María D. Busto

María D. Busto
Universidad de Burgos | UBU · Department of Biotechnology and Food Science

PhD (Valladolid University, Spain)

About

64
Publications
9,057
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1,808
Citations
Citations since 2017
10 Research Items
771 Citations
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2017201820192020202120222023020406080100120140
2017201820192020202120222023020406080100120140
2017201820192020202120222023020406080100120140

Publications

Publications (64)
Article
Full-text available
In this study, the use of spray-drying technology for encapsulating Flavourzyme® (protease-peptidase complex) was evaluated to overcome the limitations (low encapsulation efficiency and no large-scale production) of other encapsulation processes. To the best of our knowledge, spray drying has not been applied previously for the immobilization of th...
Article
Full-text available
The behavior against temperature and thermal stability of enzymes is a topic of importance for industrial biocatalysis. This study focuses on the kinetics and thermodynamics of the thermal inactivation of Lipase PS from B. cepacia and Palatase from R. miehei. Thermal inactivation was investigated using eight inactivation models at a temperature ran...
Article
Full-text available
Monoacylglycerols (MAG) containing polyunsaturated fatty acids (PUFA) have been synthesized by glycerolysis of anchovy oil using Lipozyme RM-IM in a solvent-free system. The experimental conditions, reaction temperature , substrate molar ratio (glycerol/triacylglycerol, Gly/TAG) and enzyme concentration were studied. A response surface methodology...
Article
An enzymatic membrane reactor (EMR) is assembled for the immobilization of naringinase on a polyethersulfone ultrafiltration membrane, based on fouling-induced method. The effects of molecular weight cut-off, membrane configuration, applied pressure, enzyme concentration and pH are studied in terms of permeate rate, immobilization efficiency, and b...
Article
The objective of this investigation was the optimization of the immobilization of naringinase from Penicillium decumbens by entrapment in poly(vinyl alcohol) and polyethylene glycol cryogels. First, the effect of the characteristics of the matrix components on immobilization efficiency and mechanical stability of the biocatalyst obtained, was deter...
Article
Full-text available
In this work, barley seed coating with urease and phosphatase, free and immobilized in humic acids, was studied as a tool to enhance the use efficiency of N and P by plants. The seeds were encapsulated in gels of Ca-alginate by a method that allows gelation of the polymer, containing enzymes, from the surface of seeds outward. The coating process d...
Article
The treatment of must with glucose oxidase (GOX) is a biotechnological alternative to the production of reduced-alcohol wines. Nevertheless, the low pH of must is a limiting factor for the enzyme's activity. This study reports on a simple immobilization method that uses enzyme encapsulation to overcome that limiting factor. The encapsulation of GOX...
Article
The production of mono- and diacylglycerols rich in polyunsaturated fatty acids is achieved in this study, by solvent-free glycerolysis of anchovy oil with lipase PS-DI from Burkholderia cepacia. Attention is focused on the oxidative stability of the reaction products, determined in terms of induction time (It). The effects of glycerol/triacylglyce...
Article
The applicability of qPCR in olive-oil authentication depends on the DNA obtained from the oils and the amplification primers. Therefore, four olive-specific amplification systems based on the trnL gene were designed (A-, B-, C- and D-trnL systems). The qPCR conditions, primer concentration and annealing temperature, were optimized. The systems wer...
Article
The traceability of olive oil is an unresolved issue that remains a challenge. In this field, DNA-based techniques are very powerful tools for discrimination that are less negatively influenced by environmental conditions than other techniques. More specifically, quantitative real time PCR (qPCR) achieves a high degree of sensitivity, although the...
Article
BACKGROUND Most of the N fertilizers added to the soil are not efficiently used by plants and are lost to the atmosphere or leached from the soil, causing environmental pollution and increasing cost. The barley seed encapsulation in calcium alginate gels containing free or immobilized urease was investigated to enhance plant utilization of soil N.R...
Article
This study describes the design of a suitable DNA isolation method from commercial vegetable oils for the application of DNA markers for food safety and traceability. Firstly, a comparative study was made of eight methods for the recovery of high quality DNA from olive, sunflower and palm oils, and a CTAB-based method was selected. In order to opti...
