Gianna Palmieri - Academia.edu (original) (raw)

Papers by Gianna Palmieri

Journal of Proteome Research, Dec 4, 2008

The specific inhibition of serine proteinases, which are crucial switches in many important physi... more The specific inhibition of serine proteinases, which are crucial switches in many important physiological processes, is of great value both for basic research and for therapeutic applications. In this study, we report the molecular cloning of the sso0767 gene from Sulfolobus solfataricus, and the functional characterization of its product, SsCEI, which represents the first archaeal phosphatidylethanolaminebinding protein (PEBP)-serine proteinase inhibitor, reported to date. SsCEI is a monomer protein with a molecular mass of 19.0 kDa and a pI of 6.7, which is able to inhibit the serine proteases R-chymotrypsin and elastase with K i values of 0.08 and 0.1 µM, respectively. Moreover SsCEI is extremely resistant to both thermal inactivation and proteolytic attack suggesting compact folding of the protein. Within the I51 family, the archaeal inhibitor shows strong similarity to the human and murine members. The threedimensional model of SsCEI revealed a general-fold and the presence of an anion-binding pocket, the hallmark of the PEBP family. Moreover SsCEI binds the cognate proteases according to a common, substrate-like standard mechanism. Point mutation experiments supported the prediction of the protease-binding site located on the surface at the C-terminal region of the protein. Interestingly, searches based on preidentified structural reactive loop motifs revealed the occurrence of a sequence (T123-N130) that is not represented in all serine-protease inhibitor families. This unique motif may provide new insights into both the inhibitor/protease binding mode and the specific biological functions of SsCEI within the PEBP family.

Journal of Protein Chemistry, Mar 1, 2001

Microbiological Research, Apr 1, 2019

This is a PDF file of an unedited manuscript that has been accepted for publication. As a service... more This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. An alternative biocontrol agent of soil-borne phytopathogens: a new antifungal compound produced by a plant growth promoting bacterium isolated from North Algeria

PLOS ONE, May 24, 2012

A novel acylpeptide hydrolase, named APEH-3 Ss , was isolated from the hypertermophilic archaeon ... more A novel acylpeptide hydrolase, named APEH-3 Ss , was isolated from the hypertermophilic archaeon Sulfolobus solfataricus. APEH is a member of the prolyl oligopeptidase family which catalyzes the removal of acetylated amino acid residues from the N terminus of oligopeptides. The purified enzyme shows a homotrimeric structure, unique among the associate partners of the APEH cluster and, in contrast to the archaeal APEHs which show both exo/endo peptidase activities, it appears to be a ''true'' aminopeptidase as exemplified by its mammalian counterparts, with which it shares a similar substrate specificity. Furthermore, a comparative study on the regulation of apeh gene expression, revealed a significant but divergent alteration in the expression pattern of apeh-3 Ss and apeh Ss (the gene encoding the previously identified APEH Ss from S. solfataricus), which is induced in response to various stressful growth conditions. Hence, both APEH enzymes can be defined as stressregulated proteins which play a complementary role in enabling the survival of S. solfataricus cells under different conditions. These results provide new structural and functional insights into S. solfataricus APEH, offering a possible explanation for the multiplicity of this enzyme in Archaea.

Journal of Bacteriology, Feb 1, 2011

In this study we gain insight into the structural and functional characterization of the Aeropyru... more In this study we gain insight into the structural and functional characterization of the Aeropyrum pernix oligopeptide-binding protein (OppA Ap) previously identified from the extracellular medium of an Aeropyrum pernix cell culture at late stationary phase. OppA Ap showed an N-terminal Q32 in a pyroglutamate form and C-terminal processing at the level of a threonine-rich region probably involved in protein membrane anchoring. Moreover, the OppA Ap protein released into the medium was identified as a "nicked" form composed of two tightly associated fragments detachable only under strong denaturing conditions. The cleavage site E569-G570 seems be located on an exposed surface loop that is highly conserved in several three-dimensional (3D) structures of dipeptide/oligopeptide-binding proteins from different sources. Structural and biochemical properties of the nicked protein were virtually indistinguishable from those of the intact form. Indeed, studies of the entire bacterially expressed OppA Ap protein owning the same N and C termini of the nicked form supported these findings. Moreover, in the middle exponential growth phase, OppA Ap was found as an intact cell membrane-associated protein. Interestingly, the native exoprotein OppA Ap was copurified with a hexapeptide (EKFKIV) showing both lysines methylated and possibly originating from an A. pernix endogenous stress-induced lipoprotein. Therefore, the involvement of OppA Ap in the recycling of endogenous proteins was suggested to be a potential physiological function. Finally, a new OppA from Sulfolobus solfataricus, SSO1288, was purified and preliminarily characterized, allowing the identification of a common structural/genetic organization shared by all "true" archaeal OppA proteins of the dipeptide/oligopeptide class.

