Transcriptome Profiling, Cloning, and Characterization of AnGlu04478, a Ginsenoside Hydrolyzing β-Glucosidase from Aspergillus niger NG1306 DOI Creative Commons
Ming-Xing Jiang, Ling Zhu,

Shuhan Xie

et al.

Current Microbiology, Journal Year: 2024, Volume and Issue: 82(1)

Published: Dec. 24, 2024

β-Glucosidase plays a pivotal role in transforming ginsenosides into specific minor ginsenosides. In this study, total from Panax notoginseng leaves were used as substrates to stimulate the growth of Aspergillus niger NG1306. Transcriptome analysis identified β-glucosidase gene, Anglu04478 (1455 bp, 484 amino acids, 54.5 kDa, pI = 5.1), participant biotransformation process. This gene was cloned and expressed Escherichia coli BL21 Transetta (DE3). The AnGlu04478 protein purified using Ni2+ column, its enzymatic properties characterized. Purified exhibited activity 32.97 U/mg when assayed against pNPG. Under optimal conditions (pH 4.5, temperature 40 °C), kinetic parameters, Km Vmax, for pNPG 1.55 mmol/L 0.014 mmol/min, respectively. Cu2+ displayed an inhibitory effect on AnGlu04478, whereas Ca2+, Co2+, ions had minimal impact. enzyme showed tolerance ethanol largely unaffected by glucose feedback inhibition. Testing with revealed selective hydrolysis at C3 position Rb1, Rb2, Rb3, Rc, metabolic pathway delineated Rb1 → GypXVII, Rb2 C–O, Rb3 C-Mx1 C-Mx, Rc C-Mc1. conversion rates varied 2.58 20.63%. With 0.5 U mg ginsenosides, incubated °C 12 h, 42.6% 10.4% 6.27% C-Mx1, 26.96% 90% Rc. These results suggest that displays substrate promiscuity β-glucosidase, thus broadening potential ginsenoside biotransformation.

Language: Английский

Comparison of the Transformation Ability of the Major Saponins in Panax notoginseng by Penicillum fimorum Enzyme and Commercial β-glucosidase DOI Creative Commons

Feixing Li,

Ruixue Zhang,

Dongmei Lin

et al.

Microorganisms, Journal Year: 2025, Volume and Issue: 13(3), P. 495 - 495

Published: Feb. 23, 2025

Ginsenosides with less sugar groups, which are called minor ginsenosides, might have a greater pharmacological activity and better adsorptive ability, but their content in nature is extremely low. In this study, strain of Penicillium fimorum strong saponin transformation ability was isolated from fresh Gastrodia elata. A comparative biotransformation experiment the major saponins Panax notoginseng root were conducted using crude enzymes P. commercial β-glucosidase to produce ginsenosides. Specifically, enzyme able transform into 13 72 h, while same 15 h. The most significant difference between these two Rb1. To best our knowledge, reported here for first time. These potential improve economic value expand methods preparing by transforming total root.

Language: Английский

Citations

0
Transcriptome Profiling, Cloning, and Characterization of AnGlu04478, a Ginsenoside Hydrolyzing β-Glucosidase from Aspergillus niger NG1306 DOI Creative Commons
Ming-Xing Jiang, Ling Zhu,

Shuhan Xie

et al.

Current Microbiology, Journal Year: 2024, Volume and Issue: 82(1)

Published: Dec. 24, 2024

β-Glucosidase plays a pivotal role in transforming ginsenosides into specific minor ginsenosides. In this study, total from Panax notoginseng leaves were used as substrates to stimulate the growth of Aspergillus niger NG1306. Transcriptome analysis identified β-glucosidase gene, Anglu04478 (1455 bp, 484 amino acids, 54.5 kDa, pI = 5.1), participant biotransformation process. This gene was cloned and expressed Escherichia coli BL21 Transetta (DE3). The AnGlu04478 protein purified using Ni2+ column, its enzymatic properties characterized. Purified exhibited activity 32.97 U/mg when assayed against pNPG. Under optimal conditions (pH 4.5, temperature 40 °C), kinetic parameters, Km Vmax, for pNPG 1.55 mmol/L 0.014 mmol/min, respectively. Cu2+ displayed an inhibitory effect on AnGlu04478, whereas Ca2+, Co2+, ions had minimal impact. enzyme showed tolerance ethanol largely unaffected by glucose feedback inhibition. Testing with revealed selective hydrolysis at C3 position Rb1, Rb2, Rb3, Rc, metabolic pathway delineated Rb1 → GypXVII, Rb2 C–O, Rb3 C-Mx1 C-Mx, Rc C-Mc1. conversion rates varied 2.58 20.63%. With 0.5 U mg ginsenosides, incubated °C 12 h, 42.6% 10.4% 6.27% C-Mx1, 26.96% 90% Rc. These results suggest that displays substrate promiscuity β-glucosidase, thus broadening potential ginsenoside biotransformation.

Language: Английский

Citations

0