Characterisation of maltase from enteropathogenic<i>Escherichia coli</i>

Olusanya, O.; Olutiola, P. O.

doi:10.1111/j.1574-6968.1986.tb01702.x

Cited by 7 publications

(3 citation statements)

References 8 publications

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“…Previously, Mn 2+ was reported to stimulate the activity of α-glucosidase from Thermotoga maritima [21], Ca 2+ was required for the catalytic efficiency of the enzyme from E. coli [22], and Mucor racemosus α-glucosidase was able to be stimulated by Al 3+ , Ca 2+ , Mg 2+ and Na + [23]. In this study, the effect of metal ions on the activity of GSJ was analyzed by preincubating the enzyme with 1 mM metal ions at 65 °C for 1 h. As shown in Figure 5, Mg 2+ and K + did not affect the enzyme activity of GSJ, Cu 2+ , Zn 2+ and Ni 2+ strongly inhibited the enzyme activity by 12.3%–38.6%, while Fe 2+ almost completely inhibited the enzyme activity.…”

Section: Resultsmentioning

confidence: 99%

Heterologous Expression of a Thermostable α-Glucosidase from Geobacillus sp. Strain HTA-462 by Escherichia coli and Its Potential Application for Isomaltose–Oligosaccharide Synthesis

et al. 2019

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Isomaltose–oligosaccharides (IMOs), as food ingredients with prebiotic functionality, can be prepared via enzymatic synthesis using α-glucosidase. In the present study, the α-glucosidase (GSJ) from Geobacillus sp. strain HTA-462 was cloned and expressed in Escherichia coli BL21 (DE3). Recombinant GSJ was purified and biochemically characterized. The optimum temperature condition of the recombinant enzyme was 65 °C, and the half-life was 84 h at 60 °C, whereas the enzyme was active over the range of pH 6.0–10.0 with maximal activity at pH 7.0. The α-glucosidase activity in shake flasks reached 107.9 U/mL and using 4-Nitrophenyl β-D-glucopyranoside (pNPG) as substrate, the Km and Vmax values were 2.321 mM and 306.3 U/mg, respectively. The divalent ions Mn2+ and Ca2+ could improve GSJ activity by 32.1% and 13.8%. Moreover, the hydrolysis ability of recombinant α-glucosidase was almost the same as that of the commercial α-glucosidase (Bacillus stearothermophilus). In terms of the transglycosylation reaction, with 30% maltose syrup under the condition of 60 °C and pH 7.0, IMOs were synthesized with a conversion rate of 37%. These studies lay the basis for the industrial application of recombinant α-glucosidase.

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Section: Resultsmentioning

confidence: 99%

Heterologous Expression of a Thermostable α-Glucosidase from Geobacillus sp. Strain HTA-462 by Escherichia coli and Its Potential Application for Isomaltose–Oligosaccharide Synthesis

et al. 2019

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“…[25] If pH optimum for the enzyme from Escherichia coli was 6.9, [25] similar optimal pH, 6.5, was obtained for another Escherichia coli maltase. [26] Nevertheless, the maltase from the yeast Saccharomyces cerevisiae had pH optimum had pH optimum between 7.0 and 7.5, [17] but for enzyme of Saccharomyces carlsbergensis the pH optimum was found to be between pH 6.7 and 6.8. [27] The purified maltase of Bacillus subtilis was relatively stable at pH values of 5.5 to 6.6, [25] and pH stability range of partially purified enzyme from Bacillus brevis was from 5.0 to 7.0.…”

Section: Influence Of Ph and Type Of Buffermentioning

confidence: 96%

Biosensor for maltose quantification and estimation of maltase activity

Emelyanova

2019

Adv Biochips

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The aim of this study was to create a laboratory model of an amperometric microbial biosensor for maltose quantification in the presence and absence of starch and to estimate the use of the model in the study of maltase activity of the culture-receptor. The biosensor for maltose was developed on the basis of a Clark-type oxygen electrode, coupled with a bioreceptor, which contained bacterial cells immobilized on the membrane. The determination of maltose concentration was based on measuring the rate of electrode current change in response to addition of the analyte. The detection limit of the biosensor was 1 µM maltose, a linear interval of standard curve was observed from 14 µM up to 1.9 mM of maltose. The microbial biosensor demonstrated good sensitivity to maltose, 36.02 nA (M•s) −1. Combination of bioreceptors on the basis of fungus and bacterium allowed of using the biosensor for quantification of maltose in the presence of starch. Changes in metabolism of the culture-receptor had an effect on the biosensor response. It indicated that the developed model was a tool of simple construction and easy-to-use in the study of maltase activity of the immobilized culture-receptor.

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“…Maltose was transported to the E. coli cell via the maltodextrin/maltose transport proteins encoded by its maltose operon (mal) (Boos and Shuman, 1998). Then, maltase or α-glucosidase could catalyze the hydrolysis of maltose to two molecules of glucose, which were further metabolized by the central metabolic pathway (Olusanya and Olutiola, 1986). Several studies have demonstrated that the expression of genes in metabolic pathways in bacteria can be regulated by different carbon sources (Yao et al, 2011;Beisel and Afroz, 2016).…”

Section: Perspective Of Recombinant Protein Production With Sugar Sup...mentioning

confidence: 99%

Effect of supplemented sugar in lysogeny broth medium on growth of Escherichia coli BL21(DE3) and recombinant protein production

2022

ANRES

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Importance of the work: Escherichia coli BL21(DE3) is among the powerful hosts for recombinant protein expression. Varying approaches have been explored to improve the production of large amounts of recombinant protein. Objectives:To examine the effect of single sugar addition on the growth of E. coli BL21(DE3) and the production ability of recombinant green fluorescence protein (GFP) as a model for this study. Materials & Methods: Bacteria were cultured in Luria-Bertani broth (LB) media with supplemental sugars as additional carbon sources (2-10 g/L). Cell growth was monitored by measuring the optical density at 600 nm. Following the inducement of recombinant GFP expression by adding isopropyl β-d-1-thiogalactopyranoside, the content of soluble GFP was quantitated using two distinct methods: fluorescence detection and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. Results: The effect of sugar on the growth of E. coli BL21(DE3) could be classified into four groups of positive, negative, positive/negative and neutral effects, with a similar trend when pET-11a was included in the bacterial cells. Then, the expression of GFP under the optimum condition was determined in LB with the addition of the positive effect sugars (xylose or maltose). The addition of xylose or maltose in LB media significantly (p < 0.05) increased both cell growth by 2.0-2.5 fold and the production of recombinant GFP by 1.5-2.0 fold than those of the control and other sugars. Main finding: This study presented a proof of concept to improve the recombinant protein production in E. coli BL21(DE3) by adding a carbon source into traditional culture media without using engineered cells.

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Characterisation of maltase from enteropathogenicEscherichia coli

Cited by 7 publications

References 8 publications

Heterologous Expression of a Thermostable α-Glucosidase from Geobacillus sp. Strain HTA-462 by Escherichia coli and Its Potential Application for Isomaltose–Oligosaccharide Synthesis

Heterologous Expression of a Thermostable α-Glucosidase from Geobacillus sp. Strain HTA-462 by Escherichia coli and Its Potential Application for Isomaltose–Oligosaccharide Synthesis

Biosensor for maltose quantification and estimation of maltase activity

Effect of supplemented sugar in lysogeny broth medium on growth of Escherichia coli BL21(DE3) and recombinant protein production

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