Immobilization of β-glucuronidase in lysozyme-induced biosilica particles to improve its stability

Song, Xiaokai; Jiang, Zhongyi; Li, Lin; Wu, Hong

doi:10.1007/s11705-014-1421-2

Cited by 24 publications

(23 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Earlier studies have highlighted the possibility of measuring the 29 Si spectrum of the material to understand the changes in compaction upon changes of experimental conditions [11,103,104,15]. However, also the organic/biological fraction of these systems is readily accessible to SSNMR [12,13,105].…”

Section: Resolution In Bioinspired Materials and Its Interpretationmentioning

confidence: 99%

“…Furthermore, there is a large number of samples that can be studied at atomic resolution only by SSNMR such as fibrils [1][2][3], membrane proteins [3][4][5][6] and large protein aggregates [7][8][9][10], plus a number of hybrid protein-material samples that greatly benefit from the study of both the protein and the material sides by means of SSNMR [11][12][13][14][15].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

NMR of sedimented, fibrillized, silica-entrapped and microcrystalline (metallo)proteins

Ravera

Schubeis

Martelli

et al. 2015

Journal of Magnetic Resonance

View full text Add to dashboard Cite

Section: Resolution In Bioinspired Materials and Its Interpretationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

NMR of sedimented, fibrillized, silica-entrapped and microcrystalline (metallo)proteins

Ravera

Schubeis

Martelli

et al. 2015

Journal of Magnetic Resonance

View full text Add to dashboard Cite

“…Chemical shift perturbations along the backbone and sidechains were linked to respective changes in conformation and proximity of residues to silica [24][25][26] . Similarly, recent highresolution MAS NMR studies of enzymes entrapped in porous silica has examined the effects of confinement on the structure and functionality of proteins [27][28][29][30] .…”

Section: Introductionmentioning

confidence: 99%

Studying the Conformation of a Silaffin-Derived Pentalysine Peptide Embedded in Bioinspired Silica using Solution and Dynamic Nuclear Polarization Magic-Angle Spinning NMR

et al. 2016

View full text Add to dashboard Cite

Smart materials are created in nature at interfaces between biomolecules and solid materials. The ability to probe the structure of functional peptides that engineer biogenic materials at this heterogeneous setting can be facilitated tremendously by use of DNP-enhanced solid-state NMR spectroscopy. This sensitive NMR technique allows simple and quick measurements, often without the need for isotope enrichment. Here, it is used to characterize a pentalysine peptide, derived from a diatom's silaffin protein. The peptide accelerates the formation of bioinspired silica and gets embedded inside the material as it is formed. Two-dimensional DNP MAS NMR of the silica-bound peptide and solution NMR of the free peptide are used to derive its secondary structure in the two states and to pinpoint some subtle conformational changes that the peptide undergoes in order to adapt to the silica environment. In addition, interactions between abundant lysine residues and silica surface are identified, and proximity of other side chains to silica and to neighboring peptide molecules is discussed.

show abstract

“…It has been widely used in removing the sugar moiety from glucuronides to manipulate the solubility to enhance the efficacy and alleviate the associated side effect (Sakurama et al 2014;Song et al 2014;Wang et al 2015). However, the low thermostability of β-glucuronidase has impeded its industrial application and the mutants with higher thermostability have been rarely reported.…”

Section: Introductionmentioning

confidence: 99%

Engineering the thermostability of β-glucuronidase from Penicillium purpurogenum Li-3 by loop transplant

Feng

Tang

Han

et al. 2016

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

In this study, we proposed a loop transplant strategy to improve the thermostability of Penicillium purpurogenum Li-3 β-glucuronidase expressed in Escherichia coli (abbreviated to PGUS-E). Firstly, three unstable surface loops of PGUS-E to be replaced were identified with regards to B-factor values and in-depth structure analysis: loops 205-211, 258-263, and 25-31. Then, based on B-factor analysis, eight stable loops for substitution were selected from two typical thermophilic glycosidases which had low homology with PGUS-E (less than 25 %). By analyzing the common features of these stable loops, it was found that they shared a common residue skeleton DXXTX(X)R, based on this, three chimera loops were also manually designed: RSQTSND, RSSTQRD, and DDQTSR. All these loops were introduced to replace the unstable loops of PGUS-E by homology structure modeling, and only mutants with increased hydrogen bonds number and good compatibility with the local mutated region were further subjected to experimental verification. By using this strategy, 10 mutants were experimentally generated, among which three mutants, M1, M3, and M8, were obtained which showed 11.8, 3.3, and 9.4 times higher half-life at 70 °C than that of wild-type (8.5 min). Finally, the MD simulation indicated that the increased hydrogen bonds, decreased flexibility of N-terminal, and increased π-π stacking interaction were responsible for the improved thermostability.

show abstract

Immobilization of β-glucuronidase in lysozyme-induced biosilica particles to improve its stability

Cited by 24 publications

References 35 publications

NMR of sedimented, fibrillized, silica-entrapped and microcrystalline (metallo)proteins

NMR of sedimented, fibrillized, silica-entrapped and microcrystalline (metallo)proteins

Studying the Conformation of a Silaffin-Derived Pentalysine Peptide Embedded in Bioinspired Silica using Solution and Dynamic Nuclear Polarization Magic-Angle Spinning NMR

Engineering the thermostability of β-glucuronidase from Penicillium purpurogenum Li-3 by loop transplant

Contact Info

Product

Resources

About