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F X Malcata

Publications and source records attributed to F X Malcata.

32 records · Page 2Linked to original sources

Development of probiotic cheese manufactured from goat milk: response surface analysis via technological manipulation.

Production of caprine milk has been rising steadily, partially because of its good nutritional value; the possibility of improving nutritional benefits by adding probiotic species such as Bifidobacterium lactis and Lactobacillus acidophilus was assessed. The manufacturing process of a traditional semi-hard goat cheese was technologically modified to optimize the process. The amount of starter inoculum, the concentration of salt, the addition of a protein hydrolysate, and the ripening time were varied to improve the microbiological, biochemical, and sensory properties of the cheese. Bifidobacterium lactis was able to grow slightly (up to 3 x 10(8) cfu/g), but growth was dependent on the physicochemical characteristics of the cheese. Lactobacillus acidophilus did not grow substantially in any of the experimental cheeses, and maximum numbers did not exceed 6 x 10(7) cfu/g. Concentrations of lactic acid and acetic acid increased throughout cheese manufacture, indicating that production of these acids was uncoupled from growth. Viability of the probiotic strains during ripening was sufficient to yield numbers that were above the accepted threshold (10(6) cfu/g) for a probiotic effect. Both strains contributed significantly to ripening, especially in the formation of low molecular mass peptides and amino acids, but lipolysis was not greatly affected. Statistical analyses using response surface methodology indicated that the manufacture of goat cheese could be optimized by the addition of 0.30% (vol/wt) milk hydrolysate, 3 x 10(7) of viable B. lactis and 7 x 10(6) of viable L. acidophilus cells/ml of milk, respectively, 3.50% (wt/wt) salt, and ripening for 70 d.

Acetic Acid↗

Growth enhancement of Bifidobacterium lactis Bo and Lactobacillus acidophilus Ki by milk hydrolyzates.

The determination of the best conditions of preparation of a (tentatively) probiotic starter culture that might be suitable for cheese making composed solely of Bifidobacterium lactis Bo and Lactobacillus acidophilus Ki is critical if a consistently reliable acid production is to be achieved, especially because bifidobacteria have stringent requirements for growth. Therefore, we determined whether B. lactis Bo and L. acidophilus Ki required or benefitted from the addition of milk hydrolyzates (brought about by proteinase or neutrase as the nitrogen source). The growth and acid production of B. lactis in milk were affected by the addition of proteinase-mediated hydrolyzate and, to a lesser extent, by neutrase-mediated hydrolyzate; a higher degree of hydrolysis of either hydrolyzate resulted in greater biomass increase and greater acid production. This result suggests that the poor growth of bifidobacteria in milk is due partially to the lack of small peptides and free amino acids. The rates of growth and acidification by B. lactis were enhanced when cocultured with L. acidophilus (1:1 inoculum ratio). Conversely, the growth rates and acid production of L. acidophilus were not positively affected by the addition of either milk hydrolyzate. Although L. acidophilus grew slowly, its proteolytic system was apparently able to generate its own nitrogen source. Nevertheless, coculture with B. lactis (1:1 inoculum ratio) led to enhanced rates of growth and acidification when compared with that of the single strain, suggesting some degree of symbiosis between the strains.

Animals↗

Microbiological characterization of Serra da Estrela cheese throughout its Appellation d'Origine Protégée region.

The purpose of this study was to assess the typical microbiological quality of the most famous Portuguese traditional cheese, Serra da Estrela, and to assess its ripening time and geographical dependence. Ninety-six experimental cheeses manufactured from sixteen batches of milk on eight dairy farms scattered over the Appellation d' Origine Protégée (AOP) region were qualitatively and quantitatively evaluated microbiologically at various ripening times. Viable counts were performed after inoculation on appropriate selective media for aerobic mesophiles and proteolytic and lipolytic microflora, as well as lactococci, lactobacilli, species of Enterobacteriaceae, lactic streptococci, staphylococci, and yeasts. Members of the Enterobacteriaceae and lactic acid bacteria were the predominant microbial groups on all dairy farms throughout maturation; the latter are probably the microbial group responsible for most proteolytic and lipolytic breakdown in Serra da Estrela cheese. The microbial groups whose numbers were most affected by dairy-to-dairy variation were species of Enterobacteriaceae staphylococci, and enterococci, which are the most critical groups in terms of health hazards. It is therefore suggested that tighter control should be implemented at the level of choice of raw materials, in milk-handling practices, and in general throughout the manufacturing process in attempts to standardize production and consistently reduce microbiological risks (even though the distinctiveness of a few final organoleptic characteristics may somehow be reduced.

Animals↗

Bioreactors with immobilized lipases: state of the art.

