Publications by authors named "Francesco Grieco"

41 Publications

Effect of Soil Management and Training System on Negroamaro Wine Aroma.

Foods 2021 Feb 19;10(2). Epub 2021 Feb 19.

Council for Agricultural Research and Economics-Research Center for Viticulture and Enology (CREA-VE), Via Casamassima 148, 70010 Turi, Italy.

This study aimed to assess the impact of two soil managements and training systems on yield and wine aroma compounds of Negroamaro variety grown in a warm climate region (southern Italy). Cover crop (CC) and soil tillage (ST) as soil management, whilst bilateral Guyot (BG) and monolateral Guyot (MG) as training systems were compared. Free and bound volatile fractions were evaluated by GC-MS. ST and CC as well as BG and MG significantly affected yield parameters. In particular, yield was higher in ST and BG than in CC and MG, respectively; moreover, it was found to be positively influenced by interaction between BG and ST. Regarding aroma compounds, significant interactions between soil management and training system factors were observed. In case of free volatiles, the most positive interaction was found between BG and ST, whereas, for bound volatiles, the best interaction was represented by MG with both soil tillage and cover crop. Vine leaf area and development over vine growth stages along with water stress levels played an important role in determining the aroma profile as well as yield parameters. In conclusion, the training system significantly interacted with soil management and affected most of important aroma compounds in Negroamaro wine.
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http://dx.doi.org/10.3390/foods10020454DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7922071PMC
February 2021

Influence of Non- on Wine Chemistry: A Focus on Aroma-Related Compounds.

Molecules 2021 Jan 26;26(3). Epub 2021 Jan 26.

CNR-Institute of Sciences of Food Production (ISPA), via Prov. le, Lecce-Monteroni, 73100 Lecce, Italy.

Wine fermentation processes are driven by complex microbial systems, which comprise eukaryotic and prokaryotic microorganisms that participate in several biochemical interactions with the must and wine chemicals and modulate the organoleptic properties of wine. Among these, yeasts play a fundamental role, since they carry out the alcoholic fermentation (AF), converting sugars to ethanol and CO together with a wide range of volatile organic compounds. The contribution of , the reference organism associated with AF, has been extensively studied. However, in the last decade, selected strains received considerable commercial and oenological interest due to their specific pro-technological aptitudes and the positive influence on sensory quality. This review aims to highlight the inter-specific variability within the heterogeneous class of non- in terms of synthesis and release of volatile organic compounds during controlled AF in wine. In particular, we reported findings on the presence of model non- organisms, including spp. and , in combination with . The evidence is discussed from both basic and applicative scientific perspective. In particular, the oenological significance in different kind of wines has been underlined.
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http://dx.doi.org/10.3390/molecules26030644DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7865429PMC
January 2021

and Table Grapes: A Review of the Main Physical, Chemical, and Bio-Based Control Treatments in Post-Harvest.

Foods 2020 Aug 19;9(9). Epub 2020 Aug 19.

Department of the Sciences of Agriculture, Food and Environment, University of Foggia, Via Napoli 25, 71122 Foggia, Italy.

Consumers highly appreciate table grapes for their pleasant sensory attributes and as good sources of nutritional and functional compounds. This explains the rising market and global interest in this product. Along with other fruits and vegetables, table grapes are considerably perishable post-harvest due to the growth of undesired microorganisms. Among the microbial spoilers, represents a model organism because of its degrading potential and the huge economic losses caused by its infection. The present review provides an overview of the recent primary physical, chemical, and biological control treatments adopted against the development of in table grapes to extend shelf life. These treatments preserve product quality and safety. This article also focuses on the compliance of different approaches with organic and sustainable production processes. Tailored approaches include those that rely on controlled atmosphere and the application of edible coating and packaging, as well as microbial-based activities. These strategies, applied alone or in combination, are among the most promising solutions in order to prolong table grape quality during cold storage. In general, the innovative design of applications dealing with hurdle technologies holds great promise for future improvements.
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http://dx.doi.org/10.3390/foods9091138DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7555317PMC
August 2020

Effect of Co-Inoculation of , and for the Industrial Production of Negroamaro Wine in Apulia (Southern Italy).

Microorganisms 2020 May 13;8(5). Epub 2020 May 13.

Institute of Sciences of Food Production, National Research Council, 73100 Lecce, Italy.

The employment of multi-species starter cultures has growing importance in modern winemaking for improving the complexity and wine attributes. The assessment of compatibility for selected species/strains at the industrial-scale is crucial to assure the quality and the safety associated with fermentations. An aspect particularly relevant when the species belong to non-Saccharomyces, Saccharomyces spp. and malolactic bacteria, three categories with different biological characteristics and oenological significance. To the best of our knowledge, the present report is the first study regarding the utilization of a combined starter culture composed of three strains of non-Saccharomyces, Saccharomyces cerevisiae and Lactobacillus plantarum for production of wine at the industrial scale. More in-depth, this work investigated the oenological potential of the autochthonous characterized strains from the Apulian region (Southern Italy), Candida zemplinina (syn. Starmerella bacillaris) 35NC1, S. cerevisiae (NP103), and L. plantarum (LP44), in co-inoculation following a complete scale-up scheme. Microbial dynamics, fermentative profiles and production of volatile secondary compounds were assessed in lab-scale micro-vinification tests and then the performances of the mixed starter formulation were further evaluated by pilot-scale wine production. The above results were finally validated by performing an industrial-scale vinification on 100HL of Negroamaro cultivar grape must. The multi-starter formulation was able to rule the different stages of the fermentation processes effectively, and the different microbial combinations enhanced the organoleptic wine features to different extents. The findings indicated that the simultaneous inoculation of the three species affect the quality and quantity of several volatile compounds, confirming that the complexity of the wine can reflect the complexity of the starter cultures. Moreover, the results underlined that the same mixed culture could differently influence wine quality when tested at the lab-, pilot- and industrial-scale. Finally, we highlighted the significance of employment non-Saccharomyces and L. plantarum, together with S. cerevisiae, autochthonous strains in the design of custom-made starter culture formulation for typical regional wine production with pronounced unique quality.
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http://dx.doi.org/10.3390/microorganisms8050726DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7285497PMC
May 2020

New Insights into the Oenological Significance of : Impact of Selected Autochthonous Strains on the Volatile Profile of Apulian Wines.

