Enzymatic Cellulose Hydrolysis: Enzyme Reusability and Visualization of β-Glucosidase Immobilized in Calcium Alginate

Tsai, Chien Tai; Meyer, Anne S.

doi:10.3390/molecules191219390

Cited by 59 publications

(23 citation statements)

References 37 publications

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“…More recently, these enzymes have gained commercial significance because of their biotechnological application in processes related to preparation of plant-based foods, e.g., conversion of phytoestrogen glucosides in fruits and vegetables to aglycone moieties, detoxification of cassava, and degradation of bitter compounds in citrus fruit juices and unripe olives (5,6). Many mesophilic and thermophilic BGs have been cloned, purified, and characterized during the past 2 decades (5,(7)(8)(9)(10). In contrast to mesophilic or thermophilic homologues, BGs from organisms adapted to cold environments have potential application in fields that include food processing to prevent spoilage or changes of nutrient content and taste, detergents, textiles, and bioremediation of polluted soils and wastewaters at low to moderate temperatures (11)(12)(13).…”

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confidence: 99%

Molecular Structural Basis for the Cold Adaptedness of the Psychrophilic β-Glucosidase BglU in Micrococcus antarcticus

Miao

Hou

Fan

et al. 2016

Appl Environ Microbiol

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bPsychrophilic enzymes play crucial roles in cold adaptation of microbes and provide useful models for studies of protein evolution, folding, and dynamic properties. We examined the crystal structure (2.2-Å resolution) of the psychrophilic ␤-glucosidase BglU, a member of the glycosyl hydrolase 1 (GH1) enzyme family found in the cold-adapted bacterium Micrococcus antarcticus. Structural comparison and sequence alignment between BglU and its mesophilic and thermophilic counterpart enzymes (BglB and GlyTn, respectively) revealed two notable features distinct to BglU: (i) a unique long-loop L3 (35 versus 7 amino acids in others) involved in substrate binding and (ii) a unique amino acid, His299 (Tyr in others), involved in the stabilization of an ordered water molecule chain. Shortening of loop L3 to 25 amino acids reduced low-temperature catalytic activity, substrate-binding ability, the optimal temperature, and the melting temperature (T m ). Mutation of His299 to Tyr increased the optimal temperature, the T m , and the catalytic activity. Conversely, mutation of Tyr301 to His in BglB caused a reduction in catalytic activity, thermostability, and the optimal temperature (45 to 35°C). Loop L3 shortening and H299Y substitution jointly restored enzyme activity to the level of BglU, but at moderate temperatures. Our findings indicate that loop L3 controls the level of catalytic activity at low temperatures, residue His299 is responsible for thermolability (particularly heat lability of the active center), and long-loop L3 and His299 are jointly responsible for the psychrophilic properties. The described structural basis for the cold adaptedness of BglU will be helpful for structure-based engineering of new cold-adapted enzymes and for the production of mutants useful in a variety of industrial processes at different temperatures.T he ␤-glucosidases (BGs; ␤-D-glucoside glycohydrolases, EC 3.2.1.21) are a widely occurring group of enzymes that cleave the carbohydrate moiety of short-chain oligosaccharides (two to six degrees of polymerization), alkyl, and aryl ␤-glucosides (1, 2). BGs have been classified into 135 glycoside hydrolase (GH) families, based on amino acid sequence similarity. GHs can also be classified into two major types, retaining and inverting enzymes, according to changes in anomeric configuration during hydrolytic reactions. Early studies of BGs were focused on their role in degrading the plant polymers cellulose and xylan (3, 4). More recently, these enzymes have gained commercial significance because of their biotechnological application in processes related to preparation of plant-based foods, e.g., conversion of phytoestrogen glucosides in fruits and vegetables to aglycone moieties, detoxification of cassava, and degradation of bitter compounds in citrus fruit juices and unripe olives (5, 6). Many mesophilic and thermophilic BGs have been cloned, purified, and characterized during the past 2 decades (5, 7-10). In contrast to mesophilic or thermophilic homologues, BGs from organisms adapted to cold...

