scholarly journals Purification and Characterization of Lipase Produced by Leuconostoc mesenteroides Subsp. mesenteroides ATCC 8293 Using an Aqueous Two-Phase System (ATPS) Composed of Triton X-100 and Maltitol

Molecules ◽  
2018 ◽  
Vol 23 (7) ◽  
pp. 1800 ◽  
Author(s):  
Nurfadhilah Eko Sukohidayat ◽  
Mohammad Zarei ◽  
Badlishah Baharin ◽  
Mohd Manap
Keyword(s):  
Phase System ◽  
Two Phase ◽  
Aqueous Two Phase System ◽  
Process Characteristics ◽  
Sds Page ◽  
Ph Range ◽  
Lipase Purification ◽  
Triton X ◽  
First Time

Purification of lipase produced by L. mesenteroides subsp. mesenteroides ATCC 8293 was conducted for the first time using a novel aqueous two-phase system (ATPS) composed of Triton X-100 and maltitol. The partitioning of lipase was optimized according to several parameters including pH, temperature, and crude load. Results showed that lipase preferentially migrated to the Triton X-100 rich phase and optimum lipase partitioning was achieved in ATPS at TLL of 46.4% and crude load of 20% at 30 °C and pH 8, resulting in high lipase purification factor of 17.28 and yield of 94.7%. The purified lipase showed a prominent band on SDS-PAGE with an estimated molecular weight of 50 kDa. The lipase was stable at the temperature range of 30–60 °C and pH range of 6–11, however, it revealed its optimum activity at the temperature of 37 °C and pH 8. Moreover, lipase exhibited enhanced activity in the presence of non-ionic surfactants with increased activity up to 40%. Furthermore, results exhibited that metals ions such as Na+, Mg2+, K+ and Ca2+ stimulated lipase activity. This study demonstrated that this novel system could be potentially used as an alternative to traditional ATPS for the purification and recovery of enzymes since the purified lipase still possesses good process characteristics after undergoing the purification process.

Characterization of alcohol/salt aqueous two-phase system for optimal separation of gallic acids

2021 ◽  
Vol 131 (5) ◽  
pp. 537-542
Author(s):  
Hui Suan Ng ◽  
Phei Er Kee ◽  
Hip Seng Yim ◽  
Joo Shun Tan ◽  
Yin Hui Chow ◽  
...  
Keyword(s):  
Phase System ◽  
Two Phase ◽  
Aqueous Two Phase System ◽  
Optimal Separation

scholarly journals Characterization of a Robust and pH-Stable Tannase from Mangrove-Derived Yeast Rhodosporidium diobovatum Q95

Marine Drugs ◽  
2020 ◽  
Vol 18 (11) ◽  
pp. 546
Author(s):  
Jie Pan ◽  
Ni-Na Wang ◽  
Xue-Jing Yin ◽  
Xiao-Ling Liang ◽  
Zhi-Peng Wang
Keyword(s):  
Gallic Acid ◽  
Tannic Acid ◽  
Specific Activity ◽  
Maximum Activity ◽  
Sds Page ◽  
Rhodosporidium Diobovatum ◽  
Bacteria And Fungi ◽  
Ph Range ◽  
First Time

Tannase plays a crucial role in many fields, such as the pharmaceutical industry, beverage processing, and brewing. Although many tannases derived from bacteria and fungi have been thoroughly studied, those with good pH stabilities are still less reported. In this work, a mangrove-derived yeast strain Rhodosporidium diobovatum Q95, capable of efficiently degrading tannin, was screened to induce tannase, which exhibited an activity of up to 26.4 U/mL after 48 h cultivation in the presence of 15 g/L tannic acid. The tannase coding gene TANRD was cloned and expressed in Yarrowia lipolytica. The activity of recombinant tannase (named TanRd) was as high as 27.3 U/mL. TanRd was purified by chromatography and analysed by SDS-PAGE, showing a molecular weight of 75.1 kDa. The specific activity of TanRd towards tannic acid was 676.4 U/mg. Its highest activity was obtained at 40 °C, with more than 70% of the activity observed at 25–60 °C. Furthermore, it possessed at least 60% of the activity in a broad pH range of 2.5–6.5. Notably, TanRd was excellently stable at a pH range from 3.0 to 8.0; over 65% of its maximum activity remained after incubation. Besides, the broad substrate specificity of TanRd to esters of gallic acid has attracted wide attention. In view of the above, tannase resources were developed from mangrove-derived yeasts for the first time in this study. This tannase can become a promising material in tannin biodegradation and gallic acid production.

scholarly journals Purification and characterization of a collagenase from Penicillium sp. UCP 1286 by polyethylene glycol-phosphate aqueous two-phase system

2017 ◽  
Vol 133 ◽  
pp. 8-14 ◽  
Author(s):  
Maria Carolina de Albuquerque Wanderley ◽  
José Manoel Wanderley Duarte Neto ◽  
Wendell Wagner Campos Albuquerque ◽  
Daniela de Araújo Viana Marques ◽  
Carolina de Albuquerque Lima ◽  
...  
Keyword(s):  
Polyethylene Glycol ◽  
Phase System ◽  
Purification And Characterization ◽  
Two Phase ◽  
Aqueous Two Phase System ◽  
Penicillium Sp

