• BMB Reports
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• v.32 no.6
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• pp.567-572
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• 1999

We have previously shown that a DNA fragment is responsible for the anticoagulatory effect of an earthworm, Lumbricus rubellus. The anticoagluant increased the activated partial thromboplastin time (APTT) and also inhibited the thrombin activity observed with either N-${\alpha}$-p-tosyl-L-arginine methyl ester (TAME) or H-D-phenyl-alanyl-L-pipecoil-L-arginine-p-nitroanilide (S-2238). Since trypsin digestion of the anticoagulant further increased the APTT, the possible presence of a regulatory protein for the anticoagulatory DNA was investigated by digesting the anticoagulant with trypsin and isolating the DNA fragment with C4-reversed phase HPLC. The DNA fragment lacking a regulatory protein was eluted in the flow-through fraction, and analyzed with thrombin and activated factor X. Activated factor X activity was more strongly inhibited than thrombin activity. For DNA digestion, we treated the anticoagulant with DNase and purified the DNA-binding protein with a FPLC Resource-S cation exchange column. The regulatory protein, with an $M_r$ of 55.0 kDa, reduced the anticoagulatory effect of the DNA fragment.

• BMB Reports
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• v.32 no.6
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• pp.573-578
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• 1999

Treatment of Hafnia alvei aspartase with limited tryptic digestion resulted in a marked increase in enzymatic activity. The activation required a few minutes to attain maximum level and, thereafter, the activity gradually decreased to complete inactivation. The degree of cleavage associated with the activation was extremely small as judged by SDS-PAGE. Upon activation, the optimum pH and temperature were essentially unchanged. When trypsin-activated enzyme was denatured in 4 M guanidine-HCI followed by removal of the denaturant by dilution, the restoration of activity was similar (40%) to that of the native enzyme, indicating a degree of stability. The $pK_a$ obtained on the acidic side and the $pK_b$ obtained on the basic side of trypsin-activated aspartase were 6.6 and 8.6, respectively, the same as those of the native aspartase, indicating that aspartase may exist in a stable conformation after limited tryptic digestion. These results indicate that the activation of H. alvei may be mediated by a conformational change away from the active site of individual subunits.

• Applied Biological Chemistry
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• v.12
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• pp.7-11
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• 1969

Mucor-rennin, the crystalline milk-clotting enzyme, isolated from Mucor pusillus var. Lindt, has an acid protease activity. The optimum pH for the digestion of k-casein is 4.5, while that for hemoglobin digestion is 4.0. The skim milk solution was easily clotted acidic solution than alkalin solution, and the milk clotting activated by Ca ion. The enzyme was heat stable against heat from pH 4.0 to 6.0 but was more stable at pH 5.0. The activity of the enzyme at pH 5.0 did not decrease at 30 C for 15 days and the activity was not effected by sodium propionate and salicilic acid. Therefore, the enzyme of liguid type could store for a long time and could be transported from Erzyme production Co. to Manufacture of cheese Co. by adding the antiesptic and by adjusting pH to 5.0.

• Journal of Korean Society on Water Environment
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• v.21 no.5
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• pp.490-495
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• 2005

