• Asian-Australasian Journal of Animal Sciences
• /
• v.16 no.5
• /
• pp.714-718
• /
• 2003

This study was conducted to determine the effect of DL-methionine (Met) and DL-methionine hydroxy analogue (MHA) supplementation on the growth performance, contents of serum amino acids and activities of digestive protease in broiler chicks from 0 to 3 weeks old . There were three treatments in this study: (1) control (basal diet), (2) 0.24% Met supplementation and (3) 0.368% MHA supplementation. The results showed that Met and MHA supplementation did not significantly (p>0.05) improve feed efficiency and body weight gain for broilers from 0-1, 0-2 and 0-3 weeks of age. The serum levels of homocystine, methionine and taurine were significantly (p<0.05) higher with supplementation of Met or MHA than with control. The pepsin activity of proventriculus was increased with (p<0.05) Met supplementation at 21 days of age and with MHA supplementation at 7 and 14 days of age. The trypsin activity was also increased (p<0.05) with MHA supplementation at 7 days of age. The chymotrypsin activity in pancreas and the dipeptidase activity in small intestinal mucosa and content were not affected (p>0.05) by Met or MHA supplementation.

• Current Research on Agriculture and Life Sciences
• /
• v.32 no.2
• /
• pp.63-66
• /
• 2014

The effects of a newly developed flower thinning formulation (FTF) on the vitality of the honey bee Apis mellifera were examined by measuring the activities of various digestive enzymes in adult worker bees. First, direct spraying of the FTF solution did not cause any behavioral changes or lethal effects for the honey bees based on 24 h observation. Second, oral ingestion of a sugar solution containing the FTF did not produce any significant change in the activities of amylase, proteinases, lipase, or acetylcholine esterase (AChE) in the worker bees 6 h or 24 h after treatment. Meanwhile, a commercial formulation containing sulfur compounds showed slightly reduced activities for several digestive enzymes and AChE, although no behavioral disturbance. Thus, the results of the present study suggest that the FTF is not toxic for honey bees, in terms of contact and ingestion. Therefore, this newly developed FTF can be used for flower thinning without any detrimental effects on pollinating insects.

• Preventive Nutrition and Food Science
• /
• v.18 no.4
• /
• pp.273-279
• /
• 2013

Protease widely exists in the digestive tract of animals and humans, playing a very important role in protein digestion and absorption. In this study, a high protease-producing strain Planomicrobium sp. L-2 was isolated and identified from the digestive tract of Octopus variabilis. The strain was identified by physiological and biochemical experiments and 16S rDNA sequences analysis. A protease was obtained from the strain Planomicrobium sp. L-2 through ammonium sulfate precipitation, dialysis and enrichment, DEAE-Sephadex A50 anion-exchange chromatography, and Sephadex G-100 gel chromatography. The molecular weight and properties of the protease were characterized, including optimum temperature and pH, thermal stability, protease inhibitions and metal ions. According to our results, the protease from Planomicrobium sp. L-2 strain designated as F1-1 was obtained by three-step separation and purification from crude enzyme. The molecular weight of the protease was 61.4 kDa and its optimum temperature was $40^{\circ}C$. The protease F1-1 showed a broad pH profile for casein hydrolysis between 5.0~11.0. No residual activity was observed after incubation for 40 min at $60^{\circ}C$ and 60 min at $50^{\circ}C$. F1-1 protease was inhibited by $Mn^{2+}$, $Hg^{2+}$, $Pb^{2+}$, $Zn^{2+}$, and $Cu^{2+}$ ions, as well as PMSF, indicating that the protease F1-1 was a serine protease. Additionally, research basis provided by this study could be considered for industrial application of octopus intestinal proteases.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.25 no.2
• /
• pp.181-185
• /
• 2012

Chymotrypsin serine protease is one of the main digestive proteases in the midgut of and is involved in various essential processes. In a previous study, a gene encoding a chymotrypsin-like protease, Hi-SP1, was cloned from the larvae of Hermetia illucens and characterized. In this study, we produced the recombinant chymotrypsin-like protease Hi-SP1 in Escherichia coli cells. The molecular weight of the recombinant Hi-SP1 was estimated to be approximately 26 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western-blotting. Chymotrypsin activity was detected when AAPF was used as the substrate. Examination of the effects of temperature and pH revealed that the proteolytic activity of recombinant Hi-SP1 decreased markedly at temperatures above $30^{\circ}C$, and the optimum pH was found to be 10.0.

