CURRICULUM
AUTORES:
Rodríguez Vico, F.
TITULO:
Reconstitución de la 3-hidroxi-3-metilglutaril-CoA reductasa en vesículas
fosfolipídicas.
REF.:
Public. Univ. Granada. ISBN 84-388-0811-7. Dep. GR- 946/1988.
CLAVE: LIBRO
AUTORES:
Rodríguez-Vico, F., Castillo, M., López, J.M., Zafra, M.F., García-Peregrín,
E.
TITULO:
Induction of neonatal hipercholesterolemia by saturated fat; Changes in
lipoprotein profile.
REF.:
"PHYSIOLOGIC BASIS OF PERINATAL CARE" 129-132 (Medina, J.M. &
Quero, J. ed), Salvat Publishers, Barcelona, 1992
AUTORES:
Aguilera, J, García-Molina,V, Rodríguez-Vico, F, Arce, V., Linares, A., García-Peregrín,
E.
TITULO:
Age-related changes in the mevalonate metabolism in vivo in chick kidneys.
REF. :
Arch. Inter. Physiol. Biochim., 96; 121-126. (1.988).
AUTORES:
Martínez-Cayuela, M., Rodríguez-Vico, F., Faus, M.J., Gil, A.
TITULO:
Partial purification
and intracellular localization of cherimoya (annona
cherimolia Mill.) polyphenoloxidase.
REF.:
J. Plant Physiol., 133; 660-663, (1989)
AUTORES:
Rodríguez-Vico, F., Martínez-Cayuela, M., Ramírez, H., García-Peregrín, E.
TITULO:
A procedure for eliminating interferences in the Lowry method of protein
determination
REF.:
Anal. Biochemistry, 1983; 275-278, (1989)
AUTORES:
Castillo, M., Burgos, C., Rodríguez-Vico, F., Zafra, M.F., García-Peregrín,
E.
TITULO:
Effects of clofibrate on the main regulatory enzimes of Cholesterogenesis.
REF.: Life
Sciences, 46; 397-403, (1.990)
AUTORES:
Castillo, M., Burgos,
F., Rodríguez-Vico, F., Zafra, M., García-Peregrín, E.
TITULO:
Acyl-CoA: Cholesterol
acyltransferase in eel (Anguilla anguilla)
liver: effect of lipid content of diet.
REF.:
Comp. Biochem. Physiol., 98B; 143-146, (1.991)
AUTORES:
Rodríguez-Vico, F., Martínez-Cayuela, M., Zafra, M., García-Peregrín,
E.
TITULO:
A procedure for the
simultaneous determination of lipid and protein in biomem. and other biol.
samples.
REF.:
Lipids, 26; 77-80, (1.991)
AUTORES:
Rodríguez-Vico, F., López, J., Castillo, M., Zafra, M., García-Peregrín, E.
TITULO:
Characterization of chick serum lipoproteins isolated by density gradient
ultracentrifugation.
REF.:
Arch. Inter. Pgysiol. Biochim. Biophys., 100; 19-22, (1.992)
AUTORES:
Zafra, M.F., Castillo, M., Rodríguez-Vico, F., García-Peregrín, E
TITULO:
Induction in Gallus Domesticus of experimental hypercholesterolemia by saturated
fat. Effects of cholesterogenic
enzyme activity.
REF.:
Arch. Inter. Physiol. Biochim. Biophys., 100;133-136, (1.992).
AUTORES:
Burgos, C., Castillo, M., Rodríguez-Vico, F., Zafra, M.F., García-Peregrín,
E.
TITULO:
Influence of protein/lipid ratio of diet on cholesterol synthesis and
esterification in eel liver.
REF.:
Arch. Inter. Physiol. Biochim. Biophys. 101:53-55, (1993).
AUTORES:
Castillo, M., Zafra, M.F., Rodríguez-Vico, F., López, J.M., García-Peregrín,
E.
TITULO:
Changes in the chick lipoprotein profile during
postnatal development.
REF.:
Biochemical Archives, 8:183-190, (1992).
AUTORES:
Rodríguez-Vico, F., Castillo, M., Zafra, M.F., López, J.M., García-Peregrín,
E.
TITULO:
Effect of dietary coconut oil on lipoprotein composition of Young Chick.
REF.:
Comp. Biochem. Physiol. vol. 106A, 4: 799-802, (1993).
AUTORES:
Las Heras-Vázquez, F.J., Clemente Jiménez, J.M., Rodríguez-Vico, F.
TITULO:
RAPD fingerprinting of pepper (Capsicum
annum L.) breedingg lines.
REF.:
Capsicum and Eggplant Newsletter, (1996) 15:37-40.
ABSTRACT:
AUTORES :
Las Heras-Vázquez, FJ, Clemente-Jiménez, JM, Riado-Abad, J., García-Maroto,
F, Rodríguez-Vico, F.
TITULO :An
example of the practical use of RAPD in pepper breeding.
REF.:
Proc. of The National Pepper Conference, 47-49
(1996)
AUTORES: Rodriguez-Vico,
F., Soler, A., Martinez-Cayuella, M., Ramirez,
H., Garcia-Peregrin, E.
TITULO:
Effects of free fatty acidds and phospholipids hydrogcarbon chains and polar
heads on soluble and reconstituded 3-hydroxy-3methylglutaryl CoA reductase.
REF.:
Biochemical Archives. (1998) 14:59-66
ABSTRACT:
We have studied the possible regulation of solubilized and reconstituted 3-hydroxy-3-metylglutaryl-CoA
reductase by free fatty acids and by both hydrocarbon chains and polar heads of
different phospholipids. Polyunsaturated (20:4, 18:3, and 18:2) and
monounsaturated (18:1) fatty acids showed a clear inhibitory effect on
microsomal reductase suggesting that this effect appears to depend upon the
integrity of microsomal membrane, since in solubilized reductase no appreciable
inhibition could be found. Hoowever, no significant differences were observed in
the specific activities recovered in the presence of different choline
derivatives containing different fatty acid chains. Phospholipid polar heads had
also no effect on reconstituted and solubilized reductase activity. Our results
suggest that phospholipid hydrocarbon chains and polar heads, as a part of the
different types of phospholipids, don ot participate in controlling the activity
of 3-hydroxy-3-methylglutaryl-CoA reductase, while free fatty acids produced a
significant inhibition of reductase activity.
AUTHORS:
FJ Las Heras Vázquez, F Gómez-Mercado,
JL Guil Guerrero, I Rodríguez-García, F García-Maroto
TITLE:
Genetic relationships and population structure within of the endemic Sideritis
pusilla (Lamiaceae) assessed using RAPDs
JOURNAL:
Botanical Journal of the Linnean Society (1999) 129: 345-358
ABSTRACT:
RAPD analysis has been performed for 15 plant populations, covering eight
diVerent infraspecific taxa of Sideritis pusilla (Lange) Pau, member of the
Lamiaceae endemic to southeastern Spain, and their putative parental species
Sideritis hirsuta L. and Sideritis leucantha Cav. Genetic distances, together
with the presence of numerous fixed molecular markers diVerentiating S. pusilla
from its putative ancestors, indicate that it should be considered as a true
species. Cladistic and populational analysis led to the allocation of S. pusilla
populations into three major groups—osteoxylla, flavovirens and pusilla/almeriensis—which
include taxa previously described as ranging from the varietal to the specific
level. Low genetic diVer-entiation among groups revealed by a reduced number of
specific molecular markers justify their assignment under the infraspecific
range. Moreover, the existence of both morphological and biogeographical
diVerences, supports a status for these groups as subspecies of S. pusilla.
Highly significant (P<0.002) variance partitioning data (AMOVA) extracted
from the analysis of individuals within S. pusilla populations show that, of the
total genetic diversity, 68.8% was attributable to individual diVerences within
populations, 19.9% to populational diVerences within groups, and only 11.3% to
divergence between groups. This distribution is in agreement with the
outcrossing nature of these plants. Comparative analysis of variance within
populations reveals a reduced genetic variation for the osteoxylla group, thus
supporting its previous consideration as an endangered taxon.
AUTORES:
Ortiz-Salmerón, E., Yazed ,Z., Clemente-Jimenez, J.M., Las Heras-Vázquez, F.J.,
Rodríguez-Vico, F., Barón, C., García-Fuentes, L.
TITULO:
Thermodynamic analysis of the binding of glutathione S-transferase over a range
of temperatures.
REF.
Eur J Biochem. 2001 268(15):4307-4314.
ABSTRACT:
The binding properties of a glutathione S-transferase (EC 2.5.1.18) from
Schistosoma japonicum to substrate glutathione (GSH) has been investigated by
intrinsic fluorescence and isothermal titration calorimetry (ITC) at pH 6.5 over
a temperature range of 15-30 degrees C. Calorimetric measurements in various
buffer systems with different ionization heats suggest that protons are released
during the binding of GSH at pH 6.5. We have also studied the effect of pH on
the thermodynamics of GSH-GST interaction. The behaviour shown at different pHs
indicates that at least three groups must participate in the exchange of protons.
Fluorimetric and calorimetric measurements indicate that GSH binds to two sites
in the dimer of 26-kDa glutathione S-transferase from Schistosoma japonicum (SjGST).
