Genetic and biochemical characterization of genes involved in hyaluronic acid synthesis in Streptococcus zooepidemicus

Zhang, Yali, Luo, Kailai, Zhao, Qiushuang, Qi, Zhengliang, Nielsen, Lars Keld and Liu, Hao (2016) Genetic and biochemical characterization of genes involved in hyaluronic acid synthesis in Streptococcus zooepidemicus. Applied Microbiology and Biotechnology, 100 8: 3611-3620. doi:10.1007/s00253-016-7286-1


Author Zhang, Yali
Luo, Kailai
Zhao, Qiushuang
Qi, Zhengliang
Nielsen, Lars Keld
Liu, Hao
Title Genetic and biochemical characterization of genes involved in hyaluronic acid synthesis in Streptococcus zooepidemicus
Formatted title
Genetic and biochemical characterization of genes involved in hyaluronic acid synthesis in Streptococcus zooepidemicus
Journal name Applied Microbiology and Biotechnology   Check publisher's open access policy
ISSN 1432-0614
0175-7598
Publication date 2016-04
Sub-type Article (original research)
DOI 10.1007/s00253-016-7286-1
Open Access Status Not Open Access
Volume 100
Issue 8
Start page 3611
End page 3620
Total pages 10
Place of publication Heidelberg, Germany
Publisher Springer
Collection year 2017
Language eng
Formatted abstract
The biosynthetic pathway for hyaluronic acid (HA) has been proposed; however, a thorough genetic and functional analysis is required to further elucidate the roles of genes involved in HA production. Previously, we developed a markerless gene-deletion system for Streptococcus zooepidemicus and confirmed that hasA is essential for HA synthesis. Here, we constructed a comprehensive set of deletion mutants and investigated the roles of ten additional predicted genes in the HA synthetic pathway. Phenotypic assays revealed that all ten genes play a role in cell growth and/or HA synthesis. As expected, the deletion of hasA or hasB abolished HA production with little effect on growth, while the deletion of genes that are also required for peptidoglycan biosynthesis (hasE, glmM, and glmS) significantly reduced cell growth and HA production. Either of the glmU homologues (hasD and gcaD) was sufficient for optimal growth and the mucoid phenotype, while no double mutant could be isolated. Of the two UDP-glucose pyrophosphorylase (UGPase) paralogues, the operon-encoded hasC1 was responsible for 65 % of the activity, while hasC2 was responsible for the remaining 35 %. The deletion of hasC1 had no effect on cell growth and caused only a moderate decrease in the UDP-glucose level and HA production. The deletion of both hasC1 and hasC2 resulted in a severe growth defect and negligible UDP-glucose accumulation, HA production, and pyrophosphorylase activity. Of the two phosphoglucomutase paralogues, pgm1 and pgm2, the former is responsible for around 10 % of activity, while the latter is responsible for 90 %. The deletion of pgm1 showed no apparent effect on HA synthesis and growth, while the deletion of pgm2 resulted in the abolishment of HA synthesis and a significantly slower growth. These results should guide the metabolic engineering of S. zooepidemicus to improve HA productivity and quality.
Keyword Gene function
Hyaluronic acid
Metabolic engineering
Streptococcus zooepidemicus
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: HERDC Pre-Audit
Australian Institute for Bioengineering and Nanotechnology Publications
 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 0 times in Thomson Reuters Web of Science Article
Scopus Citation Count Cited 0 times in Scopus Article
Google Scholar Search Google Scholar
Created: Tue, 02 Feb 2016, 00:36:51 EST by System User on behalf of Learning and Research Services (UQ Library)