Asn57 N-glycosylation promotes the degradation of hemicellulose by β-1,3–1,4-glucanase from Rhizopus homothallicus

Zha, Zi-Qian; You, Shuai; Hu, Yanghao; Zhang, Fang; Chen, Yiwen; Wang, Jun

doi:10.1007/s11356-022-19959-5

Cited by 3 publications

(2 citation statements)

References 50 publications

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“…Group C: Positive control group PSCs were directedly treated with 10 ug/mL exogenous bovine trypsinogen (S27969, Yuanye, Shanghai, China) [ 6 , 18 ] for 6 h.…”

Section: Methodsmentioning

confidence: 99%

The Rabep1-Mediated Endocytosis and Activation of Trypsinogen to Promote Pancreatic Stellate Cell Activation

Yao

Luo

et al. 2022

Biomolecules

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Background: The pathogenesis of chronic pancreatitis is still unclear. Trypsinogen activation is an active factor in acute pancreatitis that has not been studied in the occurrence of chronic pancreatitis. Methods: Immunofluorescence was used to detect the location and expression of trypsinogen in chronic pancreatitis and normal tissues. Microarray and single-cell RNA-seq (scRNA-seq) were used to screen core genes and pathways in pancreatic stellate cells (PSCs). Western blotting and immunofluorescence were used to verify trypsinogen expression in PSCs after silencing Rabep1. Immunofluorescence and flow cytometry were used to validate trypsinogen activation and PSC activation after intervening in the endocytosis pathway. Results: Endocytosed trypsinogen was found in PSCs in CP clinical samples. Bioinformatic analysis showed that Rabep1 is a core gene that regulates trypsinogen endocytosis through the endocytosis pathway, verified by Western blot and immunofluorescence. Immunofluorescence and flow cytometry analyses confirmed the activation of trypsinogen and PSCs through the endocytosis pathway in PSCs. Conclusion: This study discovered a new mechanism by which trypsinogen affects the activation of PSCs and the occurrence and development of CP. Through communication between pancreatic acinar cells and PSCs, trypsinogen can be endocytosed by PSCs and activated by the Rabep1 gene.

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“…Group C: Positive control group PSCs were directedly treated with 10 ug/mL exogenous bovine trypsinogen (S27969, Yuanye, Shanghai, China) [ 6 , 18 ] for 6 h.…”

Section: Methodsmentioning

confidence: 99%

The Rabep1-Mediated Endocytosis and Activation of Trypsinogen to Promote Pancreatic Stellate Cell Activation

Yao

Luo

et al. 2022

Biomolecules

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“…The target protein was puri ed using a 5 cm × 1 cm 2 Ni-NTA a nity column, and the protein m purity was veri ed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The protein concentration was measured according to the Bradford method using bovine serum albumin (BSA) as the standard protein (Zha et al 2022).…”

Section: Cloning and Expression Of Wild-type Eccslg And Its Mutantsmentioning

confidence: 99%

Rational Design of Disulfide Bonds Increases Thermostability of a β-1,3-1,4-Glucanase from Paenibacillus

Wang

liang

et al. 2023

Preprint

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β-1,3 − 1,4-gluconases can specifically hydrolyze the adjacent β-1,4 glycoside bond of β-1,3 in β-glucan, which is widely used in food, brewing and feed industries. Its sources include bacteria, fungi, and plant endosperm cell walls, most β-1,3 − 1,4-glucanases lose their activity when the temperature exceeds 65 ℃. In this study, we selected and modified the β-1,3 − 1,4-glucanase (PlicA) gene from Paenibacillus and expressed it in Escherichia coli BL21 (DE3). Adding disulfide bonds by rational design increased the optimal temperature of the enzyme from 55 ℃ to 80 ℃, and temperature stability was also improved. The optimum pH of the modified β-1,3 − 1,4-gluconanase (Eccsl69) was 9.0–10.0. The enzyme activity in 16.9 U/mL of Eccsl69 was measured at 540 nm with 0.8% gluan as the substrate, and a nickel column purified specific enzyme activity of 320 U/mg was determined. The Km and Vmax values of Eccsl69 using barley β-glucan as substrate were 1.5 mg/ml and 8.3 mol/min·mg. The structure of the β-1,3 − 1,4-glucanase Eccsl69 tended to be stable after molecular dynamics simulation for approximately 20 ns. The enzyme was successfully applied in the pulping and papermaking field for the first time, and the pulp freeness was adjusted from 55.0 °SR to 47 °SR, which enhanced water filtration. This study provides a successful strategy for improving the heat resistance of Eccsl69, which is promising for its application in pulping and paper making industries.

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The functional role of N-link glycosylation in a novel cellobiohydrolase II (LsCel6A) from a white-rot fungus Lentinus sp. WR2

Wang,

Shyur

2024

International Journal of Biological Macromolecules

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Asn57 N-glycosylation promotes the degradation of hemicellulose by β-1,3–1,4-glucanase from Rhizopus homothallicus

Cited by 3 publications

References 50 publications

The Rabep1-Mediated Endocytosis and Activation of Trypsinogen to Promote Pancreatic Stellate Cell Activation

The Rabep1-Mediated Endocytosis and Activation of Trypsinogen to Promote Pancreatic Stellate Cell Activation

Rational Design of Disulfide Bonds Increases Thermostability of a β-1,3-1,4-Glucanase from Paenibacillus

The functional role of N-link glycosylation in a novel cellobiohydrolase II (LsCel6A) from a white-rot fungus Lentinus sp. WR2

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