Fernando García-Carreño

Fernando García-Carreño
Centro de Investigaciones Biológicas del Noroeste | CIBNOR · Biochemistry

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120
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5,054
Citations
Citations since 2017
15 Research Items
1784 Citations
2017201820192020202120222023050100150200250300
2017201820192020202120222023050100150200250300
2017201820192020202120222023050100150200250300
2017201820192020202120222023050100150200250300

Publications

Publications (120)
Article
Gene edition using the CRISPR/Cas9 system has been achieved in few crustaceans, but in penaeid shrimps, several hurdles exist, restraining the potential of CRISPR‐Cas9 technology in shrimp Aquaculture. A key step for gene editing is the efficient introduction of the CRISPR‐Cas9 system into the cells. Although microinjection is an effective method f...
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In arthropods, the cleavage of specific proteins by peptidases has pivotal roles in multiple physiological processes including oogenesis, immunity, nutrition, and parasitic infection. These enzymes are also key players in the larval development, and well-described triggers of molting and metamorphosis. In this work the peptidase complement througho...
Article
Background: Decapods belong to Pancrustacea, all are of ecological and economic importance since many species are profitable either harvested in the wild or produced in aquafarming. The description of biochemical physiology is paramount for both, ecology and economy. Being protein the most expensive ingredient in fabricated feeds, explaining food p...
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MicroRNAs are a class of small non‐coding RNAs that regulate gene expression at post‐transcriptional level. Regulatory RNAs were discovered in the nematode Caenorhabditis elegans, since then, their biological functions have been studied on different animals, including marine organisms. Several species of penaeid shrimp are important in ecology and...
Article
Soybean meal is a recurrent ingredient in shrimp feed formulations; although economically convenient, the downside is the antinutritional molecules often present in its active forms. Recently, a midgut gland proteolytic activity adjustment to compensate the effect of increasing concentrations of soybean trypsin inhibitor (SBTI) delivered in shrimp...
Article
Se evaluó el efecto de las enzimas de vísceras de sardina Monterey en el tratamiento del agua de cola producida por sardinas crinuda y Monterey y, se compararon con enzimas de un extracto comercial. Además, se aplicó una centrifugación complementaria al ensayo. La enzima comercial tuvo mayor actividad proteasa que la enzima de víscera de sardina (0...
Article
Pancrustacea, including both crustaceans and insects, is the most abundant and diverse clade of Animalia and occupy all types of environments. They are studied because of their ecological, agricultural, health and economic issues, some being delicatessens. Crustaceans evolved during the Cambrian, thanks to adaptations, such as macrophagy, that allo...
Article
Suitable peptidases for biotechnological applications are those active at low temperature, in organic solvents, detergents or proteolytic additives. American lobster cathepsin D1 (CD1) is an enzyme highly efficient at 5–50 °C and at pH 2.5–5.5. We assessed the effect of common industrial additives on CD1 activity. CD1 was isolated from lobster gast...
Article
Cathepsin D is an aspartic endopetidase with typical characteristics of lysosomal enzymes. Cathepsin D activity has been reported in the gastric fluid of clawed lobsters where it acts as an extracellular digestive enzyme. Here we investigate whether cathepsin D is unique in clawed lobsters or, instead, common in decapod crustaceans. Eleven species...
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Nick Lane (1967) is a biochemist and English writer who collaborates as a researcher and speaker in evolutionary biochemistry in University College London. He has published four scientifi c-dissemination books that have won diverse prizes, as “Oxygen”, which has been reviewed in this journal. The book has been described as “one of the deepest and m...
Article
During shrimp larval development, changes occur in molecular components. Enzyme activity and mRNA expression of proteinases were assayed in Penaeus vannamei during larval development, which consists of 5 nauplius stages, 3 protozoeal stages, 3 mysis stages, and 12 postlarval stages. Trypsin activity reached a maximum at the beginning of postlarval...
Article
During shrimp larval development, changes occur in molecular components. Enzyme activity and mRNA expression of proteinases were assayed in Penaeus vannamei during larval development, which consists of 5 nauplius stages, 3 protozoeal stages, 3 mysis stages, and 12 postlarval stages. Trypsin activity reached a maximum at the beginning of postlarval...