Italian Journal of Food Safety, Aug 11, 2022

Fresh fishery products are highly perishable foods mainly due to their high-water content and hig... more Fresh fishery products are highly perishable foods mainly due to their high-water content and high level of pH which act as promoters of spoilage processes. In these matrices, the deterioration phenomena are the result of the action of oxidative, and enzymatic processes due in part to the presence of specific microorganisms. Indeed, the microbial communities responsible for spoilage are a small fraction of the flora detectable in the fish and are known as specific spoilage organisms (SSOs). In the last decades, the scientific community has worked to achieve the ambitious goal of reducing the impact of microbial deterioration on food losses through innovative solutions, including antimicrobial packaging. The goal of this study was to evaluate the efficacy of an active polypropylene (PP)based packaging functionalized with the antimicrobial peptide 1018K6 to extend the shelf life of dolphinfish burgers (Coryphaena hippurus) by evaluating its effect on sensorial and microbiological profile. The microbiological results showed an evident antimicrobial activity of the active packaging against hygiene indicator microorganisms and SSOs, recording a reduction of about 1 Log (CFU/g) of their concentrations compared to those of the control groups. Furthermore, a significant influence of functionalized packaging on the organoleptic characteristics was noted, accentuating the differences in freshness between the two experimental groups. This work confirmed the hypothesis of considering antimicrobial packaging as a potential tool capable of slowing down surface microbial replication and, therefore, extending the shelf-life and improving the health and hygiene aspect of fresh fish products.

International Journal of Molecular Sciences

Since penicillin was discovered, antibiotics have been critical in the fight against infections. ... more Since penicillin was discovered, antibiotics have been critical in the fight against infections. However, antibiotic misuse has led to drug resistance, which now constitutes a serious health problem. In this context, antimicrobial peptides (AMPs) constitute a natural group of short proteins, varying in structure and length, that act against certain types of bacterial pathogens. The antimicrobial peptide 1018-K6 (VRLIVKVRIWRR- NH2) has significant bactericidal and antibiofilm activity against Listeria monocytogenes isolates, and against different strains and serotypes of Salmonella. Here, the mechanism of action of 1018-K6 was explored further to understand the peptide–membrane interactions relevant to its activity, and to define their determinants. We combined studies with model synthetic membranes (liposomes) and model biological membranes, assessing the absorption maximum and the quenching of 1018-K6 fluorescence in aqueous and lipid environments, the self-quenching of carboxyfluo...

Enzyme and microbial technology, Feb 1, 1994

A new procedure was developed for enzyme immobilization by entrapment in copper alginate gel. The... more A new procedure was developed for enzyme immobilization by entrapment in copper alginate gel. The mechanical properties of the copper alginate gel were characterized and compared with those of the most widely used calcium alginate. The system was applied to the immobilization of a fungal phenol oxidase. Optimal conditions for enzyme immobilization were set up: the system immobilized 85% of the enzyme, and the remaining 15% was recovered in the aqueous immobilization medium. The stability and activity of the immobilized enzyme were studied. After immobilization, the enzyme was active in a wider pH range, the temperature of its optimal activity was shifted to lower values, and the possibility of storage at 4 degrees C was greatly improved. The immobilized enzyme generally increased the rate of oxidation of various substrates. The results indicate a potential use of this system for the construction of bioreactors to be used in the detoxification of polluted waste waters.

Methods in molecular biology, 2022

International Journal of Molecular Sciences, 2021

APEH is a ubiquitous and cytosolic serine protease belonging to the prolyl oligopeptidase (POP) f... more APEH is a ubiquitous and cytosolic serine protease belonging to the prolyl oligopeptidase (POP) family, playing a critical role in the processes of degradation of proteins through both exo- and endopeptidase events. Endopeptidase activity has been associated with protein oxidation; however, the actual mechanisms have yet to be elucidated. We show that a synthetic fragment of GDF11 spanning the region 48–64 acquires sensitivity to the endopeptidase activity of APEH only when the methionines are transformed into the corresponding sulphoxide derivatives. The data suggest that the presence of sulphoxide-modified methionines is an important prerequisite for the substrates to be processed by APEH and that the residue is crucial for switching the enzyme activity from exo- to endoprotease. The cleavage occurs on residues placed on the C-terminal side of Met(O), with an efficiency depending on the methionine adjacent residues, which thereby may play a crucial role in driving and modulating A...

Antioxidants

Superoxide dismutase (SOD) is a fundamental antioxidant enzyme that neutralises superoxide ions, ... more Superoxide dismutase (SOD) is a fundamental antioxidant enzyme that neutralises superoxide ions, one of the main reactive oxygen species (ROS). Extremophile organisms possess enzymes that offer high stability and catalytic performances under a wide range of conditions, thus representing an exceptional source of biocatalysts useful for industrial processes. In this study, SODs from the thermo-halophilic Aeropyrum pernix (SODAp) and the thermo-acidophilic Saccharolobus solfataricus (SODSs) were heterologously expressed in transgenic tomato cell cultures. Cell extracts enriched with SODAp and SODSs showed a remarkable resistance to salt and low pHs, respectively, together with optimal activity at high temperatures. Moreover, the treatment of tuna fillets with SODAp-extracts induced an extension of the shelf-life of this product without resorting to the use of illicit substances. The results suggested that the recombinant plant extracts enriched with the extremozymes could find potentia...

