Aspergillus oryzae alpha-amylase partition in potassium phosphate-polyethylene glycol aqueous two-phase systems

Porfiri, María Cecilia; Picó, Guillermo; Romanini, Diana; Farruggia, Beatriz

doi:10.1016/j.ijbiomac.2011.03.003

Cited by 20 publications

(10 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It was verified that α-amylase has a greater affinity for that phase than the other proteins, since the value found for selectivity was greater than one (4.974). Porfiri et al (2011) assessed the partitioning of α-amylase from Aspergillus oryzae in PEG-phosphate systems and also noted that the enzyme has a greater affinity for the upper phase when employing polyethylene glycol with a molecular weight of 2000.…”

Section: Partition Of α-Amylase Produced By Solid State Fermentation mentioning

confidence: 99%

Study of Alpha-Amylase Obtained by Solid State Fermentation of Cassava Residue in Aqueous Two-Phase Systems

Pereira

Fontan

Franco

et al. 2018

Braz. J. Chem. Eng.

View full text Add to dashboard Cite

The α-amylase enzyme was produced by solid state fermentation (SSF) and its partition behavior in Aqueous Two-Phase Systems (ATPS) was determined using systems consisting of polyethylene glycol (PEG) and potassium phosphate buffer. To determine the best conditions for α-amylase production, a central composite design was constructed for two independent variables: fermentation time and moisture content, and for the partition study, a face-centered central composite design (CCF) with four independent variables: PEG molecular weight, pH, temperature and partition time. It was indicated that the best enzyme production conditions were a fermentation time of 35.0 hours and 45.7% moisture content. From the results obtained, it was found that increasing the polymer molecular weight promotes a reduction of the enzyme concentration in the upper phase, increasing its concentration in the lower phase. It was also observed that the partition coefficients increased with increasing pH values and a reduction in temperature.

show abstract

Section: Partition Of α-Amylase Produced By Solid State Fermentation mentioning

confidence: 99%

Study of Alpha-Amylase Obtained by Solid State Fermentation of Cassava Residue in Aqueous Two-Phase Systems

Pereira

Fontan

Franco

et al. 2018

Braz. J. Chem. Eng.

View full text Add to dashboard Cite

show abstract

“…Additionally, the protein shift from the bottom into the top phase occurs due to an increasing salting-out effect in the salt-rich bottom phase as well as a rising shielding of the proteins' surface charges caused by Na + and Cl − ions [13,84]. Furthermore, NaCl has the ability of modifying the ordered water structure around the hydrophobic chain, like the ethylene group of PEG or the hydrophobic protein surface area exposed to solvent [70,85,86]. A loss of this ordered water occurs with a protein transfer from the bottom into the top phase, thus facilitating the protein-PEG interactions and increasing its K [2,20,85,86].…”

Section: Effect Of Nacl Concentrationmentioning

confidence: 99%

“…Furthermore, NaCl has the ability of modifying the ordered water structure around the hydrophobic chain, like the ethylene group of PEG or the hydrophobic protein surface area exposed to solvent [70,85,86]. A loss of this ordered water occurs with a protein transfer from the bottom into the top phase, thus facilitating the protein-PEG interactions and increasing its K [2,20,85,86]. Thereby, an increasing hydrophobic difference between the phases is resulting in a decrease or replacement of the bound amount of water by NaCl in order to maintain a constant final system composition in terms of the phaseforming component concentrations [20,67,80].…”

Section: Effect Of Nacl Concentrationmentioning

confidence: 99%

Evaluation of Driving Forces for Protein Partition in PEG-Salt Aqueous Two- Phase Systems and Optimization by Design of Experiments

Glyk¹,

Solle²,

Scheper³

et al. 2016

J Chromatogr Sep Tech

View full text Add to dashboard Cite

“…It is one of low cost and high-efficient purification methods (Asenjo and Andrews 2011). Recently, many enzymes (α-amylase, α-galactosidase, lipase, proteinase), recombinant proteins, and antibody have been purified using this method (Azevedo et al 2009;Porfiri et al 2011;Gu et al 2012;Loc et al 2013;Ramakrishnan et al 2016).…”

Section: Introductionmentioning

confidence: 99%

Purification and characterizations of a novel recombinant Bacillus velezensis endoglucanase by aqueous two-phase system

Liu

Guo

et al. 2018

Bioresour. Bioprocess.

View full text Add to dashboard Cite

Background: Cellulases played an important role in the production of bioenergy and bio-products. Cellulases from bacteria with some special characteristics drew great attention due to its fast growth speed, wide adaption to harsh environment, and production of multi-function cellulases.Results: An endoglucanase gene egls from Bacillus velezensis A4 was cloned and expressed in Escherichia coli BL21 (DE3). The recombinant enzyme Egls was partially purified using aqueous two-phase system. The highest recovery rate of the enzyme was 90.39% at PEG 4000 (25% w/w), phosphate buffer 8.08% (w/w) (pH 6.0), and NaCl (5% w/w). The enzyme molecular weight was 55 KD estimated by zymogram. The optimal pH and temperature of recombinant enzyme Egls were pH 6.0 and 55 °C, respectively. The enzyme was stable at pH range of 5.0-7.0 at 55 °C for 60 min. The enzyme exhibited K m , V max , K cat values as 63.38 mg/ml, 55.6 mg/min, and 3.93 × 10 3 /S, respectively. The addition of 10 mM of Mg 2+ , Mn 2+ , or 5% (w/w) of Triton-X 100 in the reaction system enhanced the enzyme activity significantly. The enzyme showed both endoglucanase and exoglucanase activity. Conclusions:An endoglucanase gene egls from B. velezensis A4 was cloned and expressed in E. coli BL21 (DE3). The recombinant enzyme Egls was purified by aqueous two-phase system and characterized. The enzyme can be applied for the efficient pretreatment of lignocellulosic biomass for bioenergy and bio-products production.

show abstract

Aspergillus oryzae alpha-amylase partition in potassium phosphate-polyethylene glycol aqueous two-phase systems

Cited by 20 publications

References 15 publications

Study of Alpha-Amylase Obtained by Solid State Fermentation of Cassava Residue in Aqueous Two-Phase Systems

Study of Alpha-Amylase Obtained by Solid State Fermentation of Cassava Residue in Aqueous Two-Phase Systems

Evaluation of Driving Forces for Protein Partition in PEG-Salt Aqueous Two- Phase Systems and Optimization by Design of Experiments

Purification and characterizations of a novel recombinant Bacillus velezensis endoglucanase by aqueous two-phase system

Contact Info

Product

Resources

About