Chapter
Conventional methods to reduce the bitterness in citrus juices involve not only the removal of the bitter compounds (naringin and limonin), but also bioactive components (organic acids, flavonoids, limonoids, etc.) from the juice. In this regard, biotechnological methods, either isolated enzymes or whole cells, have been suggested as an alternative...
Article
A new spectrophotometric end-point method to evaluate lipase activity in aqueous media is described. This procedure is based on blocking the enzyme reaction by adding chloroform:isoamyl alcohol (24:1) as a denaturating agent, and removing the precipitate by centrifugation. Emulsifier screening showed that the chemical component with the least effec...
Article
A large portion of the soluble P fertilizers introduced into soil reacts with soil components forming insoluble P products, which are not efficiently taken up by plants. Barley seed (Hordeum vulgare L. 'Volley') encapsulation in Ca-alginate gels containing phosphatase free or immobilized with soil humates (humate-phosphatase complexes, HPC) was inv...
Article
Full-text available
Coating barley seeds with free and immobilized alkaline phosphatase was investigated as a potential means to enhance plant utilization of accumulated soil phosphorus (P). Two coating techniques were studied: film-coating and pelleting. The highest phosphatase activity retention in the coating layer, ranging from 0·48 to 0·67, was observed when seed...
Article
Antioxidant capacity, radical scavenging activity, as well as protective effect on lipoperoxidation, glutathione oxidation and DNA damage, were evaluated in grapefruit juice subjected to bitterness removal by naringinase or by physical adsorption with Amberlite®IRA-400. The results showed a reduction in naringin content for the naringinase-treated...
Article
The experiment described in this paper introduce to students to immobilisation of β-glucosidase, induced in Aspergillus niger cultures, by entrapment method in calcium alginate gel. Additionally, the determination of apparent kinetic parameters, the pH/enzyme activity curves and the study of the thermal stability permits students to check and under...
Article
This study was designed to evaluate and compare antioxidant capacity and radical scavenging activity of naringin and its aglycone by different in vitro assays. The effects of flavanones on lipid peroxidation, glutathione (GSH) oxidation and DNA cleavage were also assessed. The results showed that naringenin exhibited higher antioxidant capacity and...
Article
An alkaline phosphatase (EC 3.1.3.1) from Escherichia coli ATCC27257 was immobilized by copolymerization with resorcinol. The phosphatase-polyresorcinol complex synthesized retained about 74% of the original enzymatic activity. The pH and temperature profile of the immobilized and free enzyme revealed a similar behavior. Kinetic parameters were det...
Article
Neutrase, a commercial preparation of Bacillus subtilis , was covalently immobilized on alginate-glutaraldehyde beads. Immobilization conditions and characterization of the immobilized enzyme were investigated. Central composite design and response surface methods were employed to evaluate the effects of immobilization parameters, such as glutarald...
Article
One way to study the state in which stabilized extracellular enzymes persist and are active in the soil is by extraction from the soil, with subsequent fractionation of enzyme–organomineral complexes and characterization of such complexes. In order to investigate the location and characteristics of soil β-glucosidase, three soil fractions were obta...
Article
In this paper, a long-term investigation of the enzyme hydrolysis of pectin, using a free enzyme membrane reactor (FEMR), is presented. Production of crude enzymes (polygalacturonase and pectin lyase), in Aspergillus niger CECT 2088 cultures grown on an agricultural subproduct (apple pomace) as a low cost inducer, was also studied. Observing the re...
Article
Naringinase, induced from Aspergillus niger CECT 2088 cultures, was immobilized into a polymeric matrix consisting of poly(vinyl alcohol) (PVA) hydrogel, cryostructured in liquid nitrogen, to obtain biocatalytically active beads. The effects of matrix concentration, enzyme load and pH on immobilization efficiency were studied. Between 95% and 108%...
Article
Optimisation of pectin hydrolysis using pectolytic enzymes produced by Aspergillus niger CECT 2088 was carried out. The effect of enzyme concentration (0.018–0.072%, w/v), substrate concentration (0.004–0.896%, w/v), pH (4.49–5.91), temperature (32.2–67.8 °C) and reaction time (18.2–71.8 min) on the enzymatic process was studied. The experiments we...
