The Experts below are selected from a list of 4521 Experts worldwide ranked by ideXlab platform

Daniel Lemos - One of the best experts on this subject based on the ideXlab platform.

  • Species specific in vitro protein digestion (pH-stat) for fish: method development and application for juvenile rainbow trout (Oncorhynchus mykiss), cobia (Rachycentron canadum), and Nile tilapia (Oreochromis niloticus)
    Aquaculture, 2014
    Co-Authors: F. Yasumaru, Daniel Lemos
    Abstract:

    Abstract Aqua feed manufacture requires flexible formulations and effective methods to screen suitable feed ingredients. In vitro digestion may assist in the characterization and quality control of protein in feedstuffs for fish species once standardized species-specific digestive enzyme Extracts are available. This study aimed to develop a species-specific in vitro enzymatic method to assess protein digestion in fish under the pH-stat concept. Two carnivorous (rainbow trout, Oncorhynchus mykiss , and cobia, Rachycentron canadum ) and one omnivorous (Nile tilapia, Oreochromis niloticus ) fish species were used as models. Crude digestive enzyme Extracts were recovered from Stomach and pyloric caeca or intestine of individuals of different weight groups, feeding status, and farming systems. The hydrolytic capacity of the species-specific enzyme Extracts was standardized on purified protein substrates and measured as degree of protein hydrolysis (DH) in the pH-stat assay. A group of twenty-four feed ingredients, including fish meals and by-products of plant and animal origin, was assessed for DH using the recovered enzymes from Stomach and pyloric caeca/intestine. Ingredients were hydrolyzed with fish (i) Stomach Extract, (ii) pyloric caeca/intestine Extract or (iii) Stomach enzymes followed by pyloric caeca/intestine Extract. Among plant by-products, cotton seed meal presented the highest DH with Stomach plus pyloric caeca/intestine enzymes, followed by soy protein concentrate and soybean meals. Blood meals were the land animal by-product with higher DH outputs compared to poultry by-product meals and feather meals. No significant difference was observed among the DHs of fish meals. The significance of measuring the DH with Stomach enzyme Extract is still not well understood but, overall, the pre-hydrolysis of feedstuffs with Stomach enzymes increased pyloric caeca/intestine DH value. For cage and pond farmed Nile tilapia, ingredient DHs followed the same trend, describing a significant correlation and a high determination coefficient regression. Routine use of the method may yet depend on the prompt availability of more practical sources of enzymes. The determination of the degree of protein hydrolysis by the in vitro pH-stat with species-specific enzymes has shown to be a precise method that may be a useful tool to rank feed ingredients, and also an accessory method in the quality control of feedstuffs.

  • method development and application for juvenile rainbow trout (Oncorhynchus mykiss), cobia (Rachycentron canadum), and Nile tilapia (Oreochromis niloticus)
    2014
    Co-Authors: F. Yasumaru, Daniel Lemos
    Abstract:

    article i nfo Aqua feed manufacture requires flexible formulations and effective methods to screen suitable feed ingredients. Invitrodigestionmayassistinthecharacterizationandqualitycontrolofproteininfeedstuffsfor fishspeciesonce standardized species-specific digestive enzyme Extracts are available. This study aimed to develop a species-specific in vitro enzymatic method to assess protein digestion in fish under the pH-stat concept. Two carnivorous (rainbow trout, Oncorhynchus mykiss, and cobia, Rachycentron canadum) and one omnivorous (Nile tilapia, Oreochromis niloticus) fish species were used as models. Crude digestive enzyme Extracts were recovered from Stomach and py- loric caeca or intestine of individuals of different weight groups, feeding status, and farming systems. The hydrolytic capacity of the species-specific enzyme Extracts was standardized on purified protein substrates and measured as degree of protein hydrolysis (DH) in the pH-stat assay. A group of twenty-four feed ingredients, including fish meals and by-products of plant and animal origin, was assessed for DH using the recovered enzymes from Stomach and pyloric caeca/intestine. Ingredients were hydrolyzed with fish (i) Stomach Extract, (ii) pyloric caeca/intestine Extract or (iii) Stomach enzymes followed by pyloric caeca/intestine Extract. Among plant by-products, cotton seed meal presentedthe highestDHwithStomach plus pyloric caeca/intestine enzymes, follow- ed by soy protein concentrate and soybean meals. Blood meals were the land animal by-product with higher DH outputs compared to poultry by-product meals and feather meals. No significant difference was observed among the DHs of fish meals. The significance of measuring the DH with Stomach enzyme Extract is still not well under- stood but, overall, the pre-hydrolysis of feedstuffs with Stomach enzymes increased pyloric caeca/intestine DH value. For cage and pond farmed Nile tilapia, ingredient DHs followed the same trend, describing a significant correlation and a high determination coefficient regression. Routine use of the method may yet depend on the prompt availability of more practical sources of enzymes. The determination of the degree of protein hydrolysis by the in vitro pH-stat with species-specific enzymes has shown to be a precise method that may be a useful tool to rank feed ingredients, and also an accessory method in the quality control of feedstuffs.