• Journal of Animal Science and Technology
• /
• v.65 no.1
• /
• pp.32-56
• /
• 2023

This review explores the factors that improve meat protein digestibility and applies the findings to the development of home meal replacements with improved protein digestion rates in older adults. Various methods improve the digestion rate of proteins, such as heat, ultrasound, high pressure, or pulse electric field. In addition, probiotics aid in protein digestion by improving the function of digestive organs and secreting enzymes. Plant-derived proteases, such as papain, bromelain, ficin, actinidin, or zingibain, can also improve the protein digestion rate; however, the digestion rate is dependent on the plant enzyme used and protein characteristics. Sous vide processing improves the rate and extent of protein digestibility, but the protein digestion rate decreases with increasing temperature and heating time. Ultrasound, high pressure, or pulsed electric field treatments degrade the protein structure and increase the proteolytic enzyme contact area to improve the protein digestion rate.

• Journal of Aquaculture
• /
• v.22 no.1
• /
• pp.105-111
• /
• 2009

In this study, we carried out an experiment for estimation the larval digestibility in aspects which digestive enzymatic activities and nutrition of the rotifers, Brachionus rotundiformis. Thus we enhanced the digestive enzymatic activity through the addition of starch for the increase of digestibility of rotifer (starch-rotifer), and compared with the feed efficiency through rearing of the olive flounder, Paralichthys olivaceus used rotifer lipid-enriched with Algamac $2000^{(R)}$ (CE-rotifer). The digestive enzyme activities (except for TG-lipase), total protein contents, total essential amino acid, essential amino acids (methionin and phenylalanine) of starch-rotifer (the rotifer used a starch as additive, and enriched not) was assayed significantly higher than CE-rotifer (P<0.05). And total lipid, lipid classes (except for sterol) and fatty acids as DHA and EPA showed higher in CE-rotifer than starch-rotifer (P<0.05). But, sterol contents and ST/TG ratio were shown significantly higher in starch-rotifer (P<0.05). The flounder larvae supplied the two rotifers showed standard length and body weight that not significantly differed with ranges $3.72{\sim}3.79\;mm$ and $32.9{\sim}37.8\;mg$/larva on 6 days after hatching (DAH), respectively (P>0.05). However, these of 12 DAH showed the values of significantly higher to $5.94{\pm}0.249\;mm$, $144.0{\pm}23.86\;mg$/larva and $26.2{\pm}12.13%$ in standard length, body weight and survival in CE-flounder than that of starch-flounder (P<0.05). The hydrolytic enzymatic activities of flounder larvae severally supplied the two rotifers showed the significantly higher activities in acidic -amylase, neutral -amylase, TG-lipase, lysozyme and acidic phosphatase in starch-flounder on 5 DAH (P<0.05). But neutral $\alpha$-amylase, three proteases and two phosphatases of CE-flounder on 11 DAH showed the significantly higher activities than that of starch-flounder (P<0.05). Therefore, for the flounder, Paralichthys olivaceus larvae just depleted yolk was more beneficial to supply the feed, rotifer, enhanced the digestibility than to supply the feed lipid-enriched for aspect of larval digestibility up to 6 DAH, thereafter nutrition of absorption due to the development of digestive organs suggested that enrichment effect appeared with larval somatic growth. Consequently, investigation more detailed about the larval digestive physiological and nutritional requirement variations after 6 DAH will be necessary, thereafter.

