Functional expression and characterization of a glucuronoyl esterase from the fungus &lt;i&gt;Neurospora crassa&lt;/i&gt;: identification of novel consensus sequences containing the catalytic triad

Huynh, Hung Quang; Arioka, Manabu

doi:10.2323/jgam.2016.03.004

Cited by 16 publications

(9 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…However, AaGE1 and PcGE1 exhibited only a 22 and 28% reduction in activity respectively after incubation for 6 h at 50 °C. Thus, they showed better stability than GEs from other mesophilic organisms, such as NcGE from Neurospora crassa (Huynh and Arioka 2016) or Cip2 from Hypocrea jecorina (Li et al 2007). …”

Section: Discussionmentioning

confidence: 99%

Characterisation of three fungal glucuronoyl esterases on glucuronic acid ester model compounds

Hüttner

Klaubauf

Vries

et al. 2017

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

The glucuronoyl esterases (GEs) that have been identified so far belong to family 15 of the carbohydrate esterases in the CAZy classification system and are presumed to target ester bonds between lignin alcohols and (4-O-methyl-)d-glucuronic acid residues of xylan. Few GEs have been cloned, expressed and characterised to date. Characterisation has been done on a variety of synthetic substrates; however, the number of commercially available substrates is very limited. We identified novel putative GEs from a wide taxonomic range of fungi and expressed the enzymes originating from Acremonium alcalophilum and Wolfiporia cocos as well as the previously described PcGE1 from Phanerochaete chrysosporium. All three fungal GEs were active on the commercially available compounds benzyl glucuronic acid (BnGlcA), allyl glucuronic acid (allylGlcA) and to a lower degree on methyl glucuronic acid (MeGlcA). The enzymes showed pH stability over a wide pH range and tolerated 6-h incubations of up to 50 °C. Kinetic parameters were determined for BnGlcA. This study shows the suitability of the commercially available model compounds BnGlcA, MeGlcA and allylGlcA in GE activity screening and characterisation experiments. We enriched the spectrum of characterised GEs with two new members of a relatively young enzyme family. Due to its biotechnological significance, this family deserves to be more extensively studied. The presented enzymes are promising candidates as auxiliary enzymes to improve saccharification of plant biomass.Electronic supplementary materialThe online version of this article (doi:10.1007/s00253-017-8266-9) contains supplementary material, which is available to authorized users.

show abstract

Section: Discussionmentioning

confidence: 99%

Characterisation of three fungal glucuronoyl esterases on glucuronic acid ester model compounds

Hüttner

Klaubauf

Vries

et al. 2017

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

show abstract

“…The ascomycete GEs, Trichoderma reesei GE (Cip2, [12]) and Podospora anserina GE (PaGE1, [27]) clustered in Sub-group 5. Sub-group 8 consisted of the GEs from the ascomycete fungi Myceliophthora thermophila (StGE1, [28]) and Neurospora crassa (NcGE, [29]), whereas Sub-group 1 consisted of a second GE from M. thermophila (StGE2, [30]). No characterized GE belongs to Sub-group 2, 3, 6 and 7.…”

Section: Genome Mining and Phylogenetic Analysis Of Novel Fungal Gesmentioning

confidence: 99%

Fungal glucuronoyl esterases: Genome mining based enzyme discovery and biochemical characterization

et al. 2018

View full text Add to dashboard Cite

4-O-Methyl-d-glucuronic acid (MeGlcA) is a side-residue of glucuronoarabinoxylan and can form ester linkages to lignin, contributing significantly to the strength and rigidity of the plant cell wall. Glucuronoyl esterases (4-O-methyl-glucuronoyl methylesterases, GEs) can cleave this ester bond, and therefore may play a significant role as auxiliary enzymes in biomass saccharification for the production of biofuels and biochemicals. GEs belong to a relatively new family of carbohydrate esterases (CE15) in the CAZy database (www.cazy.org), and so far around ten fungal GEs have been characterized. To explore additional GE enzymes, we used a genome mining strategy. BLAST analysis with characterized GEs against approximately 250 publicly accessible fungal genomes identified more than 150 putative fungal GEs, which were classified into eight phylogenetic sub-groups. To validate the genome mining strategy, 21 selected GEs from both ascomycete and basidiomycete fungi were heterologously produced in Pichia pastoris. Of these enzymes, 18 were active against benzyl d-glucuronate demonstrating the suitability of our genome mining strategy for enzyme discovery.

