Enzyme kinetics of fungal glucuronoyl esterases on natural lignin-carbohydrate complexes

Mosbech, Caroline; Holck, Jesper; Meyer, Anne S.; Agger, Jane Wittrup

doi:10.1007/s00253-019-09797-w

Cited by 23 publications

(22 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Active site architecture and ligand-induced oxyanion hole. Consistent with the proven role in interfacial biocatalysis of derivatives of large LCC substrates 5,18 , the active site of CuGE is readily accessible at the surface of the protein. The crystal structures (resolution 1.39-1.73 Å) of CuGE complexes with aldouronic acids representing the carbohydrate portion of the natural substrate (XU m4 X, Fig.…”

Section: Resultsmentioning

confidence: 65%

“…The inactive variant ΔS270A is destabilized by ca 3°C compared to variants with an intact catalytic triad, implying that some stabilizing interactions have been lost, but the overall conclusion is that CuGE is a highly stable enzyme with T i 's in the range of 72.2-75.8°C for all variants. Previous studies have shown that both WT and ΔWT CuGE are fully active on both soluble and insoluble substrates 18 .…”

Section: Resultsmentioning

confidence: 98%

“…These background signals result in a low signal-to-noise ratio in the actual binding experiment, but the estimated K d value (5 µM) imply a strong interaction between the enzyme and this insoluble substrate in comparison to those observed for soluble ligands. Previous kinetic studies with WT CuGE on the same substrate 18 revealed a K M of~220 µM and if K M is taken as a measure of substrate affinity, the higher value compared to K d measured by ITC suggests non-productive binding for instance "Out" Fig. 4 Active site, crystallographic complexes, and XU m4 X-recognition.…”

Section: Resultsmentioning

confidence: 99%

“…CuGE variants. WT CuGE has a modular architecture with a Nterminal carbohydrate-binding module 1 (CBM1) domain 17,18 connected to the catalytic domain by a proline-rich linker region. The structure of CuGE is potentially flexible and together with heterogenous N-and O-glycosylation 17 it could represent an obstacle for high-resolution structural studies of the full-length enzyme (WT).…”

Section: Resultsmentioning

confidence: 99%

“…CE15-B comprises mostly fungal enzymes, including CuGE, CiP2, and StGE2, but also enzymes of bacterial origin. Variations in the identity and position of the catalytic acid among members of CE15 have been discussed previously 6,7,[13][14][15]18,27,28 . However, with our direct reference to the α/β-hydrolase superfamily we have been able to obtain a clear division of CE15 enzymes into two subgroups with fundamentally different properties encoded in their structural arrangements.…”

Section: Resultsmentioning

confidence: 99%

See 4 more Smart Citations

The structural basis of fungal glucuronoyl esterase activity on natural substrates

et al. 2020

Self Cite

View full text Add to dashboard Cite

Structural and functional studies were conducted of the glucuronoyl esterase (GE) from Cerrena unicolor (CuGE), an enzyme catalyzing cleavage of lignin-carbohydrate ester bonds. CuGE is an α/β-hydrolase belonging to carbohydrate esterase family 15 (CE15). The enzyme is modular, comprised of a catalytic and a carbohydrate-binding domain. SAXS data show CuGE as an elongated rigid molecule where the two domains are connected by a rigid linker. Detailed structural information of the catalytic domain in its apo-and inactivated form and complexes with aldouronic acids reveal well-defined binding of the 4-O-methyl-aD -glucuronoyl moiety, not influenced by the nature of the attached xylo-oligosaccharide. Structural and sequence comparisons within CE15 enzymes reveal two distinct structural subgroups. CuGE belongs to the group of fungal CE15-B enzymes with an open and flat substrate-binding site. The interactions between CuGE and its natural substrates are explained and rationalized by the structural results, microscale thermophoresis and isothermal calorimetry.

show abstract

Section: Resultsmentioning

confidence: 65%

Section: Resultsmentioning

confidence: 98%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

See 3 more Smart Citations

The structural basis of fungal glucuronoyl esterase activity on natural substrates

et al. 2020

Self Cite

View full text Add to dashboard Cite

show abstract

The coordinated action of glucuronoyl esterase and α‐glucuronidase promotes the disassembly of lignin–carbohydrate complexes

et al. 2021

View full text Add to dashboard Cite

Glucuronoxylans represent a significant fraction of woody biomass, and its decomposition is complicated by the presence of lignin–carbohydrate complexes (LCCs). Herein, LCCs from birchwood were used to investigate the potential coordinated action of a glucuronoyl esterase (TtCE15A) and two α‐glucuronidases (SdeAgu115A and AxyAgu115A). When supplementing α‐glucuronidase with equimolar quantities of TtCE15A, total MeGlcpA released after 72 h by SdeAgu115A and AxyAgu115A increased from 52% to 67%, and 61% to 95%, respectively. Based on the combined TtCE15A and AxyAgu115A activities, ~ 34% of MeGlcpA in the extracted birchwood glucuronoxylan was occupied as LCCs. Notably, insoluble LCC fractions reduced soluble α‐glucuronidase concentrations by up to 70%, whereas reduction in soluble TtCE15A was less than 30%, indicating different tendencies to adsorb onto the LCC substrate.

show abstract

Esterases as emerging biocatalysts: Mechanistic insights, genomic and metagenomic, immobilization, and biotechnological applications

Rafeeq

Hussain

Shabbir

et al. 2021

Biotech and App Biochem

View full text Add to dashboard Cite

Esterase enzymes are a family of hydrolases that catalyze the breakdown and formation of ester bonds. Esterases have gained a prominent position in today's world's industrial enzymes market. Due to their unique biocatalytic attributes, esterases contribute to environmentally sustainable design approaches, including biomass degradation, food and feed industry, dairy, clothing, agrochemical (herbicides, insecticides), bioremediation, biosensor development, anticancer, antitumor, gene therapy, and diagnostic purposes. Esterases can be isolated by a diverse range of mammalian tissues, animals, and microorganisms. The isolation of extremophilic esterases increases the interest of researchers in the extraction and utilization of these enzymes at the industrial level. Genomic, metagenomic, and immobilization techniques have opened innovative ways to extract esterases and utilize them for a longer time to take advantage of their beneficial activities. The current study discusses the types of esterases, metagenomic studies for exploring new esterases, and their biomedical applications in different industrial sectors.

show abstract

Enzyme kinetics of fungal glucuronoyl esterases on natural lignin-carbohydrate complexes

Cited by 23 publications

References 29 publications

The structural basis of fungal glucuronoyl esterase activity on natural substrates

The structural basis of fungal glucuronoyl esterase activity on natural substrates

The coordinated action of glucuronoyl esterase and α‐glucuronidase promotes the disassembly of lignin–carbohydrate complexes

Esterases as emerging biocatalysts: Mechanistic insights, genomic and metagenomic, immobilization, and biotechnological applications

Contact Info

Product

Resources

About