• Food Science of Animal Resources
• /
• v.32 no.5
• /
• pp.652-658
• /
• 2012

This study was carried out to identify the ACE (Angiotensin converting enzyme) inhibitory activity of casein hydrolysates for development of anti-hypertensive hydrolysates. Sodium caseinate was treated with six kinds of commercial proteases such as Flavourzyme, Protamex, Neutrase 1.5, Alcalase, Protease M, and Protease S for 8 h individually, and was then treated with the enzyme combination for 4 h at $45^{\circ}C$. The hydrolysate which had the highest ACE inhibitory effect was then hydrolysed successively with three digestive enzymes: pepsin, trypsin, and ${\alpha}$-chymotrypsin, at $37^{\circ}C$ for 4 h under conditions mimicking those of the gastrointestinal tract. UF (ultra filtration) treatment was applied to one of the secondary hydrolysates to determine ACE inhibitory activity. When sodium caseinate was hydrolysed by commercial proteases, the degree of hydrolysis (DH) showed 2.54 to 4.25% and after secondary hydrolysis, DH showed 4.30 to 5.22%. ACE inhibitory activity and $IC_{50}$ values decreased, and inhibition rates increased during hydrolysis. Protamex treatment showed the lowest $IC_{50}$ value ($516{\mu}g/mL$) and Flavourzyme hydrolysate showed the highest $IC_{50}$value ($866{\mu}g/mL$). As the first hydrolysate was treated with Flavourzyme, the ACE inhibitory activity increased. Neutrase hydrolysate had the highest activity with an $IC_{50}$ value ($282{\mu}g/mL$). When Neutrase plus Flavourzyme treatment was hydrolyzed by digestive enzymes, the $IC_{50}$ value ($597{\mu}g/mL$) was decreased statistically (p<0.05). As Neutrase plus Flavourzyme hydrolysate is treated by UF with MW cut-off 10,000, permeate showed $273{\mu}g/mL$ of $IC_{50}$ value, showed no difference, but retentate which has over MW 10,000 showed statistically different $IC_{50}$ value, $635{\mu}g/mL$ (p<0.05).

• Journal of Nutrition and Health
• /
• v.19 no.6
• /
• pp.363-373
• /
• 1986

This study was devised to elucidate whether soused shrimp exhibits a digestive action on boiled pork meats. and the mechanism by which sousing with a high concentration of sodium chloride preserves nutrients in foods for a prolonged pe\ulcornerriod. Protease was isolated from soused shrimp using a combination of ammonium sulfate fractionation. DEAE - cellulose ion exchange chromatography and gel filtra\ulcornertion. The isolated protease had specific activity of 1.560 units. 210 purification fo\ulcornerld with an yield of 38%. Its optimum pH and temperature were 8.0 and $43^{\circ}C$ respectively. The molecular weight of the enzyme was 35.000. The Km value of the enzyme for casein was 1.6 x $10^{-6}$ M The e=yme required the presence of cu\ulcornerpric ion to exhibit its full activity. Eighty eight percent of the enzyme activity was in\ulcornerhibited by 3.5M NaCI showing a reversibly linear decrease of the enzyme activity as NaCI concentration increased. The nature of the inhibition by NaCl was rever\ulcornersible and noncompetitive. The protease activity in soused shrimp was well preser\ulcornerved with the elapse of time at least in part due to NaCI induced suppression of autodigestion. The enzyme was denatured by acid easily. i.e. 1% of the original activity remained after staying at pH 2 for 10 minutes. which is within the norm\ulcorneral range of pH of the human stomach. Soused shrimp was observed to be one of those containing the highest protease activity compared with the other soused foo\ulcornerds such as soused oyster. squid. clam. and Pollack intestine with respect to spec\ulcornerific activities of dialized 1:4 whole homogenates(w/v) in 5 mM sodium phospha\ulcornerte - 2.4 mM j3 - mercaptoethanol buffer. pH 8.0. Casein and boiled meats including pork, beef, and chicken appeared to be the good substrates for the protease. Casein was the best. Therefore. the ingestion of boiled meats including pork together with soused sh\ulcornerrimp would help digestion of boiled pork in human not only by increasing appe\ulcornertite also by the direct proteolytic digestion of boiled meats by soused shrimp to\ulcorner some extent. And a high concentration of sodium chloride inhibited the protease activity reversibly in a remarkable degree, which ensued in a significant retardat\ulcornerion of autodigestion of protein in foods by proteases, and hereby contributed to the preservation of foods for an extended period.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.31 no.6
• /
• pp.875-881
• /
• 1998

