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

PARTICIPATION IN BIOTRANS 2023

Several members of GLICOENZ are participating the renowned Biotrans2023 congress held in La Rochelle, France. This congress offers an overview of the latest advancements in the fields of biocatalysis and biotransformations, bringing together innovative and interdisciplinary strategies to overcome scientific and technological challenges.

Marina Minguet, Fadia Cervantes, and Eglė Narmontaitė presented their work on hyaluronidases, tagatose, and fructooligosaccharides, respectively.

                               Eglė Narmontaitė

Characterization of a novel invertase from Trichoderma sp., a producer of a wide range of fructooligosaccharides.

Eglė Narmontaitė, Francisco J. Plou, María Fernández-Lobato

                             Fadia V. Cervantes

Fully biocatalytic synthesis of D-tagatose from whey permeate as a raw material.

Fadia V. Cervantes, Sawssan Neifar, Zoran Merdzo, Antonio O. Ballesteros, Maria Fernandez-Lobato, Samir Bejar, and Francisco J. Plou

                                                                                                                                                                                                                                                                                                                                                    

                       Marina Minguet-Lobato

New Hyaluronic Acid Degrading Enzymes from Fungi

Marina Minguet-Lobato, Fadia V. Cervantes, David Fernández-Polo, María Fernández-Lobato, Francisco J. Plou

Elimination of D-Glucose in carbohydrate syrups published in ACS Food Science & Technology

 

During the synthesis of many bioactive carbohydrates, D-glucose is released as a side-product of the transglycosylation process. It is desirable to remove it due to its caloric contribution and its effect on caries and diabetes.

In this work, we have investigated the use of immobilized Komagataella phaffii (formerly Pichia pastoris) for elimination of D-glucose and D-fructose in several sugar syrups. K. phaffii cells were immobilized in calcium alginate beads to facilitate the separation of the yeast cells from the reaction medium and reuse of the biocatalyst. The immobilized yeasts were successfully reutilized for at least 20 cycles  to remove D-glucose and D-fructose in a FOS syrup, without affecting the concentration of oligosaccharides. Excellent selectivity was also found for elimination of D-glucose in IMOS syrups. 

The methodology is versatile and easy to scale-up, as demonstrated in the removal of D-glucose and D-fructose  for the purification of heteroglucooligosaccharides synthesized by Metschnikowia reukaufii α-glucosidase. In addition, D-glucose was selectively removed by K. phaffii beads in the presence of D-galactose for at least 20 cycles of 150 min and applied to GOS purification.

Ref.: "Reuse of Immobilized Komagataella phaffii Cells for the Elimination of d-Glucose in Syrups of Bioactive Carbohydrates". Fadia V. Cervantes, David Fernandez-Polo, Zoran Merdzo, Noa Miguez, Martin Garcia-Gonzalez, Antonio O. Ballesteros, Maria Fernandez-Lobato, and Francisco J. Plou. ACS Food Science & Technology (2022), https://doi.org/10.1021/acsfoodscitech.2c00008