Papain-catalyzed hydrolysis of and amino acid incorporation into BSA and zein substrates in low water organic media

Ferjancic-Biagini, A.; Gaertner, Hubert; Puigserver, Antoine

doi:10.1021/jf00031a027

Cited by 7 publications

(6 citation statements)

References 17 publications

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“…[31][32][33][34][35][36][37] Moreover, they often produce very short peptide fragments that could not be clearly separated, therefore, hindering further analysis of the protein structure, particularly in middle-down proteomics, which focuses on peptide fragments of around 10 kDa. [36][37][38] Hydrolysis using trypsin in aqueous alkaline solutions has also been reported, but this only produced small amounts of peptide fragments. [32,39] In view of these findings, it is clear that Chemistry-A European Journal artificial proteases that can selectively hydrolyze insoluble proteins under mild conditions, and produce larger and clearly distinguishable fragments, would facilitate the study of their structure in proteomics analysis.…”

Section: Hydrolysis Of Zeinmentioning

confidence: 99%

“…[30] The enzymatic hydrolysis of zein has been previously reported, but many of the currently existing procedures typically require highly alkaline conditions, the use of organic solvents and/or reducing and alkylating agents to break disulfide bonds and facilitate hydrolysis. [31][32][33][34][35][36][37] Moreover, they often produce very short peptide fragments that could not be clearly separated, therefore, hindering further analysis of the protein structure, particularly in middle-down proteomics, which focuses on peptide fragments of around 10 kDa. [36][37][38] Hydrolysis using trypsin in aqueous alkaline solutions has also been reported, but this only produced small amounts of peptide fragments.…”

Section: Hydrolysis Of Zeinmentioning

confidence: 99%

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Expanding the Scope of Polyoxometalates as Artificial Proteases towards Hydrolysis of Insoluble Proteins

Parac‐Vogt

Savić

Marcano

2021

Chemistry A European J

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Despite the enormous importance of insoluble proteins in biological processes, their structural investigation remains a challenging task. The development of artificial enzyme‐like catalysts would greatly facilitate the elucidation of their structure since currently used enzymes in proteomics largely lose activity in the presence of surfactants, which are necessary to solubilize insoluble proteins. In this study, the hydrolysis of a fully insoluble protein by polyoxometalate complexes as artificial proteases in surfactant solutions is reported for the first time. The hydrolysis of zein as a model protein was investigated in the presence of Zr(IV) and Hf(IV) substituted Keggin‐type polyoxometalates (POMs), (Et2NH2)10[M(α‐PW11O39)2] (M = Zr or Hf), and different concentrations of the anionic surfactant sodium dodecyl sulfate (SDS). Selective hydrolysis of the protein upon incubation with the catalyst was observed, and the results indicate that the hydrolytic selectivity and activity of the POM catalysts strongly depends on the concentration of surfactant. The molecular interactions between the POM catalyst and zein in the presence of SDS were explored using a combination of spectroscopic techniques which indicated competitive binding between POM and SDS towards the protein. Furthermore, the formation of micellar superstructures in ternary POM/surfactant/protein solutions has been confirmed by conductivity and Dynamic Light Scattering measurements.

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Section: Hydrolysis Of Zeinmentioning

confidence: 99%

Section: Hydrolysis Of Zeinmentioning

confidence: 99%

Expanding the Scope of Polyoxometalates as Artificial Proteases towards Hydrolysis of Insoluble Proteins

Parac‐Vogt

Savić

Marcano

2021

Chemistry A European J

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show abstract

“…[30] The enzymatic hydrolysis of zein has been previously reported, but many of the currently existing procedures typically require highly alkaline conditions, the use of organic solvents and/or reducing and alkylating agents to break disulfide bonds and facilitate hydrolysis. [31][32][33][34][35][36][37][38] Moreover, they often produce very short peptide fragments that could not be clearly separated therefore hindering further analysis of the protein structure. [37,38] Hydrolysis using trypsin in aqueous alkaline solutions has also been reported, but this only produced small amounts of two peptide fragments.…”

Section: Hydrolysis Of Zein In the Presence Of Different Concentratio...mentioning

confidence: 99%

“…[31][32][33][34][35][36][37][38] Moreover, they often produce very short peptide fragments that could not be clearly separated therefore hindering further analysis of the protein structure. [37,38] Hydrolysis using trypsin in aqueous alkaline solutions has also been reported, but this only produced small amounts of two peptide fragments. [32,39] In view of these findings, it is clear that artificial proteases that can selectively hydrolyze insoluble proteins under mild conditions, and produce larger and clearly distinguishable fragments, would facilitate the study of their structure in proteomics analysis.…”

Section: Hydrolysis Of Zein In the Presence Of Different Concentratio...mentioning

confidence: 99%

Expanding the Scope of Polyoxometalates as Artificial Proteases towards Hydrolysis of Insoluble Proteins

Savić

Marcano

Parac-Vogt

2021

Preprint

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Despite the enormous importance of insoluble proteins in biological processes, their structural investigation remains a challenging task. Development of artificial enzymatic catalysts would greatly facilitate elucidation of their structure as currently used enzymes in proteomics largely lose activity in the presence of surfactants, which are necessary to solubilize insoluble proteins. In this study the hydrolysis of a fully insoluble protein by a polyoxometalate complex as an artificial protease in surfactant solutions is reported for the first time. The hydrolysis of zein as a model protein was investigated in the presence of Zr(IV)-substituted Keggin-type polyoxometalate (POM), (Et2NH2)10[Zr(α-PW11O39)2], and different concentrations of the anionic surfactant sodium dodecyl sulfate (SDS). The selective hydrolysis of the protein upon incubation with the catalyst was observed, and the results indicate that hydrolytic selectivity and activity of the POM catalysts strongly depends on the concentration of surfactant. The molecular interactions between the POM catalyst and zein in the presence of SDS were explored using a combination of spectroscopic techniques which indicated competitive binding between POM and SDS towards the protein. The formation of micellar superstructures in tertiary POM/surfactant/protein solutions has been confirmed by electrical conductivity and Dynamic Light Scattering.

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“…Papain catalyzed synthesis of a protected dipeptide BocGly-PheOMe in the presence of L-Cysteine has been carried out in an aqueous organic two-phase system consisting of different solvents like carbon tetrachloride, trichloroethylene, toluene and benzene [34]. Mercaptoethanol was used as an antioxidant in papain catalyzed hydrolysis and amino acid incorporation into BSA and Zein (a protein from corn) in low water organic media [35]. Mercaptoethanol has also been used for papain stabilization during the synthesis of LeuEnkaphalin precursors in ethyl acetate saturated with Mes/NaOH buffer and Tris/HCl buffer [36,37].…”

Section: Page 2 Ofmentioning

confidence: 99%

Efficacy of L-Cysteine as an Anti-Oxidant in Papain Catalyzed Synthesis of Oligopeptides in Organic Solvent System

Srinivasan¹,

Kapila²,

Forciniti³

et al. 2017

Biochem Anal Biochem

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Papain-catalyzed hydrolysis of and amino acid incorporation into BSA and zein substrates in low water organic media

Cited by 7 publications

References 17 publications

Expanding the Scope of Polyoxometalates as Artificial Proteases towards Hydrolysis of Insoluble Proteins

Expanding the Scope of Polyoxometalates as Artificial Proteases towards Hydrolysis of Insoluble Proteins

Expanding the Scope of Polyoxometalates as Artificial Proteases towards Hydrolysis of Insoluble Proteins

Efficacy of L-Cysteine as an Anti-Oxidant in Papain Catalyzed Synthesis of Oligopeptides in Organic Solvent System

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