Pengpeng Li

Effect of heat shock protein 90 against ROS-induced phospholipid oxidation

The antioxidative role of Hsp90 purified from duck muscle on phospholipid was investigated in this study. Lipid oxidation was assessed by ferric thiocyanate method, and particle size, zeta potential measurements and transmission electron microscopy were employed to detect physical and chemical changes during oxidation of liposomes. Increase of the peroxidation products, decrease of the particle size and disruption of the spherical unity of the liposomes were detected when liposome was exposed to hydrogen peroxide (H2O2) oxidation, and Hsp90 was shown to protect liposome against oxidation. To further test the protection of Hsp90 on cell membrane, the Hsp90 transformed E. coli cell model was built. The Hsp90-containing E. coli showed higher survival and better membrane integrity as compared to the control upon H2O2 exposure. It is expected that Hsp90 plays an important role in the prevention of phospholipids oxidation in meat and has potential use as a food antioxidant.

Effect of ultrasound assisted extraction on the physicochemical and functional properties of collagen from soft-shelled turtle calipash

The aim of the present study was to evaluate the physicochemical and functional properties of acid-soluble collagen by ultrasound assisted extraction (UASC) from calipash of soft-shelled turtle (Pelodiscus sinensis). The results showed the collagen content was increased by 16.3% in UASC over the collagen from the conventional extraction (ASC). Both ASC and UASC contained a moderate amount of imino acid (197 and 216 residues/1000 residues, respectively) and hydrophobic amino acid (353 and 391 residues/1000 residues, respectively) in amino acid composition. Fourier transform infrared (FTIR) spectroscopy and X-ray diffraction (XRD) analyses confirmed that the ultrasound treatment did not disrupt the triple-stranded helical structures in UASC. UASC had higher thermal stability compared with ASC by viscosity and differential scanning calorimetry (DSC) measurements, therefore, UASC might have the advantage to be used. In dynamic elastic behavior measurement, UASC showed a larger elasticity than ASC. With a mild modification by ultrasound, UASC had superior functional properties to ASC, including water/oil absorption capacity, water-holding capacity, emulsifying properties and foaming properties. These results suggested that UASC from the soft-shelled turtle calipash had a potential to be used widely in food, medicine, cosmetics and biomedical materials.

Interaction of Hsp90 with phospholipid model membranes

Heat shock protein 90 (Hsp90) is an essential molecular chaperone with versatile functions in cell homeostatic control under both normal and stress conditions. Hsp90 has been found to be expressed on the cell surface, but the mechanism of Hsp90 association to the membrane remains obscure. In this study, the direct interaction of Hsp90 and phospholipid vesicles was characterized, and the role of Hsp90 on membrane physical state was explored. Using surface plasmon resonance (SPR), we observed a strong interaction between Hsp90 and different compositions of lipid. Hsp90 had a preference to bind with more unsaturated phospholipid species and the affinity was higher with negatively charged lipids than zwitterionic lipids. Increasing the mole fraction of cholesterol in the phospholipid led to a decrease of binding affinity to Hsp90. Circular dichroism (CD) spectroscopy of Hsp90 in PC membranes showed more α-helix structure than in aqueous buffer. The differential scanning calorimeter (DSC) and fluorescence polarization results showed Hsp90 could affect the transition temperature and fluidity of the bilayer. We postulate from these results that the association between Hsp90 and membranes may involve both electrostatic and hydrophobic force, and constitute a possible mechanism that modulates membrane lipid order during thermal fluctuations.

Synergistic antibacterial mechanism of the Lactobacillus crispatus surface layer protein and nisin on Staphylococcus saprophyticus

SlpB, a surface layer protein isolated from Lactobacillus crispatus, has the potential to enhance the antimicrobial activity of nisin. Previous research indicated that, when combined with nisin, SlpB acted synergistically to inhibit Staphylococcus saprophyticus growth, thus extending the shelf life of chicken meat. In order to understand how SlpB enhances the antibacterial activity of nisin, electron microscopy, confocal laser scanning microscopy, flow cytometry and transmembrane electrical potential analysis were used to study cell wall organization and cell membrane integrity. No remarkable bacteriolytic effects were observed, indicating that cell death could not be attributed to cell lysis, although SlpB caused dramatic modifications of cell wall, thereby altering cell shape. The combination of SlpB and nisin also induced the release of ATP or UV-absorbing materials, as well as sudden dissipation of the transmembrane electrical potential by compromising membrane integrity. Considering that SlpB led to structural disorganization of the cell wall, and nisin access is enhanced to form a stable pore, cell death is a predictable outcome. SlpB significantly enhanced the effect of nisin at half of the minimum inhibitory concentration, which resulted in cell death by destroying the cell wall and cell membrane, therefore providing a new, feasible approach in food preservation.

