Manuela Poletto Klein

• Doctor in Food Engineering
• Professor at Federal University of Health Sciences of Porto Alegre

Milk whey (MW) and milk whey permeate (MWP) constitute the main part of co-products derived from milk processing. The use of the transgalactosylation activity of β-galactosidases is an excellent alternative to add value to these co-products since this is the route for the synthesis of galactooligosaccharides (GOS), a valuable prebiotic sugar. Hereb...

Olive leaves (OL) are by-products of olive cultivation that are usually discarded, but due to the presence of bioactive compounds beneficial to health, they could be used. Thus, the objective of this paper is to develop a cracker from an olive leaf flour (OLF) and to verify the sensory effect of its use. Crackers were made from 5 formulations: F0 (...

The oxidation activity of laccase on a broad range of substrates has attracted great interest in the development of technologies for industrial and environmental applications. In this work, a crude laccase from Marasmiellus palmivorus was immobilized onto chitosan-coated magnetic particles. The support was developed and characterized by X-ray diffr...

In this study, a commercial enzyme cocktail designed for fruit juice clarification was immobilized on chitosan beads activated with glutaraldehyde and used for setting-up continuous packed-bed and fluidized-bed reactors. Firstly, the enzyme cocktail concentration in the immobilization process was tested to get maximum biocatalyst activity in the re...

Pectin lyase (from Rohapect 10 L) was immobilized on glutaraldehyde supports at low ionic strength at pH 5, 6.5 or 8 and later incubated at pH 8 for 48 h. The activity recovery of the biocatalysts versus pectin was quite low, under 10% for all of the immobilized biocatalyst at 20 °C. However, a high stabilization was found when the enzyme was immob...

Magnetic-chitosan particles were prepared following three different protocols enabling the preparation of particles with different sizes - nano (Nano-CMag, Micro (Micro-CMag) and Macro (Macro-CMag) - and used for pectinase immobilization and clarification of grape, apple and orange juices. The particle size had a great effect in the kinetic paramet...

Polygalacturonase (PG) from Aspergillus niger was immobilized using glyoxyl, vinylsulfone or glutaraldehyde-activated supports. The use of supports pre-activated with glutaraldehyde presented the best results. The immobilization of PG on glutaraldehyde-supports was studied under different conditions: at pH 5 for 24 h; at pH 5, 6.5 or 8 for 3 h and...

Rohapect 10L is an enzyme cocktail commercialized for juice clarification. Here, we characterized the activity and stability of five enzymatic activities present in this cocktail: total pectinase (PE), polygalacturonase (PG), pectin lyase (PL), pectin methyl esterase (PME), and total cellulase (CE) activities. All these enzyme activities have the m...

In this work, we studied the effect of some variables in the reaction between genipin and chitosan, to prepare a support for enzyme immobilization. Reaction temperature, pH, time and genipin concentration were evaluated for support activation and β-galactosidase from Aspergillus oryzae was used as the enzyme model. The immobilized enzyme was charac...

For optimization of biochemical processes in food and pharmaceutical industries, the evaluation of enzyme inactivation kinetic models is necessary to allow their adequate use. Kinetic studies of thermal inactivation of β-galactosidase from Aspergillus oryzae were conducted in order to critically evaluate mathematical equations presented in the lite...

In the present study, we prepared two different magnetic biocatalysts of pectinase and cellulase: carrier-free magnetic CLEAs (CLEA-MP*) and immobilization on glutaraldehyde-activated magnetite (Enz-Glu-MP*). The biocatalysts were compared to their magnetic properties, immobilization parameters, stability and grape juice clarification. Enz-Glu-MP*...

Enzyme-assisted extraction in liquid-liquid two-phase aqueous system was applied for the first time in order to extract genipin from genipap. The effect of different commercial enzymes, their concentrations, and extraction parameters were investigated. Moreover, chitosan gels were prepared, crosslinked with glutaraldehyde or genipin and characteriz...

In this work, the synergistic effects of the combination of Pectinex Ultra Clear (PUC), presenting high pectinase activity, and Lallzyme Beta (LB), presenting cellulase activity were tested for grape juice extraction from Vitis labrusca L. variety Concord. The variables time, temperature, enzyme concentration and PUC/LB ratio were analyzed with the...

Combined cross-linked enzyme aggregates (combi-CLEAs) are a novel prospect for immobilization of a mixture of enzymes and the present study addresses the preparation, characterization and application of pectinases–cellulases combi-CLEAs for grape juice clarification. Initially, 8 enzymatic preparations were tested for turbidity reduction in grape j...

