Crowded milieu tuning the stability and activity of stem bromelain

Bhakuni, Kavya; Venkatesu, Pannuru

doi:10.1016/j.ijbiomac.2017.12.060

Cited by 11 publications

(2 citation statements)

References 42 publications

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“…Figure S2 portrays the intrinsic fluorescence of SB in IL, DES, and HIF. The maximum fluorescence intensity (I max ) of native SB (control) lies at 348 nm when excited at 295 nm, 41 while in the presence of all of the studied concentrations of IL (i.e. 0.15− 0.75 M) there is a red shift (from 345 to 365 nm) in 0.75 M IL in the wavelength maxima (λ max ), which explicitly conveys that the tryptophan (Trp) amino acid residue has been realigned from the hydrophobic pocket to hydrophilic solvent environment (Figure S2a).…”

Section: ■ Introductionmentioning

confidence: 99%

A Vibrant Accelerating Strategic Solvent System for Sustainability and Functionality for Proteins: Unvarying Structure of Stem Bromelain in Hybrid Ionic Fluids

Nakka

Sindhu

Chahar

et al. 2023

ACS Sustainable Chem. Eng.

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Very recently, a new class of cosolvents, hybrid ionic fluids (HIFs), have been designed by combining an ionic liquid (IL) and a deep eutectic solvent (DES) with a common anion. The product HIF formed is devoid of the deleterious effects of one component and has augmented advantageous characteristics of the other component, and is emerging as a fascinating sustainable solvent media and a new environmentally friendly solvent for biomolecules. Herein, we present the effects of 1-ethyl-3-methylimidazolium chloride ([Emim][Cl]), DES (synthesized using 1:2 choline chloride and glycerol), and their respective HIF on the structural stability and enzymatic activity of stem bromelain (SB). The spectroscopic results explicitly elucidated that [Emim][Cl] interacts favorably with SB and affects the structural as well as thermal stability of SB, whereas the stability and activity of the enzyme were increased in DES. Interestingly, the results obtained for HIF were the intermediate of the two (IL and DES) as the enzymatic activity of SB in the presence of HIF is maintained at all studied concentrations. Moreover, with the help of TEM and AFM studies, the changes in morphologies of SB in the presence and in the absence of IL, DES, and HIF were ascertained, and it was found that the morphology of SB in the presence of HIF and DES was similar to that of SB in buffer (control), indicating that DES and HIF preserve the native structure of SB. In other words, the morphology of SB has changed in the presence of IL. Apparently, the morphological results are in accordance with the spectroscopic results; thus, HIF significantly enhances the structural stability and enzymatic activity of SB, eventually confirming the suitability of HIF media as a potential and ecofriendly prolonged storage system for SB’s native structure.

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Section: ■ Introductionmentioning

confidence: 99%

A Vibrant Accelerating Strategic Solvent System for Sustainability and Functionality for Proteins: Unvarying Structure of Stem Bromelain in Hybrid Ionic Fluids

Nakka

Sindhu

Chahar

et al. 2023

ACS Sustainable Chem. Eng.

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“…This may be evaluated experimentally by using concentrated solutions of model "crowding agents" such as polyethylene glycol (PEG), dextran, Ficoll (branched carbohydrate derivative), or inert proteins [5]. Most of the enzymatic studies carried out in vitro under macromolecular crowding (MC) conditions have been monitored either by UV-visible spectroscopy, fluorescence, or far UV-circular dichroism spectroscopy [6][7][8][9]. Some other studies have used isothermal titration calorimetry and turbidimetric methods to study the effect of crowders on enzymatic activity [10,11].…”

Section: Introductionmentioning

confidence: 99%

Polyethylene glycol crowding effect on hyaluronidase activity monitored by capillary electrophoresis

et al. 2020

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To mimic the activity of hyaluronidase in natural environment, the hydrolysis of hyaluronic acid (HA) by hyaluronidase was investigated for the first time in the presence of crowding agents using capillary electrophoresis (CE) as a simple and reliable technique for conducting enzymatic assay. Polyethylene glycol (PEG) 6000 was selected as a model crowder and the hyaluronic acid degradation catalyzed by bovine testes hyaluronidase (BTH) was carried out at different PEG concentrations (0%, 10%, and 17%). After optimization of the CE analytical method and enzymatic assay, the degradation products were monitored at different HA concentrations. At 10% of PEG and 0.3 mg mL −1 of HA, the activity of the enzyme was significantly reduced showing inconvenient interactions of PEG with the hyaluronidase blocking the release of hydrolysis products. A similar reduction of hyaluronidase activity was observed at 1 mg mL −1 of HA due to the presumable formation of the BTH-substrate complex. The experimental curves obtained by CE also evidence that the overall kinetics are governed by the hydrolysis of hexasaccharide intermediates. Finally, the effect of PEG on hyaluronidase activity was evaluated in the presence of natural or synthetic inhibitors. Our results show a significant difference of the inhibitors' affinity toward hyaluronidase in the presence of PEG. Surprisingly, the presence of the crowding agent results in a loss of the inhibition effect of small polycyclic inhibitors, while larger charged inhibitors were less affected. In this work, CE analyses confirm the importance of mimicking the cellular environment for the discovery and development of reliable inhibitors.

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Does Microsecond Active-Site Dynamics Primarily control Proteolytic Activity of Bromelain? Clues from Single Molecular Level Study with a Denaturant, a Stabilizer and a Macromolecular Crowder

et al. 2022

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Crowded milieu tuning the stability and activity of stem bromelain

Cited by 11 publications

References 42 publications

A Vibrant Accelerating Strategic Solvent System for Sustainability and Functionality for Proteins: Unvarying Structure of Stem Bromelain in Hybrid Ionic Fluids

A Vibrant Accelerating Strategic Solvent System for Sustainability and Functionality for Proteins: Unvarying Structure of Stem Bromelain in Hybrid Ionic Fluids

Polyethylene glycol crowding effect on hyaluronidase activity monitored by capillary electrophoresis

Does Microsecond Active-Site Dynamics Primarily control Proteolytic Activity of Bromelain? Clues from Single Molecular Level Study with a Denaturant, a Stabilizer and a Macromolecular Crowder

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