Enhanced catalytic efficiency of CotA-laccase by DNA shuffling

Ouyang, Fengju; Zhao, Min

doi:10.1080/21655979.2019.1621134

Cited by 19 publications

(9 citation statements)

References 26 publications

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“…In studies attempting to improve the catalytic activity of xylanases, most of the work were limited to modifying on the catalytic domain [ 17 ], with little attention paid to the non-catalytic domain. However, modifications performed by site-directed mutagenesis have not achieved substantial improvements in the catalytic efficiency of xylanases [ 29 ].…”

Section: Discussionmentioning

confidence: 99%

Fusion of a proline-rich oligopeptide to the C-terminus of a ruminal xylanase improves catalytic efficiency

et al. 2022

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Xylanases are widely used in the degradation of lignocellulose and are important industrial enzymes. Therefore, increasing the catalytic activity of xylanases can improve their efficiency and performance. In this study, we introduced the C-terminal proline-rich oligopeptide of the rumen-derived XynA into XylR, a GH10 family xylanase. The optimum temperature and pH of the fused enzyme (XylR-Fu) were consistent with those of XylR; however, its catalytic efficiency was 2.48-fold higher than that of XylR. Although the proline-rich oligopeptide did not change the enzyme hydrolysis mode, the amount of oligosaccharides released from beechwood xylan by XylR-Fu was 17% higher than that released by XylR. This increase may be due to the abundance of proline in the oligopeptide, which plays an important role in substrate binding. Furthermore, circular dichroism analysis indicated that the proline-rich oligopeptide might increase the rigidity of the overall structure, thereby enhancing the affinity to the substrate and catalytic activity of the enzyme. Our study shows that the proline-rich oligopeptide enhances the catalytic efficiency of GH10 xylanases and provides a better understanding of the C-terminal oligopeptide-function relationships. This knowledge can guide the rational design of GH10 xylanases to improve their catalytic activity and provides clues for further applications of xylanases in industry.

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Section: Discussionmentioning

confidence: 99%

Fusion of a proline-rich oligopeptide to the C-terminus of a ruminal xylanase improves catalytic efficiency

et al. 2022

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“…A random mutagenesis library for directed evolution of RuBisCO was constructed by DNA shuffling and error-prone-PCR. 50 Highthroughput selection of RuBisCO mutants is to create RuBisCO-dependent E. coli (RDE), which can be realized by using a minimal medium with arabinose and CO 2 to feed engineered E. coli. By coupling the cell growth with the phenotype of RuBisCO, the screening of RuBisCO mutants with high activity and great phenotype would be more efficient.…”

Section: Fixation Efficiency Of the Cbb Cycle In Heterotrophic Microo...mentioning

confidence: 99%

“…coli with a minimal medium supplemented for selecting RuBisCO mutants that has improved kinetic and biophysical properties. A random mutagenesis library for directed evolution of RuBisCO was constructed by DNA shuffling and error-prone-PCR . High-throughput selection of RuBisCO mutants is to create RuBisCO-dependent E .…”

Section: Strategies For Enhancing the Carbon Fixation Efficiency Of T...mentioning

confidence: 99%

Recent Advances in Engineering Heterotrophic Microorganisms for Reinforcing CO₂ Fixation Based on Calvin–Benson–Bassham Cycle

Feng

Gao

et al. 2023

ACS Sustainable Chem. Eng.

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Global climate change caused by greenhouse gas emission is the fundamental challenge facing mankind. Biological carbon dioxide fixation has recently attracted much attention. The Calvin–Benson–Bassham (CBB) cycle is a major carbon fixation pathway in the biosphere, and it is the most intensively studied because of its prominence and key role in nature as the primary pathway for net CO2 fixation. It was recently shown that expressing the enzymes ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) and phosphoribulokinase (PRK) in Escherichia coli, Saccharomyces cerevisiae, Methylobacterium extorquens, Kluyveromyces marxianus, and Pichia pastoris could achieve CO2 fixation for the production of value-added chemicals. In addition, expressing the synthetic CBB cycle allowed strains to grow autotrophically on CO2. Here, we review the current understanding of the CBB cycle for CO2 fixation, summarize research progress in the assembly of the CBB cycle in heterotrophic microorganisms or developing synthetic autotrophs, and present the strategies to enhance carbon fixation efficiency of the CBB cycle in heterotrophic microorganisms. Recent progress in such applications in reinforcing CO2 fixation for the improved production of various value-added chemicals is also summarized. The challenges encountered in this process and future prospects are further discussed.

