scholarly journals Three phase partitioning as a rapid and efficient method for purification of plant-esterase from wheat flour

2020 ◽  
Vol 22 (2) ◽  
pp. 42-49
Author(s):  
Liang Dong ◽  
Linxin He ◽  
Danqun Huo
Keyword(s):  
Wheat Flour ◽  
Single Step ◽  
Phase Partitioning ◽  
Surface Method ◽  
Three Phase ◽  
High Efficient ◽  
New Type ◽  
Purification Plant ◽  
Very High

AbstractThree-phase partitioning (TPP) was used to purify plant-esterase from wheat flour. Effect of various process parameters has been evaluated and plant-esterase was purified to 11.35-fold by optimized single step TPP system (50%, (w/v) (NH4)2SO4 saturation, 1:1 (v/v) ratio of crude extract: t-butanol at pH 4).The enzyme was found to be exclusively partitioned in the aqueous phase. Using TPP system, plant-esterase quickly purified to homogeneity with very high purity and activity. On the basis of single factor research, purification process was optimized by using response surface method, established a new type of high efficient purification plant-esterase method. To the best of our knowledge, this is the first report for purification and characterization of plant-esterase by using three phase partitioning (TPP). The results indicated that, TPP is a simple, quick, economical and very attractive process for purification of plant-esterase compared to conventional chromatographic protocols.

Three phase partitioning for simultaneous purification of aloe polysaccharide and protein using a single-step extraction

2015 ◽  
Vol 50 (3) ◽  
pp. 482-486 ◽  
Author(s):  
Zhi-Jian Tan ◽  
Chao-Yun Wang ◽  
Yong-Jian Yi ◽  
Hong-Ying Wang ◽  
Wan-Lai Zhou ◽  
...  
Keyword(s):  
Single Step ◽  
Phase Partitioning ◽  
Three Phase

Three-phase partitioning and immobilization of Bacillus methylotrophicus Y37 cellulase on organo-bentonite and its kinetic and thermodynamic properties

Clay Minerals ◽  
2020 ◽  
Vol 55 (2) ◽  
pp. 120-131
Author(s):  
Yonca Avci Duman ◽  
A. Uğur Kaya ◽  
Çiğdem Yağci
Keyword(s):  
Immobilized Enzyme ◽  
Maximum Velocity ◽  
Catalytic Performance ◽  
Covalent Immobilization ◽  
Single Step ◽  
Free Enzyme ◽  
Phase Partitioning ◽  
Bacillus Methylotrophicus ◽  
Three Phase ◽  
First Time

AbstractIn this study, for the first time Bacillus methylotrophicus Y37 cellulase was purified and recovered in a single step by three-phase partitioning (TPP). The optimal purification parameters for TPP were 40% ammonium sulfate saturation (m/v) with a 1.0:1.0 (v/v) ratio of crude extract:t-butanol, which gave 5.8-fold purification with 155% recovery of cellulase. Non-covalent immobilization of the partitioned cellulase was performed using bentonite as a support material. The activity observed in the 20th experiment was 100%. The optimal pH values and temperatures determined for the free enzyme and the immobilized enzyme were 5.0 and 6.0 and 45°C and 50°C, respectively. The Arrhenius activation energy (Ea) of the immobilized enzyme was lower than that of the free enzyme, whereas the Michaelis–Menten constant (Km) and maximum velocity (Vm) of the immobilized enzyme increased. The turnover number (kcat) and the catalytic performance (kcat/Km) demonstrated the improved catalytic properties of the immobilized enzyme compared to the free enzyme. Immobilization of cellulase is thermodynamically preferred.

Application of three-phase partitioning to the purification and characterization of polyphenol oxidase from antioxidant rosemary (Rosmarinus officinalis L.)

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Yonca Yuzugullu Karakus ◽  
Busra Kahveci ◽  
Arda Acemi ◽  
Gulden Kocak
Keyword(s):  
Enzyme Activity ◽  
Polyphenol Oxidase ◽  
Cetyltrimethylammonium Bromide ◽  
Rosmarinus Officinalis ◽  
Phase Partitioning ◽  
Three Phase ◽  
Diammonium Salt ◽  
The Common ◽  
Rosmarinus Officinalis L

AbstractPolyphenol oxidase (PPO) has been purified from the rosemary plant (Rosmarinus officinalis L.) through three-phase partitioning (TPP) and has been biochemically characterized. The optimized TPP consisted of 50% (w/v) ammonium sulfate and equal volumes of crude extract and tert-butanol prepared at pH 6.5 and room temperature. Using this system, PPO was purified 14-fold, with 230% recovery of activity from the middle phase. The partitioned enzyme had a molecular mass of 53 kDa. The highest enzyme activity was detected at 30 °C and pH 7.0 against catechol. In substrate specificity tests, the enzyme displayed activity towards catechol, 4-methylcatechol, caffeic acid, hydroquinone, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), pyrogallol, syringaldezine, and 3,4-dihydroxy-L-phenylalanine but no activity towards L-tyrosine. The enzyme was inhibited by the common PPO inhibitors; salicylhydroxamic acid (SHAM), cetyltrimethylammonium bromide (CTAB), polyvinylpyrrolidone (PVP), and the organic solvent dimethyl sulfoxide (DMSO). Enzyme activity increased in the presence of the organic solvents acetone, ethanol, and methanol.

scholarly journals Extraction and characterization of anti-oxidative polysaccharide–protein complexes from Corbicula fluminea through three-phase partitioning

RSC Advances ◽  
2017 ◽  
Vol 7 (18) ◽  
pp. 11067-11075 ◽  
Author(s):  
Yao-Yao Wang ◽  
Wen-Yi Qiu ◽  
Zhen-Bin Wang ◽  
Hai-Le Ma ◽  
Jing-Kun Yan
Keyword(s):  
Protein Complexes ◽  
Corbicula Fluminea ◽  
Phase Partitioning ◽  
Three Phase

In this study, a simple, efficient, and green bioseparation technology called three-phase partitioning (TPP) was employed to extract and separate polysaccharide–protein complexes (PSP) from Corbicula fluminea.

Sign in / Sign up

Export Citation Format

Share Document