Immobilization and topochemical mechanism of a new β-amylase extracted from Pergularia tomentosa

Lahmar, Imen Ben; Radeva, Greta; Marinkova, Dessislava; Velitchkova, Maya; Belghith, Hafedh; Abdallah, Ferjani Ben; Yotova, Lyubov; Belghith, Karima

doi:10.1016/j.procbio.2017.09.007

Cited by 14 publications

(6 citation statements)

References 45 publications

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“…In contrast homotetramers have been reported from sweet potato and bindweed [VanDamme et al 2001;Chandrika et al 2013] while existence of five forms of -amylase in ungerminated and germinated rice seeds had previously been reported by [Matsui 1977]. The estimated molecular weight of the enzyme was the same order of magnitude as values that had been reported for β-amylase from Bacillus subtilis Isolated from Kolanut Weevil (39.4 kDa) [Femi-Ola 2013], Bacillus subtilis β-amylase isolated from peels of Cassava barley (34.67 kDa) [Adeyanju et al 2012], higher than 6.5kDa reported for sweet potato [Manisha et al 2016] and 24kDa for Pergularia tomentosa [ImenLahmar et al 2018], but lower than that of Chinese yam tuber (56kDa) [Chiba 2006],Glycine max seed (57kDa) [Chandrika et al 2013] hedge bindweed (55 kDa) [Vandamme et al 2001] and soybean (57 kDa) [Ajele 1997] and millet (58 kDa) [Yamasaki 2003].…”

Section: Resultssupporting

confidence: 70%

Biochemical Characterization of purified β-amylase from Dioscorea alata (water yam)

Fadunsin¹,

Ebuehi²,

Akande³

et al. 2020

Preprint

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Water yam (Dioscorea alata) β-amylase was isolated and purified to apparent homogeneity. The enzyme was found to be monomeric with a molecular weight of 30.1 kDa based on SDS-PAGE and 31.0 kDa based on non-denaturation gel filtration. The enzyme readily hydrolyzed soluble starch, amylose, amylopectin and glycogen. The enzyme was stable over a wide range of pHs (4 - 8). The enzyme has a Km, of 2.24 mg/ml for soluble starch. Activation energy (Ea) for catalysis by β-amylase at 25 0C was 6.45 kcal/mol. β-amylase Contains high content of both acidic and hydrophobic amino acid but low in arginine and histidine. The activation energy (Ea), half-life, free energy change (ΔG‡), enthalpy change (ΔH‡), and entropy change (ΔS‡) for inactivation were 13.92 kcal/mol, 41.25 min, 20.89 kcal/mol, 13.30 kcal/mol and -24.25 cal/mol/K, respectively. The binding profile of β-amylase with epicatechin, quercetin, rutin and gallic acid were all spontaneous with stoichiometric ratio of 1:1. Hydrophobic bonding played a major role in stabilizing the β-amylase-ligand complex. Sodium alginate immobilization of the enzyme improved the Optimum temperature from 40 to 50 0C and changed the optimum pH from 5.0 to 6.0 with retainment up to 67 % activity after 5 cycles of usage. The enzyme would be of importance in manufacturing company based on the kinetics and other parameters reported in this study.

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Section: Resultssupporting

confidence: 70%

Biochemical Characterization of purified β-amylase from Dioscorea alata (water yam)

Fadunsin¹,

Ebuehi²,

Akande³

et al. 2020

Preprint

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“…Prolonged exposure to elevated temperatures induces conformational changes that affect enzyme 3D structure [ 66 ]. Nevertheless, this is not common; it is more frequent during short time exposures.…”

