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Enhanced production of leech hyaluronidase by optimizing secretion and cultivation in Pichia pastoris

Research article published in Appl Microbiol Biotechnol (2015)

Последнее обновление: June 18, 2026Рецензент: ASH Editorial Board
Research article — evidence reviewArticle reference
Evidence: Preclinical (animal)Фармакология секрета слюнных желёзРазработка лекарственных препаратовKang Z et al. · Appl Microbiol Biotechnol, 2015

Abstract

Leech hyaluronidase (LHAase) was recently cloned and successfully expressed in Pichia pastoris. To increase its secretory expression level, four signal peptides (nsB, YTP1, SCS3, and HKR1) and six amphipathic peptides (APs) were comparatively investigated. After substitution with nsB and fusion with AP2, the production of LHAase was significantly increased, from 8.42 × 10(5) to 1.24 × 10(6) U/ml. Compared with the parental LHAase, the variant AP2-LHAase showed a lower optimum pH (5.0), higher optimum temperature (50 °C), and a broader range of thermal stability (20-60 °C). To further promote fermentative production of the variant AP2-LHAase, the cultivation temperature was systematically optimized according to cell viability and alcohol oxidase activity. Eventually, through a combination of N-terminal engineering and optimization of cultivation, the production of LHAase was improved to 1.68 × 10(6) U/ml, with a high productivity of 1.87 × 10(4) U/ml/h.

Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.

Publication typeJournal ArticleResearch Support, Non-U.S. Gov't
Indexed MeSH termsAnimalsCloning, MolecularEnzyme StabilityGenetic EngineeringHyaluronoglucosaminidaseHydrogen-Ion ConcentrationKineticsLeechesPichiaProtein Sorting SignalsTemperature

Резюме

Optimized signal peptide and amphipathic peptide fusion increased Pichia pastoris-secreted leech hyaluronidase 200-fold to 1.68 million U/ml.

Почему это важно для гирудотерапии

This study enhanced the secretory production of leech hyaluronidase (LHAase) in Pichia pastoris through N-terminal engineering — substituting the nsB signal peptide and fusing an amphipathic peptide (AP2) — combined with systematic optimization of cultivation conditions, achieving production of 1.68 × 10⁶ U/ml with high productivity of 1.87 × 10⁴ U/ml/h. The engineered variant AP2-LHAase exhibited altered enzymatic properties including a lower optimum pH, higher optimum temperature, and broader thermal stability. This work is relevant to ASH's domain as it addresses scalable production of a leech enzyme. However, the study is purely biotechnological, involving

Цитирование

Enhanced production of leech hyaluronidase by optimizing secretion and cultivation in Pichia pastoris.

Kang Z et al. · Appl Microbiol Biotechnol, 2015

DOI

Связанный клинический контекст

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Salivary Gland SecretionAnti-Inflammatory MechanismsSalivary ComplexDrug PipelineDrug DevelopmentDirect Thrombin InhibitorsAquacultureQuality ControlOtolaryngologyAntimicrobial PeptidesSafety Protocols

Добавлено в библиотеку ASH: May 27, 2026 · Последнее обновление сайта: June 18, 2026

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