Differential expression of heat shock protein 90, 70, 60 in chicken muscles postmortem and its relationship with meat quality

Objective The aim of this study was to investigate the expression of heat shock protein (HSP) 90, 70, and 60 in chicken muscles and their possible relationship with quality traits of meat. Methods The breast muscles from one hundred broiler chickens were analyzed for drip loss and other quality parameters, and the levels of heat shock protein (HSP) 90, 70, and 60 were determined by immunoblots. Results Based on the data, chicken breast muscles were segregated into low (drip loss≤5%), intermediate (5%<drip loss<9.5%) and high (drip loss≥9.5) drip loss groups. The expression of HSP90 and HSP60 were significantly lower in the high drip loss group compared to that in the low and intermediate drip loss group (p<0.05), while HSP70 was equivalent in abundance in all groups (p>0.05). Conclusion Results of this study suggests that higher levels of HSP90 and HSP60 may be advantageous for maintenance of cell function and reduction of water loss, and they could act as potential indicator for better water holding capacity of meat.

Class III bacteriocin Helveticin-M causes sublethal damage on target cells through impairment of cell wall and membrane

Helveticin-M, a novel Class III bacteriocin produced by Lactobacillus crispatus exhibited an antimicrobial activity against Staphylococcus aureus, S. saprophyticus, and Enterobacter cloacae. To understand how Helveticin-M injured target cells, Helveticin-M was cloned and heterologously expressed in Escherichia coli. Subsequently, the cell wall organization and cell membrane integrity of target cells were determined. The mechanism of cellular damage differed according to bacterial species. Based on morphology analysis, Helveticin-M disrupted the cell wall of Gram-positive bacteria and disorganized the outer membrane of Gram-negative bacteria, therefore, altering surface structure. Helveticin-M also disrupted the inner membrane, as confirmed by leakage of intracellular ATP from cells and depolarization of membrane potential of target bacteria. Based on cell population analysis, Helveticin-M treatment caused the increase of cell membrane permeability, but the cytosolic enzymes were not influenced, indicating that it was the sublethal injury. Therefore, the mode of Helveticin-M action is bacteriostatic rather than bactericidal.

Interaction of heat shock protein 90 B1 (Hsp90B1) with liposome reveals its potential role in protection the integrity of lipid membranes

Heat shock proteins of 90kDa (Hsp90) are molecular chaperones essential for protein homeostasis. Besides chaperone activity, Hsp90 exhibits other cellular functions at membranes, yet how it interacts with membranes remains elusive. We report here that Hsp90B1 interacts with phospholipid membranes. We first cloned the full-length open reading frame of Hsp90B1 from Anas platyrhnchos (ApHsp90B1), and the gene was then heterologously expressed and purified. SPR analysis show the purified ApHsp90B1 interacts with phospholipid membranes with high affinity (KD 176±25nM), and the interaction occurs over a wide range of pH, which is especially distinct under acidic conditions. Tryptophan fluorescence and far-UV CD spectra studies find that the interaction of ApHsp90B1 with phospholipid membrane induces microenvironment changes of tryptophan residues and conformational change of some regions in ApHsp90B1, which might be the reason of its increased ATPase activity upon addition phospholipid vesicles. Importantly, the interaction of ApHsp90B1 with phospholipid vesicles significantly reduces lipolysis of the membrane phospholipid, suggesting that the interaction of Hsp90B1 with membrane could preserve membrane integrity. The present study therefore demonstrates for the first time that Hsp90B1 exhibits high affinity for phospholipid membrane and suggest Hsp90B1 play an important role in membrane-stabilizing via its interaction with membrane phospholipids.

Lipolysis and Lipid Oxidation during Processing of Chinese Traditional Dry-Cured White Amur Bream (Parabramis pekinensis)

ABSTRACT Lipolysis and lipid oxidation of intramuscular lipids during the dry-curing process of white amur bream (Parabramis pekinensis) were studied. All lipolytic enzyme activities decreased (p < 0.05) at the end of the process. The lipoxygenase activity increased during wet-salting (p < 0.05) and then decreased until the end of the process (p < 0.05). The daily variation of free fatty acids was correlated with those of the peroxide value (r = 0.976, p < 0.01) and thiobarbituric acid-reactive substances (r = 0.922, p < 0.01), respectively. These results suggest that lipolysis promotes lipid oxidation during the processing of dry-cured fish.