Article
Capillary electrophoresis using an untreated fused-silica capillary and a low-pH buffer containing urea and hydroxypropyl methyl cellulose was used to follow the proteolytic action of neutral protease of Bacillus subtilis on isolated casein fractions and whole milk casein (CN) samples. Electrophoretic analysis showed that this protease caused inten...
Article
The effect of the ripening time on the proteolytic process in cheeses made from ewe's milk during a 139-day ripening period was monitored by the use of capillary electrophoresis of pH 4.6 insoluble fraction. Totals of 18 and 21 peaks were recognized and matched in the electropherograms obtained with a fused-silica capillary and a neutral capillary...
Article
Pectinlyase, present in different commercial pectinases used in juice technology, was immobilized on alginate beads. The optimal conditions were: 0.17 g alginate ml(-1), 1.2% (w/v or v/v) enzyme concentration and acetic-HCl/glycine-HCl buffer at pH 3.6 or tris-HCl/imidazole buffer at pH 6.4. Maximum percentage of immobilization (10.6%) was obtained...
Article
The effect of the ripening time on the proteolytic process in cheeses manufactured from mixtures of cow's and ewe's milk during a 167-day ripening period was monitored by capillary electrophoresis of the pH 4.6-insoluble fraction. Totals of 21 and 16 peaks were recognized and matched in the electropherograms obtained with a fused-silica capillary a...
Article
The kinetic and operational behavior of a free enzyme membrane reactor (FEMR) were analyzed. High conversion (83%) of the pectin could be achieved with a substrate-to-enzyme ratio of 23.3 at 46°C and pH 4.8, with a transmembrane pressure of 34.5 kPa and recycling flow rate of 36 L/h. The FEMR showed a 1st-order deactiva-tion profile with a calculat...
Article
Kinetic properties and thermal inactivation of polygalacturonase (PG) assayed in commercial preparations (Rapidase C80, Pectinase CCM and Pectinex 3XL) were studied. The PG activities in all samples followed Michaelis–Menten kinetics and their catalytic efficiencies were calculated. The optimum pH for enzyme activity was 4.7 for Pectinex 3XL and 4....
Article
Fractional factorial screening design and response surface methodology were applied to optimize the entrapment of glucose oxidase in liposomes by the dehydration–rehydration vesicle (DRV) method. Phosphatidylcholine from different sources, cholesterol:phosphatidylcholine (Ch:PC) and enzyme:lipid (E:L) ratios, buffer pH, sonication frequency and tre...
Article
Kinetic properties and thermal inactivation of pectinlyase (PL) were assayed in commercial pectinase preparations (Rapidase C80, Pectinase CCM, Pectinex 3XL and Grindamyl 3PA) by using apple pectin as substrate. The PL activity of Rapidase C80 showed substrate inhibition, while the other enzyme preparations followed typical Michaelis–Menten kinetic...
Article
Central composite design and response surface methods were applied to optimise and assess the robustness of a capillary zone electrophoresis method for the separation of bovine caseins. Buffer pH, running voltage and polymeric additive buffer concentration were the parameters selected for this study. Successful results were obtained at 18.5 kV, wit...
Article
Alkaline phosphatase (EC 3.1.3.1) extracted from Escherichia coli ATCC27257 was immobilised by co-flocculation with soil humates in the presence of Ca2+. The effects of time, temperature, pH and concentration of enzyme and support on immobilisation were studied. Between 58 and 92% of the added phosphatase was strongly bound to the humates, dependin...
Article
In order to optimize the enzymatic hydrolysis of three celluloses (carboxymethyl cellulose, powdered cellulose and microgranular cellulose), the effect of varying enzyme and substrate concentration, temperature and pH was studied. The best experimental conditions found to degrade the celluloses were 48 h of incubation, an enzyme concentration of fo...
Article
Three methods of wheat straw treatment (with NaOH, H2O2 and butylamine) and its subsequent saccharification by Trichoderma reesei cellulases and Aspergillus niger β-glucosidase are reported. The treatment of straw with NaOH for 1 h in the autoclave (120°C) caused a great loss of the hemicellulose content and a partial removal of lignin, provoking a...
Article
Full-text available
Mature pea seeds contain two major lipoxygenases (LOX) isoenzymes designated LOX-2 and LOX-3. The thermal inactivation of crude pea LOX and the recombinant LOX (rLOX) were studied. Heat-inactivation plots for crude extracts of pea LOX were linear from which thermodynamic activation parameters, ΔH#, ΔS# sand ΔG# have been estimated. The enzymatic ac...
