Proteases as Powerful Catalysts for Organic Synthesis

Illanes, Andrés; Guzmán, Fanny; Barberis, S

doi:10.1002/9783527631780.ch8

Cited by 4 publications

(4 citation statements)

References 290 publications

(227 reference statements)

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“…The direct effect of the organic solvent on the enzyme could be the main reason of deactivation. The literature reports that other enzymes in almost anhydrous organic solvents showed an increased structural rigidity which led to a lower reactivity (27,34). The substrate solvation (into the microenvironment of the enzyme active site) does not seem to be the deactivation cause because as the reaction progresses the amount of water produced also increases.…”

Section: Enzymatic Synthesis Of Peptidementioning

confidence: 99%

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Biosynthesis of a Novel Antibacterial Dipeptide, Using Proteases From South American Native Fruits, Useful as a Food Preservative

et al. 2021

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Antiacanthain and granulosain are the partially purified proteolytic extracts from the South American native fruits of Bromelia antiacantha (Bertol. ) and Solanum granuloso leprosum, respectively. The aim of this work was to compare the ability of both soluble and immobilized antiacanthain and granulosain f or the synthesis of Z-Tyr-Val-OH, a novel antibacterial dipeptide, in different reaction systems formed by almost anhydrous organic solvents (Xw: 1 × 10−5) and several percentages of immiscible organic solvents in 100 mM Tris(hydroxymethyl)aminomethane hydrochloride buffer pH 8.0. Soluble antiacanthain in half of the 24 different organic biphasic media showed higher catalytic potential than in 100 mM Tris(hydroxymethyl)aminomethane hydrolchloride buffer pH 8.0. Soluble granulosain showed lower catalytic potential in all liquid-liquid biphasic media than in the same buffer. However, 50% (v/v) ethyl ethanoate in 100 mM Tris(hydroxymethyl)aminomethane hydrolchloride buffer pH 8.0 allowed to express the highest catalytic potential of both soluble enzymes. In 50% v/v ethyl ethanoate, soluble antiacanthain and granulosain catalyzed the synthesis of Z-Tyr-Val-OH with 72 ± 0.15 and 60 ± 0.10% maximal peptide yields, respectively. Multi-point immobilization in glyoxyl-silica did not lead to better peptide yields than soluble enzymes, in that liquid-liquid biphasic medium under the same reaction conditions. Soluble and glyoxyl-silica immobilized antiacanthain in almost anhydrous ethyl ethanoate (Xw: 1 × 10−5) were able to retain 17.3 and 45% of the initial proteolytic activity of antiacanthain in 100 mM Tris hydrolchloride buffer pH 8.0, respectively, at 40°C under agitation (200 rpm). Soluble and glyoxyl-silica immobilized granulosain were inactivated under the same reaction conditions. Glyoxyl-silica immobilized antiacanthain showed to be a robust biocatalyst in almost anhydrous ethyl ethanoate (Xw: 1 × 10−5), eliciting the best peptide yield (75 ± 0.13%). The synthesis reaction of Z-Tyr-Val-OH could not proceed when soluble antiacanthain was used under the same conditions. Both peptidases only catalyzed the synthesis reaction under kinetic control, using activated acyl donor substrates. Finally, this work reports a novel broad-spectrum antibacterial peptide that significantly decreased (p ≤ 0.05) the specific growth rates of Gram positive and Gram negative microorganisms at very low concentrations (≥15 and 35 μg/ml, respectively); contributing with a new safe food preservative of applying for different food systems.

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Section: Enzymatic Synthesis Of Peptidementioning

confidence: 99%

“…Enzyme purification techniques show some restrictions related to the scaling up and increasing production costs to be applied to industrial process ( 34 ). Consequently, partially purified extracts were selected for this study.…”

Section: Introductionmentioning

confidence: 99%

Biosynthesis of a Novel Antibacterial Dipeptide, Using Proteases From South American Native Fruits, Useful as a Food Preservative

et al. 2021

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“…Proteases as papain, bromelain and a-chymotrypsin, have been used as catalysts for the co-oligomerization of hydrophobic amino acid ethyl esters, namely L-leucine ethyl ester with L-glutamic acid diethyl ester hydrochloride or L-phenylalanine ethyl ester (Li et al, 2008;Qin et al, 2013a;Uyama et al, 2002;Viswanathan et al, 2010). In the last years, there is significant progress in proteasecatalyzed methods to synthesize oligopeptides (Illanes et al, 2010;Kobayashi and Makino, 2009;Li et al, 2008;Qin et al, 2013b). The enzymatic-triggered self-assembly based on the oligomerization of Lys-Leu ethyl esters (KL-ethyl ester) catalysed by achymotrypsin was introduced by Gross and co-workers (Qin et al, 2013a(Qin et al, , 2013b.…”