Article
The aim of this work was to study mechanisms of homeostasis of the digestive system of the whiteleg shrimp Litopenaeus vannamei when fed anti-nutritional soybean trypsin inhibitor (SBTI). In a previous work, we tested if the shrimp could respond against SBTI in feed with the increase in the digestive proteolytic activity. This time we tested the ti...
Article
The aim of this study was to assess the regulatory process of digestive peptidases of crustaceans in the presence of soybean trypsin inhibitor (SBTI). This naturally occurring inhibitor in soybean meal was used to inhibit the activity of digestive serine peptidases of the whiteleg shrimp Litopenaeus vannamei. In vitro, SBTI inhibited the total prot...
Article
The American lobster [Homarus americanus (Edwards, 1837)] is an omnivore decapod with generalist and opportunistic feeding habits, in which gastric fluid cysteine and aspartic peptidases are the main proteolytic enzymes. Using in vitro assays, the network of concerted acting peptidases belonging to aspartic and cysteine catalytic mechanisms that hy...
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Crustaceans are a diverse group, distributed in widely variable environmental conditions for which they show an equally extensive range of biochemical adaptations. Some digestive enzymes have been studied by purification/characterization approaches. However, global analysis is crucial to understand how digestive enzymes interplay. Here, we present...
Article
In vitro assays used porcine or bovine trypsin as models of exogenous enzymes to determine functioning in the presence of enzymatic extracts from the digestive gland of whiteleg shrimp Penaeus vannamei. Using electrophoresis and zymograms, when enzymes from the shrimp were mixed in the absence of protein substrate, they hydrolysed the trypsin from...
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A chymotrypsin was purified from the gastric juice of California spiny lobster (Panulirus interrutpus), using preparative electrophoresis and affinity chromatography on agarose-p-aminobenzamidine. The molecular mass was estimated by polyacrylamide gel electrophoresis (SDS-PAGE) under denaturing conditions to be 28 kDa. Chymotrypsin activity was tot...
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Marine viruses are ubiquitous, extremely diverse, and outnumber any form of life in the sea. Despite their ecological importance, viruses in marine environments have been largely ignored by the academic community, and only those that have caused substantial economic losses have received more attention. Fortunately, our current understanding on mari...
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Over the long course of evolution and under the selective pressure exerted by pathogens and parasites, animals have selectively fixed a number of defense mechanisms against the constant attack of intruders. The immune response represents a key component to optimize the biological fitness of individuals. Two decades ago, prevention and control of di...
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Waste materials derived from shrimp processing signify 40 to 45 % of the weight of the catch or harvest. For Biotechnology, waste materials are secondary material, as contrast of raw materials, to generate value-added products, such as chitin, protein, and pigments. We used the pH-shift and enzymatic autohydrolysis processes to obtain protein conce...
Article
To test the efficacy of adding enzyme supplements to feeds, an in vitro study was conducted by mixing bovine trypsin or proteinases from the gastric juice of the Cortez swimming crab Callinectes bellicosus with an enzyme extract from the digestive gland of the whiteleg shrimp Penaeus vannamei. Enzymes alone and mixtures were tested for hydrolysing...
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Aspartic proteinases in the gastric fluid of clawed lobsters Homarus americanus and Homarus gammarus were isolated to homogeneity by single-step pepstatin-A affinity chromatography; such enzymes have been previously identified as cathepsin D-like enzymes based on their deduced amino acid sequence. Here, we describe their biochemical characteristics...
Article
Digestive and intracellular lipases were studied in the digestive gland of whiteleg shrimp Penaeus vannamei. A partial sequence of the intracellular lipase was obtained from the digestive gland cDNA. The digestive and intracellular lipase mRNAs were detected differentially in different body parts of shrimp; digestive lipase mRNA is exclusively foun...
Article
Cathepsin B is a cystein proteinase scarcely studied in crustaceans. Its function has not been clearly described in shrimp species belonging to the sub-order Dendrobranchiata, which includes the white shrimp Litopenaeus vannamei and other species from the Penaeidae family. Studies on vertebrates suggest that these lysosomal enzymes intracellularly...
