Enhancing the thermostability of β-glucuronidase from T. pinophilus enables the biotransformation of glycyrrhizin at elevated temperature

Feng, Xudong; Liu, Xiaofei; Jia, Jintong; Chen, Xinyi; Jiang, Tianyue; Li, Chun

doi:10.1016/j.ces.2019.04.020

Cited by 7 publications

(3 citation statements)

References 37 publications

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“…5,6 In addition, a large amount of free enzymes cannot be recovered after the reaction, which remains a major difficulty in industrial production. 7,8 Hence, immobilized enzymes have been confirmed to be a useful strategy to improve the activity, stability, and selectivity of enzymes to achieve wider applications in the biochemical and medical industries. 9−12 Metal−organic frameworks (MOFs) are a kind of poriferous carriers formed with metal ions and organic ligands through coordination bonds.…”

Section: Introductionmentioning

confidence: 99%

“…Enzyme is a kind of powerful biocatalyst evolved from nature. The vigorous development of biotechnology represented by enzyme catalysis has brought new opportunities for the sustainable development of the chemical industry. , As a highly efficient biocatalyst, natural enzymes have excellent specificity of the substrate, selectivity, and eco-friendly reaction conditions, imposing important impacts on biocompatibility, biodegradability, and renewability. , Although the stability of enzymes has been improved through protein engineering, the sensibility of enzymes leads to the lack of catalytic activity under extreme conditions. , In addition, a large amount of free enzymes cannot be recovered after the reaction, which remains a major difficulty in industrial production. , Hence, immobilized enzymes have been confirmed to be a useful strategy to improve the activity, stability, and selectivity of enzymes to achieve wider applications in the biochemical and medical industries. − …”

Section: Introductionmentioning

confidence: 99%

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Highly Selective Entrapment of His-Tagged Enzymes on Superparamagnetic Zirconium-Based MOFs with Robust Renewability to Enhance pH and Thermal Stability

Wei

Cheng

et al. 2021

ACS Biomater. Sci. Eng.

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Metal–organic frameworks (MOFs), as a kind of poriferous nanoparticle, are promising candidates for enzyme immobilization to enhance their stability and reusability. However, most MOFs could not specifically immobilize enzymes and regenerate easily, which inevitably leads to serious high consumption and environmental pollution. In this study, renewable and magnetic MOFs were first constructed to specially immobilize His-tagged enzymes from the cell lysates without purification. The immobilized β-glucuronidase exhibited wider pH adaptability and temperature stability. The relative activity of immobilized β-glucuronidase was still maintained at ∼80% after eight cycles. Importantly, after simple treatment, the immobilization capacity of regenerated MOFs after simple treatment was restored to more than 90% in the first three times. The specific magnetic MOFs were proven to be an efficient and renewable platform for one-step immobilization and purification of His-tagged enzymes, showing great potential in industrial applications of nanotechnology and biocatalysis.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Highly Selective Entrapment of His-Tagged Enzymes on Superparamagnetic Zirconium-Based MOFs with Robust Renewability to Enhance pH and Thermal Stability

Wei

Cheng

et al. 2021

ACS Biomater. Sci. Eng.

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“…This indicated that the introduction of the ISD of EcGUS into PGUS and AtGUS also significantly increased enzyme thermostability. The reported thermostability of β‐glucuronidase in this study was higher than that of an engineered β‐glucuronidase from Talaromyces pinophilus Li‐93 ( t 1/2 of 175 min at 55°C), whose thermostability was also tested with natural substrate GL hydrolysis (Feng et al, 2019).…”

Section: Resultsmentioning

confidence: 72%

Improving the activity and thermostability of GH2 β‐glucuronidases via domain reassembly

Liu

Jing

et al. 2021

Biotech & Bioengineering

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Glycoside hydrolase family 2 (GH2) enzymes are generally composed of three domains: TIM‐barrel domain (TIM), immunoglobulin‐like β‐sandwich domain (ISD), and sugar‐binding domain (SBD). The combination of these three domains yields multiple structural combinations with different properties. Theoretically, the drawbacks of a given GH2 fold may be circumvented by efficiently reassembling the three domains. However, very few successful cases have been reported. In this study, we used six GH2 β‐glucuronidases (GUSs) from bacteria, fungi, or humans as model enzymes and constructed a series of mutants by reassembling the domains from different GUSs. The mutants PGUS‐At, GUS‐PAA, and GUS‐PAP, with reassembled domains from fungal GUSs, showed improved expression levels, activity, and thermostability, respectively. Specifically, compared to the parental enzyme, the mutant PGUS‐At displayed 3.8 times higher expression, the mutant GUS‐PAA displayed 1.0 time higher catalytic efficiency (kcat/Km), and the mutant GUS‐PAP displayed 7.5 times higher thermostability at 65°C. Furthermore, two‐hybrid mutants, GUS‐AEA and GUS‐PEP, were constructed with the ISD from a bacterial GUS and SBD and TIM domain from fungal GUSs. GUS‐AEA and GUS‐PEP showed 30.4% and 23.0% higher thermostability than GUS‐PAP, respectively. Finally, molecular dynamics simulations were conducted to uncover the molecular reasons for the increased thermostability of the mutant.

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Endophytic fungi from Dongxiang wild rice (Oryza rufipogon Griff.) show diverse catalytic potential for converting glycyrrhizin

et al. 2022

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Enhancing the thermostability of β-glucuronidase from T. pinophilus enables the biotransformation of glycyrrhizin at elevated temperature

Cited by 7 publications

References 37 publications

Highly Selective Entrapment of His-Tagged Enzymes on Superparamagnetic Zirconium-Based MOFs with Robust Renewability to Enhance pH and Thermal Stability

Highly Selective Entrapment of His-Tagged Enzymes on Superparamagnetic Zirconium-Based MOFs with Robust Renewability to Enhance pH and Thermal Stability

Improving the activity and thermostability of GH2 β‐glucuronidases via domain reassembly

Endophytic fungi from Dongxiang wild rice (Oryza rufipogon Griff.) show diverse catalytic potential for converting glycyrrhizin

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