In this work, the effects of eight different enzymatic preparations were compared in terms of the extraction of bioactive compounds as well as the yield and quality of grape juice from Vitis labrusca L. variety Concord. Juices were prepared by enzymatic treatment and the used preparations were characterized through five enzymatic activities: total...

We studied the dynamics of ethanol production on lactose-hydrolyzed whey (LHW) and lactose-hydrolyzed whey permeate (LHWP) in batch fluidized-bed bioreactors using single and co-cultures of immobilized cells of industrial strains of Saccharomyces cerevisiae and non-industrial strains of Kluyveromyces marxianus. Although the co-culture of S. cerevis...

Abstract In order to develop safer processes for the food industry, we prepared a chitosan support with the naturally occurring crosslinking reagent, genipin, for enzyme. As application model, it was tested the immobilization of β-D-galactosidase from Aspergillus oryzae. Chitosan particles were obtained by precipitation followed by adsorption of th...

Thin films containing silver nanoparticles homogeneously dispersed, with narrow size distribution below 10 nm, were synthesized on flat glass surface, by using an ionic silsesquioxane as stabilizer and crosslinking agent. The films can be prepared without previous functionalization of substrate surfaces and without addition of other components. The...

In this work, β-fructofuranosidase and β-fructosyltransferase were covalently immobilized on chitosan spheres, using glutaraldehyde as a coupling agent, in order to produce invert sugar and fructooligosaccharides (FOS). Maxinvert L was used to make β-fructofuranosidase biocatalyst yielding 7000 HU/g. A partial purified β-fructosyltransferase from V...

The enzymatic synthesis of fructooligosaccharides (FOS) was carried out using a partially purified β-fructofuranosidase from the commercial enzyme preparation Viscozyme L. Partial purification of β-fructofuranosidase from Viscozyme L was done by batch adsorption using ion-exchange resin DEAE-Sepharose, showing a 6-fold increase in specific activity...

Cyclodextrin glycosyltransferase (CGTase) from Thermoanaerobacter sp. was covalently immobilized on glutaraldehyde-activated chitosan spheres and used in a packed bed reactor to investigate the continuous production of β-cyclodextrin (β-CD). The optimum temperatures were 75°C and 85°C at pH 6.0, respectively for free and immobilized CGTase, and the...

β-d-Galactosidase from Kluyveromyces lactis was immobilized on glutaraldehyde-activated chitosan and used in a packed-bed reactor for the continuous hydrolysis of lactose and the synthesis of galactooligosaccharides (GOS). The biocatalyst was tested for its optima pH and temperature, thermal stability in the presence of substrate and products, and...

Silver metal nanoparticles were immobilized in chitosan/carboxymethylcellulose/BMI.BF4(1-n-butyl-3-methylimidazolium tetrafluoroborate ionic liquid) (CS/CMC/IL) to form polymeric membrane with 20 μm thickness. The CS/CMC/IL polymeric membrane was prepared using a simple solution blending method. Irregularly shaped Ag(0) nanoparticles with monomodal...

Invertase (E.C.3.2.1.26) from Saccharomyces cerevisiae was covalently immobilized on chitosan nanoparticles prepared by the ionotropic gelation method and activated with glutaraldehyde. The support was characterized and it was studied its load capacity, the influence of the presence of substrate during immobilization, and determined the biocatalyst...

The effect of the support size on the properties of enzyme immobilization was investigated by using chitosan macroparticles and nanoparticles. They were prepared by precipitation and ionotropic gelation, respectively, and were characterized by Fourier transform infrared (FTIR) spectroscopy, differential scanning calorimetry (DSC), transmission elec...

β-Galactosidase was immobilized in an ionic liquid-cellulose film that presents properties as increased flexibility and formability. A polyamine was added during the preparation of the film and their presence was detected by infrared analysis. Glutaraldehyde was used to activate the films in order to generate conditions for enzyme immobilization th...

The effect of using β-galactosidase on the crystallization of lactose in milk sweet has been studied. The concentrations of lactase ranged from 0 to 0.4 g/L of processed milk. The product was subjected to sensorial analysis (crystallization degree ranking test using a specific scale) by trained tasters after 30, 60, 90 e 180 days of storage. The ra...

The survival of food pathogens during frozen storage of chicken hamburgers was evaluated. Hamburgers were contaminated with Escherichia coli (ECHC), Staphylococcus aureus (SAFH) and Salmonella Enteritidis (SE86), separately, and stored at −18C for 28 days. Results demonstrated reductions of 0.63 log 10 cfu/g, 0.73 log 10 cfu/g and 0.27 log 10 most...