Improving the Thermostability of α-Glucosidase from Xanthomonas campestris through Proline Substitutions Guided by Semi-rational Design

Chen, Luyi; Jiang, Kunyao; Zhou, Yaoyao; Zhu, Linjiang; Chen, Xiaolong

doi:10.1007/s12257-022-0129-2

Cited by 5 publications

(2 citation statements)

References 39 publications

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“…Semi-rational design is a hybrid approach combining the benefits of rational design and directed evolution [ 52 , 53 ]. This approach uses existing libraries (like rational design) to select a specific promising sequence, but the specific amino acid related to the targeted function is unknown [ 54 , 55 ]. The mutation of single amino acids or a combination of them generates new mutants to be screened and evaluated like in directed evolution to identify the desirable variant in the library [ 54 ].…”

Section: Strategies and Challenges For The Development Of Engineered ...mentioning

confidence: 99%

Bioengineered Enzymes and Precision Fermentation in the Food Industry

Boukid¹,

Ganeshan²,

Wang³

et al. 2023

IJMS

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Enzymes have been used in the food processing industry for many years. However, the use of native enzymes is not conducive to high activity, efficiency, range of substrates, and adaptability to harsh food processing conditions. The advent of enzyme engineering approaches such as rational design, directed evolution, and semi-rational design provided much-needed impetus for tailor-made enzymes with improved or novel catalytic properties. Production of designer enzymes became further refined with the emergence of synthetic biology and gene editing techniques and a plethora of other tools such as artificial intelligence, and computational and bioinformatics analyses which have paved the way for what is referred to as precision fermentation for the production of these designer enzymes more efficiently. With all the technologies available, the bottleneck is now in the scale-up production of these enzymes. There is generally a lack of accessibility thereof of large-scale capabilities and know-how. This review is aimed at highlighting these various enzyme-engineering strategies and the associated scale-up challenges, including safety concerns surrounding genetically modified microorganisms and the use of cell-free systems to circumvent this issue. The use of solid-state fermentation (SSF) is also addressed as a potentially low-cost production system, amenable to customization and employing inexpensive feedstocks as substrate.

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Section: Strategies and Challenges For The Development Of Engineered ...mentioning

confidence: 99%

Bioengineered Enzymes and Precision Fermentation in the Food Industry

Boukid¹,

Ganeshan²,

Wang³

et al. 2023

IJMS

View full text Add to dashboard Cite

show abstract

“…Protein thermostability is one of the major factors affecting industrial applicability and engineering thermostability of proteins that has been rigorously studied [ 1 , 2 , 3 ]. The statistical studies of thermophilic and mesophilic proteins have attempted to understand major structural features that govern protein thermostability.…”

Section: Introductionmentioning

confidence: 99%

Statistical Analysis of the Role of Cavity Flexibility in Thermostability of Proteins

Hong,

Yoon,

et al. 2024

Polymers

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Conventional statistical investigations have primarily focused on the comparison of the simple one-dimensional characteristics of protein cavities, such as number, surface area, and volume. These studies have failed to discern the crucial distinctions in cavity properties between thermophilic and mesophilic proteins that contribute to protein thermostability. In this study, the significance of cavity properties, i.e., flexibility and location, in protein thermostability was investigated by comparing structural differences between homologous thermophilic and mesophilic proteins. Three dimensions of protein structure were categorized into three regions (core, boundary, and surface) and a comparative analysis of cavity properties using this structural index was conducted. The statistical analysis revealed that cavity flexibility is closely related to protein thermostability. The core cavities of thermophilic proteins were less flexible than those of mesophilic proteins (averaged B’ factor values, −0.6484 and −0.5111), which might be less deleterious to protein thermostability. Thermophilic proteins exhibited fewer cavities in the boundary and surface regions. Notably, cavities in mesophilic proteins, across all regions, exhibited greater flexibility than those in thermophilic proteins (>95% probability). The increased flexibility of cavities in the boundary and surface regions of mesophilic proteins, as opposed to thermophilic proteins, may compromise stability. Recent protein engineering investigations involving mesophilic xylanase and protease showed results consistent with the findings of this study, suggesting that the manipulation of flexible cavities in the surface region can enhance thermostability. Consequently, our findings suggest that a rational or computational approach to the design of flexible cavities in surface or boundary regions could serve as an effective strategy to enhance the thermostability of mesophilic proteins.

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A Bibliometric Analysis: Current Perspectives and Potential Trends of Enzyme Thermostability from 1991–2022

Zhang

Wang

et al. 2023

Appl Biochem Biotechnol

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Improving the Thermostability of α-Glucosidase from Xanthomonas campestris through Proline Substitutions Guided by Semi-rational Design

Cited by 5 publications

References 39 publications

Bioengineered Enzymes and Precision Fermentation in the Food Industry

Bioengineered Enzymes and Precision Fermentation in the Food Industry

Statistical Analysis of the Role of Cavity Flexibility in Thermostability of Proteins

A Bibliometric Analysis: Current Perspectives and Potential Trends of Enzyme Thermostability from 1991–2022

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