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“…Such a computer-driven directed evolution led to the synthesis of laccases with enhanced activity from Pleurotus ostreatus [ 97 , 98 ]. Moreover, Ouyant and Zhao [ 99 ] reported improved laccase activity from the yeast mutant 5E29 with two amino acid substitutions. Directed evolution proved to be a successful technique also for the production of laccases from non-native hosts.…”

Section: Engineering New Ligninolytic Enzymesmentioning

confidence: 99%

Genome-based engineering of ligninolytic enzymes in fungi

et al. 2021

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Background Many fungi grow as saprobic organisms and obtain nutrients from a wide range of dead organic materials. Among saprobes, fungal species that grow on wood or in polluted environments have evolved prolific mechanisms for the production of degrading compounds, such as ligninolytic enzymes. These enzymes include arrays of intense redox-potential oxidoreductase, such as laccase, catalase, and peroxidases. The ability to produce ligninolytic enzymes makes a variety of fungal species suitable for application in many industries, including the production of biofuels and antibiotics, bioremediation, and biomedical application as biosensors. However, fungal ligninolytic enzymes are produced naturally in small quantities that may not meet the industrial or market demands. Over the last decade, combined synthetic biology and computational designs have yielded significant results in enhancing the synthesis of natural compounds in fungi. Main body of the abstract In this review, we gave insights into different protein engineering methods, including rational, semi-rational, and directed evolution approaches that have been employed to enhance the production of some important ligninolytic enzymes in fungi. We described the role of metabolic pathway engineering to optimize the synthesis of chemical compounds of interest in various fields. We highlighted synthetic biology novel techniques for biosynthetic gene cluster (BGC) activation in fungo and heterologous reconstruction of BGC in microbial cells. We also discussed in detail some recombinant ligninolytic enzymes that have been successfully enhanced and expressed in different heterologous hosts. Finally, we described recent advance in CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-Cas (CRISPR associated) protein systems as the most promising biotechnology for large-scale production of ligninolytic enzymes. Short conclusion Aggregation, expression, and regulation of ligninolytic enzymes in fungi require very complex procedures with many interfering factors. Synthetic and computational biology strategies, as explained in this review, are powerful tools that can be combined to solve these puzzles. These integrated strategies can lead to the production of enzymes with special abilities, such as wide substrate specifications, thermo-stability, tolerance to long time storage, and stability in different substrate conditions, such as pH and nutrients.

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Enhanced catalytic efficiency of CotA-laccase by DNA shuffling

Cited by 19 publications

References 26 publications

Fusion of a proline-rich oligopeptide to the C-terminus of a ruminal xylanase improves catalytic efficiency

Fusion of a proline-rich oligopeptide to the C-terminus of a ruminal xylanase improves catalytic efficiency

Recent Advances in Engineering Heterotrophic Microorganisms for Reinforcing CO₂ Fixation Based on Calvin–Benson–Bassham Cycle

Genome-based engineering of ligninolytic enzymes in fungi

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Enhanced catalytic efficiency of CotA-laccase by DNA shuffling

Cited by 19 publications

References 26 publications

Fusion of a proline-rich oligopeptide to the C-terminus of a ruminal xylanase improves catalytic efficiency

Fusion of a proline-rich oligopeptide to the C-terminus of a ruminal xylanase improves catalytic efficiency

Recent Advances in Engineering Heterotrophic Microorganisms for Reinforcing CO2 Fixation Based on Calvin–Benson–Bassham Cycle

Genome-based engineering of ligninolytic enzymes in fungi

Contact Info

Product

Resources

About

Recent Advances in Engineering Heterotrophic Microorganisms for Reinforcing CO₂ Fixation Based on Calvin–Benson–Bassham Cycle