This review attempts to provide an updated compilation of studies reported in the literature pertaining to reactors containing lipases in immobilized forms, in a way that helps the reader direct a bibliographic search and develop an integrated perspective of the subject. Highlights are given to industrial applications of lipases (including control and economic considerations), as well as to methods of immobilization and configurations of reactors in which lipases are used. Features associated with immobilized lipase kinetics such as enzyme activities, adsorption properties, optimum operating conditions, and estimates of the lumped parameters in classical kinetic formulations (Michaelis-Menten model for enzyme action and first-order model for enzyme decay) are presented in the text in a systematic tabular form.

Amino Acid Sequence↗

STADEERS: a software package for the statistical design of experiments pertaining to the estimation of parameters in rate expressions that describe enzyme-catalyzed processes.

This paper describes a computational algorithm (STADEERS--STAtistical Design of Experiments in Enzyme ReactorS) for the statistical design of biochemical engineering experiments. The type of experiment that qualifies for this package involves a batch reaction catalyzed by a soluble enzyme where the activity of the enzyme decays with time. Assuming that both the catalytic action and the deactivation of the enzyme obey known rate expressions, the present code is helpful in the process of obtaining estimates of the kinetic parameters by providing as output the times at which samples should be withdrawn from the reacting mixture. Starting D-optimal design is used as a basis for the statistical approach. This BASIC code is a powerful tool when fitting a rate expression to data because it increases the effectiveness of experimentation by helping the biochemical kineticist obtain data points with the largest possible informational content.

Algorithms↗

Kinetics and mechanisms of reactions catalysed by immobilized lipases.

This review focuses on the kinetics and mechanisms of reactions catalysed by immobilized lipases. The effects of pH, temperature, and various substances on the catalytic properties of immobilized lipases and on the processes by which they are deactivated are reviewed and discussed.

Amino Acid Sequence↗

Starting D-optimal designs for batch kinetics studies of enzyme-catalyzed reactions in the presence of enzyme deactivation.

This paper describes a strategy for the starting experimental design of experiments required by general research in the field of biochemical kinetics. The type of experiments that qualify for this analysis involve batch reactions catalyzed by soluble enzymes where the activity of the enzyme decays with time. Assuming that the catalytic action of the enzyme obeys a Michaelis-Menten rate expression and that the deactivation of the enzyme follows a first-order decay, the present analysis employs the dimensionless, integrated form of the overall rate expression to obtain a criterion (based on the maximization of the determinant of the derivative matrix) that relates the a priori estimates of the parameters with the times at which samples should be withdrawn from the reacting mixture. The analysis indicates that the initial concentration of substrate should be as large as possible, and that the samples should be taken at times corresponding to substrate concentrations of approximately 2/3, 1/4, and I/epsilon of the initial concentration (where epsilon should be as large as possible).

Enzyme Inhibitors↗

HERSIM: a microcomputer program designed to compute the limits of conversion for real homogeneous isothermal enzymic reactors.

HERSIM is a program written in BASIC designed to aid the investigator interested in determining the substrate conversion in a real homogeneous isothermal enzymic reactor, for various kinetic equations. The program runs after tracer data relative to a Dirac impulse to the reactor have been entered, and computes the two limits of real conversion: total segregation and maximum mixedness. The kinetic constants of the reacting system are input as data, and the variation of conversion with reactor temperature between given limits is computed as accurately as requested.

Algorithms↗

Computer aided design of pellets for fixed-bed reactors performing Michaelis-Menten reactions.

The problem of designing the pellet characteristics for existing fixed-bed reactors has been studied. The reactor is assumed to be a unidimensional, plug-flow, heterogeneous type. Three particle shapes, two intraparticle mass transfer mechanisms, two kinds of film resistance to mass transfer and two poisoning patterns have been considered. An economic balance to both product market price and pellet market cost was taken into account. An interactive program (HIMER) containing all these contributions was thus developed, enabling the investigator to follow the reactor performance along the flow coordinate.

Algorithms↗

Transfer of carbon dioxide within cultures of microalgae: plain bubbling versus hollow-fiber modules.