On the other hand, noncooperativity for substrate binding to SjGST was detected
over a temperature range of 15-30 degrees C. Among thermodynamic parameters,
whereas DeltaG degrees remains practically invariant as a function of
temperature, DeltaH and DeltaS degrees both decrease with an increase in
temperature. While the binding is enthalpically favorable at all temperatures
studied, at temperatures below 25 degrees C, DeltaG degrees is also favoured by
entropic contributions. As the temperature increases, the entropic contributions
progressively decrease, attaining a value of zero at 24.3 degrees C, and then
becoming unfavorable. During this transition, the enthalpic contributions become
progressively favorable, resulting in an enthalpy-entropy compensation. The
temperature dependence of the enthalpy change yields the heat capacity change (DeltaCp
degrees ) of -0.238 +/- 0.04 kcal per K per mol of GSH bound.
AUTORES:
Ortiz-Salmerón, E., Zeyad, Y., Clemente-Jimenez, J.M., Las Heras-Vázquez, F.J.,
Rodríguez-Vico, F., Barón, C., García-Fuentes, L.
TITULO:
A calorimetric study of S-alkylglutathiones to Glutathione S-transferase.
REVISTA:
Biochim Biophys Acta. 2001 1548(1):106-113.
ABSTRACT: The binding of three competitive glutathione analogue inhibitors (S-alkylglutathione derivatives) to glutathione S-transferase from Schistosoma japonicum, SjGST, has been investigated by isothermal titration microcalorimetry at pH 6.5 over a temperature range of 15--30 degrees C. Calorimetric measurements in various buffer systems with different ionization heats suggest that no protons are exchanged during the binding of S-alkylglutathione derivatives. Thus, at pH 6.5, the protons released during the binding of substrate may be from its thiol group. Calorimetric analyses show that S-methyl-, S-butyl-, and S-octylglutathione bind to two equal and independent sites in the dimer of SjGST. The affinity of these inhibitors to SjGST is greater as the number of methylene groups in the hydrocarbon side chain increases. In all cases studied, Delta G(0) remains invariant as a function of temperature, while Delta H(b) and Delta S(0) both decrease as the temperature increases. The binding of three S-alkylglutathione derivatives to the enzyme is enthalpically favourable at all temperatures studied. The temperature dependence of the enthalpy change yields negative heat capacity changes, which become less negative as the length of the side chain increases.
AUTORES:
Martinez-Rodrigez, S.,
Las Heras-Vazquez, F.J., Clemente-Jimenez, J.M., Mingorance-Cazorla, L.,
Rodriguez-Vico, F..
TITULO: Complete conversion of D,L-5-monosubsitituted hydantoins with a low velocity of chemical racemization into D-amino acids using whole cells of recombinant Escherichia coli.
REVISTA: Biotechnology Progress 18:1201-1206, (2002)
ABSTRACT:
A reaction system
was developed for the production of D-amino acids from D,L-5-monosubstituted
hydantoins with a very slow rate of spontaneous racemization. For this purpose
the D-hydantoinase and D-carbamoylase from Agrobacterium radiobacter NRRL B11291
were cloned in separate plasmids and expressed in Escherichia coli. The third
enzyme, hydantoin racemase, was cloned from Agrobacterium tumefaciens C58. The
hydantoin racemase amino acid sequence was significantly similar to those
previously described. A reaction system consisting of recombinant Escherichia
coli whole cell biocatalysts containing separately expressed D-hydantoinase, D-carbamoylase,
and hydantoin recemase showed high substrate specificity and was effective
toward both aliphatic and aromatic D,L-5-monosubstituted hydantoins. After
optimizing reaction conditions (pH 8 and 50 degrees C), 100% conversion of D,L-5-(2-methylthioethyl)-hydantoin
(15 mM) into D-methionine was obtained in 30 min.
AUTORES:
Las Heras-Vazquez, F.J., Mingorance-Cazorla, L., Clemente-Jimenez, J.M., Rodriguez-Vico, F.
TITULO:
Identification of yeast species from orange by RFLP and sequence analysis of the
5.8S rRNA gene and the two internal transcribed spacers.
REF.:
FEMS Yeast Research
3(1):3-9 (2003)
ABSTRACT: Yeast
isolates from orange fruit and juice in a spontaneous fermentation were
identified and classified by two molecular techniques. The first was analysis of
the restriction pattern generated from the polymerase chain reaction (PCR)-amplified
5.8S rRNA gene and the two internal transcribed spacers (ITS)using specific
primers.The second technique was sequence analysis of the ITS regions using the
same two primers.Nine different restriction profiles were obtained from the size
of the PCR products and the restriction analyses with three endonucleases (Cfo
I,Hae III and Hin fI).These groups were identified as Candida tropicalis ,Clavispora
lusitaniae ,Hanseniaspora uvarum , Pichia anomala ,Pichia fermentans ,Rhodotorula
mucilaginosa ,Saccharomyces cerevisiae ,Saccharomyces unisporus ,and
Trichosporon asahii. Checking against identification according to
morphological,physiological and biochemical trait corroborated the molecular
identification.A total concordance was found in the identification with PCR-restriction
fragment length polymorphism of the ITS region after analysing certified yeast
strains from two different culture collections.Consequently,a rapid and reliable
identification of the yeast populations was achieved by using molecular
techniques.
AUTORES:
Mingorance-Cazorla, L., Clemente-Jiménez, J.M., Martinez-Rodriguez, S., Las
Heras-Vázquez, F.J., Rodriguez-Vico, F.
TITULO:
“Contribucion of different natural yeast to the aroma of two fermented
beverages.”
REF.:
.: World Journal of Microbiology and Biotechnology 19:297-304 (2003)
ABSTRACT:
The aroma formation in the fermentation of two types of natural musts by 12
different yeasts has been analysed. In grape must fermentation Pichia fermentans
CECT 11773, Clavispora lusitaniae OJ6 and Pichia anomala OJ5 produced the best
balance between concentrations of ethyl acetate and high alcohols. When orange
juice was fermented with the 12 yeasts, Pichia fermentans CECT 11773,
Rhodotorula mucilaginosa OJ2 and Hanseniaspora uvarum CECT 10885 produced a good
beverage with low alcoholic grade. For both types of natural musts Pichia
fermentans CECT 11773 increased the presence of higher alcohols and ethyl
acetate. After using this strain both alcoholic beverages obtained the highest
evaluation in the sensory analysis.
AUTORES:
Clemente-Jiménez, J., Las Heras-Vázquez, F.J., Martinez-Rodriguez, S.,
Mingorance-Cazorla, L., de la Escalera-Hueso, S., Rodriguez Vico, F.
TITULO:
Study parameters affecting the catalytic performance of recombinant
hydantoinase from Agrobacterium
REVISTA:
Biotechnology letters 25:1067-1073 (2003)
ABSTRACT; The D-hydantoinase gene of a wild strain of Agrobacterium tumefaciens BQL9 was isolated and studied. It had 99.78% nucleotide sequence identity with other available Agrobacterium genes. The resulting amino acid sequence showed two important substitutions affecting two a-helixes in the secondary structure of the protein. The union of Mn2+ to the protein proved to be essential for activating the enzyme and was independent of the temperature. D-hydantoinase only was inactivated in the presence of 70 mM EDTA and at temperatures over 40 ºC. The enzyme showed both hydantoinase and pyrimidinase activity, but only with the D-enantiomers of the substrates. Activity was greater in substrates with apolar groups in the number 5 carbon atom of the hydantoin. The native structure of the N-terminal end of this D-hydantoinase proved to be indispensable to its enzymatic activity.
AUTORES:
Las Heras-Vázquez, F.J., Martinez-Rodriguez, S., Mingorance-Cazorla, L.,
Clemente-Jiménez, J.M., Rodriguez Vico, F.
TITULO:
Overxpression and characterization of hydantoin racemase from Agrobacterium
tumefaciens C58
REF.: Biochemical and Biophysical Research Communications 303:541-547 (2003)
ABSTRACT;Hydantoin
racemase enzyme together with a stereoselective hydantoinase and a
stereospecific D-carbamoylase guarantee the total conversion from D,L-5-monosubstituted
hydantoins with a low velocity of racemization to optically pure D-amino acids.
In this work we have cloned and expressed the hydantoin racemase gene from two
strains of Agrobacterium tumefaciens,
C58 and LBA4404, in Escherichia coli
BL21. The recombinant protein was purified in a one-step procedure by using
immobilized cobalt affinity chromatography and showed an apparent molecular mass
of 32,000 Da in SDS-gel electrophoresis. Size exclusion chromatography analysis
determined a molecular mass of about 100,000 Da, suggesting that the native
enzyme is a tetramer. The optimal conditions for hydantoin racemase activity
were pH 7.5 and 55 ºC with L-5-ethylhydantoin as substrate. Enzyme activity was
slightly affected by the addition of Ni2+ and Co2+, and
strongly inhibited by Cu2+ and Hg2+. No effect on enzyme
activity was detected with Mn2+, EDTA or DTT. Kinetic studies showed
the preference of the enzyme for hydantoins with short rather than long
aliphatic side chains or hydantoins with aromatic rings.
AUTORES:
Andujar-Sanchez, M., Clemente-Jiménez, J.M., Las Heras-Vázquez, F.J.,
Rodriguez-Vico, F., Camara-Artigas, A., Jara-Perez V.
