Effect of deglycosylation on the properties of thermophilic invertase purified from the yeast Candida guilliermondii MpIIIa

Plascencia-Espinosa, Miguel Ángel; Santiago-Hernández, Alejandro; Pavón-Orozco, Patricia; Vallejo-Becerra, Vanessa; Trejo-Estrada, Sergio Rubén; Sosa‐Peinado, Alejandro; Benítez‐Cardoza, Claudia G.; Hidalgo-Lara, María Eugenia

doi:10.1016/j.procbio.2014.05.022

Cited by 18 publications

(18 citation statements)

References 36 publications

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“…Similar to INV3a-N, rInvHJ14 remained approximately 50% of its highest activity in the presence of 2045.0 mM sucrose. The big differences between INV3a-N and rInvHJ14 are that INV3a-N is an acidic and thermophilic invertase7. Together, rInvHJ14 may be a potential candidate for enzymatic hydrolysis of sucrose in the production of high-quality inverted syrups.…”

Section: Discussionmentioning

confidence: 99%

Characterization of a novel low-temperature-active, alkaline and sucrose-tolerant invertase

Zhou

Gao

et al. 2016

Sci Rep

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A glycoside hydrolase family 32 invertase from Bacillus sp. HJ14 was expressed in Escherichia coli. The purified recombinant enzyme (rInvHJ14) showed typical biochemical properties of low-temperature-active and alkaline enzymes: (i) rInvHJ14 was active and stable in the range of pH 7.0–9.5 with an apparent pH optimum of 8.0; (ii) rInvHJ14 was most active but not stable at 30–32.5 °C, with 19.7, 48.2 and 82.1% of its maximum activity when assayed at 0, 10 and 20 °C, respectively, and the Ea, ΔG* (30 °C), Km (30 °C) and kcat (30 °C) values for hydrolysis of sucrose by rInvHJ14 was 47.6 kJ mol−1, 57.6 kJ mol−1, 62.9 mM and 746.2 s−1, respectively. The enzyme also showed strong sucrose tolerance. rInvHJ14 preserved approximately 50% of its highest activity in the presence of 2045.0 mM sucrose. Furthermore, potential factors for low-temperature-active and alkaline adaptations of rInvHJ14 were presumed. Compared with more thermostable homologs, rInvHJ14 has a higher frequency of glycine residues and a longer loop but a lower frequency of proline residues (especially in a loop) in the catalytic domain. The catalytic pockets of acid invertases were almost negatively charged while that of alkaline rInvHJ14 was mostly positively charged.

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Section: Discussionmentioning

confidence: 99%

Characterization of a novel low-temperature-active, alkaline and sucrose-tolerant invertase

Zhou

Gao

et al. 2016

Sci Rep

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“…For example, it increased the INVA and INVB activities by 80 and 20%, respectively (Pérez de los Santos et al, 2016). This activating effect has been observed for the invertase from the yeast Candida guilliermondii (Plascencia-Espinosa et al, 2014). Mn 2+ enhanced its activity by up to 277% for invertase from Anthene phoenicis (Rustiguel et al, 2015).…”

Section: Gspinv Characterizationmentioning

confidence: 76%

“…Invertases from several strains, such as Zymomonas mobilis and Aspergillus niger, have been expressed heterologously in K. pastoris (Veana et al, 2014;Pérez de los Santos et al, 2016). The GspInv productivity was considerably higher than those of most invertases expressed heterologously in K. pastoris ( Supplementary Table S3; Hsieh et al, 2006;Plascencia-Espinosa et al, 2014). Furthermore, GspInv is electrophoretically homogenous on an SDS-PAGE gel after ultrafiltration, which means no additional purification step is necessary to purify GspInv and decrease production costs.…”

Section: Expression Of Gspinv In K Pastoris and Purificationmentioning

confidence: 99%

“…Furthermore, Cu 2+ oxidizes cysteine residues in the protein and cause structural changes and alteration in the protein activity. The substrate specificity and action mode of GspInv were investigated by incubating the enzyme with sucrose, trehalose, cellobiose, maltose, isomaltose, raffinose, melezitose, stachyose, and inulin at pH 5.0 and 45 • C. Similar to most of yeast invertases, which are active only against sucrose and raffinose (Plascencia-Espinosa et al, 2014), GspInv released fructose from sucrose and raffinose and did not exhibit hydrolysis activity toward other substrates ( Supplementary Table S4), suggesting that GspInv is invertase (Zhang et al, 2015). The kinetic constants of GspInv on sucrose were tested at optimal conditions.…”