Optimization and validation of an environmentally friendly method for zinc determination in new and used lubricating oil samples

2016 ◽  
Vol 8 (48) ◽  
pp. 8435-8442 ◽  
Author(s):  
Mariana F. B. Mota ◽  
Ednilton M. Gama ◽  
Guilherme D. Rodrigues ◽  
Letícia M. Costa
Keyword(s):  
Environmentally Friendly ◽  
Lubricating Oil ◽  
Phase System ◽  
Two Phase ◽  
Aqueous Two Phase System ◽  
Used Lubricating Oil ◽  
Zinc Determination ◽  
First Time ◽  
Environmentally Friendly Method

This work presents the optimization and validation of a new environmentally friendly method for zinc quantification in new and used lubricating oil samples, using, for the first time, the extracting agent diphenylthiocarbazone (dithizone) associated with the aqueous two-phase system (ATPS).

scholarly journals Characterization of Amyloid β Fibrils with An Aqueous Two-Phase System: Implications of Fibril Formation

2010 ◽  
Vol 17 (0) ◽  
pp. 121-128 ◽  
Author(s):  
Toshinori SHIMANOUCHI ◽  
Naoya SHIMAUCHI ◽  
Keiichi NISHIYAMA ◽  
Huong Thi VU ◽  
Hisashi YAGI ◽  
...  
Keyword(s):  
Amyloid Β ◽  
Fibril Formation ◽  
Phase System ◽  
Two Phase ◽  
Aqueous Two Phase System

Phase Diagram of Aqueous Two-Phase System (ATPS) Composed of Polyethylene Glycol (PEG) and Gelatin

2012 ◽  
Vol 554-556 ◽  
pp. 286-294 ◽  
Author(s):  
Chao Ma ◽  
Xiao Dong Chen ◽  
Yin Qiu Kong ◽  
Li Ming Che
Keyword(s):  
Phase Diagram ◽  
Aqueous Solution ◽  
Phase Separation ◽  
Polyethylene Glycol ◽  
Solid Content ◽  
Phase System ◽  
Dense Phase ◽  
Two Phase ◽  
Aqueous Two Phase System ◽  
First Time

Phase diagram of aqueous two-phase system (ATPS) composed of polyethylene glycol (PEG) and gelatin is of paramount importance for the application of such system in microencapsulation of bioactive compounds. Phase separation of PEG/gelatin aqueous solution was investigated in the present work and the phase diagram of resultant ATPS was reported for the first time. The results show that phase separation will happen if the solid content of PEG/gelatin aqueous solution is higher than a critical value, resulting in an ATPS. The resultant ATPS consists of a low-density phase enriched in PEG and a dense phase enriched in gelatin. The phase compositions of the resultant ATPS were determined accurately using the method developed. The phase diagrams obtained show that higher solid content is required for the phase separation of PEG10,000/gelatin aqueous solution when compared with that of PEG20,000/gelatin one. And PEG is found to be more hydrophilic when compared with gelatin.

Extraction and Biochemical Characterization of Peptidases from Giant Catfish Viscera by Aqueous Two-Phase System

2015 ◽  
Vol 39 (4) ◽  
pp. 429-438 ◽  
Author(s):  
Sunantha Ketnawa ◽  
Oscar Martinez-Alvarez ◽  
Soottawat Benjakul ◽  
Saroat Rawdkuen
Keyword(s):  
Biochemical Characterization ◽  
Phase System ◽  
Two Phase ◽  
Aqueous Two Phase System ◽  
Giant Catfish

scholarly journals A Novel Aqueous Micellar Two-Phase System Composed of Surfactant and Sorbitol for Purification of Pectinase Enzyme fromPsidium guajavaand Recycling Phase Components

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Mehrnoush Amid ◽  
Fara Syazana Murshid ◽  
Mohd Yazid Manap ◽  
Muhaini Hussin
Keyword(s):  
Psidium Guajava ◽  
Bottom Phase ◽  
Phase System ◽  
The Novel ◽  
Two Phase ◽  
High Purification ◽  
Novel Method ◽  
Triton X ◽  
First Time ◽  
Economical Alternative

A novel aqueous two-phase system composed of a surfactant and sorbitol was employed for the first time to purify pectinase fromPsidium guajava. The influences of different parameters, including the type and concentration of the surfactant and the concentration and composition of the surfactant/sorbitol ratio, on the partitioning behavior and recovery of pectinase were investigated. Moreover, the effects of system pH and the crude load on purification fold and the yield of purified pectinase were studied. The experimental results indicated that the pectinase was partitioned into surfactant-rich top phase, and the impurities were partitioned into the sorbitol-rich bottom phase with the novel method involving an ATPS composed of 26% (w/w) Triton X-100 and 23% (w/w) sorbitol at 54.2% of the TLL crude load of 20% (w/w) at pH 6.0. The enzyme was successfully recovered by this method with a high purification factor of 15.2 and a yield of 98.3%, whereas the phase components were also recovered and recycled at rates above 96%. This study demonstrated that this novel ATPS method can be used as an efficient and economical alternative to the traditional ATPS for the purification and recovery of the valuable enzyme.

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