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LUND UNIVERSITY LIBRARIES

Screening and Characterization of Glucuronoyl Esterase Candidates from the CE15 Family

Wang, Junyang LU (2026) KBTM01 20261
Biotechnology (MSc)
Biotechnology (M.Sc.Eng.)
Abstract
This study aimed to screen and characterize unknown glucuronoyl esterases (GEs) belonging to bacterial carbohydrate esterase family 15 (CE15), which were CE15_6nl, CE15_13nl, and CE15_15nsp. First, purified proteins were obtained by heterologous expression in E. coli induced by IPTG. Next, nanoDSF assays were performed to determine the thermostability of CE15 enzymes under different pH conditions, CE15_6nl and CE15_15nsp were most stable under pH 7 and pH 6.5, respectively, whereas CE15_13nl was more stable under alkaline conditions. Moreover, kinetics parameters were determined by coupled continuous assays on three model substrates. CE15_15nsp exhibited the highest catalytic efficiency and substrate affinity among the three enzymes. All... (More)
This study aimed to screen and characterize unknown glucuronoyl esterases (GEs) belonging to bacterial carbohydrate esterase family 15 (CE15), which were CE15_6nl, CE15_13nl, and CE15_15nsp. First, purified proteins were obtained by heterologous expression in E. coli induced by IPTG. Next, nanoDSF assays were performed to determine the thermostability of CE15 enzymes under different pH conditions, CE15_6nl and CE15_15nsp were most stable under pH 7 and pH 6.5, respectively, whereas CE15_13nl was more stable under alkaline conditions. Moreover, kinetics parameters were determined by coupled continuous assays on three model substrates. CE15_15nsp exhibited the highest catalytic efficiency and substrate affinity among the three enzymes. All candidates showed a preference for long substrates. To explore the action of single CE15 enzyme and CE15-GH30 synergy in the hydrolysis of natural biomass such as corn bran, HPAEC-PAD analysis was conducted. The results suggested that CE15 enzymes promoted the release of xylose and xylo-oligosaccharides, however, no synergistic effect between CE15 and GH30 could be detected. In total, CE15 enzymes show promising potential in lignocellulosic biomass processing, but further analytical experiments are required. (Less)
Popular Abstract
Fossil fuels are a major driver of climate change and environmental pollution, causing substantial impacts on ecosystems and human health. Lignocellulosic biomass is a promising alternative to fossil fuel. However, the treatment of lignocellulosic biomass remains challenging for industrial application partially due to the recalcitrance of lignin-carbohydrate complexes (LCCs). Since 2006, glucuronoyl esterase (GEs) from carbohydrate esterase family 15 (CE15) have been discovered, and it has been shown that they were able to hydrolyze ester bonds between lignin and carbohydrates. Compared with fungal GEs, bacterial GEs have been reported to show wider substrate specificity, but many bacterial GEs remain uncharacterized.
Therefore, this... (More)
Fossil fuels are a major driver of climate change and environmental pollution, causing substantial impacts on ecosystems and human health. Lignocellulosic biomass is a promising alternative to fossil fuel. However, the treatment of lignocellulosic biomass remains challenging for industrial application partially due to the recalcitrance of lignin-carbohydrate complexes (LCCs). Since 2006, glucuronoyl esterase (GEs) from carbohydrate esterase family 15 (CE15) have been discovered, and it has been shown that they were able to hydrolyze ester bonds between lignin and carbohydrates. Compared with fungal GEs, bacterial GEs have been reported to show wider substrate specificity, but many bacterial GEs remain uncharacterized.
Therefore, this study focused on screening and characterizing unknown bacterial GEs. Three bacterial GEs were obtained and characterized. Their stability under different conditions was determined. Two of them were most stable under neutral or slightly acidic conditions, while another CE15 enzyme showed higher stability under alkaline conditions. Moreover, these enzymes showed GE activity on different substrates even on natural biomass. Bacterial GEs have the potential to assist in the degradation of lignocellulosic biomass. (Less)
Please use this url to cite or link to this publication:
author
Wang, Junyang LU
supervisor
organization
course
KBTM01 20261
year
type
H2 - Master's Degree (Two Years)
subject
keywords
biotechnology, glucuronoyl esterase, lignocellulosic biomass, corn bran
language
English
id
9230717
date added to LUP
2026-09-14 15:04:37
date last changed
2026-09-14 15:04:37
@misc{9230717,
  abstract     = {{This study aimed to screen and characterize unknown glucuronoyl esterases (GEs) belonging to bacterial carbohydrate esterase family 15 (CE15), which were CE15_6nl, CE15_13nl, and CE15_15nsp. First, purified proteins were obtained by heterologous expression in E. coli induced by IPTG. Next, nanoDSF assays were performed to determine the thermostability of CE15 enzymes under different pH conditions, CE15_6nl and CE15_15nsp were most stable under pH 7 and pH 6.5, respectively, whereas CE15_13nl was more stable under alkaline conditions. Moreover, kinetics parameters were determined by coupled continuous assays on three model substrates. CE15_15nsp exhibited the highest catalytic efficiency and substrate affinity among the three enzymes. All candidates showed a preference for long substrates. To explore the action of single CE15 enzyme and CE15-GH30 synergy in the hydrolysis of natural biomass such as corn bran, HPAEC-PAD analysis was conducted. The results suggested that CE15 enzymes promoted the release of xylose and xylo-oligosaccharides, however, no synergistic effect between CE15 and GH30 could be detected. In total, CE15 enzymes show promising potential in lignocellulosic biomass processing, but further analytical experiments are required.}},
  author       = {{Wang, Junyang}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{Screening and Characterization of Glucuronoyl Esterase Candidates from the CE15 Family}},
  year         = {{2026}},
}