Article
β-D-Glucosidase was immobilised by entrapment in two different matrices (calcium alginate and polyacrylamide gels), in order to compare how the immobilisation could stabilise the enzyme towards thermal and proteolytic deactivation. While the enzyme trapped in polyacrylamide gel showed an optimum temperature for activity at 10°C lower than that of t...
Article
beta-D-Glucosidase was immobilised by entrapment in two different matrices (calcium alginate and polyacrylamide gels), in order to compare how the immobilisation could stabilise the enzyme towards thermal and proteolytic deactivation. While the enzyme trapped in polyacrylamide gel showed an optimum temperature for activity at 10 degrees C lower tha...
Article
Immobilisation of β-glucosidase, isolated from Aspergillus niger, by entrapment in both calcium alginate and polyacrylamide gels was studied. A retention of 66% of initial activity was observed in the alginate beads prepared with 3% (w/v) alginate, 0.2 m CaCl2 and 1 h of treatment. The maximum β-glucosidase activity in polyacrylamide gels (∼55%) wa...
Article
Endo-β-glucanase (endo-β-1,4-glucano-glucanase EC 3.2.1.4), isolated from Trichoderma reesei, was immobilized in calcium alginate beads, retaining 75% of its original activity. The polyanionic moiety surrounding the immobilized enzyme displaced the pH-activity profile to alkaline regions with respect to that of the free enzyme. The enzyme was inhib...
Article
The thermal and proteolytic stability of free and immobilized β-d-glucosidase, isolated from Pseudomonas pickettii and Aspergillus niger, were determined. The optimal temperatures of soluble and entrapped β-glucosidase extracted from P. pickettii were 40 and 50°C, respectively. In contrast, the optimal temperature of enzyme isolated from A. niger r...
Article
β-Glucosidase and carboxymethyl cellulase were induced in and extracted from Aspergillus niger and Trichoderma reesei cultures and cross-linked with glutaraldehyde or complexed with a soil polyphenolic structure (humates). The concentration of the bifunctional reagent (2·5 or 5·0%) determined the activity of the immobilised β-glucosidase while the...
Article
Crosslinked polyacrilamide gels was used to immobilize alkaline phosphatase. The maximum activity of entrapped enzyme (about 47.6%) was achieved at crosslinked agent concentration of 0.4 w/v percent and acrylamide concentration of 25 w/v percent. The matrix enzyme permeability was minimal at that cross linking degree. The apparent Michaelis constan...
Article
Amorphous cellulose induced the synthesis of endoglucanase in Trichoderma reesei to a greater extent than did cellobiose, lactose, sucrose or other commercial celluloses. In contrast, none of these carbohydrates was able to induce significant levels of endoglucanase activity in Aspergillus niger or Pseudomonas pickettii. Both endoglucanase and β-gl...
Article
About 50–60% β-d-glucosidase (isolated from Pseudomonas pickettii and Aspergillus niger) was immobilized in calcium alginate gel beads by entrapment, retaining 15–26% of original activity. The pH values for maximum activitiy of the immobilized bacterial and fungal enzymes appeared to be 5.0 and 3.0 respectively, very similar to those of the free en...
Article
The production of β-glucosidase by soil enrichment cultures grown on cellobiose and carboxymethylcellulose and several experimental conditions were investigated in this work. The highest rates of induced enzyme activity were found in the soil cultures in which 0.4% cellobiose at pH 8.0 was used as the sole C and energy source. A bacterial strain, p...
Article
Humic compounds with -glucosidase activity were extracted from soil using tetrasodium pyrophosphate as extracting solution. Mixture of soil samples with 0.01 M pyrophosphate in a ratio of 1:5 (w/v), adjustment to pH 7.0–7.3, extraction for 18 h with reciprocating agitation, and bacteriological filtration after centrifugation were the optimum condit...
Article
Some characteristics of a 2,2-dichloropropionate dehalogenase induced in a bacterial strain capable of degrading high concentrations of the herbicide dalapon were studied. Polyacrilamide gel electrophoresis of the crude cell free extracts identified only one type of dehalogenase. The single enzymatic protein showed activity against a variety of chl...
Article
Full-text available
Different factors (agitation time, type of buffer, enzyme solution/soil ratio, enzyme solution concentration) affecting alkaline phosphatase immobilisation by a rendzina soil were studied. Appreciable quantities (>48%) of commercial active phosphatase were immobilised by the soil colloidal particles in a process well adjusted to a Langmuir isotherm...

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