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CRISPRi-assisted metabolic engineering of cyanobacteria for photosynthetic hyaluronic acid from CO2
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CRISPRi-assisted metabolic engineering of cyanobacteria for photosynthetic hyaluronic acid from CO2
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Journal Article
CRISPRi-assisted metabolic engineering of cyanobacteria for photosynthetic hyaluronic acid from CO2
2025
Overview
Background
Hyaluronic acid (HA) is widely used in pharmaceuticals, medicine, and cosmetics. Sustainable production has shifted to microbial fermentation using engineered GRAS strains. Diverse carbon sources and CO
2
conversion via engineered microorganisms enhance HA production. Herein we applied advances in CRISPR technologies and tools to optimize metabolic pathway by redirecting carbon portioning in cyanobacterium
Synechoccous elongatus
PCC 7942, demonstrating enhanced HA production.
Results
S. elongatus
PCC 7942 lacking hyaluronan synthase (HAS) required pathway engineering for HA production. By expressing heterologous Class I HAS, a modular gene expression system was employed, incorporating
hasB
and
hasC
for the HA-GlcA module and
glmU
,
glmM
, and
glmS
for the GlcNAc module. This approach resulted in construction of four engineered cyanobacterial strains. Optimizing metabolic pathway involving the HA-GlcA and GlcNAc modules led to SeHA220 (wild-type with HA-GlcA and GlcNAc modules) producing 2.4 ± 0.85 mg/L HA at 21 d, a 27.5-fold increase compared to the control. Targeting F6P and G6P metabolic nodes via CRISPR interference to repress
zwf
and
pfk
genes further improved production, with SeHA226 (SeHA220 with a gene repression module) achieving 5.0 ± 0.3 mg/L HA from CO
2
at 15 d. Notably, SeHA226 produced photosynthetic HA with a molecular weight (Mw) of 4.2 MDa, comparable to native producers, emphasizing the importance of precursor balance and growth conditions.
Conclusions
This study engineered cyanobacteria for efficient HA biosynthesis using modular gene expression and CRISPR-interference systems. Optimizing heterologous metabolic pathway was key to achieving high-molecular-weight photosynthetic HA production from CO
2
. The findings provide insights into tunable HA production, with future efforts aimed at scaling up photosynthetic HA production for larger-scale applications.
Publisher
BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
Subject