Since there are very limited numbers of thermophilic anaerobic digesters being operated, it is often difficult to start up a new one using sludge from an existing reactor as a seed. However, for obvious reasons it seems few attempts have been made to compare the start-up performance of thermophilic anaerobic digestion using different sources of seed sludges. The purpose of this study was to evaluate the start-up performance of anaerobic digestion using aerobic Waste Activated Sludge (WAS) from a plant and mesophilic Anaerobic Digested Sludge (ADS) as the seed source at both mesophilic ($35^{\circ}C$) and thermophilic ($55^{\circ}C$) temperatures. In this study, two experiments were conducted. First, thermophilic anaerobic reactors were seeded with WAS (VSS = 4,400 mg/L) and ADS (VSS = 14,500 mg/L) to investigate start-up performance with a feed of acetate as well as propionate. The results show that WAS started to produce $CH_4$ soon after acetate feeding without a lag time, while ADS had a lag time of 10 days. When the feed was changed to propionate, WAS removed propionate down to below the detection limit of 10 mg/L, while ADS removed little propionate and produced little $CH_4$. Second, in order to further compare the methanogenic activity of WAS and ADS, both mesophilic and thermophilic reactors were operated. WAS acclimated to anaerobic conditions shortly and after acclimating it produced more $CH_4$ than ADS. WAS at mesophilic temperature biodegraded acetate at the same rate as for thermophilic. However WAS at mesophilic temperature biodegraded propionate at a much faster rate than at thermophilic. WAS as the seed source of anaerobic digestion resulted in much better performance than ADS at both mesophilic and thermophilic temperatures for both acetate and propionate metabolism.

• Journal of Aquaculture
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• v.5 no.1
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• pp.1-7
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• 1992

Digestion rates of protein, carbohydrate, and lipid in the two kinds of extruding processed soybean meals, heated (H-SBM) and raw (R-SBM), were tested for evaluate the effectiveness of soybean meal in the rainbow trout diet. The relation between digestion rate of protein and trypsin inhibitor (TI) was also determined. The protein digestion rate of both H-SBM and R-SBM were increased up to $95{\%}$ by extruding process compared to the none treated soybean meal. The digestion rate of carbohydrate in R-SBM was increased by extruding process whereas the one in H-SBM was not. The activity of trypsin inhibitor was almost diminished by the extruding process and digestion rate of dietary protein was improved. However, the reason of this improvement was not clear whether caused by the deactivation of trypsin inhibitor.

• Fisheries and Aquatic Sciences
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• v.18 no.4
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• pp.341-347
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• 2015

We evaluated the biological activities of five water extracts of tissue of the mottled anemone Urticina crassicornis. Most extracts exhibited broad-spectrum antimicrobial activity as determined by ultrasensitive radial diffusion assay (URDA) against gram-positive and -negative bacteria, including a fish pathogen, Aeromonas hydrophila, but no activity against fungi. The activity of the extracts was abolished by tryptic digestion, indicating that protein compounds were responsible for the antimicrobial activity. Furthermore, in a 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical-scavenging activity assay, only the visceral tissue extract showed activity. However, no extract had hemolytic activity against human red blood cells. Consequently, this study suggests the water-soluble extract of mottled anemone to be a promising source of proteinaceous antimicrobial compounds that can be utilized for development of novel antibiotics.

• Food Science of Animal Resources
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• v.33 no.4
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• pp.493-500
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• 2013

Gelatin is a collagen-containing thermohydrolytic substance commonly incorporated in cosmetic and pharmaceutical products. This study investigated the antioxidant activity of gelatin by using different reagents, such as 2,2-azinobis-(3-ethylbenzothiazoline- 6-sulfonic acid) (ABTS), 2,2-di (4-tert-octylphenyl)-1-picrylhydrazyl (DPPH), and oxygen radical absorbance capacity-fluorescein (ORAC-FL) in a porcine gelatin hydrolysate obtained using gastrointestinal enzymes. Electrophoretic analysis of the gelatin hydrolysis products showed extensive degradation by pepsin and pancreatin, resulting in an increase in the peptide concentration (12.1 mg/mL). Antioxidant activity, as measured by ABTS, exhibited the highest values after 48-h incubation with pancreatin treatment after pepsin digestion. Similar effects were observed at 48 h incubation, that is, 61.5% for the DPPH assay and 69.3% for the ABTS assay. However, the gallic acid equivalent (GE) at 48 h was $87.8{\mu}M$, whereas $14.5{\mu}M$ GE was obtained using the ABTS and DPPH assays, indicating about sixfold increase. In the ORACFL assay, antioxidant activity corresponding to $45.7{\mu}M$ of trolox equivalent was found in the gelatin hydrolysate after 24 h hydrolysis with pancreatin treatment after pepsin digestion, whereas this activity decreased at 48 h. These antioxidant assay results showed that digestion of gelatin by gastrointestinal enzymes prevents oxidative damage.

• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.26-35
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• 2017

The hydrolysis of proteins constitutes an invaluable tool, granting access to a variety of peptide fragments with potentially interesting biological properties. Therefore, a hemoglobin (Hb) hydrolysate of Crocodylus siamensis was generated by digestion under acidic conditions. The antibacterial and antioxidant activities of the Hb hydrolysate were assessed in comparison with intact Hb. A disc diffusion assay revealed that the Hb hydrolysate exhibited antibacterial activity against eight strains of gram-positive bacteria and showed a higher efficacy than intact Hb. Moreover, the antioxidant activity of intact Hb and its hydrolysate was evaluated using ABTS and DPPH radical scavenging assays. The Hb hydrolysate exhibited free radical scavenging rates of 6-32%, whereas intact Hb showed a slightly higher activity. In addition, non-toxicity to human erythrocytes was observed after treatment with quantities of Hb hydrolysate up to $10{\mu}g$. Moreover, active fragmented Hb (P3) was obtained after purifying the Hb hydrolysate by reversed-phase HPLC. Scanning electron microscopy demonstrated the induction of bacterial cell membrane abnormalities after exposure to P3. Antibacterial and antioxidant activities play crucial roles for supporting the wound healing activity. Consequently, an in vivo mice excisional skin wound healing assay was carried out to investigate the effects of intact Hb treatment on wound healing in more detail. The results clearly demonstrate that intact Hb is capable of promoting 75% wound closure within 6 days. These findings imply that intact Hb of C. siamensis and its acid hydrolysate may serve as valuable precursors for food supplementary products benefitting human health.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.369.2-369.2
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• 2002

Rumex crispus (Polygonaceae) is a well known perennial plant. which is called So-Ri-Jaeng-Yi. growing in the field and on the roadside. It has been used as a Korean Folk medicine in treating of acute and chronic cutaneous disease. cathartics, fever and jaundice. Also, the seed of this plant has been used as only a folk medicine for the treatment of digestion problems, liver diseases and many sorts of tumor. So we examined analgesic activity, anti-inflammatory activities and hepatoprotective activity using MeOH extraction and BuOH fraction in this plant. (omitted)

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.1
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• pp.113-128
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• 1999

Protozoa can represent as half of the total rumen microbial biomass. Around 10 genera are generally present on the same time in the rumen. Based on nutritional aspects they can be divided in large entodiniomorphs, small entodiniomorphs and isotrichs. Their feeding behaviour and their enzymatic activities differ considerably. Many comparisons between defaunated and refaunated animals were carried out during the last two decades to explain the global role of protozoa at the ruminal or animal levels. It is now generally considered that a presence of an abundant protozoal population in the rumen has a negative effect on the amino acid (AA) supply to ruminants and contribute to generate more methane but, nevertheless, protozoa must not be considered as parasites. They are useful for numerous reasons. They stabilise rumen pH when animal are fed diets rich in available starch and decrease the redox potential of rumen digesta. Because cellulolytic bacteria are very sensitive to these two parameters, protozoa indirectly stimulate the bacterial cellulolytic activity and supply their own activity to the rumen microbial ecosystem. They could also supply some peptides in the rumen medium which can stimulate the growth of the rumen microbiota, but this aspect has never been considered in the past. Their high contribution to ammonia production has bad consequences on the urinary nitrogen excretion but means also that less dietary soluble nitrogen is necessary when protozoa are present. Changes in the molar percentages of VFA and gases from rumen fermentations are not so large that they could alter significantly the use of energy by animals. The answer of animals to elimination of protozoa (defaunation) depends on the balance between energy and protein needs of animals and the supply of nutrients supplied through the diet. Defaunation is useful in case of diets short in protein nitrogen but not limited in energy supply for animals having high needs of proteins.