Production optimization of invertase by Lactobacillus brevis Mm-6 and its immobilization on alginate beads

Awad, Ghada E. A.; Amer, Hassan; El-Gammal, Eman W.; Helmy, Wafaa A.; Esawy, Mona A.; Elnashar, Magdy M. M.

doi:10.1016/j.carbpol.2012.12.039

Cited by 42 publications

(37 citation statements)

References 31 publications

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“…Current biocatalysts comprising entrapped S. cerevisiae cells or the immobilized invertase exhibit substrate inhibition kinetics and behave rather no stable at temperature exceeding 50°C [2]. Immobilized biocatalysts using other microorganisms or enzymes have shown technical limitations such as low thermal stability, decreased catalytic efficiency, substrate inhibition, incomplete sucrose inversion, or by-products formation [2,13,14,16,30,39-41]. The immobilized whole-cell biocatalyst developed in this work fully hydrolyzed cane sugar in a highly concentrated solution (2.04 M; 70%, w/v) during repeated batches or continuous operation at 60°C without a remarkable loss of the invertase activity.…”

Section: Resultsmentioning

confidence: 99%

Complete sucrose hydrolysis by heat-killed recombinant Pichia pastoris cells entrapped in calcium alginate

et al. 2014

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BackgroundAn ideal immobilized biocatalyst for the industrial-scale production of invert sugar should stably operate at elevated temperatures (60-70°C) and high sucrose concentrations (above 60%, w/v). Commercial invertase from the yeast Saccharomyces cerevisiae is thermolabile and suffers from substrate inhibition. Thermotoga maritima β-fructosidase (BfrA) is the most thermoactive and thermostable sucrose-hydrolysing enzyme so far identified and allows complete inversion of the substrate in highly concentrated solutions.ResultsIn this study, heat-killed Pichia pastoris cells bearing N-glycosylated BfrA in the periplasmic space were entrapped in calcium alginate beads. The immobilized recombinant yeast showed maximal sucrose hydrolysis at pH 5–7 and 90°C. BfrA was 65% active at 60°C and had no activity loss after incubation without the substrate at this temperature for 15 h. Complete inversion of cane sugar (2.04 M) at 60°C was achieved in batchwise and continuous operation with respective productivities of 4.37 and 0.88 gram of substrate hydrolysed per gram of dry beads per hour. The half-life values of the biocatalyst were 14 and 20 days when operated at 60°C in the stirred tank and the fixed-bed column, respectively. The reaction with non-viable cells prevented the occurrence of sucrose fermentation and the formation of by-products. Six-month storage of the biocatalyst in 1.46 M sucrose (pH 5.5) at 4°C caused no reduction of the invertase activity.ConclusionsThe features of the novel thermostable biocatalyst developed in this study are more attractive than those of immobilized S. cerevisiae cells for application in the enzymatic manufacture of inverted sugar syrup in batch and fixed-bed reactors.

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Section: Resultsmentioning

confidence: 99%

Complete sucrose hydrolysis by heat-killed recombinant Pichia pastoris cells entrapped in calcium alginate

et al. 2014

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“…Invertase is one of the most widely used enzymes in the food industry [2][3][4]; moreover, it has been extensively studied as a model enzyme for describing many of their kinetic principles and for its role on human health [2]. This enzyme is used to catalyze the conversion of sucrose to a mixture of glucose and fructose, which is known as inverted sugar syrup.…”

Section: Introductionmentioning

confidence: 99%

“…β-Fructosidases from lactic acid bacteria and bifidobacteria (LAB&B) sources are able to efficiently hydrolyze sucrose [3,[7][8][9][10]. Enzymes from a microbial source are more useful than those ones obtained from plants because is easier to genetically manipulate microbes than plants.…”

Section: Introductionmentioning

confidence: 99%

Lactobacillus plantarum WCFS1 β-Fructosidase: Evidence for an Open Funnel-Like Channel Through the Catalytic Domain with Importance for the Substrate Selectivity

Mendoza-Llerenas

Pérez

Gómez‐Sandoval

et al. 2016

Appl Biochem Biotechnol

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β-Fructosidase, a glycoside hydrolase of a biotechnologically important strain, was studied for its biochemical, physicochemical, and three-dimensional structure characteristics. This enzyme was heterologously expressed in Escherichia coli as a C-terminal His-tagged protein (SacB). β-Fructosidase catalyzes the cleavage of glycoside bonds toward certain carbohydrates with β-fructofuranosyl linkages; however, SacB exhibited selectivity toward sucrose and an optimum activity at pH 6.0-6.5 and 37 °C. In such optimum enzymatic activity conditions, the SacB was commonly observed as a monodisperse protein by dynamic light scattering (DLS). As β-fructosidase belongs to glycoside hydrolase family 32 (GH32), a β-sandwich and a five-bladed β-propeller domain are typical predicted folds in its structure. Docking and molecular dynamic simulations revealed for the first time a funnel-like channel perfectly exposed in the β-propeller domain of the Lactobacillus plantarum β-fructosidase (this allows the interaction between its entire catalytic triad and substrates that are larger than sucrose). In contrast, SacB showed a closed central tunnel collaterally induced by its His-tag.

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“…Pectinase (EC 3.1.1.11) represents for pectic enzymes in brewing (pectolyase, pectozyme and polygalacturonase) have been used in fruit juice extraction and clarification [3]. β-fructofuranosidase namely invertase (EC 3.2.1.26) is an predominant enzyme used in food (confectionery) industry for catalyzing the hydrolysis of terminal nonreducing β-fructofuranoside residues of β-fructofuranosides where fructose is preferred over sucrose due to its sweetness [4].…”

Section: Introductionmentioning

confidence: 99%

Isolation and Screening of Fungal Isolates for Multienzyme Production Through Submerged and Solid State Fermentations

Garlapati¹

2015

J Bioprocess Biotech

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Production optimization of invertase by Lactobacillus brevis Mm-6 and its immobilization on alginate beads

Cited by 42 publications

References 31 publications

Complete sucrose hydrolysis by heat-killed recombinant Pichia pastoris cells entrapped in calcium alginate

Complete sucrose hydrolysis by heat-killed recombinant Pichia pastoris cells entrapped in calcium alginate

Lactobacillus plantarum WCFS1 β-Fructosidase: Evidence for an Open Funnel-Like Channel Through the Catalytic Domain with Importance for the Substrate Selectivity

Isolation and Screening of Fungal Isolates for Multienzyme Production Through Submerged and Solid State Fermentations

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