Section: Oligopeptides Characterizationmentioning

confidence: 99%

α-Chymotrypsin catalysed oligopeptide synthesis for hair modelling

Martins

Silva

Cavaco-Paulo

2019

Journal of Cleaner Production

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Conventional chemical treatments imply the use of harsh chemicals which can cause serious adverse effects on consumers' health, hair fibre and environment. In this work was reported for the first time the modelling shape of Asian hair by oligopeptide coating synthesized by a protease. a-Chymotrypsin from Bovine pancreas catalysed the oligomerization of the tri-peptides, KCL (Lysine-Cysteine-Leucine) ethyl ester and KSL (Lysine-Serine-Leucine) ethyl ester in aqueous media. The synthesis reaction gave rise to oligopeptides with DP ¼ 6, which were characterized by mass spectra (MALDI-TOF). The in situ synthesis reaction onto the hair surface, under controlled pH (8.5e9) and temperatures between 25 C and 55 C, formed a coating layer inducing changes in the hair shape from straight to curly. Qualitative and quantitative characterization of coated hair was performed by image visualization and by perming efficiency assessment. The developed strategy revealed to be a safer and cleaner alternative to the conventional chemical treatments generally used for hair shape modification. Low cost associated to the catalyst and lower side effects to the hair fibre, health consumer and environment might lead to the development of a cost-effective and sustainable approach in the hair care industry.

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“…Synthesis of peptides will not proceed efficiently in aqueous medium where the hydrolytic potential of the enzyme prevails. The design of reaction media is a major challenge for peptide synthesis, since proteases, different from lipases, are not structurally conditioned to act in such environments ( Illanes et al, 2009 ; Origone et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

Enzymatic synthesis of new antimicrobial peptides for food purposes

et al. 2023

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Growing consumer awareness of the potential negative health effects of synthetic antibiotics has prompted the search for more natural preservatives that can improve the safety and quality of food. In this study we report the enzymatic synthesis of N-α-[Carbobenzyloxy]-Ile-Gln (Z-IQ) which is the precursor of Ile-Gln (IQ), a new antibacterial dipeptide, using an aqueous–organic biphasic system formed by 50% (v/v) ethyl acetate in 0.1 M Tris – HCl buffer pH 8. A partially purified proteolytic extract from the fruits of Solanum granuloso leprosum, named granulosain, proved to be a robust biocatalyst for the synthesis of Z-IQ, eliciting 71 ± 0.10% maximal peptide yield in the above described conditions. After cleaving and purifying IQ dipeptide, antimicrobial activity was assayed against Staphylococcus aureus ATCC 25923, Staphylococcus hominis A17771, and Staphylococcus aureus C00195, and MIC values between 118 ± 0.01 μg/mL and 133.7 ± 0.05 μg/mL were obtained. In addition, IQ showed MIC of 82.4 ± 0.01 μg/mL and 85.0 ± 0.00 μg/mL against Escherichia coli ATCC 25922 and Escherichia coli A17683, respectively. IQ did not show inhibitory activity against single-drug resistance (SDR) strains, such as Klebsiella oxytoca A19438 (SDR) and Pseudomonas aeruginosa C00213 (SDR), and against multidrug-resistant Enterococcus faecalis I00125 (MDR). IQ also caused growth inhibition of Helicobacter pylori NCTC 11638 and three wild-type H. pylori strains, which are sensitive to AML, MTZ, LEV and CLA (H. pylori 659), resistant to LEV (H. pylori 661 SDR), and resistant to MTZ (H. pylori 662 SDR). Finally, this study contributes with a new dipeptide (IQ) that can be used as an antimicrobial agent for food preservation or as a safe ingredient of functional foods.

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Proteases as Powerful Catalysts for Organic Synthesis

Cited by 4 publications

References 290 publications

Biosynthesis of a Novel Antibacterial Dipeptide, Using Proteases From South American Native Fruits, Useful as a Food Preservative

Biosynthesis of a Novel Antibacterial Dipeptide, Using Proteases From South American Native Fruits, Useful as a Food Preservative

α-Chymotrypsin catalysed oligopeptide synthesis for hair modelling

Enzymatic synthesis of new antimicrobial peptides for food purposes

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