Giovanni Mita - Academia.edu (original) (raw)

Papers by Giovanni Mita

Frontiers in Microbiology

International Journal of Systematic and Evolutionary Microbiology

Molecular Biology Reports, 2011

Applied Microbiology and Biotechnology - APPL MICROBIOL BIOTECHNOL, 2008

A full length cDNA encoding an extracellular laccase was isolated by reverse transcription polyme... more A full length cDNA encoding an extracellular laccase was isolated by reverse transcription polymerase chain reaction from the mycelia of the mushroom Pleurotus eryngii. The isolated sequence, denoted Ery3, encodes for a mature laccase isoenzyme of 531 amino acid residues with a predicted molecular weight of 56.6 kDa. All sequence motifs, being the signature sequences used to identify the laccases, were found in the Ery3 protein sequence. The Ery3 cDNA was expressed in Saccharomyces cerevisiae and the effects of copper concentration and cultivation temperature were investigated. S. cerevisiae cells were immobilized in calcium alginate gel and the optimal immobilization parameters for the enhanced production of laccase were determined. The immobilization was most effective with 3% sodium alginate, 0.1 M calcium chloride and an initial biomass of 4.5 × 108 cells. The enzyme yield obtained with immobilized cells (139 mU ml−1) showed a 1.6-fold increase compared to the highest yield obtained with free cells. The alginate beads showed good stability and retained 84% capacity of enzyme production after seven repeated cycles of batch fermentation. The immobilization system proved to increase the proteolytic stability of the recombinant Ery3 protein. To our knowledge, this is the first report on S. cerevisiae whole-cell immobilization for recombinant laccase production.

Environmental and Experimental Botany, 2009

Annals of Microbiology, 2011

Plant Cell Reports, 1986

Nineteen chlorate-resistant variants were isolated after mutagenesis from cells of Medicago coeru... more Nineteen chlorate-resistant variants were isolated after mutagenesis from cells of Medicago coerulea. The level of nitrate reductase activity was variable in these lines and ranged from 100% to less than 5% of the wild type level. Xanthine dehydrogenase was not affected in any of those variants tested. Methylammonium-resistant variants were also isolated from the same type of cells. They show a different regulation of nitrogen utilization. In particular, the enzymatic level of nitrate reductase which, in wild type cells, is sensitive to ammonium repression, is much less affected in the variants. Differences were also seen in the regulation of other functions of the nitrogen-utilizing pathway: xanthine dehydrogenase and, possibly, adenine uptake.

Journal of Biotechnology, 2010

Plant Biology, 2010

Plants respond to environmental stimuli, such as heat shock, by re-programming cellular activity ... more Plants respond to environmental stimuli, such as heat shock, by re-programming cellular activity through differential gene expression, mainly controlled at the transcription level. The current study refers to two sunflower small heat shock protein (sHSP) genes arranged in tandem in head-to-head orientation and linked by a 3809 bp region. These genes exhibit only slight structural differences in the coding portion. They code for cytosolic class I sHSPs and are named HaHSP17.6a and HaHSP17.6b according to the molecular weight of the putative proteins. The genomic organization of these genes is consistent with the idea that many HSP genes originate from duplication events; in this case, probably an inversion and duplication occurred. The HaHSP17.6a and HaHSP17.6b genes are characterized by different expression levels under various heat stress conditions; moreover, their expression is differently induced by various elicitors. The differential regulation observed for HaHSP17.6a and HaHSP17.6b genes differs from previous observations on duplicated sHSP genes in plants.

Plant Biosystems, 2011

The objective of this study was to investigate at the molecular level the heat shock response in ... more The objective of this study was to investigate at the molecular level the heat shock response in olive by identifying sequences coding for heat shock proteins (HSPs). Young twigs of Olea europaea trees (cv Cellina di Nardò) were subjected to different temperature treatments in order to induce the expression of heat shock genes. In order to identify genes induced by heat treatment, we used a PCR-based approach to amplify specific HS cDNAs. Search for low molecular weight HSP sequences was performed in public domain databases in order to design specific primers based upon multisequence alignments. By this approach, we isolated the first full length cDNA encoding a low-molecular weight HSP from O. europaea. This is a class I low molecular weight HSP of 18.3 kDa, which is highly expressed in young twigs subjected to heat stress.

