27-40 °C for Saplp and 45 °C for Sapp1p, respectively. At physiological temperature, both the proteinases are stable. The characterization of Saps of clinical isolates ofC. parapsilosisandC. lusitaniaeshould contribute to design of novel drugs specifically targeted on proteinases.

Characterization of a Laccase from White Rot Fungus Cerrena unicolor

2013 ◽  
Vol 442 ◽  
pp. 120-124
Author(s):  
Bei Bei Zhang ◽  
Yong Jian Shen ◽  
Chang Xu ◽  
Ying Li ◽  
Ming Hu Jiang
Keyword(s):  
Dye Decolorization ◽  
White Rot ◽  
White Rot Fungus ◽  
Optimum Temperature ◽  
Complex Chemical ◽  
Chemical Structures ◽  
Cerrena Unicolor ◽  
Sds Page ◽  
Optimum Ph

Dyes are usually difficult to be decolorized due to their complex chemical structures. In this work, laccase was purified from the white rot fungus Cerrena unicolor to evaluate its application in dye decolorization. SDS-PAGE analysis showed the purified laccase to be a monomeric protein of 63.7 kDa. The optimum pH for the oxidation of 2,2-azinobis-(3-ethylbenzthiaoline-6-sufonic acid) (ABTS) was 2.2 and the optimum temperature was 50°C. The activity of the purified enzyme was strongly inhibited by sodium azide and partially inhibited by cysteine, dithiothreitol. The Km values of the purified laccase for the substrate ABTS, syringaldazine and 2,6-dimethoxyphenol were 0.217, 0.306 and 0.199 mmol/L.

scholarly journals Immunohistochemical and immunochemical characterization of a new endothelial cell-specific antigen.

1986 ◽  
Vol 34 (2) ◽  
pp. 209-214 ◽  
Author(s):  
J U Alles ◽  
K Bosslet
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibody ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Specific Antigen ◽  
Similar Molecular Weight ◽  
Immunochemical Characterization ◽  
Sds Page

A new monoclonal antibody (moab BW 200) of IgG3 kappa-isotype was generated which recognizes an epitope located on an antigen molecule restricted to human neoplastic and non-neoplastic endothelial cells. The molecular weight of the antigen was determined using immunoprecipitation analysis followed by SDS-PAGE. Despite its similar molecular weight to FVIII-RAG, the antigen detected by moab BW 200 was shown to be different from FVIII-RAG.

Sinapine Esterase I. Characterization of Sinapine Esterase from Cotyledons of Raphanus sativus

1979 ◽  
Vol 34 (9-10) ◽  
pp. 715-720 ◽  
Author(s):  
Gerhild Nurmann ◽  
Dieter Strack
Keyword(s):  
Enzyme Activity ◽  
Esterase Activity ◽  
Raphanus Sativus ◽  
Sinapic Acid ◽  
Inhibitory Effect ◽  
Ph Optimum ◽  
Diisopropyl Fluorophosphate ◽  
E 600 ◽  
Red Radish

Abstract From cotyledons of Raphanus sativus (red radish) an esterase activity which catalyzes the hy­drolysis of sinapine into sinapic acid and choline has been isolated. The enzyme, which has a near absolute specificity, is not analogous with any esterase described in the literature. The reaction has a pH optimum of 8.5 and the apparent Km is 1.95 × 10-5 m. The enzyme is relatively insensi­tive to both physostigmine (eserine) {Ki = 1.73 × 10-4 m) and neostigmine (Ki = 2 .1 3 × 10-4 ᴍ). Diisopropyl fluorophosphate (DFP) showed no inhibition and diethyl p-nitrophenylphosphate (E 600) only a slight inhibitory effect at 10-5 ᴍ, respectively. Choline (10-2 ᴍ) was inhibitory but acetylcholine (10-2 ᴍ) stimulated the enzyme activity.

Characterization of ��-Glucosidase Produced by the White Rot Fungus Flammulina velutipes

2015 ◽  
Vol 25 (1) ◽  
pp. 57-65 ◽  
Author(s):  
Julieta Mallerman ◽  
Leandro Papinutti ◽  
Laura Levin
Keyword(s):  
White Rot ◽  
Flammulina Velutipes ◽  
White Rot Fungus
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