show abstract

“…The following media was prepared with distillated water: LBL (1% tryptone, 0.5% yeast extract, and 0.5% NaCl, w/v) (Huynh & Arioka, ), MM agar medium (10 mM NaNO 3 , 10 mM KH 2 PO 4 , 7 mM KCl, 2 mM MgSO 4 , 2 ml/l of Hutner's trace metals, and 1.0% glucose, w/v), YPD (1% peptone, 1% yeast extract, and 2% glucose, w/v), BMGY (100 mmol/l potassium phosphate buffer solution, pH 6.0, 1.34% YNB, 1% yeast extract, 2% peptone, and 1% glycerol, w/v), BMMY (100 mmol/l potassium phosphate buffer solution, pH 6.0, 1.34% YNB, 1% yeast extract, and 2% peptone, w/v), and autoclaved at 120°C for 20 min. Zero point two mg/l of biotin and 1% (v/v) methanol was added in BMMY medium before induction.…”

Section: Methodsmentioning

confidence: 99%

Identification and characterization of thermostable endo‐polygalacturonase II B from Aspergillus luchuensis

et al. 2020

View full text Add to dashboard Cite

Endo‐polygalacturonase II B (PgaB) from Aspergillus luchuensis was orthologous to endo‐polygalacturonase from Aspergillus niger with mutant sites Thr42Ser and Glu52Ala. Mature pgaB gene was cloned from the genomic DNA of A. luchuensis and secreted expressed with over 90% purity in Pichia Pastoris and reached 1.0 g/L after 144 hr culture. The recombinant PgaB was further purified by Ni‐NTA chromatography. Using polygalacturonic acid (PGA) as substrate, the optimal condition for PgaB activity was 40°C and pH 4.5, respectively. Km and Vmax of PgaB were 0.19 mmol/l and 103.58 μmol min−1 mg−1, respectively. The relative activity of PgaB remained more than 60% and 40% of maximum activity at 50 and 60°C for 7 hr. PgaB increased the light transmittance by 85% and showed high efficiency in juice clarification. The main product was galacturonic acid oligosaccharides with degrees of polymers (DP) 1–3. The PgaB is a potential pectinolytic enzyme in food industries. Practical applications Endo‐polygalacturonase II B (PgaB) was identified from Aspergillus luchuensis, a filamentous fungus widely used in food and beverage fermentation in East Asia. PgaB still kept its most activity at 60°C for 7 hr. Polygalacturonic acid (PGA) can be digested effectively by the PgaB and the main products are galacturonic acid oligosaccharides with degrees of polymers (DP) 1–3. PgaB shows high efficiency in juice clarification. The PgaB is a potential pectinolytic enzyme for the applications in food industries.

show abstract

Functional expression and characterization of a glucuronoyl esterase from the fungus <i>Neurospora crassa</i>: identification of novel consensus sequences containing the catalytic triad

Cited by 16 publications

References 19 publications

Characterisation of three fungal glucuronoyl esterases on glucuronic acid ester model compounds

Characterisation of three fungal glucuronoyl esterases on glucuronic acid ester model compounds

Fungal glucuronoyl esterases: Genome mining based enzyme discovery and biochemical characterization

Identification and characterization of thermostable endo‐polygalacturonase II B from Aspergillus luchuensis

Contact Info

Product

Resources

About