TITULO:
“Thermodynamics of glutatione binding to the tyrosine 7 to phenylalanine
mutant of Glutatione S- Transferase from Schistosoma japonicum”.
REF.: International Journal of Biological Macromoleculars 32(3-5):77-82 (2003).
ABSTRACT:
The binding of glutathione (GSH) to the tyrosine 7 to phenylalanine mutant
of Schistosoma japonicum glutathione
S-transferase (SjGST-Y7F) has been studied by isothermal titration calorimetry (ITC).
At pH 6.5 and 25 .C this mutant shows a higher affinity for glutathione than wild type
enzyme despite an almost complete loss of activity in the presence of 1-chloro-2,4-dinitrobenzene
(CDNB) as second substrate. The enthalpy change upon binding of GSH is more
negative for the mutant than for the wild type GST (SjGST). Changes in
accessible solvent areas (ASA) have been calculated based on enthalpy and heat
capacity changes. ASA values indicated the burial of apolar surfaces of protein
and ligand upon binding. A more negative Cp
value has been obtained for the
mutant enzyme, suggesting a more hydrophobic interaction, as may be expected
from the change of a tyrosine residue to phenylalanine.
AUTORES:
Zeyad Yassin, Emilia Ortiz-Salmerón, M. José Clemente-Jiménez, Carmen Barón
and Luis García-Fuentes
TITULO:
Role of mutation Y6F on the binding properties of Schistosoma japonicum
glutathione S-transferase
REF.:
International Journal of Biological Macromolecules 32(3-5):67-75
(2003).
ABSTRACT:
There has
been some speculation about the salt independence of Schistosoma japonicum glutathione
S-transferase (Sj26GST, EC.2.5.1.18), but this aspect has not been carefully
studied before. To establish the basis for a further development of this dependence, we have performed a methodical study of
the influence of some important ions and their concentration on the binding
properties of glutathione to Sj26GST by means of isothermal calorimetry and
fluorescence quenching. Salts like NaCl, Na2 SO4 and MgSO4 do not change
practically the affinity of the protein for its substrate, whilst MgCl2 has the
effect of decreasing the affinity as its concentration rises. However, the enthalpy change is not
affected by all the salts studied, and so, the entropy change is the causal
factor in dropping the affinity. We also looked at the conformational stability
of the protein under different conditions to check the structural changes they
provide, and found that the
unfolding parameters are practically not affected by the salt concentration. We
discuss the results in terms of the chaotropic nature of the ions implied.
AUTORES:
Yassin Z., Clemente-Jimenez, J.M., Téllez-Sanz R., Garcia-Fuente L.
TITULO:
Salt influence on glutathione Schistosoma japonicum glutathione S-transferase
binding.
REF.: International Journal of Biological Macromolecules 31:155-162 (2003)
ABSTRACT:
There has been some speculation about the salt independence of Schistosoma japonicum glutathione S-transferase (Sj26GST, EC. 2.5.1.18), but this aspect has not been carefully studied before. To establish the basis for a further development of this dependence, we have performed a methodical study of the influence of some important ions and their concentration on the binding properties of glutathione to Sj26GST by means of isothermal calorimetry and fluorescence quenching. Salts like NaCl, Na2SO4 and MgSO4 do not change practically the affinity of the protein for its substrate, whilst MgCl2 has the effect of decreasing the affinity as its concentration rises. However, the enthalpy change is not affected by all the salts studied, and so, the entropy change is the causal factor in dropping the affinity. We also looked at the conformational stability of the protein under different conditions to check the structural changes they provide, and found that the unfolding parameters are practically not affected by the salt concentration. We discuss the results in terms of the chaotropic nature of the ions implied.
AUTORES:
Clemente-Jiménez, J.M., Mingorance-Cazorla, L., Martinez-Rodriguez, S., Las
Heras-Vázquez, F.J., Rodriguez-Vico, F.
TITULO: “Molecular characterisation and oenological properties of wine yeast isolated during spontaneous fermentation of six varieties of grape must.”
REF.:
Food Microbiology 21(2): 149-155 (2004)
ABSTRACT: Fermentation by naturally occurring yeasts may produce wines of complex oenological properties that are unique to a specific region. The present work analyses the population dynamics of the yeasts during spontaneous fermentation of six varieties of grape must from the ‘‘Valle del Andarax’’ area (Spain). In this study we identified members of the genera Candida, Hanseniaspora, Issatchenkia, Metschnikowia, Pichia and Saccharomyces by PCR-RFLP of the ITS region. The capability of these yeasts to ferment grape must of the Macabeo variety was studied, and the volatile profile of the wine from each microvinification has been determined. Of all the yeasts isolated Candida stellata and Saccharomyces cerevisiae were able to consume virtually all the initial glucose, producing ethanol contents typical of table wines. The best profile of higher alcohols was given by Saccharomyces cerevisiae followed by Hanseniaspora uvarum, Issatchenkia orientalis and Candida stellata. Metschnikowia pulcherrima and Pichia fermentans showed the highest production of ethyl caprilate and 2-phenyl ethanol, compounds associated with pleasant aromas.
TITULO:
Molecular cloning purification and biochemical characterization of hydantoin
racemase from legume symbiont Siorhizobium meliloti
AUTORES:
Martinez-Rodriguez, S., Las Heras-Vázquez, F.J., Mingorance-Cazorla, L.,
Clemente-Jiménez, J.M., Rodriguez-Vico, F.
REF.:
Applied Enviromental Microbiology 70:625-630 (2004)
ABSTRACT:
Hydantoin racemase from Sinorhizobium meliloti was functionally expressed in
Escherichia coli. The native form of the enzyme was a homo-tetramer of 100 kDa.
The optimum temperature and pH for the enzyme were 40 ºC and pH 8.5,
respectively. The enzyme showed slight preference for hydantoins with short
rather than long aliphatic side chains or those with aromatic rings. Substrates,
which showed no detectable activity towards the enzyme, were found to exhibit
competitive inhibition.
TÍTULO:
A monomer form of the glutathione S-transferase Y7F mutant from Schistosoma
japonicum at acidic pH.
AUTORES:
Andujar-Sanchez, M., Clemente-Jiménez, J.M., Rodriguez-Vico, F., Las Heras-Vázquez,
F.J., Jara-Perez V., Camara-Artigas, A.,
REF:
Biochemical and Biophysical Research Communications 314(1):6-10 (2004).
ABSTRACT:
Dissociation and unfolding of homodimeric glutathione S-transferase Y7F mutant
from Schistosoma japonicum (SjGST-Y7F) were investigated at equilibrium using
urea as denaturant. The conserved residue Tyr7 plays a central role in the
catalytic mechanism and the mutation Tyr–Phe yields an inactive enzyme that is
able to bind the substrate GSH with a higher binding constant than the wild type
enzyme. Mutant SjGST-Y7F is a dimer at pH 6 or higher and a stable monomer at pH
5 that binds GSH (K value of 1.2_105 _6.4_103M_1 at pH 6.5 and 6.3_104
_1.25_103M_1 at pH 5). The stability of the SjGST-Y7F mutant was studied by urea
induced unfolding techniques (DGW ¼ 13:86 _ 0:63 kcal mol_1 at pH 6.5 and DGW
¼ 11:22 _ 0:25 kcal mol_1 at pH 5) and the monomeric form characterized by
means of size exclusion chromatography, fluorescence, and electrophoretic
techniques.
TITULO:
“Biochemical characterization of a novel hydantoin racemase from
Agrobacterium tumefaciens C58.”
AUTORES:
Martinez-Rodriguez, S., Las Heras-Vázquez, F.J., Clemente-Jiménez, J.M.,
Rodriguez-Vico, F.
REF.:
Biochimie 86(2): 77-81 (2004).
ABSTRACT:
A novel hydantoin racemase gene of Agrobacterium tumefaciens C58 (AthyuA2) has
been cloned and expressed in Escherichia coli BL21. The recombinant protein was
purified in a one-step procedure and showed an apparent molecular mass of 27,000
Da in SDS-gel electrophoresis. Size exclusion chromatography analysis determined
a molecular mass of approximately 100,000 Da, suggesting that the native enzyme
is a tetramer. The optimum pH and temperature for hydantoin racemase activity
were 7.5 and 55 °C, respectively, with L-5-ethylhydantoin as substrate. Enzyme
activity was strongly inhibited by Cu2+ and Hg2+. No effect on enzyme activity
was detected with any other divalent cations, EDTA or DTT, suggesting that it is
not a metalloenzyme. Kinetic studies showed the preference of the enzyme for
hydantoins with short rather than long aliphatic side chains or hydantoins with
aromatic rings.
TITULO:
“Cloning of D-specific Hydantoin utilization genes from Arthrobacter
crystallopoietes.”
AUTORES:
Werner, M., Las Heras-Vázquez, F.J., Fritz, C., vielhauer, O., Siemann-Herzberg,
Altenbuchner, J., Syldatk, C.
REF.:
Engineering of Life Sciences 4(6):563-572 (2004).