In attempts to improve the metabolic efficiency in closed photosynthetic reactors, availability of light and CO(2) are often considered as limiting factors, as they are difficult to control in a culture. The carbon source is usually provided via bubbling of CO(2)-enriched air into the culture medium; however, this procedure is not particularly effective in terms of mass transfer. Besides, it leads to considerable waste of that gas to the open atmosphere, which adds to operation costs. Increase in the interfacial area of contact available for gas exchange via use of membranes might be a useful alternative; microporous membranes, in hollow-fiber form, were tested accordingly. Two hollow-fiber modules, different in both hydrophilicity and outer surface area, were tested and duly compared, in terms of mass transfer, versus traditional plain bubbling. Overall volumetric coefficients (K(L)a) for CO(2) transfer were 1.48 x 10(-2) min(-1) for the hydrophobic membrane, 1.33 x 10(-2) min(-1) for the hydrophilic membrane, and 7.0 x 10(-3) min(-1) for plain bubbling. A model microalga, viz. Nannochloropsis sp., was cultivated using the two aforementioned membrane systems and plain bubbling. The produced data showed slight (but hardly significant) increases in biomass productivity when the hollow-fiber devices were used. However, hollow-fiber modules allow recirculation of unused CO(2), thus reducing feedstock costs. Furthermore, such indirect way of supplying CO(2) offers the additional possibility for use of lower gas pressures, as no need to counterbalance hydrostatic heads exists.

Carbon Dioxide↗

Structural and functional stabilization of L-asparaginase via multisubunit immobilization onto highly activated supports.

A new protocol for the stabilization of the quaternary structure of multimeric enzymes has been attempted using as model enzyme (tetrameric) L-asparaginase from Escherichia coli. Such strategy is based upon multisubunit covalent immobilization of the enzyme onto activated supports (agarose-glutaraldehyde). Supports activated with different densities of reactive groups were used; the higher the density of groups, the higher the stabilization attained. However, because of the complexity of that enzyme, even the use of the highest densities of reactive groups was not enough to encompass all four subunits in the immobilization process. Therefore, a further chemical intersubunit cross-linking with aldehyde-dextran was pursued; these derivatives displayed a fully stabilized multimeric structure. In fact, boiling the modified enzyme derivative in the presence of sodium dodecyl sulfate and beta-mercaptoethanol did not lead to release of any enzyme subunit into the medium. Such a derivative, prepared under optimal conditions, retained ca. 40% of the intrinsic activity of the free enzyme and was also functionally stabilized, with thermostabilization enhancements of ca. 3 orders of magnitude when compared with its soluble counterpart. This type of derivative may be appropriate for extracorporeal devices in the clinical treatment of acute leukemia and might thus bring about inherent advantages in that all subunits are covalently bound to the support, with a longer half-life and a virtually nil risk of subunit release into the circulating blood stream.

Asparaginase↗

Caseinolytic activity of fruit extract from Opuntia ficus-indica on bovine, caprine, and ovine sodium caseinates.

The rates and extents of hydrolysis of alpha(S)- and beta-caseins from bovine, caprine, and ovine sodium caseinates produced by an enzymatic extract of the fruit of Opuntia ficus-indica, (L.) Miller were evaluated and compared with those produced by a commercial animal rennet. A mechanistic model based on a pseudo-first-order enzymatic reaction, in the presence of first-order deactivation of the enzyme, was postulated and successfully fitted to the experimental data. The animal rennet exhibited higher enzymatic efficiency than the fruit extract, irrespective of the source (i.e., bovine, caprine, or ovine) and the type (i.e., alpha(S)- or beta-casein) of substrate. The enzymatic efficiency (k(cat)/K(m)) for alpha(S)-casein ranged from 72 to 220 and from 43 to 65 L g(-1) h(-1), and for beta-casein from 242 to 742 and from 55 to 164 L g(-1) h(-1), for the animal rennet and the enzymatic extract of O. ficus-indica, respectively. Finally, it was observed that beta-casein from caprine and ovine caseinates was degraded by O. ficus-indica faster than its alpha(S) counterpart, but the reverse was observed for bovine caseinate.

Animals↗

Adsorption of protein from several commercial lipase preparations onto a hollow-fiber membrane module.

Ten commercially available crude preparations of lipase from various microbial sources were adsorbed from aqueous buffers at several initial concentrations onto a bundle of hydrophobic hollow fibers made of poly(propylene) at pH 7.0 and 40 degrees C. The kinetics of adsorption were evaluated from measurements at various times of the protein content of the supernatant solution (using BSA as equivalent) in a well-mixed reservoir placed in series with the hollow fiber module. Preliminary tracer experiments have indicated that the module and the tank can be simulated as a system consisting of a plug flow reactor in series with a continuous stirred tank reactor. A mechanistic model based on the hydrodynamic assumptions associated with this system coupled with the postulation of two reversible first-order steps for the adsorption of protein was successfully fitted to the experimental data via nonlinear regression analysis. The statistical significance of the model was checked using tests for lack of fit. This work is useful in predicting the time period required to immobilize a (crude) lipase by adsorption onto a hydrophobic hollow fiber module, a configuration which has proved successful in the recent past for the performance of lipase-catalyzed reactions.

Adsorption↗