scholarly journals Utilization of Integrative Technique for Partial Recovery of Proteases from Soil Microbes

2018 ◽  
Author(s):  
Said Nawab ◽  
Muhammad Aasim ◽  
Haris Saddique ◽  
Fazal Rabi ◽  
Waqar Ali ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Partition Coefficient ◽  
Cost Effective ◽  
Maximum Activity ◽  
Partial Purification ◽  
Phase System ◽  
Partial Recovery ◽  
Two Phase ◽  
Peg 4000 ◽  
Partitioning Behavior

Aqueous two-phase system (ATPS) is an efficient, cost effective, fast, simple and ecofriendly method for the recovery of biomolecules. In the present study, an ATPS composed of polyethylene glycol and ammonium sulphate (NH4)2SO4 was used for the partial purification of proteases from microbial source. The effects of different parameters such as molecular weight of PEG (4000, 6000 and 10000), concentration of PEG (15, 17.5 and 20 %) and concentration of (NH4)2SO4 (7.5, 8.3, 9.1 and 9.9 %) on the partitioning behavior of proteases at room temperature were investigated. Generally, increasing the concentration of PEG and (NH4)2SO4 moved the protease to the top i.e., polymer-rich phase. Increasing the molecular weight of PEG from 4000 to 10000 the partition coefficient decreased subsequently. The highest partition coefficient i.e., 3.32 and maximum activity i.e., 16.06 soxhlet unit was found in an optimum system composed of 20 % PEG 4000 and 9.9 % (NH4)2SO4.

scholarly journals Response Surface Methodology for the Evaluation of Magnetite Partitioning Behavior in Polyethylene Glycol-Based Aqueous Biphasic System

2019 ◽  
Vol 43 (6) ◽  
pp. 2807-2813 ◽  
Author(s):  
Behnia Shahrokhi ◽  
Mohsen Pirdashti ◽  
Mozhgan Managhebi
Keyword(s):  
Molecular Weight ◽  
Response Surface Methodology ◽  
Polyethylene Glycol ◽  
Response Surface ◽  
Polynomial Equation ◽  
Weight Percent ◽  
Phase System ◽  
Two Phase ◽  
Aqueous Biphasic ◽  
Partitioning Behavior

Abstract Response surface methodology in the framework of central composite design has successfully applied to polyethylene glycol (PEG)-based aqueous two-phase system in order to evaluate its efficiency in extracting Fe from iron ores with respect to PEG molecular weight, PEG concentration and salt concentration factors. Furthermore, ANOVA and multiple regression analysis have provided a second-order polynomial equation that is highly capable of predicting the experimental results. The PEG molecular weight and weight percent of the salt in the feed had a major and minor effects on the iron ion partition coefficient, respectively.

Interphase distribution of 2-furylethylenes

1985 ◽  
Vol 50 (8) ◽  
pp. 1642-1647 ◽  
Author(s):  
Štefan Baláž ◽  
Anton Kuchár ◽  
Ernest Šturdík ◽  
Michal Rosenberg ◽  
Ladislav Štibrányi ◽  
...  
Keyword(s):  
Partition Coefficient ◽  
Partition Coefficients ◽  
Transport Rate ◽  
Phase System ◽  
Two Phase ◽  
Interphase Distribution ◽  
Distribution Kinetics ◽  
System 1 ◽  
Kinetics Of

The distribution kinetics of 35 2-furylethylene derivatives in two-phase system 1-octanol-water was investigated. The transport rate parameters in direction water-1-octanol (l1) and backwards (l2) are partition coefficient P = l1/l2 dependent according to equations l1 = logP - log(βP + 1) + const., l2 = -log(βP + 1) + const., const. = -5.600, β = 0.261. Importance of this finding for assesment of distribution of compounds under investigation in biosystems and also the suitability of the presented method for determination of partition coefficients are discussed.

scholarly journals Purification of microbial b-galactosidase from Kluyveromyces fragilis by bioaffinity partitioning

1999 ◽  
Vol 30 (4) ◽  
pp. 324-331 ◽  
Author(s):  
Maria Estela da Silva ◽  
Telma Teixeira Franco
Keyword(s):  
Partition Coefficient ◽  
Second Step ◽  
Step Method ◽  
Kluyveromyces Fragilis ◽  
Two Phase ◽  
Rich Phase ◽  
Extraction And Purification ◽  
Peg 4000 ◽  
Aqueous Two Phase Systems ◽  
Phase Systems

This work investigated the partitioning of b-galactosidase from Kluyveromyces fragilis in aqueous two-phase systems (ATPS) by bioaffinity. PEG 4000 was chemically activated with thresyl chloride, and the biospecific ligand p-aminophenyl 1-thio-b-D-galactopyranoside (APGP) was attached to the activated PEG 4000. A new two-step method for extraction and purification of the enzyme b-galactosidase from Kluyveromyces fragilis was developed. In the first step, a system composed of 6% PEG 4000-APGP and 8% dextran 505 was used, where b-galactosidase was strongly partitioned to the top phase (K = 2,330). In the second step, a system formed of 13% PEG-APGP and 9% phosphate salt was used to revert the value of the partition coefficient of b-galactosidase (K = 2 x 10-5) in order to provide the purification and recovery of 39% of the enzyme in the bottom salt-rich phase.