Section: Discussionmentioning

confidence: 99%

Recombinant laccase rPOXA 1B real-time, accelerated and molecular dynamics stability study

et al. 2021

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Background Laccases (EC 1.10.3.2) are multi-copper oxidoreductases with great biotechnological importance due to their high oxidative potential and utility for removing synthetic dyes, oxidizing phenolic compounds, and degrading pesticides, among others. Methods A real-time stability study (RTS) was conducted for a year, by using enzyme concentrates from 3 batches (L1, L3, and L4). For which, five temperatures 243.15, 277.15, 298.15, 303.15, 308.15, and 313.15 K were assayed. Using RTS data and the Arrhenius equation, we calculated the rPOXA 1B accelerated stability (AS). Molecular dynamics (MD) computational study results were very close to those obtained experimentally at four different temperatures 241, 278, 298, and 314 K. Results In the RTS, 101.16, 115.81, 75.23, 46.09, 5.81, and 4.83% of the relative enzyme activity were recovered, at respective assayed temperatures. AS study, showed that rPOXA 1B is stable at 240.98 ± 5.38, 277.40 ± 1.32 or 297.53 ± 3.88 K; with t1/2 values of 230.8, 46.2, and 12.6 months, respectively. Kinetic and thermodynamic parameters supported the high stability of rPOXA 1B, with an Ed value of 41.40 KJ mol− 1, a low variation of KM and Vmax, at 240.98 ± 5.38, and 297.53 ± 3.88 K, and ∆G values showing deactivation reaction does not occur. The MD indicates that fluctuations in loop, coils or loops with hydrophilic or intermediate polarity amino acids as well as in some residues of POXA 1B 3D structure, increases with temperature; changing from three fluctuating residues at 278 K to six residues at 298 K, and nine residues at 314 K. Conclusions Laccase rPOXA 1B demonstrated experimentally and computationally to be a stable enzyme, with t1/2 of 230.8, 46.2 or 12.6 months, if it is preserved impure without preservatives at temperatures of 240.98 ± 5.38, 277.40 ± 1.32 or 297.53 ± 3.88 K respectively; this study could be of great utility for large scale producers.

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“…tomentosa belongs to the Asclepiadaceae family (Lahmar et al, 2022). It is a wild perennial plant that grows in the Mediterranean regions, and in arid and semi-arid regions and it is 50 cm high (Al-Hinai et al, 2018;Lahmar et al, 2018;Lahmar et al, 2022). It is classified as a poisonous plant with a distinctive odor (Al-Hinai et al, 2018;Ads et al, 2021).…”

Section: Introduction Introduction Introduction Introductionmentioning

confidence: 99%

Phytochemical profile, antioxidant and anti-inflammatory activities of crude latex (Pergularia tomentosa L.) in Algerian Saharan

SEGUENI,

CHOUIKH,

TLILI

2023

Not Sci Biol

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This study aimed to evaluate the dry crude latex extract from P. tomentosa, endemic to the Sahara, by determining the total content of polyphenols and flavonoids, and detecting the plant compounds by HPLC chromatography. Also, Antioxidant activity was measured through three tests: (BCB) beta-carotene/linoleic acid bleaching assay, (DPPH) radical scavenging and (FRAP) ferric reducing ability assay. In addition, anti-inflammatory activity. The results showed that the dry crude latex extract of P. tomentosa showed moderate content of polyphenols and flavonoids in the dry crude latex extract of P. tomentosa. Besides, qualitative HPLC analysis led to the detection of a group of phenolic compounds of this extract that have therapeutic properties. Also, the studied extract had strong activity in the beta-carotene/linoleic acid bleaching test and the values obtained were very close to the reference gallic acid. The results also indicated significant antioxidant activities in the DPPH and FRAP assay compared to ascorbic acid. In addition, the strong effect of dry crude latex extract from P. tomentosa in inhibiting protein denaturation induced by albumin hyperthermia, compared to ASPEGIC®, which is used as a reference drug in infections. In conclusion, the results indicate that the dry crude latex extract of P. tomentosa has antioxidant and anti-inflammatory activity, which confirms the use of this extract in folk medicine, which could be a first step in introducing it into therapeutic applications.

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Immobilization and topochemical mechanism of a new β-amylase extracted from Pergularia tomentosa

Cited by 14 publications

References 45 publications

Biochemical Characterization of purified β-amylase from Dioscorea alata (water yam)

Biochemical Characterization of purified β-amylase from Dioscorea alata (water yam)

Recombinant laccase rPOXA 1B real-time, accelerated and molecular dynamics stability study

Phytochemical profile, antioxidant and anti-inflammatory activities of crude latex (Pergularia tomentosa L.) in Algerian Saharan

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