Bioresource Technology, 2011

The yeast population dynamics in olive wastewaters (OMW), sampled in five mills from Salento (Apu... more The yeast population dynamics in olive wastewaters (OMW), sampled in five mills from Salento (Apulia, Southern Italy), were investigated. Three hundred yeasts were isolated in five industrial mills and identified by molecular analysis. Strains belonging to Geotrichum, Saccharomyces, Pichia, Rhodotorula and Candida were detected. Five G. candidum strains were able to grow in OMW as the sole carbon source and to reduce phenolics, chemical oxygen demand (COD) and antimicrobial compounds. One G. candidum isolate was selected for whole-cell immobilization in calcium alginate gel. The COD and phenolic reduction obtained with immobilized cells showed a 2.2- and 2-fold increase compared to the removal obtained with free cells, respectively. The immobilization system enhanced yeast oxidative activity by avoiding the presence of microbial protease in treated OMW. To our knowledge, this is the first report on G. candidum whole-cell immobilization for OMW bioremediation.

Plant Cell Reports, 1997

Sunflower suspension cell cultures were subjected to different heat treatments and the electropho... more Sunflower suspension cell cultures were subjected to different heat treatments and the electrophoretic patterns of heat-induced endocellular and secreted proteins were analyzed. In response to heat shock (3 h at 40°C), sunflower cells synthesized new polypeptides and secreted them into the medium, while the synthesis of other polypeptides was suppressed. Two major polypeptides of about 50 and 32 kDa were strongly induced. The two-dimensional electrophoretic analysis showed that the 32-kDa band is composed of at least four different polypeptides. Western blotting hybridizations of secreted proteins with various lectins were performed. The 32-kDa band gave a positive signal with concanavalin A.

Food Biophysics, 2008

In this work, purification of lactoferrin from whey was performed with high recovery rate. Lactof... more In this work, purification of lactoferrin from whey was performed with high recovery rate. Lactoferrin was then exploited in the preparation of food emulsions. Two tertiary emulsions, formed by olive oil, lecithin, chitosan, and lactoferrin, were compared: both the emulsions showed similar turbidity and stability. In the secondary emulsion formed by oil/lecithin/chitosan, the pH was increased to 9 before addition of lactoferrin. Then, lactoferrin was added, and the pH was stabilized above pH 9. Lactoferrin was found in amounts of 1 to 2.5 mg/ml in the multiple experiments. A fraction of the added lactoferrin was also present in a milky layer above the emulsion layer. This was, to our knowledge, the first study of emulsions made exploiting the interactions between lactoferrin and chitosan. It was noted that chitosan droplets remained soluble, although the hydrocolloid solubility occurs at pH lower than 5.9. These results showed the feasibility of manufacturing lactoferrin-based emulsions as functional foods.

Food Additives and Contaminants Part A-chemistry Analysis Control Exposure & Risk Assessment, 2005

Journal of Cereal Science, 1999

Journal of Cereal Science, 1998

High temperatures during grain filling are considered one of the factors that can modify dough pr... more High temperatures during grain filling are considered one of the factors that can modify dough properties and quality in wheat. In this study we analysed four Italian wheat cultivars grown under different temperature conditions to study the influence of high temperature on storage-protein-gene expression. Plants were grown both in the field and in growth cabinets, and were subjected to different thermal regimes. PolyA+ mRNAs were extracted from control and stressed plants at different stages of kernel development. Northern blot hybridisations were performed using probes for storage and heat shock proteins to monitor the expression of the relative genes under different temperature conditions. Northern analyses, performed using storage protein probes, indicated that temperature variation does not influence the synthesis of any of the storage protein mRNAs. On the contrary, the hybridisation signals obtained using heat shock probes were more intense in the stressed samples, indicating that the expression of heat shock genes is modulated by the temperature variation.

Food Additives and Contaminants Part A-chemistry Analysis Control Exposure & Risk Assessment, 2005

International Journal of Microbiology, 2014

Plant Biology, 2010

Artemisia annua L. is a herb traditionally used for treatment of fevers. The glandular trichomes ... more Artemisia annua L. is a herb traditionally used for treatment of fevers. The glandular trichomes of this plant accumulate, although at low levels, artemisinin, which is highly effective against malaria. Due to the great importance of this compound, many efforts have been made to improve knowledge on artemisinin production both in plants and in cell cultures. In this study, A. annua suspension cultures were established in order to investigate the effects of methyl jasmonate (MeJA) and miconazole on artemisinin biosynthesis. Twenty-two micro molar MeJA induced a three-fold increase of artemisinin production in around 30 min; while 200 μm miconazole induced a 2.5-fold increase of artemisinin production after 24 h, but had severe effects on cell viability. The influence of these treatments on expression of biosynthetic genes was also investigated. MeJA induced up-regulation of CYP71AV1, while miconazole induced up-regulation of CPR and DBR2.