Microorganisms 2020 Apr 26;8(5). Epub 2020 Apr 26.

CNR-Institute of Sciences of Food Production (ISPA), via Prov.le, Lecce-Monteroni, 73100 Lecce, Italy.

In this investigation, we explored the oenological significance of (syn. ) isolates from Apulian grape musts. Moreover, we provide the first evidence of the impact of different strains on the wine aromatic properties tested as monocultures. We described the diversity of strains isolated from grapes and the variability of 'volatile' phenotypes associated with this intraspecific variability. Thirty-three isolates were characterized at strain level by PCR-based approach and, among these, 16 strains were identified and then tested by microfermentation tests carried out in grape must. Analyzed strains were low producers of acetic acid and hydrogen sulphide, not able to decarboxylate a panel of representative amino acids, whereas they showed fructophilic character and significant glycerol production. Volatile profiles of produced wines were investigated by gas chromatography-mass spectrometry. The Odor Activity Values of all molecules were calculated and 12 compounds showed values above their odor thresholds. Two selected strains (35NC1 and 15PR1) could be considered as possible starter cultures since they were able to positively affect the sensory properties of obtained wine. This report firstly supplies evidence on the strain-specific impact of different strains on the final aroma of produced wines.
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http://dx.doi.org/10.3390/microorganisms8050628DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7285007PMC
April 2020

Autochthonous Starter Cultures Enhance Polyphenols Content, Antioxidant Activity, and Anti-Inflammatory Response of Apulian Red Wines.

Foods 2019 Oct 4;8(10). Epub 2019 Oct 4.

National Research Council-Institute of Sciences of Food Production (ISPA), via Prov. Lecce-Monteroni, 73100 Lecce, Italy.

Several biotic and abiotic factors can influence the amount of polyphenols in grape tissues. During vinification, the temperature, presence of grape seeds and peel, and use of enzymes, can influence the extraction of polyphenols. However, little information is available on the effects of yeast strains used in the polyphenolic composition of wine. With this aim, two selected Saccharomyces cerevisiae strains, ITEM 14093 and ITEM 14077 were used to produce wine from two Italian grape cultivars, Primitivo and Negroamaro. At the end of the alcoholic fermentation, the content of total polyphenols and of particular classes of polyphenols (stilbenes, phenolic acids, flavonols, and flavanols) and the antioxidant activity were evaluated and compared in the obtained wines. We also examined the effects of red wine extracts in a culture model of vascular inflammation. The results obtained comparing wine extracts obtained by utilizing commercial or autochthonous yeast showed that wines obtained with selected yeast significantly inhibited vascular inflammation. The results are positively directed towards the healthy properties of wine drinking.
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http://dx.doi.org/10.3390/foods8100453DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6836090PMC
October 2019

Exploitation of Prunus mahaleb fruit by fermentation with selected strains of Lactobacillus plantarum and Saccharomyces cerevisiae.

Food Microbiol 2019 Dec 8;84:103262. Epub 2019 Jul 8.

CNR, Institute of Sciences of Food Production (ISPA), via Prov.le Lecce-Monteroni, 73100, Lecce, Italy. Electronic address:

The organoleptic attributes of Prunus mahaleb, a fruit representing a new source of bioactive compounds, are so pronounced that it can be consider non-edible. This study was designed to evaluate the acceptance of P. mahaleb fruits after fermentation with different Saccharomyces cerevisiae and Lactobacillus plantarum protechnological strains. Four different bacterial and one yeast strains, as single or mixed starter formulation, were used to inoculate an aqueous suspension of P. mahaleb fruits. The fermented fruits and fermentation broths were subjected to physico-chemical characterization and the organoleptic properties of both samples were also assessed by a hedonic panel. The obtained results indicated that all the employed strains were able to grow and to ferment the matrix. However, the mixed starter FG69 + Li180-7 (L. plantarum/S. cerevisiae) had the best impact on sensory characteristics of P. mahaleb fruit and fermented medium. The adopted protocol allowed us to attain edible fruits and a new fermented non-dairy drink with valuable probiotic health-promoting properties. In our knowledge, this is the first study concerning the exploitation of P. mahaleb fruits. This investigation confirmed the potential of yeasts and lactic acid bacteria co-inoculation in the design of starter tailored for this kind of food applications.
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http://dx.doi.org/10.1016/j.fm.2019.103262DOI Listing
December 2019

Early Ongoing Speciation of Within the Grape Ecosystem Revealed by the Internal Variability Among the rDNA Operon Repeats.

Front Microbiol 2018 3;9:1687. Epub 2018 Aug 3.

Institute of Sciences of Food Production (ISPA), National Research Council (CNR), Lecce, Italy.

A yeast strain was isolated during a study on vineyard-associated yeast strains from Apulia in Southern Italy. ITS and LSU D1/D2 rDNA sequences showed this strain not to belong to any known species and was described as the type strain of , a close relative of . Several secondary peaks appeared in the sequences, suggesting internal heterogeneity among the copies of the rDNA. This hypothesis was tested by sequencing single clones of the marker region. The analyses showed different levels of variability throughout the operon with differences between the rRNA encoding genes and the internally transcribed regions. and share high frequency variants, i.e., variants frequently found in many clones, whereas there is a large variability of the low frequency polymorphisms, suggesting that the mechanism of homogenization is more active with the former than with the latter type of variation. These findings indicate that low frequency variants are detected in Sanger sequencing as secondary peaks whereas in Next Generation Sequencing (NGS) of metagenomics DNA would lead to an overestimate of the alpha diversity. For the first time in our knowledge, this investigation shed light on the variation of the copy number of the rDNA cistron during the yeast speciation process. These polymorphisms can be used to investigate on the processes occurring in these taxonomic markers during the separation of fungal species, it being a genetic process highly frequent in the complex microbial ecosystem existing in grape, must and wine.
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http://dx.doi.org/10.3389/fmicb.2018.01687DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6085423PMC
August 2018

Selection of indigenous yeast strains for the production of sparkling wines from native Apulian grape varieties.