Section: Gspinv Characterizationmentioning

confidence: 99%

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Identification and Immobilization of an Invertase With High Specific Activity and Sucrose Tolerance Ability of Gongronella sp. w5 for High Fructose Syrup Preparation

et al. 2020

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Invertases catalyze the hydrolysis of sucrose into fructose and glucose and can be employed as an alternative in producing high fructose syrup. In this study, we reported the heterologous expression of an invertase gene (GspInv) of Gongronella sp. w5 in Komagataella pastoris. GspInv activity reached 147.6 ± 0.4 U/mL after 5 days of methanol induction. GspInv is invertase with a high specific activity of 2,776.1 ± 124.2 U/mg toward sucrose. GspInv showed high tolerance to sucrose (IC 50 = 1.2 M), glucose (IC 50 > 2 M), fructose (IC 50 = 1.5 M), and a variety of metal ions that make it an ideal candidate for high fructose syrup production. A carbohydratebinding module was sequence-optimized and fused to the N-terminus of GspInv. The fusion protein had the highest immobilization efficiency at room temperature within 1 h adsorption, with 1 g of cellulose absorption up to 8,000 U protein. The celluloseimmobilized fusion protein retained the unique properties of GspInv. When applied in high fructose syrup preparation by using 1 M sucrose as the substrate, the sucrose conversion efficiency of the fused protein remained at approximately 95% after 50 h of continuous hydrolysis on a packed bed reactor. The fused protein can also hydrolyze completely the sucrose in sugarcane molasses. Our results suggest that GspInv is an unusual invertase and a promising candidate for high fructose syrup preparation.

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“…The effect of metal ion on sucrose hydrolysis was investigated by several authors [10][11][12][13][14][15]. Bagal-Kestwal et al [10] studied the effect of several metal ions such as Hg 2+ , Pb 2+ , Ag + and Cd 2+ on sucrose hydrolysis.…”

Section: Introductionmentioning

confidence: 99%

Inhibition Effect of Ca2+ Ions on Sucrose Hydrolysis Using Invertase

Hargono

Jos

Abdullah

et al. 2019

Bull. Chem. React. Eng. Catal.

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Fermentable sugar for bioethanol production can be produced from molasses due to its high sucrose content but Ca2+ ions found in the molasses may affect the hydrolysis. Therefore, this paper was focused to study the effect of Ca2+ ions as CaO on sucrose hydrolysis using invertase and to obtain the kinetic parameters. The kinetic parameters (KM and Vmax) were obtained using a Lineweaver-Burk plot. The value of KM and Vmax parameters were 36.181 g/L and 21.322 g/L.h, respectively. The Ca2+ ions act as competitive inhibitor in sucrose hydrolysis using invertase. Therefore, the inhibition mechanism was followed the competitive inhibition mechanism. The value of inhibition constant was 0.833 g/g. These parameters were obtained from the non-substrate inhibition process because this study used the low substrate concentrations which means the fermentable sugar production was low. Hence, there were still more challenges to studying the simultaneous effect of substrate and Ca2+ on sucrose hydrolysis to produce high fermentable sugar. Copyright © 2019 BCREC Group. All rights reserved

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Effect of deglycosylation on the properties of thermophilic invertase purified from the yeast Candida guilliermondii MpIIIa

Cited by 18 publications

References 36 publications

Characterization of a novel low-temperature-active, alkaline and sucrose-tolerant invertase

Characterization of a novel low-temperature-active, alkaline and sucrose-tolerant invertase

Identification and Immobilization of an Invertase With High Specific Activity and Sucrose Tolerance Ability of Gongronella sp. w5 for High Fructose Syrup Preparation

Inhibition Effect of Ca2+ Ions on Sucrose Hydrolysis Using Invertase

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