To develop functional food material with angiotensin converting enzyme (ACE) inhibitory peptides, muscle protein of anchovy, Engraulis japonica was hydrolyzed during 48 hrs by digestive pretenses such as pepsin, trypsin, $\alpha$-chymotrypsin, and commercial proteases such as papain, bromelain, complex enzyme, Elavourzyme, Novozym, Neutrase, Protamex and Alcalase. The only $50\%$ ethanol soluble hydrolysates were tested for inhibitory activity against ACE and yield of $50\%$ ethanol soluble peptide-nitrogen ($ESPN_{50}$). ACE inhibition effects and yield of $ESPN_{50}$ occurred as hydrolysis time increased to 8 hrs, Among those pretenses tested, hydrolysates by Alcalase and $\alpha$-chymohypsin had greater ACE inhibitory activity (80 and $74\%$, reipectively) with eletated levels of $ESPN_{50}$ (48 and 58 mg/ml, respectively), while Protamex hydrolysates had greater ACE inhibitory activities ($73\%$) with reduced levels of $ESPN_{50}$ (7.2mg/ml) than others. Amino acid compositions of $50\%$ ethanol solubles obtained from those hydrolysates were rich in glutamic acid, aspartic acid, cysteine and leucine.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.24 no.5
• /
• pp.273-288
• /
• 1991

To elucidate the physiological and biochemical differences between chondrichthyes and osteichthyes, the properties of the specific digestive enzymes in cat-shark, Cephaloscyllium umbratile, and mackerel, Scomber japonicus, were studied. Homogenous trypsin proved through the disc-electrophoresis, SDS-PAG electrophoresis and gel filtration was obtained from the pancreas of cat-shark by $50-70\%$ saturated ammonium sulphate fractionation, DEAE-Sephadex A-50 column chromatography, benzamidine-Sepharose 6B affinity chromatography and Sephadex G-75-120 gel filtration. Two types of trypsins were also obtained from the pyloric caeca of mackerel by $30-70\%$ saturated ammonium sulphate fractionation and the slightly modified procedure from the method adopted in the purification of cat-shark trypsin. The two trypsins, designated trypsin A and B, were proved their homogeneity by disc- and SDS-PAG electrophoresis and gel filtration. The molecular weights of the trypsins were estimated to be 31,700 for cat-shark trypsin, 30,000 for mackerel trypsin A and 29,000 for mackerel trypsin B by SDS-PAG electrophoresis, but those were estimated to be 21,500 for cat-shark trypsin, 23,700 for mackerel trypsin A and 21,500 for mackerel trypsin B by gel filtration. The trypsins exhibited their optimum conditions at pH 9.0 and on temperature ranged from $45^{\circ}C\;to\;50^{\circ}C$ for cat-shark, and at pH 8.0 and a temperature of $50^{\circ}C$ for mackerel trypsin A and B, respectively. The cat-shark trypsin was stable at pH 10.0 and the temperature below $10^{\circ}C$, whereas the mackerel trypsin A and B, were stable in the range over pH 7.0 to pH 9.0 below $10^{\circ}C$ and at pH 8.0 below $35^{\circ}C$, respectively. The mackerel trypsins were severely inhibited by some heavy metal ions such as $Ag^{2+},\;Cu^{2+}\;and\;Hg^{2+}$ compared to cat-shark trypsin. All of the enzymes were also inhibited by antipain, leupeptin, TLCK(tosyllysine chloromethyl ketone) and SBTI(soybean trypsin inhibitor) remarkably. The inhibitory effects of PMSF(phenylmethane sulphonylfluoride), DFP(diisopropyl fluorophosphate) and benzamidine were indicated that these enzymes belong to serine-proteases.