Article
Recently, several groups of researchers reported that besides serine proteinases, other classes of proteinases may be involved in crustacean's food protein digestion, including cysteine and aspartic proteinases. In this paper, a comparative study of the class and type of digestive proteinases of whiteleg shrimp Penaeus vannamei, blue shrimp Penaeus...
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In the mitochondrial F(O)F(1) ATP-synthase/ATPase complex, subunits α and β are part of the extrinsic portion that catalyses ATP synthesis. Since there are no reports about genes and proteins from these subunits in crustaceans, we analyzed the cDNA sequences of both subunits in the whiteleg shrimp Litopenaeus vannamei and their phylogenetic relatio...
Article
An aspartic proteinase was isolated from American lobster gastric fluid. The purified cathepsin D runs as a single band on native-PAGE displaying proteolytic activity on a zymogram at pH 3.0, with an isoelectric point of 4.7. Appearance of the protein in SDS-PAGE, depended on the conditions of the gel electrophoresis. SDS treatment by itself was no...
Article
An intracellular lipase present in the whiteleg shrimp Litopenaeus vannamei was detected in pleopods. The lipase from pleopods was purified and characterized by biochemical and kinetic parameters. Purified intracellular lipase has a molecular mass of 196kDa, the polypeptide is assembled by two monomers, 95.26 and 63.36kDa. The enzyme lacks glycosyl...
Article
Functional properties of protein from mantle and fin of the jumbo squid Dosidicus gigas were explained based on microscopic muscle fiber and protein fractions profiles as observed in SDS-PAGE. Fin has higher content of connective tissue and complex fiber arrangement, and we observed higher hardness of fin gels as expected. Myosin heavy chain (MHC)...
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Penaeus vannamei lipase was purified from midgut gland of whiteleg shrimp. Pure lipase (E.C. 3.1.1.3) was obtained after Superdex 200 gel filtration and Resource Q anionic exchange. The pure lipase, which is a glycosylated molecule, is a monomer having a molecular mass of about 44.8 kDa, as determined by SDS-PAGE analysis. The lipase hydrolyses sho...
Article
Lipase activity of the midgut gland during larval and postlarval stages of Penaeus vannamei was assayed to determine its capability to digest lipids from feed and involvement in digesting lipids from reserves during fasting conditions. Lipase activity was detected at all larval stages, increasing from nauplii to protozoea. Lipase isoenzymes at larv...
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Acid digestive proteinases were studied in the gastric fluids of two species of clawed lobster (Homarus americanus and Homarus gammarus). An active protein was identified in both species as aspartic proteinase by specific inhibition with pepstatin A. It was confirmed as cathepsin D by mass mapping, N-terminal, and full-length cDNA sequencing. Both...
Article
Processing of sardine fishmeal generates pollution and waste with potentially useful protein. Sardine stick-water (SW) from fishmeal operation was submitted to a complementary centrifugation step followed by pH adjustment (acidic + alkaline) to recover solids for their compositional, functional and nutritional properties evaluation. Insoluble fract...
Article
There is a growing interest in adding value to the jumbo squid Dosidicus gigas fishery. This work describes two extraction procedures for processing muscle to obtain protein isolates with suitable functional properties. The effect on muscle protein solubility and protein recovery of combining freezing and grinding raw materials during storage was e...
Article
Solubility at several ionic strengths (0 to 1.0 M), pH (2 to 13) and gelling capacity of jumbo squid Dosidicus gigas muscle proteins were evaluated. Protein recovery was >90% at pH 9–12. Autohydrolysis was evaluated and affected only sarcoplasmic proteins. Folding score was 5 on all gels. Strength was higher for thermal gels prepared from squid fin...
Article
Evaluation of the effect of pH (2 to 13) and temperature (0 to 50C) on functional properties of jumbo squid proteins was performed, followed by a 2 × 3 factorial design for producing squid protein hydrolysates bearing useful functional properties. In particular, the effects of pH (8, 9 and 10) and temperature (30, 35, 40C) were evaluated. Alcalase...