Foods, 2022

Fresh fish are highly perishable, owing mainly to their moisture content, high amount of free ami... more Fresh fish are highly perishable, owing mainly to their moisture content, high amount of free amino acids and polyunsaturated fatty acids. Microorganisms and chemical reactions cause the spoilage, leading to loss in quality, human health risks and a market value reduction. Therefore, the fishing industry has always been willing to explore new technologies to increase quality and safety of fish products through a decrease of the microbiological and biochemical damage. In this context, antimicrobial active packaging is one such promising solution to meet consumer demands. The main objective of this study was to evaluate the effects of an active polypropylene-based packaging functionalized with the antimicrobial peptide 1018K6 on microbial growth, physicochemical properties and the sensory attributes of raw salmon fillets. The results showed that application of 1018K6-polypropylene strongly inhibited the microbial growth of both pathogenic and specific spoilage organisms (SSOs) on fish...

☯ These authors contributed equally to this work.

Journal of Biotechnology, 2019

Highlights  SODs are essential enzymes in all living cells, protecting against oxidant aggressio... more Highlights  SODs are essential enzymes in all living cells, protecting against oxidant aggressions;  The use of SODs in the dermo-cosmetic sector is limited due to the instability of the enzymes in skin care formulas and once exposed to environmental factors, in particular to UV radiations;  A Mn-SOD derived from the extremophilic organism Deinococcus radiodurans, once expressed in E.coli and plant cells, was characterized for its extraordinary features of resistance to high temperature and UV radiations;  A plant cell culture extract, enriched in the Deinococcus MnSOD, showed promising potentialities for skin care applications, thus it can be proposed as novel active ingredient for cosmetic-dermatological formulas

International journal of food microbiology, Jan 20, 2018

Antimicrobial peptides have received great attention for their potential benefits to extend the s... more Antimicrobial peptides have received great attention for their potential benefits to extend the shelf-life of food-products. Innate defense regulator peptide-1018 (IDR-1018) represents a promising candidate for such applications, due to its broad-spectrum antimicrobial activity, although food-isolated pathogens have been poorly investigated. Herein, we describe the design and the structural-functional characterization of a new 1018-derivative peptide named 1018-K6, in which the alanine in position 6 was replaced with a lysine. Spectroscopic analysis revealed a noticeable switch from β-sheet to helical conformations of 1018-K6 respect to IDR-1018, with a faster folding kinetic and increased structural stability. Moreover, 1018-K6 evidenced a significant antibiofilm/bactericidal efficiency specifically against Listeria monocytogenes isolates from food-products and food-processing environments, belonging to serotype 4b involved in the majority of human-listeriosis cases, with EC values...

Biological Control, 2018

Different bacterial groups in irrigation well water are strongly implicated in soil health and pl... more Different bacterial groups in irrigation well water are strongly implicated in soil health and plant development. Herein, 48 bacterial strains were isolated from agricultural well water in northern Algeria. Among them, four strains were selected based on their antifungal potential and their ability to express Plant Growth Promoting traits such as Indole Acetic Acid (IAA), hydrolytic enzymes, siderophores etc. The isolates were identified as Pseudomonas sp. (B, D and N strains) and Serratia sp. (C strain) by 16S rRNA gene sequencing. Mycelial growth inhibition against Botrytis cinerea and Aspergillus niger ranged from 60 to 90% for the four strains. Moreover, volatiles compounds emission by the isolates resulted in Plant Growth Inhibition values ranging from 13 to 50%, specifically against B. cinerea. Impressively, the strains' antifungal activity showed high inducibility as it was obtained only by the filtered supernatants from bacterial cultures previously in contact with the fungus. Finally, a greenhouse assay, carried out to determine the strains' efficacy in promoting plant growth and protecting seedlings under Pythium aphanidermatum-infected soil, revealed that the strain N markedly enhanced pea germination (+250%) and fresh weight (+43%) and tomato fresh weigh (+10%). The results constitute an attempt for better use of the bacterial functional diversity from irrigation wells in sustainable agriculture.

Progress in Biotechnology, 2002

Biomedicines

Acylpeptide hydrolase (APEH) is a serine protease involved in amino acid recycling from acylated ... more Acylpeptide hydrolase (APEH) is a serine protease involved in amino acid recycling from acylated peptides (exopeptidase activity) and degradation of oxidized proteins (endoproteinase activity). This enzyme is inhibited by dichlorvos (DDVP), an organophosphate compound used as an insecticide. The role of APEH in spermatogenesis has not been established; therefore, the aim of this study was to characterize the distribution and activity profile of APEH during this process. For this purpose, cryosections of male reproductive tissues (testis and epididymis) and isolated cells (Sertoli cells, germ cells, and spermatozoa) were obtained from adult rats in order to analyze the intracellular localization of APEH by indirect immunofluorescence. In addition, the catalytic activity profiles of APEH in the different male reproductive tissues and isolated cells were quantified. Our results show that APEH is homogeneously distributed in Sertoli cells and early germ cells (spermatocytes and round sp...