Three phase partitioning of β-galactosidase produced by an indigenous Lactobacillus acidophilus isolate

Choonia, Huzaifa S.; Lele, S. S.

doi:10.1016/j.seppur.2013.02.033

Cited by 30 publications

(23 citation statements)

References 34 publications

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“…Therefore, the partition behavior of other bioproducts in ethanol‐salt ATPS is worth to analyze, particularly due to their low phase viscosity and low cost constituents. Given that alcohols larger than ethanol have shown positive results for the recovery of enzymes, an interesting approach that could be explored for the recovery of SOD is the use of TPP systems, composed of t‐butanol and a salt, as they have proved to be mild and effective for enzyme recovery . Therefore, it was decided to analyze the fractionation of SOD in this type of systems as an additional approach in the complex ATPS experimental stage (Processing of clarified Kluyveromyces marxianus extract in ATPS for the recovery of SOD section).…”

Section: Resultsmentioning

confidence: 99%

“…The tie line lengths (TLL) and compositions of polymer‐salt and polymer‐polymer were estimated using the binodal curves presented by Zaslavsky . The tie line lengths (TLL) and compositions for the ionic liquid systems were previously reported by Montalvo‐Hernández et al and the compositions of the butanol‐salt systems were based on a previous report …”

Section: Methodsmentioning

confidence: 99%

“…Additionally, three‐phase partitioning (TPP) systems composed of t‐butanol and ammonium sulfate were introduced at this stage (Systems 22 and 23, Table ) as a potential alternative strategy for the recovery of the enzyme of interest. In the case of t‐butanol‐ammonium sulfate systems, the protocol for systems preparation was taken from a previous report . Briefly, the appropriate amount of salt was added to 700 μL of the clarified extract in order to obtain 20% and 40% w/v respectively.…”

Section: Methodsmentioning

confidence: 99%

“…In this strategy, a biphasic system constituted usually of t‐butanol and ammonium sulfate is loaded with a biological sample. Ammonium sulfate and t‐butanol push some biomolecules out of the solution and a well‐defined interface is formed, from which the solute of interest is ideally recovered and analyzed . TPP has been used successfully for the recovery of some proteins .…”

Section: Introductionmentioning

confidence: 99%

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Potential application of aqueous two‐phase systems and three‐phase partitioning for the recovery of superoxide dismutase from a clarified homogenate of Kluyveromyces marxianus

Simental-Martínez

Rito‐Palomares

Benavides

2014

Biotechnology Progress

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Superoxide dismutase (SOD; EC 1.15.1.1) is an antioxidant enzyme that represents the primary cellular defense against superoxide radicals and has interesting applications in the medical and cosmetic industries. In the present work, the partition behavior of SOD in aqueous two-phase systems (ATPS) (using a standard solution and a complex extract from Kluyveromyces marxianus as sample) was characterized on different types of ATPS (polymer-polymer, polymer-salt, alcohol-salt, and ionic liquid (IL)-salt). The systems composed of PEG 3350-potassium phosphate, 45% TLL, 0.5 M NaCl (315 U/mg, 87% recovery, and 15.1-fold purification) and t-butanol-20% ammonium sulfate (205.8 U/mg, 80% recovery and 9.8-fold purification), coupled with a subsequent 100 kDa ultrafiltration stage, allowed the design of a prototype process for the recovery and partial purification of the product of interest. The findings reported herein demonstrate the potential of PEG-salt ATPS for the potential recovery of SOD.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Potential application of aqueous two‐phase systems and three‐phase partitioning for the recovery of superoxide dismutase from a clarified homogenate of Kluyveromyces marxianus

Simental-Martínez

Rito‐Palomares

Benavides

2014

Biotechnology Progress

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“…However, β-galactosidase enzyme derived from microorganisms will generate the enzyme more potential and more effective in the aspect of technology than the enzyme from animals and plants (Grosova et al 2008). Commercially, β-galactosidase is produced from microorganisms including yeast from the genus Kluyveromyces and fungal filaments from genus Aspergillus (Choonia and Lele 2013). Until now, the price of β-galactosidase is still relatively expensive and economically problematic (Grosova et al 2008).…”

Section: Introductionmentioning

confidence: 99%

Application of Coastal and Marine Ecological Classification Standard to organize island ecosystem: the Abu Musa Island, Persian Gulf case study