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confidence: 99%

Molecular Structural Basis for the Cold Adaptedness of the Psychrophilic β-Glucosidase BglU in Micrococcus antarcticus

Miao

Hou

Fan

et al. 2016

Appl Environ Microbiol

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“…Another most common technique for immobilization of enzymes is entrapment, where a gel‐like matrix is used to entrap the enzyme; the most commonly used gel matrices are polyacrylamide and calcium alginate. Increased reusability and a threefold enhancement in thermostability was obtained for cellulase and β‐glucosidase enzymes immobilized in alginate beads . Xylanase isolated from Aspergillus flavus and Bacillus licheniformis were subjected to entrapment in alginate beads followed by glutaraldehyde crosslinking.…”

Section: Stabilization Of Cellulolytic Enzymesmentioning

confidence: 99%

Stable Cellulolytic Enzymes and Their Application in Hydrolysis of Lignocellulosic Biomass

2018

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The effective utilization of lignocellulosic biomass for production of valueadded products in bio-refineries mainly depends on their hydrolysis into constituent soluble sugars. Despite the commercial availability of lignocellulolytic enzymes, there are certain challenges in using them at an industrial scale which arise from their inherent characteristics. Hence, development of industrially potent and stable cellulolytic enzymes is of great importance. While a lot of emphasis has been given to produce the enzymes with high titre and productivity, the properties of these enzymes, which affect their performance in hydrolyzing lignocellulosic residues, have largely been ignored. The various properties that affect the hydrolysis are thermal stability, catalytic efficiency, non-specific adsorption, end-product inhibition resistance, and shear inactivation. This review discusses several approaches such as finding new enzymes from extremophiles, recombinant production of cellulolytic enzymes in industrial strains, directed evolution and mutagenesis, and formation of the enzyme aggregates that are being investigated to develop stable cellulase enzymes. The authors also suggest that for a meaningful information on the modified cellulolytic enzymes and their application in cellulose hydrolysis, they should be evaluated under practical process condition.

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“…3 This is due to the very high activity of β-glucosidase. 4 Cellulose enzyme performance can be improved by accepting β-glucosidase activity inhibitors from lichen.…”

Section: Introductionmentioning

confidence: 99%

Effect of Beta Glucosidase Inhibitor from Lichen Extract in Microcrystalline Cellulose Preparation from Water Hyacinth (Eichhornia crassipes)

Putri

Sutriyo²,

Suryadi³

2019

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Objective: Microcrystalline cellulose (MCC) is an excipient commonly used in the manufacturing of pharmaceutical preparations, especially tablet. MCC has been successfully made from water hyacinth (Eichhornia crassipes) through enzymatic hydrolysis process. This study aimed to find the effect of β-glucosidase inhibitor from lichen extract to the yield of MCC, the optimum conditions of enzymatic hydrolysis including pH and reaction time and characteristics of MCC obtained compared to reference, Avicel PH-101. Method: The study was began with extraction of cellulose enzyme and followed by determination of optimum beta-glucosidase inhibitor concentration. Then, this conditions were used for enzymatic hydrolysis of α-cellulose to MCC, followed by identification and characterization of MCC obtained and compared with Avicel PH-101. Results: The optimum concentration of inhibitor was 120 ppm. A little higher yield of MCC was obtained when inhibitor extract is used in hydrolysis alfa-cellulose to MCC. The identity of MCC obtained was similar to infrared spectrum of reference. Other characteristics of MCC obtained were powder in the form of a slightly coarse, odorless and tasteless and slightly yellowish than reference. Conclusion: Addition of extract of Beta glucosidase inhibitor did not improve the MCC yield significantly, but identity and characteristics of MCC obtained showed similarities to the microcrystalline cellulose reference (Avicel PH = 101).

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Enzymatic Cellulose Hydrolysis: Enzyme Reusability and Visualization of β-Glucosidase Immobilized in Calcium Alginate

Cited by 59 publications

References 37 publications

Molecular Structural Basis for the Cold Adaptedness of the Psychrophilic β-Glucosidase BglU in Micrococcus antarcticus

Molecular Structural Basis for the Cold Adaptedness of the Psychrophilic β-Glucosidase BglU in Micrococcus antarcticus

Stable Cellulolytic Enzymes and Their Application in Hydrolysis of Lignocellulosic Biomass

Effect of Beta Glucosidase Inhibitor from Lichen Extract in Microcrystalline Cellulose Preparation from Water Hyacinth (Eichhornia crassipes)

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