F. Yasumaru - One of the best experts on this subject based on the ideXlab platform.

  • Species specific in vitro protein digestion (pH-stat) for fish: method development and application for juvenile rainbow trout (Oncorhynchus mykiss), cobia (Rachycentron canadum), and Nile tilapia (Oreochromis niloticus)
    Aquaculture, 2014
    Co-Authors: F. Yasumaru, Daniel Lemos
    Abstract:

    Abstract Aqua feed manufacture requires flexible formulations and effective methods to screen suitable feed ingredients. In vitro digestion may assist in the characterization and quality control of protein in feedstuffs for fish species once standardized species-specific digestive enzyme Extracts are available. This study aimed to develop a species-specific in vitro enzymatic method to assess protein digestion in fish under the pH-stat concept. Two carnivorous (rainbow trout, Oncorhynchus mykiss , and cobia, Rachycentron canadum ) and one omnivorous (Nile tilapia, Oreochromis niloticus ) fish species were used as models. Crude digestive enzyme Extracts were recovered from Stomach and pyloric caeca or intestine of individuals of different weight groups, feeding status, and farming systems. The hydrolytic capacity of the species-specific enzyme Extracts was standardized on purified protein substrates and measured as degree of protein hydrolysis (DH) in the pH-stat assay. A group of twenty-four feed ingredients, including fish meals and by-products of plant and animal origin, was assessed for DH using the recovered enzymes from Stomach and pyloric caeca/intestine. Ingredients were hydrolyzed with fish (i) Stomach Extract, (ii) pyloric caeca/intestine Extract or (iii) Stomach enzymes followed by pyloric caeca/intestine Extract. Among plant by-products, cotton seed meal presented the highest DH with Stomach plus pyloric caeca/intestine enzymes, followed by soy protein concentrate and soybean meals. Blood meals were the land animal by-product with higher DH outputs compared to poultry by-product meals and feather meals. No significant difference was observed among the DHs of fish meals. The significance of measuring the DH with Stomach enzyme Extract is still not well understood but, overall, the pre-hydrolysis of feedstuffs with Stomach enzymes increased pyloric caeca/intestine DH value. For cage and pond farmed Nile tilapia, ingredient DHs followed the same trend, describing a significant correlation and a high determination coefficient regression. Routine use of the method may yet depend on the prompt availability of more practical sources of enzymes. The determination of the degree of protein hydrolysis by the in vitro pH-stat with species-specific enzymes has shown to be a precise method that may be a useful tool to rank feed ingredients, and also an accessory method in the quality control of feedstuffs.

  • method development and application for juvenile rainbow trout (Oncorhynchus mykiss), cobia (Rachycentron canadum), and Nile tilapia (Oreochromis niloticus)
    2014
    Co-Authors: F. Yasumaru, Daniel Lemos
    Abstract:

    article i nfo Aqua feed manufacture requires flexible formulations and effective methods to screen suitable feed ingredients. Invitrodigestionmayassistinthecharacterizationandqualitycontrolofproteininfeedstuffsfor fishspeciesonce standardized species-specific digestive enzyme Extracts are available. This study aimed to develop a species-specific in vitro enzymatic method to assess protein digestion in fish under the pH-stat concept. Two carnivorous (rainbow trout, Oncorhynchus mykiss, and cobia, Rachycentron canadum) and one omnivorous (Nile tilapia, Oreochromis niloticus) fish species were used as models. Crude digestive enzyme Extracts were recovered from Stomach and py- loric caeca or intestine of individuals of different weight groups, feeding status, and farming systems. The hydrolytic capacity of the species-specific enzyme Extracts was standardized on purified protein substrates and measured as degree of protein hydrolysis (DH) in the pH-stat assay. A group of twenty-four feed ingredients, including fish meals and by-products of plant and animal origin, was assessed for DH using the recovered enzymes from Stomach and pyloric caeca/intestine. Ingredients were hydrolyzed with fish (i) Stomach Extract, (ii) pyloric caeca/intestine Extract or (iii) Stomach enzymes followed by pyloric caeca/intestine Extract. Among plant by-products, cotton seed meal presentedthe highestDHwithStomach plus pyloric caeca/intestine enzymes, follow- ed by soy protein concentrate and soybean meals. Blood meals were the land animal by-product with higher DH outputs compared to poultry by-product meals and feather meals. No significant difference was observed among the DHs of fish meals. The significance of measuring the DH with Stomach enzyme Extract is still not well under- stood but, overall, the pre-hydrolysis of feedstuffs with Stomach enzymes increased pyloric caeca/intestine DH value. For cage and pond farmed Nile tilapia, ingredient DHs followed the same trend, describing a significant correlation and a high determination coefficient regression. Routine use of the method may yet depend on the prompt availability of more practical sources of enzymes. The determination of the degree of protein hydrolysis by the in vitro pH-stat with species-specific enzymes has shown to be a precise method that may be a useful tool to rank feed ingredients, and also an accessory method in the quality control of feedstuffs.