Int J Food Microbiol 2018 Nov 5;285:7-17. Epub 2018 Jul 5.

Department of Sciences of Agriculture, Food and Environment, University of Foggia, Via Napoli 25, 71122 Foggia, Italy. Electronic address:

We report the first polyphasic characterization of native Saccharomyces cerevisiae in order to select candidate strains for the design of starter cultures tailored for Apulian sparkling wines obtained from local grape variety. In addition, it is the first survey in our region that propose the selection of autochthonous starter cultures for sparkling wine i) including a preliminary tailored genotypic and technological screening, and ii) monitoring analytical contribution during secondary fermentation in terms of volatile compounds (VOCs). Furthermore, we exploit the potential contribute of autochthonous cultures throughout the productive chain, including the possible improvement of base wine. One representative strain from each cluster was characterized i) for tolerance to abiotic and biotic stressors peculiar of sparkling wine fermentation, ii) for the performances in base wine production, and iii) for the aptitudes to promote in-bottle secondary fermentation in white and rosé sparkling wines, both obtained from Apulian grape varieties. Genetic characterization led to group 164 S. cerevisiae in 16 genetic clusters based on interdelta profiles. Stress tolerance assays shown a certain correlation with fermentative attitude. Our evidences demonstrated a different fermentative behavior and release of VOCs of the different strains in association with primary and secondary fermentations and as function of wine and rosé sparkling wine. Furthermore, performances in white/rosé sparkling wines have been found to be strain-dependent characters. Overall, we propose different strains as biotechnological resources suitable to improve the quality of regional sparkling wines and to provide a driver of innovation/segmentation in the market.
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http://dx.doi.org/10.1016/j.ijfoodmicro.2018.07.004DOI Listing
November 2018

Starter cultures as biocontrol strategy to prevent Brettanomyces bruxellensis proliferation in wine.

Appl Microbiol Biotechnol 2018 Jan 30;102(2):569-576. Epub 2017 Nov 30.

Department of Agriculture, Food and Environment Sciences, University of Foggia, Via Napoli 25, 471122, Foggia, Italy.

Brettanomyces bruxellensis is a common and significant wine spoilage microorganism. B. bruxellensis strains generally detain the molecular basis to produce compounds that are detrimental for the organoleptic quality of the wine, including some classes of volatile phenols that derive from the sequential bioconversion of specific hydroxycinnamic acids such as ferulate and p-coumarate. Although B. bruxellensis can be detected at any stage of the winemaking process, it is typically isolated at the end of the alcoholic fermentation (AF), before the staring of the spontaneous malolactic fermentation (MLF) or during barrel aging. For this reason, the endemic diffusion of B. bruxellensis leads to consistent economic losses in the wine industry. Considering the interest in reducing sulfur dioxide use during winemaking, in recent years, biological alternatives, such as the use of tailored selected yeast and bacterial strains inoculated to promote AF and MLF, are actively sought as biocontrol agents to avoid the "Bretta" character in wines. Here, we review the importance of dedicated characterization and selection of starter cultures for AF and MLF in wine, in order to reduce or prevent both growth of B. bruxellensis and its production of volatile phenols in the matrix.
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http://dx.doi.org/10.1007/s00253-017-8666-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5756568PMC
January 2018

Transcriptional Analysis of sp. Capable of Degrading Ochratoxin A.

Front Microbiol 2016 9;7:2162. Epub 2017 Jan 9.

Institute of Sciences of Food Production, National Research Council Bari, Italy.

Ochratoxin A (OTA) is a nephrotoxic and potentially carcinogenic mycotoxin produced by several species of and , contaminating grapes, wine and a variety of food products. We recently isolated from OTA contaminated soil vineyard a novel free-living strain of sp. , ITEM 17016, able to degrade OTA into the non-toxic catabolic product ochratoxin α. Biochemical studies suggested that the degradation reaction proceeds via peptide bond hydrolysis with phenylalanine (Phe) release. In order to identify genes responsible for OTA degradation we performed a differential gene expression analysis of ITEM 17016 grown in the presence or absence of the toxin. Among the differentially expressed genes, six peptidases up-regulated at 6 h were identified. The degrading activity of the carboxypeptidase PJ_1540 was confirmed in a heterologous system. The enrichment analysis for Gene Ontology terms confirmed that OTA degradation proceeds through peptidase activities and revealed the over-representation of pathways related to Phe catabolism. These results indicate that Phe may represent an energy source for this sp. strain and that OTA degrading reaction triggers the modulation of further catabolic activities.
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http://dx.doi.org/10.3389/fmicb.2016.02162DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5220012PMC
January 2017

Simultaneous Alcoholic and Malolactic Fermentations by Saccharomyces cerevisiae and Oenococcus oeni Cells Co-immobilized in Alginate Beads.

Front Microbiol 2016 14;7:943. Epub 2016 Jun 14.

Unità Operativa di Lecce, Consiglio Nazionale delle Ricerche - Istituto di Scienze delle Produzioni Alimentari Lecce, Italy.

