Zhu B.,Chinese Academy of SciencesBeijing |
Liu C.,State Key Laboratory of Microbial Technology |
Liu S.,State Key Laboratory of Microbial Technology |
Cong H.,State Key Laboratory of Microbial Technology |
And 3 more authors.
Environmental Microbiology | Year: 2016
Cyclic diguanosine monophosphate (c-di-GMP) is one of the most important bacterial second messengers that controls many bacterial cellular functions including lifestyle switch between plankton and biofilm. Surface attachment defective (SadC) is a diguanylate cyclase (DGC) involved in the biosynthesis of c-di-GMP in Pseudomonas aeruginosa, an opportunistic pathogen that can cause diverse infections. Here we report the crystal structure of GGDEF domain from SadC and the critical role of the trans-membrane (TM) domain of SadC with regard to biofilm formation, exopolysaccharide production and motility. We showed that over-expression of SadC in P. aeruginosa PAO1 totally inhibited swimming motility and significantly enhanced the production of exopolysaccharide Psl. SadC lacking TM domains (SadC300-487) could not localize on cytoplasmic membrane and form cluster, lost the ability to inhibit the swimming and twitching motility, and showed the attenuated activity to promote Psl production despite that SadC300-487 was able to catalyze the synthesize of c-di-GMP in vitro and in vivo. The GGDEF domain of SadC has a typical GGDEF structure and the α-helix connected the TM domains with SadC GGDEF domain is essential for SadC to form DGC oligomers. Our data imply that membrane association of SadC promotes its DGC activity by affecting the formation of active DGC oligomers. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd. Source
Xue M.,National Glycoengineering Research Center |
Xue M.,State Key Laboratory of Microbial Technology |
Guan W.,National Glycoengineering Research Center |
Guan W.,State Key Laboratory of Microbial Technology |
And 11 more authors.
Bioorganic and Medicinal Chemistry Letters | Year: 2012
Nucleotide sugars are essential glycosyl donors for Leloir-type glycosyltransferases. The UDP-N-acetylgalactosamine pyrophosphorylase (UDP-GalNAc PP; AGX1) from Homo sapiens catalyzes the synthesis of UDP-N-acetylgalactosamine from N-acetylgalactosamine 1-phosphate and UTP. In this Letter, we systematically studied nucleotide substrate specificity of AGX1 during its uridyltransfer reaction, and described the capability of AGX1 to catalyze dUTP and dTTP to their corresponding nucleotide sugars for the first time. Furthermore, using such a eukaryotic enzyme, we synthesized dUDP-GalNAc and dTDP-GalNAc in multiple mg scale in vitro efficiently and rapidly. © 2012 Elsevier Ltd. All rights reserved. Source
Liu J.,State Key Laboratory of Microbial Technology |
Liu J.,National Glycoengineering Research Center |
Zou Y.,State Key Laboratory of Microbial Technology |
Zou Y.,National Glycoengineering Research Center |
And 14 more authors.
Bioorganic and Medicinal Chemistry Letters | Year: 2013
Nucleotide sugars are activated forms of monosaccharides and key intermediates of carbohydrate metabolism in all organisms. The availability of structurally diverse nucleotide sugars is particularly important for the characterization of glycosyltransferases. Given that limited methods are available for preparation of nucleotide sugars, especially their useful non-natural derivatives, we introduced herein an efficient one-step three-enzyme catalytic system for the synthesis of nucleotide sugars from monosaccharides. In this study, a promiscuous UDP-sugar pyrophosphorylase (USP) from Arabidopsis thaliana (AtUSP) was used with a galactokinase from Streptococcus pneumoniae TIGR4 (SpGalK) and an inorganic pyrophosphatase (PPase) to effectively synthesize four UDP-sugars. AtUSP has better tolerance for C4-derivatives of Gal-1-P compared to UDP-glucose pyrophosphorylase from S. pneumoniae TIGR4 (SpGalU). Besides, the nucleotide substrate specificity and kinetic parameters of AtUSP were systematically studied. AtUSP exhibited considerable activity toward UTP, dUTP and dTTP, the yield of which was 87%, 85% and 84%, respectively. These results provide abundant information for better understanding of the relationship between substrate specificity and structural features of AtUSP. © 2013 Elsevier Ltd. All rights reserved. Source
Sun M.-L.,State Key Laboratory of Microbial Technology |
Sun M.-L.,Shandong University |
Zhao F.,State Key Laboratory of Microbial Technology |
Zhao F.,Shandong University |
And 9 more authors.
Scientific Reports | Year: 2015
Although many kinds of exopolysaccharides (EPSs) from microorganisms have been used in industry, the exploration and utilization of EPSs from polar microorganisms is still rather rare. In this study, a flavobacterial strain, SM1127, from the Arctic brown alga Laminaria, was screened for its high EPS production (2.11 g/l) and was identified as belonging to the genus Polaribacter. The EPS secreted by strain SM1127 has a molecular mass of 220 kDa, and it mainly comprises N-acetyl glucosamine, mannose and glucuronic acid residues bound by heterogeneous linkages. Rheological studies on the aqueous EPS showed that it had a high viscosity and good shear-thinning property. Moreover, the EPS showed a high tolerance to high salinity and a wide pH range. The EPS also had good antioxidant activity. Particularly, its moisture-retention ability was superior to that of any other reported EPS or functional ingredient generally used in cosmetics. The EPS also showed a protective effect on human dermal fibroblasts at low temperature (4 °C). Safety assessment indicated that the EPS is safe for oral administration and external use. These results indicate the promising potential of the EPS from strain SM1127 in the food, cosmetic, pharmaceutical and biomedical fields. Source