Sitthipong Nalinanon - One of the best experts on this subject based on the ideXlab platform.

  • Partitioning of protease from Stomach of albacore tuna (Thunnus alalunga) by aqueous two-phase systems
    Process Biochemistry, 2009
    Co-Authors: Sitthipong Nalinanon, Soottawat Benjakul, Wonnop Visessanguan, Hideki Kishimura
    Abstract:

    Abstract Partitioning of protease from Stomach of albacore tuna using an aqueous two-phase system (ATPS) was investigated. The best ATPS conditions for protease partitioning from Stomach Extract (SE) and acidified counterpart (ASE) were 25% PEG1000–20% MgSO4 and 15% PEG2000–15% MgSO4, which increased the purity by 7.2-fold and 2.4-fold with the recovered activity of 85.7% and 89.1%, respectively. Electrophoretic study revealed that SE had a major protein with a molecular weight (MW) of 40.6 kDa, while protein with MW of 32.7 kDa was predominant in ASE and ATPS fractions. Pepsinogen in SE might be activated to pepsin by acidification and partitioning process. SE was quite stable at 0 and 4 °C up to 14 days. The loss in protease activity in ASE and selected ATPS fractions was more pronounced when storage time and temperature increased. Therefore, ATPS can be effectively used to recover and purify protease from albacore tuna Stomach.

  • tuna pepsin characteristics and its use for collagen Extraction from the skin of threadfin bream nemipterus spp
    Journal of Food Science, 2008
    Co-Authors: Sitthipong Nalinanon, Soottawat Benjakul, Wonnop Visessanguan, H Kishimura
    Abstract:

    : Pepsin from the Stomach of albacore tuna, skipjack tuna, and tongol tuna was characterized. Pepsin from all tuna species showed maximal activity at pH 2.0 and 50 degrees C when hemoglobin was used as a substrate. Among the Stomach Extract of all species tested, that of albacore tuna showed the highest activity (40.55 units/g tissue) (P < 0.05). Substrate-Native-PAGE revealed that pepsin from albacore tuna and tongol tuna consisted of 2 isoforms, whereas pepsin from skipjack tuna had only 1 form. The activity was completely inhibited by pepstatin A, while EDTA (ethylenediaminetetraacetic acid), SBTI (soybean trypsin inhibitor), and E-64 (1-(L-trans-epoxysuccinyl-leucylamino)-4-guanidinobutane) exhibited negligible effect. The activity was strongly inhibited by SDS (sodium dodecyl sulfate) (0.05% to 0.1%, w/v). Cysteine (5 to 50 mM) also showed an inhibitory effect in a concentration dependent manner. ATP, molybdate, NaCl, MgCl(2), and CaCl(2) had no impact on the activity. When tuna pepsin (10 units/g defatted skin) was used for collagen Extraction from the skin of threadfin bream for 12 h, the yield of collagen increased by 1.84- to 2.32-fold and albacore pepsin showed the comparable Extraction efficacy to porcine pepsin. The yield generally increased with increasing Extraction time (P < 0.05). All collagen obtained with the aid of tuna pepsin showed similar protein patterns compared with those found in acid-solubilized collagen. Nevertheless, pepsin from skipjack tuna caused the degradation of alpha and beta components. All collagens were classified as type I with large portion of beta-chain. However, proteins with molecular weight (MW) greater than 200 kDa were abundant in acid-solubilized collagen.

H Kishimura - One of the best experts on this subject based on the ideXlab platform.

  • tuna pepsin characteristics and its use for collagen Extraction from the skin of threadfin bream nemipterus spp
    Journal of Food Science, 2008
    Co-Authors: Sitthipong Nalinanon, Soottawat Benjakul, Wonnop Visessanguan, H Kishimura
    Abstract:

    : Pepsin from the Stomach of albacore tuna, skipjack tuna, and tongol tuna was characterized. Pepsin from all tuna species showed maximal activity at pH 2.0 and 50 degrees C when hemoglobin was used as a substrate. Among the Stomach Extract of all species tested, that of albacore tuna showed the highest activity (40.55 units/g tissue) (P < 0.05). Substrate-Native-PAGE revealed that pepsin from albacore tuna and tongol tuna consisted of 2 isoforms, whereas pepsin from skipjack tuna had only 1 form. The activity was completely inhibited by pepstatin A, while EDTA (ethylenediaminetetraacetic acid), SBTI (soybean trypsin inhibitor), and E-64 (1-(L-trans-epoxysuccinyl-leucylamino)-4-guanidinobutane) exhibited negligible effect. The activity was strongly inhibited by SDS (sodium dodecyl sulfate) (0.05% to 0.1%, w/v). Cysteine (5 to 50 mM) also showed an inhibitory effect in a concentration dependent manner. ATP, molybdate, NaCl, MgCl(2), and CaCl(2) had no impact on the activity. When tuna pepsin (10 units/g defatted skin) was used for collagen Extraction from the skin of threadfin bream for 12 h, the yield of collagen increased by 1.84- to 2.32-fold and albacore pepsin showed the comparable Extraction efficacy to porcine pepsin. The yield generally increased with increasing Extraction time (P < 0.05). All collagen obtained with the aid of tuna pepsin showed similar protein patterns compared with those found in acid-solubilized collagen. Nevertheless, pepsin from skipjack tuna caused the degradation of alpha and beta components. All collagens were classified as type I with large portion of beta-chain. However, proteins with molecular weight (MW) greater than 200 kDa were abundant in acid-solubilized collagen.

Masayoshi Sakaguchi - One of the best experts on this subject based on the ideXlab platform.

  • Protease resistance of porcine acidic mammalian chitinase under gastrointestinal conditions implies that chitin-containing organisms can be sustainable dietary resources
    Scientific Reports, 2017
    Co-Authors: Eri Tabata, Akinori Kashimura, Satoshi Wakita, Misa Ohno, Masayoshi Sakaguchi, Yasusato Sugahara, Yasutada Imamura, Shiro Seki, Hitoshi Ueda, Vaclav Matoska
    Abstract:

    Chitin, a polymer of N -acetyl-D-glucosamine (GlcNAc), is a major structural component in chitin-containing organism including crustaceans, insects and fungi. Mammals express two chitinases, chitotriosidase (Chit1) and acidic mammalian chitinase (AMCase). Here, we report that pig AMCase is stable in the presence of other digestive proteases and functions as chitinolytic enzyme under the gastrointestinal conditions. Quantification of chitinases expression in pig tissues using quantitative real-time PCR showed that Chit1 mRNA was highly expressed in eyes, whereas the AMCase mRNA was predominantly expressed in Stomach at even higher levels than the housekeeping genes. AMCase purified from pig Stomach has highest activity at pH of around 2–4 and remains active at up to pH 7.0. It was resistant to robust proteolytic activities of pepsin at pH 2.0 and trypsin and chymotrypsin at pH 7.6. AMCase degraded polymeric chitin substrates including mealworm shells to GlcNAc dimers. Furthermore, we visualized chitin digestion of fly wings by endogenous AMCase and pepsin in Stomach Extract. Thus, pig AMCase can function as a protease resistant chitin digestive enzyme at broad pH range present in Stomach as well as in the intestine. These results indicate that chitin-containing organisms may be a sustainable feed ingredient in pig diet.

  • Acidic mammalian chitinase is a proteases-resistant glycosidase in mouse digestive system
    Scientific Reports, 2016
    Co-Authors: Misa Ohno, Eri Tabata, Akinori Kashimura, Satoshi Wakita, Masahiro Kimura, Haruko Miyazaki, Kazuaki Okawa, Riho Onuki, Chiyuki Nemoto, Masayoshi Sakaguchi
    Abstract:

    Chitinases are enzymes that hydrolyze chitin, a polymer of β-1, 4-linked N -acetyl-D-glucosamine (GlcNAc). Chitin has long been considered as a source of dietary fiber that is not digested in the mammalian digestive system. Here, we provide evidence that acidic mammalian chitinase (AMCase) can function as a major digestive enzyme that constitutively degrades chitin substrates and produces (GlcNAc)_2 fragments in the mouse gastrointestinal environment. AMCase was resistant to endogenous pepsin C digestion and remained active in the mouse Stomach Extract at pH 2.0. The AMCase mRNA levels were much higher than those of four major gastric proteins and two housekeeping genes and comparable to the level of pepsinogen C in the mouse Stomach tissues. Furthermore, AMCase was expressed in the gastric pepsinogen-synthesizing chief cells. The enzyme was also stable and active in the presence of trypsin and chymotrypsin at pH 7.6, where pepsin C was completely degraded. Mouse AMCase degraded polymeric colloidal and crystalline chitin substrates in the gastrointestinal environments in presence of the proteolytic enzymes. Thus, AMCase can function as a protease-resistant major glycosidase under the conditions of Stomach and intestine and degrade chitin substrates to produce (GlcNAc)_2, a source of carbon, nitrogen and energy.