Malolactic fermentation (MLF) usually takes place after the end of alcoholic fermentation (AF). However, the inoculation of lactic acid bacteria together with yeast starter cultures is a promising system to enhance the quality and safety of wine. In recent years, the use of immobilized cell systems has been investigated, with interesting results, for the production of different fermented foods and beverages. In this study we have carried out the simultaneous immobilization of Saccharomyces cerevisiae and Oenococcus oeni in alginate beads and used them in microvinifications tests to produce Negroamaro wine. The process was monitored by chemical and sensorial analyses and dominance of starters and cell leaking from beads were also checked. Co-immobilization of S. cerevisiae and O. oeni allowed to perform an efficient fermentation process, producing low volatile acidity levels and ethanol and glycerol concentrations comparable with those obtained by cell sequential inoculum and co-inoculum of yeast and bacteria cells in free form. More importantly, co-immobilization strategy produced a significant decrease of the time requested to complete AF and MLF. The immobilized cells could be efficiently reused for the wine fermentation at least three times without any apparent loss of cell metabolic activities. This integrated biocatalytic system is able to perform simultaneously AF and MLF, producing wines similar in organoleptic traits in comparison with wines fermented following traditional sequential AF and MLF with free cell starters. The immobilized-cell system, that we here describe for the first time in our knowledge, offers many advantages over conventional free cell fermentations, including: (i) elimination of non-productive cell growth phases; (ii) feasibility of continuous processing; (iii) re-use of the biocatalyst.
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http://dx.doi.org/10.3389/fmicb.2016.00943DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4906054PMC
July 2016

Viable But Not Culturable (VBNC) state of Brettanomyces bruxellensis in wine: New insights on molecular basis of VBNC behaviour using a transcriptomic approach.

Food Microbiol 2016 Oct 13;59:196-204. Epub 2016 Jun 13.

Department of Agricultural, Food and Environmental Sciences, University of Foggia, via Napoli 25, 71122 Foggia, Italy. Electronic address:

The spoilage potential of Brettanomyces bruxellensis in wine is strongly connected with the aptitude of this yeast to enter in a Viable But Non Culturable (VBNC) state when exposed to the harsh wine conditions. In this work, we characterized the VBNC behaviour of seven strains of B. bruxellensis representing a regional intraspecific biodiversity, reporting conclusive evidence for the assessment of VBNC as a strain-dependent character. The VBNC behaviour was monitored by fluorescein diacetate staining/flow cytometry for eleven days after addition of 0.4, 0.6, 0.8, 1 and 1.2 mg/L of molecular SO2 (entrance in the VBNC state) and after SO2 removal (exit from the VBNC state). Furthermore, one representative strain was selected and RNA-seq analysis performed after exposure to 1.2 mg/L SO2 and during the recovery phase. 30 and 1634 genes were identified as differentially expressed following VBNC entrance and 'resuscitation', respectively. The results reported strongly suggested that the entrance in the SO2-induced VBNC state in B. bruxellensis is associated with both, sulfite toxicity and oxidative stress response, confirming the crucial role of genes/proteins involved in redox cell homeostasis. Among the genes induced during recovery, the expression of genes involved in carbohydrate metabolism and encoding heat shock proteins, as well as enriched categories including amino acid transport and transporter activity was observed. The evidences of a general repression of genes involved in DNA replication suggest the occurrence of a true resuscitation of cell rather than a simple regrowth.
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http://dx.doi.org/10.1016/j.fm.2016.06.007DOI Listing
October 2016

Complexity and Dynamics of the Winemaking Bacterial Communities in Berries, Musts, and Wines from Apulian Grape Cultivars through Time and Space.

PLoS One 2016 14;11(6):e0157383. Epub 2016 Jun 14.

Institute of Biomembranes and Bioenergetics, CNR, Bari, Italy.

Currently, there is very little information available regarding the microbiome associated with the wine production chain. Here, we used an amplicon sequencing approach based on high-throughput sequencing (HTS) to obtain a comprehensive assessment of the bacterial community associated with the production of three Apulian red wines, from grape to final product. The relationships among grape variety, the microbial community, and fermentation was investigated. Moreover, the winery microbiota was evaluated compared to the autochthonous species in vineyards that persist until the end of the winemaking process. The analysis highlighted the remarkable dynamics within the microbial communities during fermentation. A common microbial core shared among the examined wine varieties was observed, and the unique taxonomic signature of each wine appellation was revealed. New species belonging to the genus Halomonas were also reported. This study demonstrates the potential of this metagenomic approach, supported by optimized protocols, for identifying the biodiversity of the wine supply chain. The developed experimental pipeline offers new prospects for other research fields in which a comprehensive view of microbial community complexity and dynamics is desirable.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0157383PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4907434PMC
July 2017

The Oenological Potential of Hanseniaspora uvarum in Simultaneous and Sequential Co-fermentation with Saccharomyces cerevisiae for Industrial Wine Production.

Front Microbiol 2016 9;7:670. Epub 2016 May 9.

Consiglio Nazionale delle Ricerche-Institute of Sciences of Food Production Lecce, Italy.

In oenology, the utilization of mixed starter cultures composed by Saccharomyces and non-Saccharomyces yeasts is an approach of growing importance for winemakers in order to enhance sensory quality and complexity of the final product without compromising the general quality and safety of the oenological products. In fact, several non-Saccharomyces yeasts are already commercialized as oenological starter cultures to be used in combination with Saccharomyces cerevisiae, while several others are the subject of various studies to evaluate their application. Our aim, in this study was to assess, for the first time, the oenological potential of H. uvarum in mixed cultures (co-inoculation) and sequential inoculation with S. cerevisiae for industrial wine production. Three previously characterized H. uvarum strains were separately used as multi-starter together with an autochthonous S. cerevisiae starter culture in lab-scale micro-vinification trials. On the basis of microbial development, fermentation kinetics and secondary compounds formation, the strain H. uvarum ITEM8795 was further selected and it was co- and sequentially inoculated, jointly with the S. cerevisiae starter, in a pilot scale wine production. The fermentation course and the quality of final product indicated that the co-inoculation was the better performing modality of inoculum. The above results were finally validated by performing an industrial scale vinification The mixed starter was able to successfully dominate the different stages of the fermentation process and the H. uvarum strain ITEM8795 contributed to increasing the wine organoleptic quality and to simultaneously reduce the volatile acidity. At the best of our knowledge, the present report is the first study regarding the utilization of a selected H. uvarum strain in multi-starter inoculation with S. cerevisiae for the industrial production of a wine. In addition, we demonstrated, at an industrial scale, the importance of non-Saccharomyces in the design of tailored starter cultures for typical wines.
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http://dx.doi.org/10.3389/fmicb.2016.00670DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4860541PMC
May 2016

Use of Lactobacillus plantarum Strains as a Bio-Control Strategy against Food-Borne Pathogenic Microorganisms.