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The stickwater is a complex effluent consisting of a variety of solids, most of which should be removed to produce a new effluent that can be directly poured into the sea. In this study, stickwater was centrifuged to remove solids and the resulting liquid was then treated with chitosan at different concentrations and pH to eliminate the solids left...
Article
An immunological approach was used to elucidate whether trypsin is synthesized and stored as trypsinogen in the midgut gland of the shrimp Litopenaeus vannamei. Two peptides were constructed using sequences deduced from known shrimp genes: trypsinogen activation peptide and an internal sequence. These peptides were used as haptens to elicit antibod...
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We studied for the first time the ATP-synthase complex from shrimp as a model to understand the basis of crustacean bioenergetics since they are exposed to endogenous processes as molting that demand high amount of energy. We analyzed the cDNA sequence of two subunits of the Fo sector from mitochondrial ATP-synthase in the white shrimp Litopenaeus...
Article
Latent phenoloxidase activity of hemocyanin (Hc) in whiteleg shrimp Penaeus vannamei was assayed to determine its potential involvement in postmortem melanosis. Conversion of pure 12-mer, but not 6-mer, hemocyanin to phenoloxidase by endogenous (serine proteinases) and exogenous (SDS) effectors demonstrated the need of complex aggregation for displ...
Article
Effect of iced storage of jumbo squid mantle with fin on gelling capacity and changes in protein fractions and functional properties of jumbo squid mantle protein during storage at 0 °C were assessed. Most values of texture variables in gels did not significantly change during storage. On average, they were: strength 65.07 ± 4.71 N; elasticity 68.1...
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Food protein hydrolysis, a crucial step in digestion, is catalyzed by trypsin enzymes from the digestive apparatus of invertebrates. Trypsin appeared early in evolution and occurs in all phyla and, in the digestive systems of invertebrates, it became the most abundant proteinase. As in vertebrates, invertebrate trypsin is also present in several fo...
Article
Nine fish species, Gillichthys seta, Oligoplites saurus, Eucinostomus entomelas, Synodus scituliceps, Diplectrum pacificum, Pseudopeneus grandisquamis, Xenistius californiensis, Arius seemanni, and Orthopristis reddinig, caught as bycatch by shrimp commercial fleet of the Guaymas, Sonora, México, were processed to produce protein concentrates using...
Article
Whiteleg shrimp (Litopenaeus vannamei) is traded as a frozen food. Changes in texture of thawed muscle are a negative economic factor. This study evaluated and compared two freezing methods, cryogenic and forced convection, and two thawing methods, at 4 and 25C by histological, textural and electrophoretical tests. Insignificant interaction was fou...
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Characteristics and functional efficacy of digestive proteases of Catla catla, catla, Labeo rohita, rohu and Hypophthalmichthys molitrix, silver carp were studied. Total protease activity was significantly (P < 0.05) higher in rohu (1.219 ± 0.059 U mg protein−1 min−1) followed by silver carp (1.084 ± 0.061 U mg protein−1 min−1), and catla (0.193 ±...
Article
White shrimp (Penaeus vannamei) were fed one of seven diets for 30 days. Diets contained 85% of a reference ration and 15% of either anchovy, tuna waste meal, deboned fish meal, langostilla crab meal, soybean meal, menhaden meal A, or menhaden B as a protein replacer. At the end of the feeding trial, animals were sacrificed and 10 aminopeptidase ac...
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Crustaceans are forced to fast during molting. Several physiological, metabolic and behavioral changes have been associated with starvation. Although some of these changes have been well studied, knowledge of the dynamics of fuel reserves during the molting process is limited. To understand the effects of short-term hunger stress on energy reserves...
Article
This study describes the digestive capacities of 10-day-old white shrimp postlarvae (PL10) and how some inhibitors of proteinases affect the digestion of protein in aquafeeds by using in vitro hydrolysis techniques. Biochemical data showed eight active proteinases in the PL10 hepatopancreas extract. Enzymes belong to the metallo- and serine-protein...
Article
Chymotrypsin was isolated from the viscera of Monterey sardine by ammonium sulphate fractionation, gel filtration, and ionic exchange chromatography. The approximate molecular weight was 26,000 and its isoelectric point was about 5. Identity as chymotrypsin was established by its catalytic specificity for amide or ester bonds on the synthetic subst...
