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Biochemical Characterization on CE1 Carbohydrate Esterases with Potential Dual Esterase Activity

Nayak, Rithika Vivekananda LU (2026) KBTM01 20261
Biotechnology (MSc)
Biotechnology (M.Sc.Eng.)
Abstract
Carbohydrate esterase family 1 (CE1) enzymes are a diverse group of enzymes involved in plant biomass degradation through removal of acetyl and feruloyl substitutions from polysaccharides. In this study, several putative CE1 enzymes were characterized to find out their substrate preference, catalytic properties and possible role during biomass hydrolysis. Functional screening using pNP-acetate and pNP-ferulate showed clear differences between the analyzed enzymes. AXE2 and FAE9 were active toward both substrates, whereas ACE3 showed lower activity and appeared to hydrolyze acetyl ester substrates. Kinetic analysis showed differences between AXE2 and FAE9, where AXE2 showed stronger affinity toward ferulate (Km = 0.821±0.127 mM), while FAE9... (More)
Carbohydrate esterase family 1 (CE1) enzymes are a diverse group of enzymes involved in plant biomass degradation through removal of acetyl and feruloyl substitutions from polysaccharides. In this study, several putative CE1 enzymes were characterized to find out their substrate preference, catalytic properties and possible role during biomass hydrolysis. Functional screening using pNP-acetate and pNP-ferulate showed clear differences between the analyzed enzymes. AXE2 and FAE9 were active toward both substrates, whereas ACE3 showed lower activity and appeared to hydrolyze acetyl ester substrates. Kinetic analysis showed differences between AXE2 and FAE9, where AXE2 showed stronger affinity toward ferulate (Km = 0.821±0.127 mM), while FAE9 displayed higher catalytic turnover and catalytic efficiency toward pNP-acetate, with a kcat of 37.414 ± 3.217 min-1 and catalytic efficiency of 43.692 ± 3.495 mM-1min-1. The hydrolysis experiments performed with the GH11 xylanase Pentopan Mono BG resulted in noticeable changes in oligosaccharide profiles across several biomass substrates, particularly for arabinoxylan and xylan-rich substrates, suggesting possible synergy between the CE1 enzymes and the GH11 xylanase during biomass degradation. Hence, the findings agree with the proposed accessory role of CE1 enzymes in biomass hydrolysis and provide insight into their possible use in enzymatic biomass conversion systems. (Less)
Popular Abstract
Plants are an important renewable resource and can be used to produce biofuels, bioplastics, biochemicals and other sustainable products. However, breaking down plant material can be challenging. Plant cell walls contain complex structures that are naturally strong and resistant. One main component is xylan, which has different chemical side groups that can make it difficult for enzymes to access and break down the material.
This project focused on a group of enzymes called the CE1 carbohydrate esterases. These enzymes help remove some of the chemical modifications on plant polysaccharides, which may support the breakdown of plant biomass by other enzymes.
In this study, several CE1 enzymes were produced and tested to understand better... (More)
Plants are an important renewable resource and can be used to produce biofuels, bioplastics, biochemicals and other sustainable products. However, breaking down plant material can be challenging. Plant cell walls contain complex structures that are naturally strong and resistant. One main component is xylan, which has different chemical side groups that can make it difficult for enzymes to access and break down the material.
This project focused on a group of enzymes called the CE1 carbohydrate esterases. These enzymes help remove some of the chemical modifications on plant polysaccharides, which may support the breakdown of plant biomass by other enzymes.
In this study, several CE1 enzymes were produced and tested to understand better what kinds of substrates they work on. The enzymes were first analyzed using simple model substrates and then tested on more complex plant-derived polysaccharides. Two enzymes, AXE2 and FAE9, showed activity toward both acetyl-linked and feruloyl-linked substrates, indicating that they might have dual activity.
The results also showed that these enzymes affected how complex polysaccharides were broken down when used with a xylan degrading enzyme. This suggests that CE1 enzymes may help improve biomass degradation by making the substrate more accessible for other enzymes.
Therefore, this project enhances our understanding of how CE1 enzymes work and their potential use in future biomass conversion and biotechnological applications. (Less)
Please use this url to cite or link to this publication:
author
Nayak, Rithika Vivekananda LU
supervisor
organization
alternative title
Functional Characterization of Dual-Acting CE1 Esterases
course
KBTM01 20261
year
type
H2 - Master's Degree (Two Years)
subject
keywords
CE1 Carbohydrate Esterases, acetyl xylan esterase, ferulic acid esterase, biomass degradation, xylan, hemicellulose, enzymatic hydrolysis, kinetic characterization, HPAEC-PAD, carbohydrate-active enzymes, biotechnology
language
English
id
9230576
date added to LUP
2026-08-24 11:03:01
date last changed
2026-08-24 11:03:01
@misc{9230576,
  abstract     = {{Carbohydrate esterase family 1 (CE1) enzymes are a diverse group of enzymes involved in plant biomass degradation through removal of acetyl and feruloyl substitutions from polysaccharides. In this study, several putative CE1 enzymes were characterized to find out their substrate preference, catalytic properties and possible role during biomass hydrolysis. Functional screening using pNP-acetate and pNP-ferulate showed clear differences between the analyzed enzymes. AXE2 and FAE9 were active toward both substrates, whereas ACE3 showed lower activity and appeared to hydrolyze acetyl ester substrates. Kinetic analysis showed differences between AXE2 and FAE9, where AXE2 showed stronger affinity toward ferulate (Km = 0.821±0.127 mM), while FAE9 displayed higher catalytic turnover and catalytic efficiency toward pNP-acetate, with a kcat of 37.414 ± 3.217 min-1 and catalytic efficiency of 43.692 ± 3.495 mM-1min-1. The hydrolysis experiments performed with the GH11 xylanase Pentopan Mono BG resulted in noticeable changes in oligosaccharide profiles across several biomass substrates, particularly for arabinoxylan and xylan-rich substrates, suggesting possible synergy between the CE1 enzymes and the GH11 xylanase during biomass degradation. Hence, the findings agree with the proposed accessory role of CE1 enzymes in biomass hydrolysis and provide insight into their possible use in enzymatic biomass conversion systems.}},
  author       = {{Nayak, Rithika Vivekananda}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{Biochemical Characterization on CE1 Carbohydrate Esterases with Potential Dual Esterase Activity}},
  year         = {{2026}},
}