ABSTRACT: Arthrobacter crystallopoietes produces a D-specific hydantoinase and D-N-carbamoylase useful for the industrial production of enantiomerically pure D-amino acids. The D-hydantoinase was purified by conventional chromatographic methods. The enzyme was digested by trypsine and some of the oligopeptides sequenced. The amino acid sequence information was used to isolate a DNA fragment of the corresponding gene. By several steps of inverse PCR a region of 6011 bp was obtained from the A. crystallopoietes genome surrounding the hydantoinase gene fragment. DNA sequence analysis revealed the presence of a D-hydantoinase, the D-N-carbamoylase and a putative L-N-carbamoylase on the 6011 bp fragment. In addition, two incomplete open reading frames (ORFs) showed similarity to LacI type transcriptional regulators and transmembrane proteins responsible for the uptake of nucleosides and allantoin, respectively. The D-hydantoinase gene and the D-N-carbamoylase gene were expressed in Escherichia coli and the enzyme activity was determined. From the putative L-N-carbamoylase gene, expressed in the same way in E. coli, only insoluble protein was obtained and no carbamoylase activity was detected.
TITULO:
“Influence
of sequential yeast mixture on wine fermentation.”
AUTORES:
Clemente-Jiménez, J.M., Mingorance-Cazorla, L.,
Martinez-Rodriguez, S., Las Heras-Vázquez, F.J., Rodriguez-Vico, F.
REF.:
International
Journal of Food Microbiology 98(3): 301-308 (2005).
ABSTRACT:
The use of Pichia fermentans in pure cultures and sequential mixtures with
Saccharomyces cerevisiae has been studied to improve the aromatic compounds and
characteristics of a wine. P. fermentans has proved to be a good starter strains
for must fermentation in the winemaking industry. It has shown the same level of
sulphur tolerance and the same growth rate as S. cerevisiae. We have
demonstrated that only 2 days of must fermentation with P. fermentans in
sequential mixtures are enough to increase the following compounds in the wine
both qualitatively and quantitatively: acetaldehyde, ethyl acetate, 1-propanol,
n-butanol, 1-hexanol, ethyl caprilate, 2,3-butanediol and glycerol. Maintaining
this non-Saccharomyces strain in contact with the must for longer periods
quantitatively increases the flavour composition.
TITULO:
“Crystallographic
and Thermodynamic Analysis of the Binding of S-Octylglutathione to the Tyr 7 to
Phe Mutant of Glutathione S-Transferase from Schistosoma japonicum.”
AUTORES: Montserrat
Andújar-Sánchez, Alex W. Smith, Josefa María Clemente-Jimenez, Felipe
Rodriguez-Vico, Francisco Javier Las Heras-Vazquez, Vicente Jara-Pérez, and Ana
Cámara-Artigas.
REF.: Biochemistry
44(4): 1174-1183 (2005).
ABSTRACT:
Glutathione S-transferases are a family of multifunctional enzymes involved in
the metabolism of drugs and xenobiotics. Two tyrosine residues, Tyr 7 and Tyr
111, in the active site of the enzyme play an important role in the binding and
catalysis of substrate ligands. The crystal structures of Schistosoma japonicum
glutathione S-transferase tyrosine 7 to phenylalanine mutant [SjGST(Y7F)] in
complex with the substrate glutathione (GSH) and the competitive inhibitor S-octylglutathione
(S-octyl-GSH) have been obtained. These new structural data combined with
fluorescence spectroscopy and thermodynamic data, obtained by means of
isothermal titration calorimetry, allow for detailed characterization of the
ligand-binding process. The binding of S-octyl-GSH to SjGST(Y7F) is
enthalpically and entropically driven at temperatures below 30 degrees C. The
stoichiometry of the binding is one molecule of S-octyl-GSH per mutant dimer,
whereas shorter alkyl derivatives bind with a stoichiometry of two molecules per
mutant dimer. The SjGST(Y7F).GSH structure showed no major structural
differences compared to the wild-type enzyme. In contrast, the structure of
SjGST(Y7F).S-octyl-GSH showed asymmetric binding of S-octyl-GSH. This lack of
symmetry is reflected in the lower symmetry space group of the SjGST(Y7F).S-octyl-GSH
crystals (P6(3)) compared to that of the SjGST(Y7F).GSH crystals (P6(3)22).
Moreover, the binding of S-octyl-GSH to the A subunit is accompanied by
conformational changes that may be responsible for the lack of binding to the B
subunit..
TITULO:
“Molecular
Cloning and Biochemical Characterization of L-N-Carbamoylase from Sinorhizobium
meliloti CECT4114.”
AUTORES: Martínez-Rodríguez
S, Clemente-Jiménez JM, Rodríguez-Vico F, Las Heras-Vázquez FJ
REF.:
J. Mol. Microbiol. Biotechnol. 9:16-25 (2005).
ABSTRACT:
An N-carbamoyl-L-amino acid amidohydrolase (L-N-carbamoylase) from Sinorhizobium
meliloti CECT 4114 was cloned and expressed in Escherichia coli. The recombinant
enzyme catalyzed the hydrolysis of N-carbamoyl alpha-amino acid to the
corresponding free amino acid, and its purification has shown it to be strictly
L-specific. The enzyme showed broad substrate specificity, and it is the first
L-N-carbamoylase that hydrolyses N-carbamoyl-L-tryptophan as well as N-carbamoyl
L-amino acids with aliphatic substituents. The apparent Km values for N-carbamoyl-L-methionine
and tryptophan were very similar (0.65 +/- 0.09 and 0.69 +/- 0.08 mM,
respectively), although the rate constant was clearly higher for the L-methionine
precursor (14.46 +/- 0.30 s(-1)) than the L-tryptophan one (0.15 +/- 0.01
s(-1)). The enzyme also hydrolyzed N-formyl-L-methionine (kcat/Km = 7.10 +/-
2.52 s(-1) x mM(-1)) and N-acetyl-L-methionine (kcat/Km = 12.16 +/- 1.93 s(-1) x
mM(-1)), but the rate of hydrolysis was lower than for N-carbamoyl-L-methionine
(kcat/Km = 21.09 +/- 2.85). This is the first L-N-carbamoylase involved in the 'hydantoinase
process' that has hydrolyzed N-carbamoyl-L-cysteine, though less efficiently
than N-carbamoyl-L-methionine. The enzyme did not hydrolyze ureidosuccinic acid
or 3-ureidopropionic acid. The native form of the enzyme was a homodimer with a
molecular mass of 90 kDa. The optimum conditions for the enzyme were 60 degrees
C and pH 8.0. Enzyme activity required the presence of divalent metal ions such
as Ni2+, Mn2+, Co2+ and Fe2+, and five amino acids putatively involved in the
metal binding were found in the amino acid sequence.
TITULO: “Obtención
de D-aminoácidos ópticamente puros a partir de un sistema recombinante. Caracterización
de las enzimas involucradas en el proceso.”
AUTORES: Sergio
Martínez Rodríguez
REF.:
Libro. Servicio de Publicaciones de la Universidad de Almería.
ISBN:
84-8240-781-3 (2005)
TITULO:
“Yeast autolysis for L-amino acids production”
AUTORES: Pozo-Dengra, J., Martínez-Rodríguez, S., Martínez-Gómez, A.I., Las Heras-Vázquez, F.J., Rodríguez-Vico, F., Clemente-Jiménez, J.M.
REF.: Enzyme and Microbial Technology (2006) 40:46-50 .
ABSTRACT:
As
yeast extracts are commonly used as a source of amino acids, the present work
studies potential of one Saccharomyces cerevisiae strain and several non-Saccharomyces
yeasts to be used as amino acids source. All the strains studied were able to
grow using sugar-cane molass as medium. Pichia strains proved to be the best
biomass producers, but Yarrowia showed the highest rates and the best yield of
hydrolysis from protein to free amino acids. Yarrowia strains also proved to
contain the greatest quantity of essential amino acids. Finally, a phylogenetic
tree was obtained from the amino acid profile which agrees with the
classification of the strains.
TITULO:
“Activity
measurement of hydantoin racemase enzyme: a key for the production of optically
pure D-amino acids”
AUTORES:
Sergio Martínez-Rodríguez, Josefa María Clemente-Jiménez, Felipe Rodríguez-Vico
and Francisco Javier Las Heras-Vázquez
REF.: Capítulo de libro. R. Konno, N. Fujii, H. Homma, H. Brückner, G. Fisher, A. D'Aniello (Eds.), D-Amino Acids: A New Frontier in Amino Acids and Protein Research. Nova Science, Inc., Hauppauge, New York, USA. (2006). ISBN: 1-60021-075-9
ABSTRACT:
Optically pure D-amino acids can be enzymatically obtained from D,L-5-monosubstituted
hydantoins, a process which is cheaper and less contaminating than
chemoenzymatic production. In this enzymatic reaction, called “hydantoinase
process” (1), firstly the chemically synthesised D,L-5-monosubstituted
hydantoin ring is hydrolysed by a stereoselective hydantoinase enzyme. Further
hydrolysis of the resulting enantiospecific N-carbamoyl-D-amino acid to the free
D-amino acid is catalysed by highly enantiospecific N-carbamoyl-D-amino acid
amidohydrolase (D-carbamoylase). At the same time as D-hydantoinase hydrolyses
the enantiospecific D-5-monosubstituted hydantoin, the chemical and/or enzymatic
racemization of L-5-monosubstituted hydantoin starts.