Article
Decapod crustaceans synthesize highly active proteolytic enzymes in the midgut gland and release at least a part of them into the stomach where they facilitate the first step in peptide hydrolysis. The most common proteinases in the gastric fluid characterized so far are serine proteinases, that is, trypsin and chymotrypsin. These enzymes show high...
Article
Protease activity in the midgut gland, gastric chamber, and gastric juice from the crabs Callinectes bellicosus and Callinectes arcuatus was characterized by several methods, confirming that the composition of digestive proteases is the same in the gastric juice and the midgut gland. Gastric juice was suitable for the identification and characteriz...
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In this paper, we review the current knowledge about the usage of carbohydrates, lipids and proteins as energy source by marine crustaceans during starvation. Crustaceans are a large and diverse group including some economically important species. The efforts to culture them for human consumption has prompted the interest to understand the preferen...
Article
Based on information about isotrypsins found by substrate–SDS-PAGE in the whiteleg shrimp Litopenaeus vannamei and their segregation, an explanation for the presence of three-trypsin phenotype was sought. Isotrypsins A, B, and C depend on two loci: locus β, which is homozygous, yielding isoenzyme C, and locus α, which is heterozygous, yielding isoe...
Article
A high density lipoprotein-beta glucan binding protein (HDL-BGBP) is synthesized in the hepatopancreas of the white shrimp Litopenaeus vannamei and secreted to the hemolymph. Recently, we reported the HDL-BGBP full length cDNA sequence and found that the predicted polypeptide is larger than the mature protein and also, that it contains a long 5'- a...
Article
Trypsin from pyloric caeca of Monterey sardine was purified by fractionation with ammonium sulfate, gel filtration, affinity and ionic exchange chromatography. Fraction 102, obtained from ionic exchange chromatography, generated one band in sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and isoelectric focusing. The molecular...
Article
Protein is the major component for shrimp feeds and must be nutritionally appropriate and readily digestible. In vitro protein digestibility of feeds and feed ingredients, and their inhibitory effect on the digestive proteases of juvenile Farfantepenaeus paulensis were examined. The analysis included animal (four types of fish meal, meat meal and b...
Article
Activities of proteolytic enzymes, represented by azocasein digestion (total protease), trypsin, and chymotrypsin from the gastric fluid of the marine crab, Cancer pagurus, were evaluated for operational parameters like pH-optimum, thermal stability, and effectors such as metal ions and organic solvents. Total protease activity was 98.5 ± 4.3 E 366...
Article
Three isotrypsins from digestive gland of Penaeus vannamei were purified and characterized by molecular, biochemical and kinetic parameters. Purified isotrypsins A, B, and C are glycoproteins with molecular masses between 30.2 and 32.9 kDa, and, therefore similar to other trypsins. The isoelectric points are anionic and different among the three is...
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Enzymes responsible for digestion of food protein were evaluated and characterized in red lobster (Panulirus interruptus). Several tissues, organs, and body fluids were analyzed. The same composition of proteases was found in gastric juice, midgut gland, and intestinal contents. Using specific substrates and inhibitors, we identified several isotry...
Article
Total enzyme activity of whole viscera, and partial characterization of acidic proteases from Monterey sardine viscera are presented. Major proteolytic activity in alkali (pH 10) and minor activity in acid (pH 3) were detected. From purified acidic proteases, six fractions with high activity were selected. One fraction (42) showed one band on SDS–P...
Article
The effect of change in feed and physical manipulation on the digestive system of white shrimp Penaeus vannamei (Boone, 1931) was examined for proteases secreted by the digestive gland and excreted in feces. Organisms were fed a commercial feed containing 45% protein and separated into two groups. One group served as the control and the other was p...
Article
Regulation of the expression of trypsin encoding genes in juveniles white shrimp (Penaeus vannamei) was studied during starvation for up to 120 h. Since molting is now recognized as a physiologically important process in penaeids, specimens were selected according to molt stage. Starvation led to immediate weight loss and a sharp decrease in hepato...
