American Society of Hirudotherapy

Production of specific-molecular-weight hyaluronan by metabolically engineered Bacillus subtilis 168

Biotechnology paper published in Metabolic Engineering (2016)

Last Updated: June 18, 2026Reviewed by: ASH Editorial Board
Research article — evidence reviewArticle reference
Evidence: In vitro / laboratoryDrug DevelopmentGenomics & ProteomicsJin P et al. · Metabolic Engineering, 2016

Abstract

Low-molecular-weight hyaluronan (LMW-HA) has attracted much attention because of its many potential applications. Here, we efficiently produced specific LMW-HAs from sucrose in Bacillus subtilis. By coexpressing the identified committed genes (tuaD, gtaB, glmU, glmM, and glmS) and downregulating the glycolytic pathway, HA production was significantly increased from 1.01gL(-1) to 3.16gL(-1), with a molecular weight range of 1.40×10(6)-1.83×10(6)Da. When leech hyaluronidase was actively expressed after N-terminal engineering (1.62×10(6)UmL(-1)), the production of HA was substantially increased from 5.96gL(-1) to 19.38gL(-1). The level of hyaluronidase was rationally regulated with a ribosome-binding site engineering strategy, allowing the production of LMW-HAs with a molecular weight range of 2.20×10(3)-1.42×10(6)Da. Our results confirm that this strategy for the controllable expression of hyaluronidase, together with the optimization of the HA synthetic pathway, effectively produces specific LMW-HAs, and could also be used to produce other LMW polysaccharides.

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 termsAnimalsBacillus subtilisHyaluronic AcidHyaluronoglucosaminidaseLeechesMetabolic EngineeringRecombinant Proteins

Summary

Bacillus subtilis 168 with N-terminally engineered leech hyaluronidase (1.62 × 10^6 U/mL activity) yielded low-molecular-weight hyaluronans 2.2 × 10^3 to 1.42 × 10^6 Da from sucrose at 19.38 g/L.

Why This Matters for Hirudotherapy

This study reports metabolic engineering of Bacillus subtilis to produce specific low-molecular-weight hyaluronan (LMW-HA) from sucrose, using leech hyaluronidase as a key enzymatic tool. After N-terminal engineering, leech hyaluronidase was expressed at high levels and its activity rationally regulated via ribosome-binding site engineering, enabling controlled production of LMW-HAs across a molecular weight range of 2.20×10³–1.42×10⁶ Da. For ASH's domain, this work demonstrates a bioprocessing application of a leech-derived enzyme as a production tool. CAVEAT: This is a microbial biotechnology study with no clinical leech therapy component; the leech connection is limited to use of recombinant leech hyaluronidase, and findings have no direct therapeutic implication for hirudotherapy.

Citation

Production of specific-molecular-weight hyaluronan by metabolically engineered Bacillus subtilis 168.

Jin P et al. · Metabolic Engineering, 2016

DOI

Added to ASH library: May 27, 2026 · Site last updated: June 18, 2026

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