TITULO:
“Optimisation of two
recombinant whole cell systems for the production of optically pure D-amino
acids”
AUTORES:
A. I. Martínez-Gómez, S. Martínez-Rodríguez,
J. Pozo-Dengra, J. M. Clemente-Jiménez, F. Rodríguez-Vico and F. J. Las Heras-Vázquez.
REF.:
Capítulo de libro: “Modern
Multidisciplinary applied Microbiology” Exploiting Microbes and their
Interactions (2006). Ed. Antonio Mendez-Vilas. Wiley VCH. Capítulo de
libro. ISBN978-3-527-31611-3. Paginas
246-250.
ABSTRACT:
Two whole cell
recombinant systems for the production of optically pure D-amino acids were
optimised and compared. Each system contained three enzymes, 2 of which were D-hydantoinase
and D-carbamoylase from Agrobacterium tumefaciens BQL9. The third enzyme was
hydantoin racemase 1 for the first system, and hydantoin racemase 2 for the
second one, both from Agrobacterium tumefaciens C58. The analysis of the
induction parameters for the two whole cell systems, in Escherichia coli JM109,
showed that the optimal induction conditions were the same as regards induction
time and temperature, i.e. 8 hours and 34ºC. However optimum concentration of
the inducer, isopropyl-b-D-thiogalactosidase
(IPTG), was different (0.1 mM of IPTG for system 1 and 0.2 mM for system 2). The
production of D-amino acids was analysed after transforming four E. coli strains
with the two constructions, and E. coli strain BL21 was found to be the best
host strain for both systems. Comparison of the two systems in BL21 showed that
system 1 produced 100% conversion from the substrate D,L-5-methylthioethyl-hydantoin
(D,L-MTEH) to the product D-methionine in 180 minutes, while system 2 took twice
as long.
TITULO:
“Binding studies of
hydantoin racemase from Sinorhizobium meliloti by calorimetric and fluorescence
analysis.”
AUTORES:
Montserrat Andújar-Sánchez,
Sergio Martínez-Rodríguez, Francisco Javier Las Heras-Vázquez, Josefa María
Clemente-Jiménez, , Felipe Rodríguez-Vico, Vicente Jara-Pérez
REF.:
Biochimica et Biophysica Acta (2006) 1764(2):292-298.
ABSTRACT:
Hydantoin racemase
enzyme together with a stereoselective hydantoinase and a stereospecific D-carbamoylase
guarantee the total conversion from D,L-5-monosubstituted hydantoins with a low
velocity of racemization, to optically pure D-amino acids. Hydantoin racemase
from Sinorhizobium meliloti was
expressed in Escherichia coli.
Calorimetric and fluorescence experiments were then carried out to obtain the
thermodynamic binding parameters, DG,
DH
and DS
for the inhibitors L- and D-5-methylthioethyl-hydantoin. The number of active
sites is four per enzyme molecule (one per monomer), and the binding of the
inhibitor is entropically and enthalpically favoured under the experimental
conditions studied. In order to obtain information about amino acids involved in
the active site, four different mutants were developed in which cysteine 76 and
181 were mutated to Alanine and Serine. Their behaviour shows that these
cysteines are essential for enzyme activity, but only cysteine 76 affects the
binding to these inhibitors.
TITULO:
“Enzymatic activity assay of D-hydantoinase by isothermal titration
calorimetry. Determination of the thermodynamic activation parameters for the
hydrolysis of several substrates.”
AUTORES:
Montserrat Andujar-Sánchez, Francisco Javier Las Heras-Vázquez, Josefa María
Clemente-Jiménez, Sergio Martínez-Rodríguez, Ana María Cámara-Artigas,
Felipe Rodríguez-Vico, Vicente Jara Pérez
REF.:
J. Biochem. and Biophys. Methods (2006) 67(1): 57-66.
ABSTRACT:
Isothermal titration calorimetry (ITC) has been applied to the determination of
the activity of D-hydantoinase (EC 3.5.2.2) with several substrates by
monitoring the heat released during the reaction. The method is based on the
proportionality between the reaction rate and the thermal power (heat/time)
generated. Microcalorimetric assays carried out at different temperatures
provided the dependence of the catalytic rate constant on temperature. We show
that ITC assay is a nondestructive method that allows the determination of the
catalytic rate constant (kcat), Michaelis constant (KM), activation energy and
activation Gibbs energy, enthalpy and entropy of this reaction.
TITULO:
Thermodynamic and mutational studies of L-N-Carbamoylase from Sinorhizobium
meliloti CECT 4114 catalytic centre.
AUTORES:
Sergio Martínez-Rodríguez, Montserrat Andujar-Sánchez, Josefa María
Clemente-Jiménez, Vicente Jara Pérez, Felipe Rodríguez-Vico, Francisco Javier
Las Heras-Vázquez,
REF.:
Biochimie (2006) 88(7): 837-847.
ABSTRACT:
Purified site-directed mutants of Sinorhizobium meliloti CECT 4114 l-N-carbamoylase
(SmLcar) in which Glu132, His230, Asn279 and Arg292 were replaced have been
studied by kinetic methods and isothermal titration calorimetry (ITC). The
importance of His230, Asn279 and Arg292 residues in the recognition of N-carbamoyl-l-alpha-amino
acids has been proved. The role of Glu132 has been confirmed in substrate
hydrolysis. ITC has confirmed two Ni atoms per monomer of wild type enzyme, and
two equal and independent substrate binding sites (one per monomer). Homology
modelling of SmLcar supports the importance of His87, His194, His386, Glu133 and
Asp98 in metal binding. A comprehensive reaction mechanism is proposed on the
basis of binding experiments measured by ITC, kinetic assays, and homology of
the active centre with beta-alanine synthase from Saccharomyces kluyveri and
other enzymes.
TITULO:
Site-directed mutagenesis indicates an important role of Cysteines 76 and 181 in
the catalysis of hydantoin racemase from Sinorhizobium meliloti CECT 4114.
AUTORES: Martinez-Rodriguez, S, Andujar Sanchez M., Neira JL, Clemente-Jimenez JM, Jara-Perez V, Rodríguez-Vico, F, Las Heras-Vazquez FJ
REF.: Protein
Science (2006) 15:2729-2738.
ABSTRACT:
Hydantoin racemase enzyme plays a crucial role in the reaction cascade known as
"hydantoinase process." In conjunction with a stereoselective
hydantoinase and a stereospecific carbamoylase, it allows the total conversion
from D,L-5-monosubstituted hydantoins, with a low rate of racemization, to
optically pure D- or L-amino acids. Residues Cys76 and Cys181 belonging to
hydantoin racemase from Sinorhizobium meliloti (SmeHyuA) have been proved to be
involved in catalysis. Here, we report biophysical data of SmeHyuA Cys76 and
Cys181 to alanine mutants, which point toward a two-base mechanism for the
racemization of 5-monosubstituted hydantoins. The secondary and the tertiary
structure of the mutants were not significantly affected, as shown by circular
dichroism. Calorimetric and fluorescence experiments have shown that Cys76 is
responsible for recognition and proton retrieval of D-isomers, while Cys181 is
responsible for L-isomer recognition and racemization. This recognition process
is further supported by measurements of protein stability followed by chemical
denaturation in the presence of the corresponding compound.
TITULO:
Cloning, overexpression, purification, crystallization and preliminary
crystallographic studies of the recombinant dihydropyrimidinase from
Sinorhizobium meliloti CECT4114.
AUTORES:
Martínez-Rodríguez S, González-Ramírez; LA, Clemente-Jiménez; JM, Rodríguez-Vico
F, Las Heras-Vázquez FJ, Gavira JA, García-Ruiz JM
REF.:
Acta Cristalografica F (2006) F62:1223-1226
ABSTRACT:
Dihydropyrimidinases are involved in the reductive pathway of pyrimidine
degradation, catalysing the hydrolysis of 5,6-dihydrouracil and 5,6-dihydrothymine
to the corresponding N-carbamoyl beta-amino acids. This enzyme has often been
referred to as hydantoinase owing to its industrial application in the
production of optically pure amino acids starting from racemic mixtures of 5-monosubstituted
hydantoins. Recombinant dihydropyrimidinase from Sinorhizobium meliloti CECT4114
(SmelDhp) has been expressed, purified and crystallized. Crystallization was
performed using the counter-diffusion method with capillaries of 0.3 mm inner
diameter. Crystals of SmelDhp suitable for data collection and structure
determination were grown in the presence of agarose at 0.1%(w/v) in order to
ensure mass transport controlled by diffusion. X-ray data were collected to a
resolution of 1.85 A. The crystal belongs to the orthorhombic space group
C222(1), with unit-cell parameters a = 124.89, b = 126.28, c = 196.10 A and two
molecules in the asymmetric unit. A molecular-replacement solution has been
determined and refinement is in progress.
TITULO:
Recombinant polycistronic structure of Hydantoinase process genes in Escherichia
coli for the production of optically pure D-amino acids.
REF.: Appl
Environ Microbiol (2007).
ABSTRACT:
Two recombinant reaction systems for the production of optically pure D-amino
acids from different D,L-5-monosubstituted hydantoins have been constructed.