Front Microbiol 2016 13;7:464. Epub 2016 Apr 13.

Department of Clinical and Experimental Medicine, University of Foggia Foggia, Italy.

Lactobacillus plantarum is one of the most versatile species extensively used in the food industry both as microbial starters and probiotic microorganisms. Several L. plantarum strains have been shown to produce different antimicrobial compounds such as organic acids, hydrogen peroxide, diacetyl, and also bacteriocins and antimicrobial peptides, both denoted by a variable spectrum of action. In recent decades, the selection of microbial molecules and/or bacterial strains able to produce antagonistic molecules to be used as antimicrobials and preservatives has been attracting scientific interest, in order to eliminate or reduce chemical additives, because of the growing attention of consumers for healthy and natural food products. The aim of this work was to investigate the antimicrobial activity of several food-isolated L. plantarum strains, analyzed against the pathogenic bacteria Listeria monocytogenes, Salmonella Enteritidis, Escherichia coli O157:H7 and Staphylococcus aureus. Antagonistic activity was assayed by agar spot test and revealed that strain L. plantarum 105 had the strongest ability to contrast the growth of L. monocytogenes, while strains L. plantarum 106 and 107 were the most active microorganisms against E. coli O157:H7. The antimicrobial ability was also screened by well diffusion assay and broth micro-dilution method using cell-free supernatants (CFS) from each Lactobacillus strain. Moreover, the chemical nature of the molecules released in the CFS, and possibly underlying the antagonistic activity, was preliminary characterized by exposure to different constraints such as pH neutralization, heating, catalase, and proteinase treatments. Our data suggest that the ability of L. plantarum cultures to contrast pathogens growth in vitro depends, at least in part, on a pH-lowering effect of supernatants and/or on the presence of organic acids. Cluster analysis was performed in order to group L. plantarum strains according to their antimicrobial effect. This study emphasizes the tempting use of the tested L. plantarum strains and/or their CFS as antimicrobial agents against food-borne pathogens.
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http://dx.doi.org/10.3389/fmicb.2016.00464DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4829616PMC
May 2016

Technological properties of Lactobacillus plantarum strains isolated from grape must fermentation.

Food Microbiol 2016 Aug 4;57:187-94. Epub 2016 Mar 4.

Dipartimento di Scienze Agrarie, degli Alimenti e dell'Ambiente, Università di Foggia, via Napoli 25, 71122, Foggia, Italy; Promis Biotech srl, via Napoli 25, 71122, Foggia, Italy.

Malolactic fermentation (MLF) is a secondary fermentation in wine that usually takes place during or at the end of alcoholic fermentation. Lactobacillus plantarum is able to conduct MLF (particularly under high pH conditions and in co-inoculation with yeasts), and some strains are commercially used as MLF starter cultures. Recent evidences suggest a further use of selected L. plantarum strains for the pre-alcoholic acidification of grape must. In this study, we have carried out an integrated (molecular, technological, and biotechnological) characterization of L. plantarum strains isolated from Apulian wines in order to combine the two protechnological features (MLF performances and must acidification aptitudes). Several parameters such as sugar, pH and ethanol tolerance, resistance to lyophilisation and behaviour in grape must were evaluated. Moreover, the expression of stress gene markers was investigated and was linked to the ability of L. plantarum strains to grow and perform MLF. Co-inoculation of Saccharomyces cerevisiae and L. plantarum in grape must improves the bacterial adaptation to harsh conditions of wine and reduced total fermentation time. For the first time, we applied a polyphasic approach for the characterization of L. plantarum in reason of the MLF performances. The proposed procedure can be generalized as a standard method for the selection of bacterial resources for the design of MLF starter cultures tailored for high pH must.
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http://dx.doi.org/10.1016/j.fm.2016.03.002DOI Listing
August 2016

From grape berries to wine: population dynamics of cultivable yeasts associated to "Nero di Troia" autochthonous grape cultivar.

World J Microbiol Biotechnol 2016 Apr 29;32(4):59. Epub 2016 Feb 29.

Dipartimento di Scienze Agrarie, degli Alimenti e dell'Ambiente, Università di Foggia, Via Napoli 25, 71100, Foggia, Italy.

The aim of this work was to study the biodiversity of yeasts isolated from the autochthonous grape variety called "Uva di Troia", monitoring the natural diversity from the grape berries to wine during a vintage. Grapes were collected in vineyards from two different geographical areas and spontaneous alcoholic fermentations (AFs) were performed. Different restriction profiles of ITS-5.8S rDNA region, corresponding to Saccharomyces cerevisiae, Issatchenkia orientalis, Metschnikowia pulcherrima, Hanseniaspora uvarum, Candida zemplinina, Issatchenkia terricola, Kluyveromyces thermotolerans, Torulaspora delbrueckii, Metschnikowia chrysoperlae, Pichia fermentans, Hanseniaspora opuntiae and Hanseniaspora guilliermondii, were observed. The yeast occurrences varied significantly from both grape berries and grape juices, depending on the sampling location. Furthermore, samples collected at the end of AF revealed the great predominance of Saccharomyces cerevisiae, with a high intraspecific biodiversity. This is the first report on the population dynamics of 'cultivable' microbiota diversity of "Uva di Troia" cultivar from the grape to the corresponding wine ("Nero di Troia"), and more general for Southern Italian oenological productions, allowing us to provide the basis for an improved management of wine yeasts (with both non-Saccharomyces and Saccharomyces) for the production of typical wines with desired unique traits. A certain geographical-dependent variability has been reported, suggesting the need of local based formulation for autochthonous starter cultures, especially in the proportion of the different species/strains in the design of mixed microbial preparations.
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http://dx.doi.org/10.1007/s11274-016-2017-4DOI Listing
April 2016

Biodegradation of Ochratoxin A by Bacterial Strains Isolated from Vineyard Soils.

