Article on FOS synthesis by a Trichoderma β-fructofuranosidase

 

We report the first β-fructofuranosidase from the Trichoderma genus capable of producing fructooligosaccharides (FOS). The enzyme, identified in Trichoderma atroviride (TaINV), was heterologously expressed, purified, and biochemically characterized. TaINV mainly hydrolyzes sucrose and substrates containing β-(2→1) linkages and, importantly, also catalyzes the synthesis of FOS from all three structural series (1F-FOS, 6F-FOS, and 6G-FOS). Under optimal conditions, TaINV produced up to 252 g/L of total FOS, representing 50.3% of total sugars, with 1-kestose as the dominant product (~85%). Structural analysis using an AlphaFold model revealed conserved catalytic motifs typical of GH32 enzymes. Site-directed mutagenesis confirmed the role of the catalytic triad (Asp63, Asp201, Glu277) and identified residues influencing transfructosylation specificity. Notably, variants W60Y and N62S increased FOS production to 62.7% and 57.4%, respectively, comparable to commercial enzymes.

Overall, TaINV is a distinctive intracellular fungal β-fructofuranosidase with strong transfructosylation activity and a preference for short-chain FOS, highlighting its potential as a biocatalyst for producing prebiotic fructooligosaccharides.

Ref: Narmontaite, E., Plou, F.J. & Fernández-Lobato, M. Enhanced fructooligosaccharides synthesis by engineered Trichoderma atroviride β-fructofuranosidase. Appl Microbiol Biotechnol 110, 96 (2026). https://doi.org/10.1007/s00253-026-13748-7

Article on biovailability of phloretin and its derivatives in dynamic digestor

This study explored the bioaccessibility and bioavailability of several phloretin derivatives using an in vitro dynamic digestion model that simulates human gastrointestinal conditions. Four compounds were evaluated: phloretin aglycone, α-glucoside, α-diglucoside, and lauroyl α-glucoside.

Results showed that glucosylation improves the bioaccessibility of phloretin. Most of the α-glucoside reached the intestinal phase as the aglycone due to intestinal glycosidase activity, while phloretin and lauroyl α-glucoside tended to precipitate in the stomach, reducing their availability. In Caco-2 cell assays, phloretin α-glucoside displayed the highest bioavailability (26.8%), likely due to active transport, compared with the aglycone (2.7%). The α-diglucoside showed lower bioavailability (4.4%), and lauroyl α-glucoside was not absorbed.

Additionally, phloretin and its derivatives inhibited intestinal glucose absorption in Caco-2 cells. Lauroyl α-glucoside also showed selective antimicrobial activity against pathogenic bacteria without affecting probiotics, along with anti-inflammatory properties, highlighting the potential of these derivatives for functional food and health applications.

Ref: “Bioaccessibility of phloretin and its derivatives using a dynamic in vitro digester. Effects on glucose uptake, prebiotic potential and anti-inflammatory activity”. J.L. Gonzalez-Alfonso, L. Barahona, S. Garcia-Benlloch, M. Martinez-Ranz, B. Gracia-Gomez, M.A. Iñiguez, B. Viadel, M. Fernandez-Lobato, and F.J. Plou*. Food Res Int. 226, 118238 (2026). https://doi.org/10.1016/j.foodres.2025.118238

BIOTEC25 Biotechnology Congress

During this week, a large group from the consortium GLICOENZ has been present at the BIOTEC25 Biotechnology Congress, held at the Faculty of Biosciences of the Autonomous University of Barcelona (UAB), organized by the SEBiot - Spanish Society of Biotechnology.

It has been a very interesting congress, with high scientific quality, where we have witnessed the latest advances in biotechnology and the application of artificial intelligence tools within this field.

Two theses defended within the consortium GLICOENZ

Two members of the GLICOENZ consortium have recently successfully defended their doctoral theses at the Autonomous University of Madrid:

• Marina Minguet Lobato: "Novel Chitinases and Hyaluronidases: Discovery and Application in the Synthesis of Bioactive Oligosaccharides"
• Angela Fernández García: "Enzymatic Biotechnology: Structure and molecular mechanisms of activity in unspecific peroxygenases for the production of compounds of chemical and pharmaceutical interest"
  

Congratulations to both for the excellent work and the magnificent defense!