Each system contained three enzymes, two of which were D-hydantoinase and D-carbamoylase
from Agrobacterium tumefaciens BQL9. The third enzyme was hydantoin racemase 1
for the first system, and hydantoin racemase 2 for the second one, both from
Agrobacterium tumefaciens C58. Each system was formed by a recombinant
Escherichia coli strain with one plasmid harbouring three genes co-expressed
with one promoter in a polycistronic structure. D-carbamoylase gene was cloned
closest to the promoter in order to obtain the highest synthesis of the enzyme,
thus avoiding intermediate accumulation, which decreases the reaction rate. Both
systems were able to produce 100% conversion and 100% optically pure D-methionine,
D-leucine, D-norleucine, D-norvaline, D-aminobutyric acid, D-valine, D-phenylalanine,
D-tyrosine and D-tryptophan from the corresponding hydantoin racemic mixture.
For the production of almost all D-amino acids studied in this work, system 1
hydrolyzed the 5-monosubstituted hydantoins faster than system 2.
AUTORES:
Mingorance-Cazorla L, Molina-Ruiz JM, Clemente-Jiménez J, Las Heras-Vázquez
FJ, Rodriguez Vico F.
TITULO:
“Procedimiento para la obtencion de un producto derivado del zumo de naranja
que implica la modificacion de su composicion quimica por un proceso de
fermentacion dirigida y natural mediante el empleo de levaduras alcoholeras”
Nº DE REGISTRO:
P 9902315
AÑO: 1999
ENTIDAD
TITULAR: Mingorance-Cazorla,
L., Molina-Ruiz, J.M., Clemente-Jiménez, J.M., Las Heras- Vázquez, F.J.,
Rodriguez-Vico F.
RESUMEN:
Procedimiento para la
obtención de un producto derivado del zumo de naranja que implica la modificación
de su composición química por un proceso de fermentación dirigida y natural
mediante el empleo de levaduras alcoholeras que consiste n la extracción de
zumo con bajo contenido en aceite esencial y clarificación hasta un nivel máximo
de pulpa del 15%: pasteurización del zumo de naranja por calentamiento hasta
los 80 ºC durante 12 segundos: inoculación del zumo con un cultivo de
microorganismos: incubación del material permitiendo el intercambio de aire estéril
y la salida de CO2: adición de sacarosa y conservación
AUTORES:
Martinez-Rodriguez S.,
Las Heras-Vázquez, F.J., Mingorance-Cazorla, L., Clemente- Jiménez, J.M.,
Rodriguez-Vico F.
TITULO:
“Sistema enzimático y procedimiento para la preparación de D-aminoacidos o
derivados de los mismos”
Nº DE REGISTRO:
P 200202208
AÑO: 2002
ENTIDAD TITULAR:
Universidad de Almería.
RESUMEN:
AUTORES:
Mingorance-Cazorla, L., Clemente-Jiménez, J.M., Las Heras-Vázquez, F.J.,
Martinez- Rodriguez, S., Ferre-Carretero, F., Rodriguez-Vico, F.
TITULO:
“ Procedimiento para el incremento del aroma del vino por fermentacion
dirigida secuencial y levaduras
utilizadas en el mismo.”
Nº DE REGISTRO:
ENVIADA
AÑO: 2002
ENTIDAD
TITULAR: Universidad de
Almería
RESUMEN:
La presente invención tiene por objeto una nueva cepa de la levadura Pichia
fermentans, así como el procedimiento de obtención de la misma, y su
aplicación en la producción de vinos. En concreto atañe, por un lado, a un
procedimiento de fermentación dirigida y secuencial para la producción de
vino, y por otro, al aislamiento, caracterización y uso de una cepa de levadura
específica para la obtención de un producto fuertemente aromatizado, con sus
características organolépticas mejoradas y reproducibles
AUTORES: Rodriguez-Vico, F., Las Heras-Vázquez, F.J., Clemente-Jiménez, J.M., Martinez-Rodriguez S., Madrid-Romero, P., Mingorance-Cazorla, L.
TITULO:
“Procedimiento de fermentación dirigida y su aplicación en la obtención de
nuevas bebidas derivadas de zumo de naranja natural”
Nº DE REGISTRO:
P 2222091
AÑO: 2003
ENTIDAD TITULAR:
Universidad de Almería.
AUTORES: Ana Isabel Martínez Gómez, Sergio Martínez Rodríguez, Josefa María Clemente Jiménez, Joaquín Pozo Dengra, Pedro Madrid Romero, Francisco Javier Las Heras Vázquez., Felipe Rodríguez Vico
TITULO:
“Sistema de coexpresión enzimática para la construcción de una nueva ruta
metabólica que permite la preparación de D-amioácidos y procedimiento para
biosíntesis de los mismos.”
Nº DE REGISTRO:
Enviada
AÑO: 2006
ENTIDAD TITULAR: Universidad de Almería.
CEPA:
Xanthomonas campestris
DEPOSITO: CECT 4807
PROPIETARIOS:
Clemente-Jiménez, J.M., Las Heras-Vázquez F.J., Rodríguez-Vico F.
PROPIETARIOS:
Las Heras-Vázquez, F.J., Clemente-Jiménez, J.M., Rodriguez-Vico, F.
CEPA:
Hanseniaspora uvarum
DEPOSITO:
CECT 10.885
PROPIETARIOS:
Mingorance-Cazorla, L., Molina-Ruíz, J.M., Las Heras-Vázquez, F.J.,
Clemente-Jiménez, J.M., Rodríguez-Vico, F.
PROPIETARIOS:
Mingorance-Cazorla L., Las Heras-Vázquez F.J., Clemente-Jiménez J.M., Rodríguez-Vico,
F.
PROPIETARIOS:
Mingorance-Cazorla, L., Las Heras-Vázquez, F.J., Clemente-Jiménez, J.M., Rodríguez-Vico,
F.
-
AUTORES: Las Heras-Vázquez F.J., Mingorance-Cazorla, L.,
Clemente-Jiménez, J.M., Rodríguez-Vico, F.
ORGANISMO:
Candida tropicalis
Nº
DE ACCESO:
AF321539
- AUTORES:
Las Heras-Vázquez F.J., Mingorance-Cazorla, L., Clemente-Jiménez, J.M., Rodríguez-Vico,
F.
ORGANISMO:
Saccharomyces cerevisiae
Nº
DE ACCESO:
AF321540
- AUTORES:
Las Heras-Vázquez F.J., Mingorance-Cazorla, L., Clemente-Jiménez, J.M., Rodríguez-Vico,
F.
ORGANISMO:
Clavispora
lusitaneae
Nº DE
ACCESO: AF321541
- AUTORES:
Mingorance-Cazorla L., Las Heras-Vázquez F.J., Clemente-Jiménez, J.M., Rodríguez-Vico,
F.
ORGANISMO:
Saccharomices unisporus
Nº
DE ACCESO:
AF321542
- AUTORES:
Mingorance-Cazorla L., Las Heras-Vázquez F.J., Clemente-Jiménez, J.M., Rodríguez-Vico,
F.
ORGANISMO:
Pichia anomala
Nº DE
ACCESO: AF321543
- AUTORES:
Mingorance-Cazorla L., Las Heras-Vázquez F.J., Clemente-Jiménez, J.M.,
Rodríguez-Vico,
F.
ORGANISMO:
Rodotorula mucilaginosa
Nº DE
ACCESO: AF321544
- AUTORES:
Mingorance-Cazorla L., Las Heras-Vázquez F.J., Clemente-Jiménez, J.M., Rodríguez-Vico,
F.
ORGANISMO:
Trichosporon sp.
Nº DE
ACCESO: AF322110
- AUTORES:
Las Heras-Vázquez F.J., Mingorance-Cazorla, L., Clemente-Jiménez, J.M., Rodríguez-Vico,
F.
ORGANISMO:
Hanseniaspora uvarum
Nº DE
ACCESO: AY027507
- AUTORES:
Las Heras-Vázquez F.J., Mingorance-Cazorla, L., Clemente-Jiménez, J.M., Rodríguez-Vico,
F.
ORGANISMO:
Pichia fermentans
Nº DE
ACCESO: AY027508
- AUTORES:
Mingorance-Cazorla, L., Las Heras-Vázquez F.J., Martínez-Rodríguez, S.,
Clemente-Jiménez, J.M., Rodríguez-Vico, F.
ORGANISMO:
Candida stellata
Nº DE
ACCESO: AY235805
- AUTORES:
Las Heras-Vázquez F.J., Mingorance-Cazorla, L., Martínez-Rodríguez, S.,
Clemente-Jiménez, J.M., Rodríguez-Vico, F.
ORGANISMO:
Hanseniaspora uvarum
Nº DE
ACCESO: AY235806
- AUTORES:
Mingorance-Cazorla, L., Las Heras-Vázquez F.J., Martínez-Rodríguez, S.,
Clemente-Jiménez, J.M., Rodríguez-Vico, F.
ORGANISMO:
Issatchenkia orientalis
Nº DE
ACCESO: AY235807
- AUTORES:
Mingorance-Cazorla, L., Las Heras-Vázquez F.J., Martínez-Rodríguez, S.,
Clemente-Jiménez, J.M., Rodríguez-Vico, F.