Article
Protein food modulates the activity of proteases of the midgut gland of Penaeus vannamei. Shrimp fed with food containing 15, 30 and 50% protein exhibited differences in trypsin and chymotrypsin activity and trypsin mRNA levels. Shrimp fed with 30% protein showed higher trypsin and chymotrypsin activities than those fed 15 or 50% protein. An additi...
Article
Enzymes responsible for the digestion of food protein by juvenile green abalone (Haliotis fulgens) were studied when fed algae or a sea grass (Phyllospadix torreyi) naturally occurring in the habitat. The effect of food on the composition and activity of the enzymes was also evaluated. Acid, serine proteinases and aminopeptidases, as confirmed by p...
Article
We investigated the effect of starvation as a stimulant of the digestive system on digestive proteinase activities in the white shrimp Penaeus vannamei. The starved organisms were sampled periodically according to the molting stage and compared with a continuously fed group. Molting stage was included as an independent variable. Most analyzed varia...
Article
Proteinase (endopeptidase), trypsin and chymotrypsin activities, RNA, and DNA contents were examined throughout the ontogenetic development of cultured Litopenaeus schmitti. Whole individuals from larval and postlarval stages, and adult hepatopancreas were homogenized and assayed for quantification of enzyme activity. Proteinase activity of differe...
Article
Digestive proteinase activities of Artemesia longinaris were assayed at different stages of the molting cycle. Total proteolytic activity in the hepatopancreas was highest during postmolt. Trypsin and chymotrypsin activities were highest during intermolt. Specific inhibitors and zymograms of A. longinaris hepatopancreas extracts showed four trypsin...
Article
Though some protein sources like squid and protein hydrolysates are assumed as growth enhancers for shrimp, little is known about the biochemical basis of this phenomenon. Low, heat-dried squid (Dosidicus gigas) (SQ) and two commercial protein hydrolysates from fish (FH) and krill (Euphasia sp.) (KH) were assayed in feeding trials with Penaeus vann...
Article
Juvenile piracanjuba, Brycon orbignyanus, in the wild consume protein from both plant and animal sources. Digestion of protein in piracanjuba begins in the stomach with pepsin, at low pH, and is followed by hydrolysis at alkaline pH in the lumen of the intestine. The digestive system in piracanjuba was evaluated to characterize the enzymes responsi...
Article
Trypsin inhibitors from four species of shrimp, P. vannamei, P, monodon, P. stylirostris, and P. californiensis were characterized by test tube and electrophoretic assays. Inhibitors in the four phylogenetically related species showed different inhibitory capacity and specificity for orthologous trypsins. Inhibitors from P. californiensis showed a...
Article
Trypsin inhibitors from four species of shrimp, P. vannamei, P. monodon, P. stylirostris, and P. californiensis were characterized by test tube and electrophoretic assays. Inhibitors in the four phylogenetically related species showed different inhibitory capacity and specificity for orthologous trypsins. Inhibitors from P. californiensis showed a...
Article
The modori phenomenon from several fish species has been associated with endogenous proteinolytic enzymes. We evaluated the presence of modori-associated proteinases in Mexican flounder and Atlantic croaker, and the decrease of the modori effect by inhibition of the responsible enzymes with serine-proteinase inhibitors from legume seeds. The presen...
Article
The present study describes the activity and some characteristics of proteinases in the hepatopancreas of red shrimp Pleoticus muelleri during the different stages of the molting cycle. Proteolytic activity was highest between pH 7.5 and 8. The hepatopancreatic protein content in the premolt stage was higher than in the other stages of the molting...
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Giant squid meal from two drying processes: high-heat (from commercial flamedried) (HHSM) and low-heat (labor ator y made) (LHSM), were evaluated as protein source for shrimp Penaeus stylirrostrisin two feeding experiments. Trial I consisted of wild shrimp fed HHSM at different rates. Shrimp survival was negatively affected by commercial squid meal...
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This work provides a comparative study of the inhibitory effect of several plant protein sources on digestive pro-teases of two snappers: yellow snapper (Lutjanus argentiventris) and dog snapper (Lutjanus novemfasciatus). Seed extracts did not affect gastric proteases whereas they significantly inhibit intestinal proteases. Inhibition of alkaline p...