Optimization of Aqueous Two Phase Extraction of Proteins from Litopenaeus Vannamei Waste by Response Surface Methodology Coupled Multi-Objective Genetic Algorithm

2019 ◽  
Vol 15 (1) ◽  
Author(s):  
P. Saravana Pandian ◽  
S. Sindhanai Selvan ◽  
A. Subathira ◽  
S. Saravanan
Keyword(s):  
Genetic Algorithm ◽  
Molecular Weight ◽  
Response Surface Methodology ◽  
Response Surface ◽  
Litopenaeus Vannamei ◽  
Value Added ◽  
Phase System ◽  
Two Phase ◽  
Multi Objective ◽  
Multi Objective Genetic Algorithm

Abstract Waste generated from industrial processing of seafood is an enormous source of commercially valuable proteins. One among the underutilized seafood waste is shrimp waste, which primarily consists of head and carapace. Litopenaeus vannamei (L. vannamei) is the widely cultivated shrimp in Asia and contributes to 90 % of aggregate shrimp production in the world. This work was focused on extraction as well as purification of value-added proteins from L. vannamei waste in a single step aqueous two phase system (ATPS). Polyethylene glycol (PEG) and trisodium citrate system were chosen for the ATPS owing to their adequate partitioning and less toxic nature. Response surface methodology (RSM) was implemented for the optimization of independent process variables such as PEG molecular weight (2000 to 6000), pH (6 to 8) and temperature (25 to 45 °C). The results obtained from RSM were further validated using a Multi-objective genetic algorithm (MGA). At the optimized condition of PEG molecular weight 2000, pH 8 and temperature 35 °C, maximum partition coefficient and protein yield were found to be 2.79 and 92.37 %, respectively. Thus, L. vannamei waste was proved to be rich in proteins, which could be processed industrially through cost-effective non-polluting ATPS extraction, and RSM coupled MGA could be a potential tool for such process optimization.

Aqueous two-phase system antibody confinement enables cost-effective analysis of protein analytes by sandwich enzyme-linked immunosorbent assay with minimal optical crosstalk

The Analyst ◽  
2020 ◽  
Vol 145 (16) ◽  
pp. 5458-5465 ◽  
Author(s):  
Maia Kvas ◽  
Alyne G. Teixeira ◽  
Beatrice Chiang ◽  
John P. Frampton
Keyword(s):  
Immunosorbent Assay ◽  
Low Cost ◽  
Cost Effective ◽  
Enzyme Linked Immunosorbent Assay ◽  
Phase System ◽  
Two Phase ◽  
Aqueous Two Phase System ◽  
Optical Crosstalk ◽  
Effective Analysis ◽  
Protein Analytes

An aqueous two-phase system was used to reduce reagent volumes and optical crosstalk for a low-cost single sandwich enzyme-linked immunoassay.

scholarly journals Optimization of Aqueous Two-Phase System (ATPS) of Recombinant Bromelain by Response Surface Methodology

2018 ◽  
Vol 7 (4.30) ◽  
pp. 377
Author(s):  
Zatul Iffah Mohd Arshad ◽  
Azura Amid
Keyword(s):  
Response Surface Methodology ◽  
Enzyme Activity ◽  
Partition Coefficient ◽  
Response Surface ◽  
Potassium Phosphate ◽  
Phase System ◽  
Two Phase ◽  
Aqueous Two Phase System ◽  
Crude Sample ◽  
Face Centered

Recombinant bromelain is a protease that was partially purified using aqueous two-phase system (ATPS). The process variables (pH, PEG 6000 and potassium phosphate concentration) were optimized on enzyme activity and partition coefficient using response surface methodology (RSM) based on a face-centered central composite design (FCCCD) model. The optimum conditions for purification were at 18.47% [w/w] PEG6000 and 13% [w/w] potassium phosphate, pH 7.0 with enzyme activity was obtained as 0.272±0.0036 unit m/L, and partition coefficient as 1.394±0.093. The recombinant bromelain was preferentially partitioned into the top phase and the band was reduced in contrast to crude sample on SDS-PAGE gel.

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