ORGANISMO:
Issatchenkia terricola
Nº DE
ACCESO: AY235808
- AUTORES:
Mingorance-Cazorla, L., Las Heras-Vázquez F.J., Martínez-Rodríguez, S.,
Clemente-Jiménez, J.M., Rodríguez-Vico, F.
ORGANISMO:
Metschnikowia pulcherrima
Nº DE
ACCESO: AY235809
- AUTORES:
Las Heras-Vázquez F.J., Mingorance-Cazorla, L., Martínez-Rodríguez, S.,
Clemente-Jiménez, J.M., Rodríguez-Vico, F.
ORGANISMO:
Pichia fermentans
Nº DE
ACCESO: AY235810
- AUTORES:
Mingorance-Cazorla, L., Las Heras-Vázquez F.J., Martínez-Rodríguez, S.,
Clemente-Jiménez, J.M., Rodríguez-Vico, F.
ORGANISMO:
Saccharomyces cerevisiae
Nº DE
ACCESO: AY235811
TIPO DE PARTICIPACION: Poster.
CONGRESO:
IX Congreso de la Sociedad Española de Bioquímica.
TITULO:
Efecto del ión floruro y de distintas fosfatasas sobre la 3-hidroxi-3-metilglutaril-Coenzima
A reductasa.
AUTORES: Rodríguez-Vico,
F., Ramírez, H., García-Peregrín, E.
LUGAR DE CELEBRACION:
Tenerife (España).
AÑO: 1.984.
TIPO DE PARTICIPACION: Poster.
CONGRESO:
25 th International Conference on the Biochemistry of lipids.
TITULO:
Improvement of the modification for the assay of 3-hidroxi-3-metilglutaril-Coenzima
A reductasa.
AUTORES: Rodríguez-Vico,
F., Ramírez, H., García-Peregrín E.
LUGAR DE CELEBRACION:
Antwerp (Holanda).
AÑO: 1.984.
TIPO DE PARTICIPACION: Poster.
CONGRESO:
XII Congreso de la Sociedad Española de Bioquímica.
TITULO:
Interacción lipidoproteica de la 3-hidroxi-3-metilglutaril-Coenzima A reductasa
en sistemas reconstituidos.
AUTORES: Rodríguez-Vico
F., García-Peregrín, E., Ramírez, H.
LUGAR DE CELEBRACION:
Valencia (España).
AÑO: 1.985.
TIPO DE PARTICIPACION: Poster.
CONGRESO:
XIII Congreso de la sociedad Española de Bioquímica.
TITULO:
Efecto del colesterol sobre la actividad de la HMG-CoA reductasa constituida en
vesículas fosfolípidicas.
AUTORES: Rodríguez-Vico, F., García-Peregrín, E., Ramírez, H.
LUGAR DE CELEBRACION:
Zaragoza (España).
AÑO: 1.986.
TIPO DE PARTICIPACION: Poster.
CONGRESO:
XIV Congreso de la Sociedad Española de Bioquímica.
TITULO:
Modulación y propiedades de la HMG-CoA reductasa reconstituida en vesículas
fosfolipídicas.
AUTORES: Rodríguez-Vico, F., Martínez-Cayuela, H., Ramírez, H., García-Peregrín, E.
LUGAR DE CELEBRACION:
Málaga (España).
AÑO: 1.987.
TIPO DE PARTICIPACION: Poster.
CONGRESO:
30 th I.C.B.L.
TITULO:
Sterol activated degradation of HMG-CoA reductase is not probably the only
mechanism of membrane-medated control of enzyme.
AUTORES: Rodríguez-Vico, F., Martínez-Cayuela, M., García-Peregrín, E., Ramírez, H.
LUGAR DE CELEBRACION:
Stresa (Italia).
AÑO: 1.989.
TIPO DE PARTICIPACION: Poster.
CONGRESO:
XVI Congreso S.E.B.
TITULO:
Metabolismo del colesterol en Anguilla
anguilla: Influencia del tipo de dieta sobre las principales enzimas
reguladoras.
AUTORES:
Burgos, C., Castillo, M., Rodríguez-Vico, F., Zafra, M.F., García-Peregrín,
E.
LUGAR DE CELEBRACION:
Alicante (España).
AÑO: 1.989.
TIPO DE PARTICIPACION: Poster.
CONGRESO:
XVI Congreso S.E.B
TITULO:
Estudio comparativo de las distintas fracciones lipídicas en Toxocara
canis, T. cati y Toxocaris leonina.
AUTORES: Fatou, A., Ortega, J.E., Rodríguez-Vico, F., Valero, A., Monteoliva, M.
LUGAR DE CELEBRACION:
Alicante (España).
AÑO: 1.989.
TIPO DE PARTICIPACION: Poster.
CONGRESO:
I Congr. Int.Alimentación, Nutrición y Dietética.
TITULO:
Efecto de la suplementación de la dieta con grasas saturadas sobre la composición
de las lipoproteínas del suero.
AUTORES:
Zafra, M.F., Castillo, M., Rodríguez-Vico, F., García-Peregrín, E.
LUGAR DE CELEBRACION:
Toledo (España).
AÑO: 1.991.
TIPO DE PARTICIPACION: Poster.
CONGRESO:
32 th I.C.B.L.
TITULO:
Influence of dietary coconut oil on chick serum cholesterol and lipoprotein
composition.
AUTORES:
Zafra, M.F., Castillo, M., López, J.M., Rodríguez-Vico, F., García-Peregrín,
E.
LUGAR DE CELEBRACION:
Granada (España).
AÑO: 1.991.
TIPO DE PARTICIPACION: Poster.
CONGRESO:
32th I.C.B.L.
TITULO:
Postnatal development of chick serum lipoproteins.
AUTORES:
Castillo, M., Zafra, M.F., López, J.M., Rodríguez-Vico, F., García-Peregrín,
E.
LUGAR DE CELEBRACION:
Granada (España).
AÑO: 1.991.
TIPO DE PARTICIPACION: Poster.
CONGRESO:
13 th International congress of Biochemistry.
TITULO:
Study and preliminary characterization of a novel system for in vitro
reconstitution of soluble HMG-CoA reductase in synthetic lilposomes.
AUTORES:
Ramírez, H., Rodríguez-Vico, F., Marco, C., García-Peregrín, E.
LUGAR DE CELEBRACION:
Amsterdam, Breukelen, Holanda. AÑO: 1.992.
TIPO DE PARTICIPACION:
Ponente oral.
CONGRESO:
Int. Symposium "Physiologic basis of perinatal
care".
TITULO:
Induction of neonatal hypercholesterolemia by saturated fat: Changes in
lipoprotein profile.
AUTORES: Rodríguez-Vico, F., Castillo, M., López, J.M., Zafra, M.F., García-Peregrín,
E.
LUGAR DE CELEBRACION:
Salamanca (España). AÑO: 1.992.
TIPO DE PARTICIPACION: Poster.
TITULO:
Genetic similarities
based on RAPD bands are in accordance with the known pedigrees of pepper
breedings lines.
AUTORES:
Las Heras-Vázquez, F.J., Clemente-Jiménez, J.M., Rodríguez-Vico F.
CONGRESO: IX
Meeting on Genetics and Breeding on Capsicum and Eggplant.
LUGAR DE CELEBRACIÓN: Budapest
(Hungría)
AÑO: 1995
TIPO DE PARTICIPACION: Poster.
TITULO:
Aislamiento y analisis de marcadores moleculares de una levadura metilotrófica.
AUTORES:
Clemente-Jiménez, J.M., Cano-Sola, H., Las Heras-Vázquez, F.J., Rodríguez-Vico,
F.
CONGRESO:
XVI Congreso Sociedad Española de Microbiología.
LUGAR DE CELEBRACION:
Barcelona (España). AÑO: 1.997.
TIPO DE PARTICIPACION: Poster.
TITULO:
Caracterizacion de la alcohol oxidasa de una nueva levadura metilotrófica.
AUTORES:
Cano-Sola, H., Clemente-Jiménez, J.M., Las Heras-Vázquez, F.J., Rodríguez-Vico,
F.
CONGRESO:
XX Congreso S.E.B.
LUGAR DE CELEBRACION:
Madrid (España).
AÑO: 1.997.
TIPO DE PARTICIPACION: Poster.
TITULO:
Caracterizacion del Metabolismo de los azúcares de una nueva cepa de Hanseniospora
uvarum.
AUTORES: Mingorance-Cazorla, L., Molina-Ruíz, J.M., Las Heras-Vázquez, F.J.,
Clemente-Jiménez, J.M., Galera-Pérez, R., Rodríguez-Vico, F.
CONGRESO:
XVII Congreso S.E.M.
LUGAR DE CELEBRACION:
Granada (España).
AÑO: 1.999
TIPO DE PARTICIPACION: Poster.
TITULO:
Producción de biomasa en vinaza por Pichia
anomala.
AUTORES:
Galera-Pérez, R., Bonel, J., Aguado, J., Clemente-Jiménez, J.M., Las Heras-Vázquez,
F.J., Molina-Ruíz, J.M., Mingorance-Cazorla, L., Rodríguez-Vico, F.
CONGRESO:
XVII Congreso S.E.M.
LUGAR DE CELEBRACION:
Granada (España).
AÑO: 1.999.
TIPO DE PARTICIPACION: Poster.