Zare

Sinaei²,

Shakouri

et al. 2017

Biodiversitas

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Abstract. Saryono, Rakhmana S, Rahayu F, Ardhi A, Rusli, Pratiwi NW, Nugroho TT. 2017. Molecular identification of endophytic fungi isolated from the tuber of Dahlia variabilis and exploration of their ability in producing ß-galactosidase. Biodiversitas 18: 145-152. LBKURCC67 and LBKURCC69 were endophytic fungi isolated from the tubers of dahlia (Dahlia variabilis) in Padang Panjang, West Sumatra. This study aimed to identify the microscopic and molecular characteristics of endophytic fungi LBKURCC67 and LBKURCC69 based on ITS-rDNA region sequences and determine the ability of the isolates to produce β-galactosidase. Microscopic characteristic of LBKURCC67 and LBKURCC69 indicated that both isolates belonged to Fusarium sp. Furthermore, DNA sequence analysis based on mega BLAST program showed that LBKURCC67 was identified to be Fusarium solani with 99% homology and LBKURCC69 showed the perfect homology similarity with Fusarium oxysporum. Meanwhile, β-galactosidase activity was determined based on the amount of o-nitrophenol produced from the substrate o-nitrophenil-β-D-galactopiranoside, whilst the specific activity was indicated by activity per unit of proteins. The result showed that both fungi could produce β-galactosidase. However, the enzyme activity did not increase significantly at various production times of 2, 4, 6, 8, and 10 days. The highest enzyme activity of LBKURCC67 and LBKURCC69 were 0.163 ± 0.064 U/mL (at day 8) and 0.126 ± 0.039 U/mL (at day 2) respectively. LBKURCC69 showed higher specific β-galactosidase activity (1.886 ± 0.277 U/mg) than LBKURCC67 (1.179 ± 0.081 U/mg).

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Ultrasound-assisted three-phase partitioning of polyphenol oxidase from potato peel (Solanum tuberosum)

Niphadkar

Rathod

2015

Biotechnol Progress

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Conventional three phase partitioning (TPP) and ultrasound assisted three phase partitioning (UATPP) were optimized for achieving the maximum extraction and purification of polyphenol oxidase (PPO) from waste potato peels. Different process parameters such as ammonium sulfate (NH4)2SO4 concentration, crude extract to t-butanol ratio, time, temperature and pH were studied for conventional TPP. Except agitation speed, the similar parameters were also optimized for UATPP. Further additional parameters were also studied for UATPP viz. irradiation time at different frequencies, duty cycle and, rated power in order to obtain the maximum purification factor and recovery of PPO. The optimized conditions for conventional TPP were (NH4)2SO4 0-40% (w/v), extract to t-butanol ratio 1:1 (v/v), time 40 min and pH 7 at 30°C. These conditions provided 6.3 purification factor and 70% recovery of PPO from bottom phase. On the other hand, UATPP gives maximum purification fold of 19.7 with 98.3% recovery under optimized parameters which includes (NH4)2SO4 0-40% (w/v), crude extract to t-butanol ratio 1: 1 (v/v) pH 7, irradiation time 5 min with 25 kHz, duty cycle 40% and rated power 150W at 30°C. UATPP delivers higher purification factor and % recovery of PPO along with reduced operation time from 40 min to 5 min when compared with TPP. SDS PAGE showed partial purification of PPO enzyme with UATPP with molecular weight in the range of 26-36 kDa. Results reveal that UATPP would be an attractive option for the isolation and purification of PPO without need of multiple steps.

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Three phase partitioning of β-galactosidase produced by an indigenous Lactobacillus acidophilus isolate

Cited by 30 publications

References 34 publications

Potential application of aqueous two‐phase systems and three‐phase partitioning for the recovery of superoxide dismutase from a clarified homogenate of Kluyveromyces marxianus

Potential application of aqueous two‐phase systems and three‐phase partitioning for the recovery of superoxide dismutase from a clarified homogenate of Kluyveromyces marxianus

Application of Coastal and Marine Ecological Classification Standard to organize island ecosystem: the Abu Musa Island, Persian Gulf case study

Ultrasound-assisted three-phase partitioning of polyphenol oxidase from potato peel (Solanum tuberosum)

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