Toxins (Basel) 2015 Nov 27;7(12):5079-93. Epub 2015 Nov 27.

Institute of Sciences of Food Production, National Research Council, Unit of Lecce, via Provinciale Lecce-Monteroni, Lecce 73100, Italy.

Ochratoxin A (OTA) is a mycotoxin with a main nephrotoxic activity contaminating several foodstuffs. In the present report, five soil samples collected from OTA-contaminated vineyards were screened to isolate microorganisms able to biodegrade OTA. When cultivated in OTA-supplemented medium, OTA was converted in OTα by 225 bacterial isolates. To reveal clonal relationships between isolates, molecular typing by using an automated rep-PCR system was carried out, thus showing the presence of 27 different strains (rep-PCR profiles). The 16S-rRNA gene sequence analysis of an isolate representative of each rep-PCR profiles indicated that they belonged to five bacterial genera, namely Pseudomonas, Leclercia, Pantoea, Enterobacter, and Acinetobacter. However, further evaluation of OTA-degrading activity by the 27 strains revealed that only Acinetobacter calcoaceticus strain 396.1 and Acinetobacter sp. strain neg1, consistently conserved the above property; their further characterization showed that they were able to convert 82% and 91% OTA into OTα in six days at 24 °C, respectively. The presence of OTα, as the unique OTA-degradation product was confirmed by LC-HRMS. This is the first report on OTA biodegradation by bacterial strains isolated from agricultural soils and carried out under aerobic conditions and moderate temperatures. These microorganisms might be used to detoxify OTA-contaminated feed and could be a new source of gene(s) for the development of a novel enzymatic detoxification system.
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http://dx.doi.org/10.3390/toxins7124864DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4690114PMC
November 2015

Functional Properties of Grape and Wine Polyphenols.

Plant Foods Hum Nutr 2015 Dec;70(4):454-62

Istituto di Scienze delle Produzioni Alimentari-CNR, Unit of Lecce, via Monteroni, 73100, Lecce, Italy.

Grape berries polyphenols are mainly synthesized in the skin tissues and seeds and they are extracted during the winemaking process. These substances have a potentially positive effect, on human health, thus giving to grape and red wine "functional properties" that can contribute to prevent a number of human illness. Nevertheless, the research community is showing that the real effect is a result of a combination of different factors, notably daily intake, bioavailability, or in vivo antioxidant activity that are yet to be resolved. Viticulture and winemaking practices, determine the concentration of polyphenols in grape and wine. To date, reduced knowledge is existing on the effects of different yeast strains on the final concentration of polyphenols in red wine. We summarize the recent findings concerning the effects of polyphenols on human chronic disease and the future directions for research to increase the amount of these compounds in wine.
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http://dx.doi.org/10.1007/s11130-015-0518-1DOI Listing
December 2015

Microbial terroir and food innovation: The case of yeast biodiversity in wine.

Microbiol Res 2015 Dec 19;181:75-83. Epub 2015 Oct 19.

Dipartimento di Scienze Agrarie, degli Alimenti e dell'Ambiente, Università di Foggia, Foggia, Italy. Electronic address:

Saccharomyces and non-Saccharomyces represents a heterogeneous class in the grape/must/wine environments including several yeast genera (e.g., Saccharomyces, Hanseniaspora, Pichia, Candida, Metschnikowia, Kluyveromyces, Zygosaccharomyces, Torulaspora, Dekkera and Schizosaccharomyces) and species. Since, each species may differently contribute to the improvement/depreciation of wine qualities, it appears clear the reason why species belong to non-Saccharomyces are also considered a biotechnological resource in wine fermentation. Here, we briefly review the oenological significance of this specific part of microbiota associated with grapes/musts/wine. Moreover, the diversity of cultivable non-Saccharomyces genera and their contribute to typical wines fermentations will be discussed.
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http://dx.doi.org/10.1016/j.micres.2015.10.005DOI Listing
December 2015

New process for production of fermented black table olives using selected autochthonous microbial resources.

Front Microbiol 2015 24;6:1007. Epub 2015 Sep 24.

Consiglio Nazionale delle Ricerche-Istituto di Scienze delle Produzioni Alimentari, Unità Operativa di Lecce Lecce, Italy.

Table olives represent one important fermented product in Europe and, in the world, their demand is constantly increasing. At the present time, no systems are available to control black table olives spontaneous fermentation by the Greek method. During this study, a new protocol for the production of black table olives belonging to two Italian (Cellina di Nardò and Leccino) and two Greek (Kalamàta and Conservolea) cultivars has been developed: for each table olive cultivar, starter-driven fermentations were performed inoculating, firstly, one selected autochthonous yeast starter and, subsequently, one selected autochthonous LAB starter. All starters formulation were able to dominate fermentation process. The olive fermentation was monitored using specific chemical descriptors able to identify a first stage (30 days) mainly characterized by aldehydes; a second period (60 days) mainly characterized by higher alcohols, styrene and terpenes; a third fermentation stage represented by acetate esters, esters and acids. A significant decrease of fermentation time (from 8 to 12 months to a maximum of 3 months) and an significant improvement in organoleptic characteristics of the final product were obtained. This study, for the first time, describes the employment of selected autochthonous microbial resources optimized to mimic the microbial evolution already recorded during spontaneous fermentations.
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http://dx.doi.org/10.3389/fmicb.2015.01007DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4585182PMC
October 2015

Physico-chemical characterization of natural fermentation process of Conservolea and Kalamàta table olives and developement of a protocol for the pre-selection of fermentation starters.