TITULO:
A calorimetric study of the S-methylglutatione to glutathione S-transferase
AUTORES:
Ortiz-Salmerón, E., Yassin, Z., Clemente-Jimenez, J.M., Las Heras-Vázquez,
F.J., Rodríguez-Vico, F., Barón, C., García-Fuentes, L.
CONGRESO:
IV Congreso Iberoamericano de Biofísica
LUGAR DE CELEBRACION:
Alicante (España).
AÑO: 2.000.
TIPO DE PARTICIPACION:
Poster
TITULO:
Thermodynamic analysis of the binding of glutathione S-transferase
AUTORES:
Ortiz-Salmerón, E., Zeyad, Y., Clemente-Jimenez, J.M., Las Heras-Vázquez, F.J,
Rodríguez-Vico, F., Barón, C., García-Fuentes, L.
CONGRESO:
III European Biophysics Congress
LUGAR DE CELEBRACION:
München (Alemania)
AÑO: 2.000.
TIPO DE PARTICIPACION: Poster.
TITULO:
Identification of Yeast species from Orange by RFLP and sequence analysis of the
5.8s rRNA gene and the two internal transcribed spacers
AUTORES: Las Heras-Vazquez, F.J., Mingorance-Cazorla, L., Clemente-Jimenez, J.M., Martinez-Rodriguez,
S., Rodriguez-Vico, F.
CONGRESO:
Symposium on Physiology of Yeasts and Filamentous Fungi
LUGAR DE CELEBRACION:
Hidsgavl Castle Denmark. AÑO: 2001
TIPO DE PARTICIPACION: Poster.
TITULO:
Production of fermentation sub-products using sugar-cane molasses by different
yeast strains
AUTORES: Mingorance-Cazorla,
L., Las Heras-Vazquez, F.J., Clemente-Jimenez, J.M., Martinez-Rodriguez, S.,
Bonel, J., Aguado, J., Rodriguez-Vico,
F.
CONGRESO:
Symposium on Physiology of Yeasts and Filamentous Fungi.
LUGAR DE CELEBRACION:
Hindsgavl Castle Denmark.
AÑO: 2001.
TIPO DE PARTICIPACION: Poster.
TITULO:
Production of D,L-hydantoinase of Agrobacterium
sp. in E. coli
AUTORES: Martinez-Rodriguez,
S., Clemente-Jimenez, J.M., Las Heraz-Vazquez, F.J., Mingorance-Cazorla, L.,
Pereira-Padilla, G.P.,de la Escalera-Hueso, S., Rodriguez-Vico, F.
CONGRESO:
10th European Congress on Biotechnology
LUGAR DE CELEBRACION:
Madrid (España).
AÑO: 2001.
TIPO DE PARTICIPACION: Poster.
TITULO:
Influence of NaCl concentration on the thermodynamic parameters for the binding
of GSH to Schistosoma japonicum GST
AUTORES: Clemente-Jimenez, J.M., Las Heras-Vazquez, F.J., Rodriguez-Vico, F., Baron, C.,
Garcia-Fuentes, L., Yassin, Z., Tellez-Sanz, R.
CONGRESO:
The International Society for Biological Calorimetry
LUGAR DE CELEBRACION:
Santiago de Compostela (España). AÑO: 2001.
TIPO
DE PARTICIPACION:
Poster.
TITULO:
The tpg-gene is not sufficient for replication of the ends of linear plasmids in
Streptomyces.
AUTORES:
Altenbuchner J. Clemente Jimenez, J.M., John T. Wagner G.
CONGRESO: International Meeting on the Biology of Bacteria producing Natural Compounds
LUGAR
DE CELEBRACION:
Groningen (Holanda).
AÑO: 2003
TIPO
DE PARTICIPACION:
Poster
TITULO:
Use of chiral HPLC for biochemical characterization of a hydantoin racemase.
AUTORES:
Martinez-Rodriguez, S., Clemente-Jimenez, J.M., Martínez-Gómez, A.I., Pozo-Dengra,
J., Mingorance-Cazorla, L., Las Heraz-Vazquez, Rodriguez-Vico, F.
CONGRESO:
3rd Scientific Meeting of the Spanish Society of Chromatography and
Related Techniques. SECYTA. 3rd Waste Water Cluster European Workshop.
PUBLICACION:
Book of Abstracts pag. 172
LUGAR
DE CELEBRACION:
Almería (España)
AÑO: 2003.
TIPO
DE PARTICIPACION:
Poster
TITULO:
Estudio de las propiedades bioquímicas de dos hidantoín racemasas para su
aplicación industrial .
AUTORES:
Martinez-Rodriguez, S, Clemente-Jimenez, JM, Martínez-Gómez, A.I., Pozo-Dengra,
J., Las Heraz-Vazquez, Rodriguez-Vico, F.
CONGRESO:
XXVII Congreso SEBBM.
PUBLICACION:
Book of Abstracts pag. 236
LUGAR
DE CELEBRACION:
Lérida (España)
AÑO: 2004.
TIPO
DE PARTICIPACION:
Poster
TITULO:
Comparison of two recombinant whole cell systems for the production of optically
pure D-amino acids.
AUTORES:
Martínez-Gómez, AI, Martinez-Rodriguez
S, Pozo-Dengra, J, Clemente-Jimenez, JM, Rodríguez-Vico, F, Las Heraz-Vazquez,
FJ
CONGRESO:
1st International Conference on Environmental Industrial and Applied
Microbiology.
PUBLICACION:
Book of Abstracts pag. 290
LUGAR DE CELEBRACION:
Badajoz (España)
AÑO: 2005.
TIPO
DE PARTICIPACION:
Poster
TITULO:
Yeast autolysis for L-amino acid production .
AUTORES:
Pozo-Dengra J, Martinez-Rodriguez, S, Martínez-Gómez, AI, Las Heraz-Vazquez FJ,
Rodriguez-Vico, F. Clemente-Jimenez, JM.
CONGRESO:
1st International Conference on Environmental Industrial and Applied
Microbiology.
PUBLICACION:
Book of Abstracts pag. 434
LUGAR DE CELEBRACION:
Badajoz (España)
AÑO: 2005.
TIPO DE PARTICIPACION: Poster
TITULO: A D-hydantoinase
from Sinorhizobium meliloti CECT4114 useful for the production of optically pure
D-amino acids.
AUTORES: S. Martínez
Rodríguez, A.I. Martínez Gómez, J. Pozo Dengra, F. Rodríguez Vico, J.M.
Clemente Jiménez, F.J. Las Heras Vázquez.
CONGRESO: 1st
International Symposium on Enviromental Biocatalysis: From remediation with
enzymes to novel green process..
LUGAR DE CELEBRACION: Córdoba (España) AÑO: 2006.
TIPO DE PARTICIPACION: Poster
TITULO:
Cristalización y estudios cristalográficos preliminares de la D-hidantoinasa
de Sinorhizobium meliloti CECT4114.
AUTORES: S. Martínez Rodríguez, González-Ramíres, L.A.,
Clemente-Jiménez J.M., Otálora-Muñez F., Rodríguez Vico, Las Heras Vázquez, F.J., Gavira-Gallardo, J:A., García
Ruiz, J.M..
CONGRESO: XVII
Symposium del Grupo Especializado de Cristalografía: “Cristalografia:
conocer, entender, diseñar, preparar”.
PUBLICACION:
Book of Abstracts.
LUGAR DE CELEBRACION:
Sigüenza (España)
AÑO: 2006.
TITULO:
Obtención de D-aminoácidos ópticamente puros mediante síntesis enzimática.
CONFERENCIANTE: Las
Heras Vázquez. Francisco Javier
CONGRESO:
BIOTEC “Retos Biotecnológicos en la Venezuela del Siglo XXI”.
LUGAR DE CELEBRACION:
Valencia (Venezuela)
AÑO: 2006.
TITULO:
Aislamiento y caracterización molecular de levaduras para la mejora de la
calidad del vino
CONFERENCIANTE:
Clemente Jiménez, Josefa María
CONGRESO:
BIOTEC “Retos Biotecnológicos en la Venezuela del Siglo XXI”.
LUGAR DE CELEBRACION:
Valencia (Venezuela)
AÑO: 2006.
TITULO: Evaluación de la reacción en cadena de la polimerasa en el análisis de poblaciones y en la caracterización molecular de organismos
DIRECTOR: Felipe
Rodríguez Vico
AÑO: 1998
TITULO: Construcción de un sistema de expresión heterólogo con los genes de la D,L-hidantoinasa y de la D-carbamilasa. Evaluación de su aplicación biotecnológica a la síntesis enzimática de D-aminoácidos a partir de derivados de la D,L-hidantoina.
DIRECTOR: Felipe
Rodríguez Vico
AÑO: 2001
TITULO: Sobre-expresión de enzimas hidantoinas y carbamilasa
DIRECTORES: Felipe Rodríguez Vico y Santiago de la Escalera Hueso AÑO: 2002
TITULO: Obtención de D-aminoácidos ópticamente puros a partir de un sistema recombinante. Caracterización de las enzimas involucradas en el proceso
DIRECTORES: Felipe Rodríguez Vico. Fco Javier Las Heras Vázquez, Josefa María Clemente Jiménez
AÑO: 2005