Frontiers in Microbiology

International Journal of Systematic and Evolutionary Microbiology

Molecular Biology Reports, 2011

Applied Microbiology and Biotechnology - APPL MICROBIOL BIOTECHNOL, 2008

A full length cDNA encoding an extracellular laccase was isolated by reverse transcription polyme... more A full length cDNA encoding an extracellular laccase was isolated by reverse transcription polymerase chain reaction from the mycelia of the mushroom Pleurotus eryngii. The isolated sequence, denoted Ery3, encodes for a mature laccase isoenzyme of 531 amino acid residues with a predicted molecular weight of 56.6 kDa. All sequence motifs, being the signature sequences used to identify the laccases, were found in the Ery3 protein sequence. The Ery3 cDNA was expressed in Saccharomyces cerevisiae and the effects of copper concentration and cultivation temperature were investigated. S. cerevisiae cells were immobilized in calcium alginate gel and the optimal immobilization parameters for the enhanced production of laccase were determined. The immobilization was most effective with 3% sodium alginate, 0.1 M calcium chloride and an initial biomass of 4.5 × 108 cells. The enzyme yield obtained with immobilized cells (139 mU ml−1) showed a 1.6-fold increase compared to the highest yield obtained with free cells. The alginate beads showed good stability and retained 84% capacity of enzyme production after seven repeated cycles of batch fermentation. The immobilization system proved to increase the proteolytic stability of the recombinant Ery3 protein. To our knowledge, this is the first report on S. cerevisiae whole-cell immobilization for recombinant laccase production.

Environmental and Experimental Botany, 2009

Annals of Microbiology, 2011

Plant Cell Reports, 1986

Nineteen chlorate-resistant variants were isolated after mutagenesis from cells of Medicago coeru... more Nineteen chlorate-resistant variants were isolated after mutagenesis from cells of Medicago coerulea. The level of nitrate reductase activity was variable in these lines and ranged from 100% to less than 5% of the wild type level. Xanthine dehydrogenase was not affected in any of those variants tested. Methylammonium-resistant variants were also isolated from the same type of cells. They show a different regulation of nitrogen utilization. In particular, the enzymatic level of nitrate reductase which, in wild type cells, is sensitive to ammonium repression, is much less affected in the variants. Differences were also seen in the regulation of other functions of the nitrogen-utilizing pathway: xanthine dehydrogenase and, possibly, adenine uptake.

Journal of Biotechnology, 2010

Plant Biology, 2010

Plants respond to environmental stimuli, such as heat shock, by re-programming cellular activity ... more Plants respond to environmental stimuli, such as heat shock, by re-programming cellular activity through differential gene expression, mainly controlled at the transcription level. The current study refers to two sunflower small heat shock protein (sHSP) genes arranged in tandem in head-to-head orientation and linked by a 3809 bp region. These genes exhibit only slight structural differences in the coding portion. They code for cytosolic class I sHSPs and are named HaHSP17.6a and HaHSP17.6b according to the molecular weight of the putative proteins. The genomic organization of these genes is consistent with the idea that many HSP genes originate from duplication events; in this case, probably an inversion and duplication occurred. The HaHSP17.6a and HaHSP17.6b genes are characterized by different expression levels under various heat stress conditions; moreover, their expression is differently induced by various elicitors. The differential regulation observed for HaHSP17.6a and HaHSP17.6b genes differs from previous observations on duplicated sHSP genes in plants.

Plant Biosystems, 2011

The objective of this study was to investigate at the molecular level the heat shock response in ... more The objective of this study was to investigate at the molecular level the heat shock response in olive by identifying sequences coding for heat shock proteins (HSPs). Young twigs of Olea europaea trees (cv Cellina di Nardò) were subjected to different temperature treatments in order to induce the expression of heat shock genes. In order to identify genes induced by heat treatment, we used a PCR-based approach to amplify specific HS cDNAs. Search for low molecular weight HSP sequences was performed in public domain databases in order to design specific primers based upon multisequence alignments. By this approach, we isolated the first full length cDNA encoding a low-molecular weight HSP from O. europaea. This is a class I low molecular weight HSP of 18.3 kDa, which is highly expressed in young twigs subjected to heat stress.