Food Microbiol 2015 Apr 6;46:368-382. Epub 2014 Sep 6.

Consiglio Nazionale delle Ricerche, Istituto di Scienze delle Produzioni Alimentari, Bari, Italy.

Table olives are one of the most important traditional fermented vegetables in Europe and their world consumption is constantly increasing. Conservolea and Kalamàta are the most important table olives Greek varieties. In the Greek system, the final product is obtained by spontaneous fermentations, without any chemical debittering treatment. This natural fermentation process is not predictable and strongly influenced by the physical-chemical conditions and by the presence of microorganisms contaminating the olives. Natural fermentations of Conservolea and Kalamàta cultivars black olives were studied in order to determine microbiological, biochemical and chemical evolution during the process. Following the process conditions generally used by producers, in both cultivars, yeasts were detected throughout the fermentation, whereas lactic acid bacteria (LAB) appeared in the last staged of the process. A new optimized specific protocol was developed to select autochthonous yeast and LAB isolates that can be good candidates as starters. These microorganisms were pre-selected for their ability to adapt to model brines, to have beta-glucosidase activity, not to produce biogenic amines. Chemical compounds deriving by microbiological activities and associated to the three different phases (30, 90 and 180 days) of the fermentation process were identified and were proposed as chemical descriptors to follow the fermentation progress.
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http://dx.doi.org/10.1016/j.fm.2014.08.021DOI Listing
April 2015

Physico-chemical and microbiological characterization of spontaneous fermentation of Cellina di Nardò and Leccino table olives.

Front Microbiol 2014 28;5:570. Epub 2014 Oct 28.

Consiglio Nazionale delle Ricerche- Istituto di Scienze delle Produzioni Alimentari Bari, Italy.

Table olives are one of the most important traditional fermented vegetables in Europe and their world consumption is constantly increasing. In the Greek style, table olives are obtained by spontaneous fermentations, without any chemical debittering treatment. Evolution of sugars, organic acids, alcohols, mono, and polyphenol compounds and volatile compounds associated with the fermentative metabolism of yeasts and bacteria throughout the natural fermentation process of the two Italian olive cultivars Cellina di Nardò and Leccino were determined. A protocol was developed and applied aimed at the technological characterization of lactic acid bacteria (LAB) and yeast strains as possible candidate autochthonous starters for table olive fermentation from Cellina di Nardò and Leccino cultivars. The study of the main physic-chemical parameters and volatile compounds during fermentation helped to determine chemical descriptors that may be suitable for monitoring olive fermentation. In both the analyzed table olive cultivars, aldehydes proved to be closely related to the first stage of fermentation (30 days), while higher alcohols (2-methyl-1-propanol; 3-methyl-1-butanol), styrene, and o-cymene were associated with the middle stage of fermentation (90 days) and acetate esters with the final step of olive fermentation (180 days).
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http://dx.doi.org/10.3389/fmicb.2014.00570DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4211558PMC
November 2014

Molecular and Technological Characterization of Saccharomyces cerevisiae Strains Isolated from Natural Fermentation of Susumaniello Grape Must in Apulia, Southern Italy.

Int J Microbiol 2014 9;2014:897428. Epub 2014 Jan 9.

C.N.R.-Istituto di Scienze delle Produzioni Alimentari, Unità Operativa di Lecce, Via Provinciale Lecce-Monteroni, 73100 Lecce, Italy.

The characterization of autochthonous Saccharomyces cerevisiae strains is an important step towards the conservation and employment of microbial biodiversity. The utilization of selected autochthonous yeast strains would be a powerful tool to enhance the organoleptic and sensory properties of typical regional wines. In fact, indigenous yeasts are better tailored to a particular must and because of this they are able to praise the peculiarities of the derived wine. The present study described the biodiversity of indigenous S. cerevisiae strains isolated from natural must fermentations of an ancient and recently rediscovered Apulian grape cultivar, denoted as "Susumaniello." The yeast strains denoted by the best oenological and technological features were identified and their fermentative performances were tested by either laboratory assay. Five yeast strains showed that they could be excellent candidates for the production of industrial starter cultures, since they dominated the fermentation process and produced wines characterized by peculiar oenological and organoleptic features.
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http://dx.doi.org/10.1155/2014/897428DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3942102PMC
March 2014

Construction of a laccase chimerical gene: recombinant protein characterization and gene expression via yeast surface display.

Appl Biochem Biotechnol 2014 Mar 24;172(6):2916-31. Epub 2014 Jan 24.

CNR - Istituto di Scienze delle Produzioni Alimentari (ISPA), via Lecce-Monteroni, Lecce, 73100, Italy.

The ERY4 laccase gene from Pleurotus eryngii was expressed in Saccharomyces cerevisiae and the recombinant laccase resulted to be not biologically active. This gene was thus modified to obtain chimerical enzymes derived from the substitution of N-, C- and both N- and C-terminal regions with the corresponding regions of Ery3 laccase, another laccase isoform of P. eryngii. The chimerical isoform named 4NC3, derived from the substitution of both N- and C-terminal regions, showed the best performances in terms of enzymatic activities, affinities for different substrates and stability at a broad range of temperatures and pHs. The chimerical 4NC3 laccase isoform was displayed on the cell surface of S. cerevisiae using the N-terminal fusion with either the Pir2 or the Flo1 S. cerevisiae proteins as anchor attachment sequence. Immunofluorescence microscopy and Western blot analyses confirmed the localization of 4NC3 on the yeast cell surface. The enzyme activity on specific laccase substrates revealed that 4NC3 laccase was immobilized in active form on the cell surface. To our knowledge, this is the first example of expression of a chimerical fungal laccase by yeast cell display.
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http://dx.doi.org/10.1007/s12010-014-0734-4DOI Listing
March 2014

Biodiversity and safety aspects of yeast strains characterized from vineyards and spontaneous fermentations in the Apulia Region, Italy.

