Preparation of Cross-Linked Glucoamylase Aggregates Immobilization by Using Dextrin and Xanthan Gum as Protecting Agents

Li, Xiaodong; Wu, Jia; Jia, Dong-Chen; Wan, Yong-Hu; Yang, Na; Qiao, Min

doi:10.3390/catal6060077

Cited by 15 publications

(8 citation statements)

References 39 publications

(41 reference statements)

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“…Hence, incorporation of co-aggregants in the enzyme mixture could provide a support for the aggregates to link onto and form a more rigid CLEA structure. Following this, CLEA-elastase has been found to exhibit an enhanced stability under high temperature when compared to free elastase, which is coherent with subsequent CLEA studies [39][40][41]. The enhanced tolerance of CLEA-elastase towards high temperature is a result of increased rigidity of the structure caused by additional linkage between enzyme molecules.…”

Section: Discussionsupporting

confidence: 71%

Biochemical and Structural Characterization of Cross-Linked Enzyme Aggregates (CLEAs) of Organic Solvent Tolerant Protease

et al. 2020

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Cross-linked enzyme aggregates (CLEAs) is an immobilization technique that can be used to customize enzymes under an optimized condition. Structural analysis on any enzyme treated with a CLEA remains elusive and has been less explored. In the present work, a method for preparing an organic solvent tolerant protease using a CLEA is disclosed and optimized for better biochemical properties, followed by an analysis of the structure of this CLEA-treated protease. The said organic solvent tolerant protease is a metalloprotease known as elastase strain K in which activity of the metalloprotease is measured by a biochemical interaction with azocasein. Results showed that when a glutaraldehyde of 0.02% (v/v) was used under a 2 h treatment, the amount of recovered activity in CLEA-elastase was highest. The recovered activity of CLEA-elastase and CLEA-elastase-SB (which was a CLEA co-aggregated with starch and bovine serum albumin (BSA)) were at an approximate 60% and 80%, respectively. The CLEA immobilization of elastase strain K allowed the stability of the enzyme to be enhanced at high temperature and at a broader pH. Both CLEA-elastase and CLEA-elastase-SB end-products were able to maintain up to 67% enzyme activity at 60 °C and exhibiting an enhanced stability within pH 5–9 with up to 90% recovering activity. By implementing a CLEA on the organic solvent tolerant protease, the characteristics of the organic solvent tolerant were preserved and enhanced with the presence of 25% (v/v) acetonitrile, ethanol, and benzene at 165%, 173%, and 153% relative activity. Structural analysis through SEM and dynamic light scattering (DLS) showed that CLEA-elastase had a random aggregate morphology with an average diameter of 1497 nm.

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Section: Discussionsupporting

confidence: 71%

Biochemical and Structural Characterization of Cross-Linked Enzyme Aggregates (CLEAs) of Organic Solvent Tolerant Protease

et al. 2020

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“…This encouraging improvement verified that, apart from the hydrophobic characteristic of the PBAP coating that can induce interfacial activation of lipase, its biocompatible property also helped to maintain the lipase activity. The advantages of using biocompatible and biomimetic supports for enzyme immobilization have also been reported by others [30][31][32]. The biocompatible property also helped to maintain the lipase activity.…”

Section: Lipase Immobilization and Activity Retentionmentioning

confidence: 53%

Adsorption and Activity of Lipase on Polyphosphazene-Modified Polypropylene Membrane Surface

et al. 2016

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Abstract:In this work, poly(n-butylamino)(allylamino)phosphazene (PBAP) was synthesized and tethered on polypropylene microporous membrane (PPMM) with the aim of offering a biocompatible and, at the same time, moderately hydrophobic microenvironment to lipase for the first time. Lipase from Candida rugosa was used and the influence of membrane surface conditions on the activities of immobilized lipases was evaluated. Water contact angle measurement as well as field emission scanning electron microscopy were used to characterize the morphology of the modified membranes. The results showed an improvement in the adsorption capacity (26.0 mg/m 2 ) and activity retention (68.2%) of the immobilized lipases on the PBAP-modified PPMM. Moreover, the lipases immobilized on the modified PPMM showed better thermal and pH stability.

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“…The starch also decreases the enzyme loading and diffusion problems. The complex organic system of coaggregation form stable activity was similar to being "locked", which is constructed following Schiff's base by chemical groups of substrate, degradation products, and enzyme molecules in crosslinking and catalytic process [27]. In addition, the α-amylase active site was protected by "setup protection", which means these reactions combine active sites with optimal concentration of substrate to avoid excessive crosslinking of the amino acid residues at the α-amylase catalytic sites.…”

Section: Discussionmentioning

confidence: 99%

“…However, it has not been reported whether starch can be a protective agent during the preparation of CLEA of α-amylase (CLEA-S). We hypothesized that the addition of BSA or starch would keep the conformation of the enzyme during the crosslinking process [27]. To test this hypothesis, we prepared CLEA, CLEA-S, and CLEA-BSA from crude alpha-amylase and characterized their optimum pH, optimum temperature, kinetic parameters, microstructure, recycling, storage, and thermal stabilities and compared them with those of the enzyme in free form.…”

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confidence: 99%

Physiochemical Characterization of α-Amylase as Crosslinked Enzyme Aggregates

Bian

et al. 2018

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Starch is promising candidate material for enhancing the catalytic activity of α-amylase during the crosslinking process. To help meet industrial needs, here we tested the influence of bovine serum albumin (BSA) and starch on the performance of crosslinked α-amylase aggregates (CLEA), α-amylase-prepared as CLEA with starch (CLEA-S), and BSA (CLEA-BSA). Our results showed that the activities of CLEA, CLEA-S, and CLEA-BSA were 1.1-, 1.0-, and 0.74-fold higher than the free α-amylase, respectively. The stability of the immobilized enzyme slightly changed. After immobilization, the enzyme increased its pH and temperature ranges with the optimal pH values of 5.5, 7.5, 5.5, respectively for CLEA, CLEA-S, and CLEA-BSA, and an upper temperature limit of 50 °C for all three immobilized forms. Among the three immobilized forms, the CLEA-S was the most thermostable, losing only 3% of its initial activity during 390 min incubation at 50 °C. Our microscopic observations of CLEA-S showed that porous structures were formed and such structures could help substance diffusion. In addition, there was excellent affinity between CLEA-S and the substrate. The results suggest that CLEA-S have great potential for industrial application, including for use in starch-based alcohol fermentation.

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Preparation of Cross-Linked Glucoamylase Aggregates Immobilization by Using Dextrin and Xanthan Gum as Protecting Agents

Cited by 15 publications

References 39 publications

Biochemical and Structural Characterization of Cross-Linked Enzyme Aggregates (CLEAs) of Organic Solvent Tolerant Protease

Biochemical and Structural Characterization of Cross-Linked Enzyme Aggregates (CLEAs) of Organic Solvent Tolerant Protease

Adsorption and Activity of Lipase on Polyphosphazene-Modified Polypropylene Membrane Surface

Physiochemical Characterization of α-Amylase as Crosslinked Enzyme Aggregates

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