Method of constructing a recombinant Bacillus subtilis that can produce specific-molecular-weight hyaluronic acids
US9771607B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Jan 30, 2016 |
| Grant date | Sep 26, 2017 |
| Priority date | — |
| Expiry date | Apr 8, 2036 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Y504/0201
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The present invention relates to the field of biotechnology engineering. It provides a method of constructing a recombinant Bacillus subtilis that can produce specific-molecular-weight hyaluronic acids. By integranted expression of hasA from Streptococcus zooepidemicus and overexpression of genes of HA synthetic pathway, tuaD, glmU and glmS, high yield HA production was achieved in the recombinant strain. Additionally, introduction and functional expression of the leech hyaluronidase in the recombinant strain substantially increased the yield of HA to 19.38 g·L−1. Moreover, HAs with a broad range of molecular weights (103 Da to 106 MDa) were efficiently produced by controlling the expression level of hyaluronidase using RBS mutants with different translational strengths. The method of the present invention can be used to produce low molecular weight HAs at large scale in industrial applications.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.