Food Microbiol 2013 Dec 16;36(2):335-42. Epub 2013 Jul 16.

CNR-Institute of Sciences of Food Production (ISPA), Operative Unit of Lecce, Lecce, Italy.

This work is the first large-scale study on vineyard-associated yeast strains from Apulia (Southern Italy). Yeasts were identified by Internal Transcribed Spacer (ITS) ribotyping and bioinformatic analysis. The polymorphism of interdelta elements was used to differentiate Saccharomyces cerevisiae strains. Twenty different species belonging to 9 genera were identified. Predominant on the grape surface were Metschnikowia pulcherrima, Hanseniaspora uvarum and Aureobasidium pullulans, whereas M. pulcherrima and H. uvarum were dominant in the early fermentation stage. A total of 692 S. cerevisiae isolates were identified and a number of S. cerevisiae strains, ranging from 26 to 55, was detected in each of the eight fermentations. The strains were tested for biogenic amines (BAs) production, either in synthetic media or grape must. Two Pichia manshurica, an Issatchenkia terricola and a M. pulcherrima strains were able to produce histamine and cadaverine, during must fermentation. The production of BAs in wine must was different than that observed in the synthetic medium. This feature indicate the importance of an "in grape must" assessment of BAs producing yeast. Overall, our results suggest the importance of microbiological control during wine-making to reduce the potential health risk for consumer represented by these spoilage yeasts.
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http://dx.doi.org/10.1016/j.fm.2013.07.001DOI Listing
December 2013

Microdroplet-based multiplex PCR on chip to detect foodborne bacteria producing biogenic amines.

Food Microbiol 2013 Aug 1;35(1):10-4. Epub 2013 Mar 1.

Center for Biomolecular Nanotechnologies, Fondazione Istituto Italiano di Tecnologia @UniLe, via Barsanti 1, I-73010 Arnesano, Lecce, Italy.

The development of fast, reliable and culture-independent molecular tools to detect bacteria producing biogenic amines deserves the attention of research and ultimately of the food industry in order to protect consumers' health. Here we present the application of a simple, low-cost, fast and sensitive method to perform microdroplet-based multiplex PCR, directly on a food matrix, for the simultaneous detection of bacterial genes involved in biogenic amine biosynthesis. After inoculating wine with Lactobacillus brevis IOEB 9809, cell lysis and DNA amplification are performed in one single step, without preliminary nucleic acid extraction or purification treatments. The assay is performed in about 30 min, requiring 150 nL of starting sample and it enables the detection of down to 15 bacterial cells. With respect to traditional culture techniques, the speed, the simplicity and the cheapness of this procedure allow an effective monitoring of microbial cells during food-making and processing.
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http://dx.doi.org/10.1016/j.fm.2013.02.010DOI Listing
August 2013

AtSYP51/52 functions diverge in the post-Golgi traffic and differently affect vacuolar sorting.

Mol Plant 2013 May 19;6(3):916-30. Epub 2012 Oct 19.

Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, University of Salento, Campus Ecotekne, 73100 Lecce, Italy.

Plant sensitive factor attachment protein receptors (SNAREs) encoded by genes of the same sub-family are generally considered as redundant in promoting vesicle-associated membrane fusion events. Nonetheless, the application of innovative experimental approaches highlighted that members of the same gene sub-family often have different functional specificities. In this work, two closely related Qc-SNAREs--the AtSYP51 and the AtSYP52--are compared in their ability to influence different secretory pathways. Their role in the vesicle sorting to the central vacuole has been revised and they were found to have a novel inhibitory function. When transiently overexpressed, the SYP51 and the SYP52 distributed between the TGN and the tonoplast. Our data demonstrate that these SYPs (syntaxin of plants) act as t-SNARE when present on the membrane of TGN/PVC, whereas they behave as inhibitory or interfering SNAREs (i-SNAREs) when they accumulate on the tonoplast. Moreover, the performed functional analysis indicated that the AtSYP51 and the AtSYP52 roles differ in the traffic to the vacuole. The findings are a novel contribution to the functional characterization of plant SNAREs that reveals additional non-fusogenic roles.
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http://dx.doi.org/10.1093/mp/sss117DOI Listing
May 2013

Role of the C-terminus of Pleurotus eryngii Ery4 laccase in determining enzyme structure, catalytic properties and stability.

Protein Eng Des Sel 2013 Jan 20;26(1):1-13. Epub 2012 Sep 20.

CNR-Istituto di Scienze delle Produzioni Alimentari (ISPA), Lecce, Italy.

The ERY4 laccase gene of Pleurotus eryngii is not biologically active when expressed in yeast. To explain this finding, we analysed the role of the C-terminus of Ery4 protein by producing a number of its different mutant variants. Two different categories of ERY4 mutant genes were produced and expressed in yeast: (i) mutants carrying C-terminal deletions and (ii) mutants carrying different site-specific mutations at their C-terminus. Investigation of the catalytic properties of the recombinant enzymes indicated that each novel variant acquired different affinities and catalytic activity for various substrates. Our results highlight that C-terminal processing is fundamental for Ery4 laccase enzymatic activities allowing substrate accessibility to the enzyme catalytic core. Apparently, the last 18 amino acids in the C-terminal end of the Ery4 laccase play a critical role in enzyme activity, stability and kinetic and, in particular biochemical and structural data indicate that the K532 residue is fundamental for enzyme activation. These studies shed light on the structure/function relationships of fungal laccases and will enhance the development of biotechnological strategies for the industrial exploitation of these enzymes.
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http://dx.doi.org/10.1093/protein/gzs056DOI Listing
January 2013
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