Yeast surface display of leech hyaluronidase for the industrial production of hyaluronic acid oligosaccharides
Research article published in Eng Microbiol (2023)
Abstract
Leech hyaluronidase (LHyal) is a hyperactive hyaluronic acid (HA) hydrolase that belongs to the hyaluronoglucuronidase family. Traditionally, LHyal is extracted from the heads of leeches, but the recent development of the Pichia pastoris recombinant LHyal expression method permitted the industrial production of size-specific HA oligosaccharides. However, at present LHyal expressed by recombinant yeast strains requires laborious protein purification steps. Moreover, the enzyme is deactivated and removed after single use. To solve this problem, we developed a recyclable LHyal biocatalyst using a yeast surface display (YSD) system. After screening and characterization, we found that the cell wall protein Sed1p displayed stronger anchoring to the P. pastoris cell wall than other cell wall proteins. By optimizing the type and length of the linkers between LHyal and Sed1p, we increased the activity of enzymes displayed on the P. pastoris cell wall by 50.34% in flask cultures. LHyal-(GGGS)6-Sed1p activity further increased to 3.58 × 105 U mL-1 in fed-batch cultivation in a 5 L bioreactor. Enzymatic property analysis results revealed that the displayed LHyal-(GGGS)6-Sed1p generated the same oligosaccharides but exhibited higher thermal stability than free LHyal enzyme. Moreover, displayed LHyal-(GGGS)6-Sed1p could be recovered easily from HA hydrolysis solutions via low-speed centrifugation and could be reused at least 5 times. YSD of LHyal not only increased the utilization efficiency of the enzyme but also simplified the purification process for HA oligosaccharides. Thus, this study provides an alternative approach for the industrial preparation of LHyal and HA oligosaccharides.
Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.
Resumen
Yeast surface display of leech hyaluronidase using Sed1p anchor; displayed enzyme reaches 3.58 million U/ml and is reusable for at least 5 cycles.
Por qué esto importa para la hirudoterapia
Este estudio de biotecnología describe el desarrollo de un biocatalizador reciclable de hialuronidasa de sanguijuela (LHyal) mediante un sistema de exposición en superficie de levaduras en Pichia pastoris, alcanzando una actividad enzimática de 3,58 × 10⁵ U mL⁻¹ en cultivo alimentado por lotes (fed-batch) y permitiendo la producción industrial de oligosacáridos de ácido hialurónico de tamaño específico. La LHyal, tradicionalmente extraída de cabezas de sanguijuela, se ancló a la pared celular de la levadura mediante la proteína Sed1p, lo que resultó en una mayor estabilidad térmica y reutilización durante al menos cinco ciclos. Esto posee relevancia indirecta para el dominio ASH al demostrar aplicaciones biotecnológicas de una enzima derivada de sanguijuela, aunque el foco se centra en la producción industrial de polisacáridos. Advertencia: Se trata de un estudio de ingeniería de proteínas y bioprocesamiento industrial sin aplicación terapéutica, datos clínicos ni examen de propiedades del secretoma relevantes para la anticoagulación o la hirudoterapia.
Citación
Yeast surface display of leech hyaluronidase for the industrial production of hyaluronic acid oligosaccharides.
Liao L et al. · Eng Microbiol, 2023
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Añadido a la biblioteca ASH: May 27, 2026 · Última actualización del sitio: 18 de junio de 2026