Extraction and purification of a plant peroxidase by aqueous two-phase extraction coupled with gel filtration

Srinivas, N. D.; Rashmi, Kothary; Raghavarao, K.S.M.S.

doi:10.1016/s0032-9592(99)00030-8

Cited by 93 publications

(53 citation statements)

References 12 publications

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“…The specific activity (470 µmol min -1 mg -1 ) of E. crassipes leaf peroxidase achieved after the last step of the three-step purification in this study was considerably higher than levels obtained previously for different sources of the enzyme: 15.21 units mg -1 (Rehman et al 1999), 86 units mg -1 (Regalado et al 1996), 135.44 units mg -1 (Maciel et al 2007), 346.43 units mg -1 (Khatun et al 2012) and 349.8 units mg -1 (Srinivas et al 1999). …”

Section: Discussioncontrasting

confidence: 62%

“…Purified peroxidases have been obtained from diverse plant sources (Deepa, Arumughan 2002), such as spring cabbage, soy bean and rice leaves (Ito et al 1991), tea (Kvaratskhelia et al 1997), okra (Yemenicioglu et al 1998), Ipomoea palmetto (Srinivas et al 1999) and Copaifera longsdorffi (Maciel et al 2007). However, less attention has been directed toward sources that are naturally widely available and regarded as unwanted in the environment.…”

Section: Introductionmentioning

confidence: 99%

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Catalytic and kinetic properties of purified Eichhornia crassipes leaf peroxidase

Arise¹,

Osundahunsi²,

Farohunbi³

et al. 2016

EEB

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Peroxidase from Eichhornia crassipes leaf was purified 23.58 fold with 18.58% yield by means of (NH 4 ) 2 SO 4 precipitation, ion exchange and Sephadex G-75 gel filtration chromatography. Optimum temperature and substrate-dependent pH optimum for enzyme activity were 40 °C and pH 4.0, 9.0 and 6.0 for 2,2'-azino-bis-(3-ethylbenzthiazolin)-6-sulfonate (ABTS), guaiacol and pyrogallol, respectively. The enzyme had high pH stability and moderate thermal stability at temperatures up to 60 °C; the activation energy of inactivation of the enzyme was ~122.29 kJ mol -1 . Temperature-denaturing and spontaneous recovery were shown to be time-dependent while Ca 2+ -enhanced recovery of the denatured enzyme was in a time-dependent manner. The enzyme showed preferential affinity for ABTS over guaiacol and pyrogallol with K M values of 31.11, 21.91 and 6.45 mM respectively. It was reversibly inhibited by Pb 2+ , Hg 2+ and EDTA while urea only caused loss of ~30.40% activity after 60 min of incubation. E. crassipes leaf peroxidase has potential use for broad range of applications.

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Section: Discussioncontrasting

confidence: 62%

Section: Introductionmentioning

confidence: 99%

Catalytic and kinetic properties of purified Eichhornia crassipes leaf peroxidase

Arise¹,

Osundahunsi²,

Farohunbi³

et al. 2016

EEB

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“…Electrical interaction and repulsion between charged aqueous phase systems and the proteins affect the partitioning of system [21]. ATPS have found application in the industrial scale purification of proteins from biomass [22]. The use of ATPS in downstream processing has been focused on the extraction, separation and concentration of various biomolecules including xylanases [23], amylase [24], anyloglucosidase [25], amino acid [26], etc.…”

mentioning

confidence: 99%

“…In this regard, partitioning in ATPS provides a powerful method for separating and purifying mixtures of proteins [23][24][25]. ATPS also offers many advantages including low process time, low energy consumption and biocompatible environment to the biomolecule due to the presence of large amounts of water in the extraction systems [22]. Furthermore, ATPS can remove contaminants such as nucleic acids and undesirable proteins.…”

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confidence: 99%

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Partitioning and recovery of proteinase from tuna spleen by aqueous two-phase systems

Klomklao

Benjakul

Visessanguan

et al. 2005

Process Biochemistry

102

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Partitioning of spleen proteinase from yellowfin tuna (Thunnus albacores) in an aqueous two-phase system (ATPS) was investigated. Phase compositions including PEG molecular mass and concentration as well as types and concentration of salts affected the protein partitioning. ATPS comprising PEG1000 (15% w/w) and magnesium sulfate (20% w/w) provided the best condition for the maximum partitioning of the proteinase into the top phase and gave a highest specific activity (47.0 units/µg protein) and purification fold (6.61). The yield of 69.0% was obtained.Under the same ATPS condition used, the partitioning of proteinase of splenic extract from three tuna species involving skipjack tuna, yellowfin tuna and tongol tuna were compared. The purity of splenic extract from all tuna species increased after ATPS process. Among all species tested, yellowfin tuna showed the highest purification fold, followed by tongol tuna and skipjack tuna, respectively. SDS-substrate gel electrophoresis revealed that the band intensity of major proteinase in ATPS fraction from all tuna species slightly increased with the concomitant decrease in band intensity of other contaminating proteins, indicating the greater specific activity of splenic extract. Therefore, ATPS was an effective method for partitioning and recovery of proteinases from tuna spleen.

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Bioreactor for cultivation of red beet hairy roots and in situ recovery of primary and secondary metabolites

Neelwarne¹,

Rudrappa²

2009

Engineering in Life Sciences

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To arrive at an appropriate bioreactor design and in situ recovery of the products, red beet hairy roots were used as a model system where the levels of betalain pigments (betacyanins and betaxanthins) were followed as secondary metabolite and the peroxidase enzyme as primary metabolite. Medium volume and other kinetic parameters were found to play significant roles by way of directly affecting the biomass yield rather than a specific metabolite. The hydrodynamic stress created on the roots by large culture volume could be minimized by pulse‐feeding of medium in shake‐flasks; and by separating the biomass chamber from the aerated medium reservoir in circulatory fed‐batch bioreactor. Accordingly the bioreactor was modified to provide anchorage and air‐enrichment chamber which resulted in higher formation of both the metabolites than in shake‐flasks. Various down‐stream processing aspects such as in situ release of pigments by non‐destructive methods, followed by adsorption through a column and recovery by desorption were optimized for betalains. A strategy for simultaneous recovery of pigment and peroxidase was worked out using aqueous two phase extraction (ATPE).

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Extraction and purification of a plant peroxidase by aqueous two-phase extraction coupled with gel filtration

Cited by 93 publications

References 12 publications

Catalytic and kinetic properties of purified Eichhornia crassipes leaf peroxidase

Catalytic and kinetic properties of purified Eichhornia crassipes leaf peroxidase

Partitioning and recovery of proteinase from tuna spleen by aqueous two-phase systems

Bioreactor for cultivation of red beet hairy roots and in situ recovery of primary and secondary metabolites

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