Bioresource Technology, 2011

The yeast population dynamics in olive wastewaters (OMW), sampled in five mills from Salento (Apu... more The yeast population dynamics in olive wastewaters (OMW), sampled in five mills from Salento (Apulia, Southern Italy), were investigated. Three hundred yeasts were isolated in five industrial mills and identified by molecular analysis. Strains belonging to Geotrichum, Saccharomyces, Pichia, Rhodotorula and Candida were detected. Five G. candidum strains were able to grow in OMW as the sole carbon source and to reduce phenolics, chemical oxygen demand (COD) and antimicrobial compounds. One G. candidum isolate was selected for whole-cell immobilization in calcium alginate gel. The COD and phenolic reduction obtained with immobilized cells showed a 2.2- and 2-fold increase compared to the removal obtained with free cells, respectively. The immobilization system enhanced yeast oxidative activity by avoiding the presence of microbial protease in treated OMW. To our knowledge, this is the first report on G. candidum whole-cell immobilization for OMW bioremediation.

Plant Cell Reports, 1997

Sunflower suspension cell cultures were subjected to different heat treatments and the electropho... more Sunflower suspension cell cultures were subjected to different heat treatments and the electrophoretic patterns of heat-induced endocellular and secreted proteins were analyzed. In response to heat shock (3 h at 40°C), sunflower cells synthesized new polypeptides and secreted them into the medium, while the synthesis of other polypeptides was suppressed. Two major polypeptides of about 50 and 32 kDa were strongly induced. The two-dimensional electrophoretic analysis showed that the 32-kDa band is composed of at least four different polypeptides. Western blotting hybridizations of secreted proteins with various lectins were performed. The 32-kDa band gave a positive signal with concanavalin A.

Food Biophysics, 2008

In this work, purification of lactoferrin from whey was performed with high recovery rate. Lactof... more In this work, purification of lactoferrin from whey was performed with high recovery rate. Lactoferrin was then exploited in the preparation of food emulsions. Two tertiary emulsions, formed by olive oil, lecithin, chitosan, and lactoferrin, were compared: both the emulsions showed similar turbidity and stability. In the secondary emulsion formed by oil/lecithin/chitosan, the pH was increased to 9 before addition of lactoferrin. Then, lactoferrin was added, and the pH was stabilized above pH 9. Lactoferrin was found in amounts of 1 to 2.5 mg/ml in the multiple experiments. A fraction of the added lactoferrin was also present in a milky layer above the emulsion layer. This was, to our knowledge, the first study of emulsions made exploiting the interactions between lactoferrin and chitosan. It was noted that chitosan droplets remained soluble, although the hydrocolloid solubility occurs at pH lower than 5.9. These results showed the feasibility of manufacturing lactoferrin-based emulsions as functional foods.

Food Additives and Contaminants Part A-chemistry Analysis Control Exposure & Risk Assessment, 2005

Journal of Cereal Science, 1999

Journal of Cereal Science, 1998

High temperatures during grain filling are considered one of the factors that can modify dough pr... more High temperatures during grain filling are considered one of the factors that can modify dough properties and quality in wheat. In this study we analysed four Italian wheat cultivars grown under different temperature conditions to study the influence of high temperature on storage-protein-gene expression. Plants were grown both in the field and in growth cabinets, and were subjected to different thermal regimes. PolyA+ mRNAs were extracted from control and stressed plants at different stages of kernel development. Northern blot hybridisations were performed using probes for storage and heat shock proteins to monitor the expression of the relative genes under different temperature conditions. Northern analyses, performed using storage protein probes, indicated that temperature variation does not influence the synthesis of any of the storage protein mRNAs. On the contrary, the hybridisation signals obtained using heat shock probes were more intense in the stressed samples, indicating that the expression of heat shock genes is modulated by the temperature variation.

Food Additives and Contaminants Part A-chemistry Analysis Control Exposure & Risk Assessment, 2005

International Journal of Microbiology, 2014

Plant Biology, 2010

Artemisia annua L. is a herb traditionally used for treatment of fevers. The glandular trichomes ... more Artemisia annua L. is a herb traditionally used for treatment of fevers. The glandular trichomes of this plant accumulate, although at low levels, artemisinin, which is highly effective against malaria. Due to the great importance of this compound, many efforts have been made to improve knowledge on artemisinin production both in plants and in cell cultures. In this study, A. annua suspension cultures were established in order to investigate the effects of methyl jasmonate (MeJA) and miconazole on artemisinin biosynthesis. Twenty-two micro molar MeJA induced a three-fold increase of artemisinin production in around 30 min; while 200 μm miconazole induced a 2.5-fold increase of artemisinin production after 24 h, but had severe effects on cell viability. The influence of these treatments on expression of biosynthetic genes was also investigated. MeJA induced up-regulation of CYP71AV1, while miconazole induced up-regulation of CPR and DBR2.