Paper on dihydromyricetin alpha-glucosylation published in Org. Biomol. Chem

A new open-access paper just published in Organic & Biomolecular Chemistry explores how enzymatic glucosylation and acylation can dramatically improve the solubility and bioavailability of dihydromyricetin (DMY), a powerful health-promoting molecule found in vine tea and known for its antioxidant, anti-inflammatory, and neuroprotective effects. Novel glucosides and acyl-glucosides of DMY were synthesized, but we also showed how these modifications enhance water solubility. This strategy can convert DMY into a more effective functional ingredient for food, nutraceutical, and pharmaceutical applications

Reference: D.  Rodriguez-Garcia,  C.  Uceda,  L.  Barahona,  M.  Ruiz-Nuñez,  A.O.  Ballesteros,  T.  Desmet,  J.  Sanz-Aparicio,  M.  Fernandez-Lobato,  J.L.  Gonzalez-Alfonso,  F.J.  Plou.  "Enzymatic modification of dihydromyricetin by glucosylation and acylation, and its effect on the solubility and antioxidant activity".  Org. Biomol. Chem 23, 1136-1145 (2025). https://doi.org/10.1039/D4OB01682C 

Participation in the IV Spanish Biocatalysis Conference

The GLICOENZ consortium has participated in the IV Spanish Biocatalysis Conference, organized by CICbiomaGUNE and SEBIOT at the Aquarium of Donostia (September 5-6th, 2024). We presented our latest results on glycosylation of bioactive polyphenols:

"Molecular basis of specificity of a sucrose phosphorylase mutant in the glycosylation of polyphenols". M. Ruiz-Nuñez, J.L. Gonzalez-Alfonso, M. Moreno-Garcia, C. Uceda, M. Fernandez-Lobato, F.J. Plou and J. Sanz-Aparicio.

"Glucosylation of polyphenols with high yield and regioselectivity using a sucrose phosphorylase mutant". J.L. Gonzalez-Alfonso, D. Rodriguez-García, M. Moreno-García, C. Uceda-Domínguez, C. Alonso, A. Poveda, Z. Ubiparip, A.O. Ballesteros, T. Desmet, J. Jiménez-Barbero, L. Coderch, and F.J. Plou

Potential of alpha-glucosidase from Sch. occidentalis published in Appl. Microbiol. Biotechnol.

The α-glucosidase from Schwanniomyces occidentalis (GAM1p) was expressed in Komagataella phaffii to about 70 mg/L, and its transferase activity studied in detail. Several isomaltooligosaccharides (IMOS) were formed using 200 g/L maltose. The major production of IMOS (81.3 g/L) was obtained when 98% maltose was hydrolysed, of which 34.8 g/L corresponded to isomaltose, 26.9 g/L to isomaltotriose, and 19.6 g/L to panose. In addition, the potential of this enzyme to glycosylate 12 possible hydroxylated acceptors, including eight sugars and four phenolic compounds, was evaluated. Among them, only sucrose, xylose, and piceid (a monoglucosylated derivative of resveratrol) were glucosylated, and the main synthesised products were purified and characterised by MS and NMR. 

Ref: “Insights into the transglucosylation activity of α-glucosidase from Schwanniomyces occidentalis”. Zoran Merdzo, Egle Narmontaite, Jose L. Gonzalez-Alfonso, Ana Poveda, Jesus Jimenez-Barbero, Francisco J. Plou, María Fernández-Lobato. Applied Microbiology and Biotechnology 108, 443 (2024). https://doi.org/10.1007/s00253-024-13262-8

GLICOENZ at the IX National Congress of Industrial Microbiology and Microbial Biotechnology.

The GLICOENZ consortium has been widely represented in the IX National Congress of Industrial Microbiology and Microbial Biotechnology. 

Laura Barahona, Noa Miguez, Egle Narmontaite, María Martínez-Ranz, Mercedes Moreno presented their works on different topics of glycoenzymes and their applications.

 

GLYCOENZ-GREEN meeting

In the GLYCOENZ-GREEN Project (PID2022-136367OB-C31/2/3) funded by the State Research Agency, we are studying the application of enzymes that act on carbohydrates for the production of bioactive substances (oligosaccharides and polyphenol derivatives). This week, we had a workshop at the Institute of Catalysis and Petrochemistry, along with colleagues from the Blas Cabrera Institute of Physical Chemistry and the Severo Ochoa Center for Molecular Biology, CBMSO. It was a very productive meeting where we strengthened collaborations, and where young students enthusiastically defended their work.

Publication in Microbial Cell Factories: three new chitinases from Mestchnikowia pulcherrima

Three new chitinases from M. pulcherrima, MpChit35, MpChit38 and MpChit41, were molecularly characterized and extracellularly expressed in Pichia pastoris.The three enzymes hydrolysed colloidal chitin with optimal activity at 45 ºC and pH 4.0-4.5, The partial separation and characterization of the complex COS mixtures produced from the hydrolysis of chitin and chitosan were achieved by a new anionic chromatography HPAEC-PAD method and mass spectrometry assays. An overview of the predicted structures of these proteins and their catalytic modes of action were also presented. Depicted their high sequence and structural homology, MpChit35 acted as an exo-chitinase producing di-acetyl-chitobiose from chitin while MpChit38 and MpChit41 both acted as endo-chitinases producing tri-acetyl-chitotriose as main final product.

Microbial Cell Factories 23, 31 (2024)  https://doi.org/10.1186/s12934-024-02300-9