• Korean Journal of Food Science and Technology
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• v.13 no.3
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• pp.241-246
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• 1981

A fungal strain which produced high levels of acid protease and amylase was isolated from the atmosphere for application to the manufacture of digestive enzme preparation. This study was carried out to elucidate its microbiological characteristics, environmental conditions for production of the enzymes, and relationships between the enzyme activity and acidity. 1. The isolate was identified as a fungal strain which belonged to Aspergillus niger by the manual of Rafer and Fennel, and was found to be a strain producing high levels of acid protease and amylase. 2. The optimal pH of tile enzymes produced by the strain were: protease, 2.0;, ${\alpha}-amylase$, 4 to 5; and glucoamylase, 3 to 5. 3. The optimal culture conditions for production of the enzymes were: protease (at pH 2.5), 2 to 3 days incubation on wheat bran at $30^{\circ}C$; ${\alpha}-amylase$ and glucoamylase(at pH 3.0), 3 days incubation at $30^{\circ}C$. 4. The production of acid protease and glucoamylase was increased approximately by 20 percent when 2 percent of corn starch was added to the wheat bran medium. 5. The addition of 0.3 percent ammonium sulfate to the wheat bran medium resulted in enhancing the enzyme production, especially of acid prctease.

• Journal of Life Science
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• v.23 no.12
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• pp.1486-1494
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• 2013

A high protease-producing strain was isolated and identified from the digestive tract of octopus vulgaris by detecting a hydrolysis circle of protease and its activity. The strain was identified by morphology observation, biochemical experiments, and 16S rRNA sequence analysis. The protease obtained from the strain was purified by a three-step process involving ammonium sulfate precipitation, carboxy methyl-cellulose (CM-52) cation-exchange chromatography, and DEAE-Sephadex A50 anion-exchange chromatography. The properties of protease were characterized as well. The strain Bacillus sp. QDV-3, which produced the highest activity of protease, was isolated. On the basis of the phenotypic and biochemical characterization and 16S rRNA gene-sequencing studies, the isolate was identified as follows: domain: Bacteria; phylum: Firmicutes; class: Bacilli; order: Bacillales; family: Bacillaceae; and genus: Bacillus. The isolate was shown to have a 99.2% similarity with Bacillus flexus. A high active protease designated as QDV-E, with a molecular weight of 61.6 kDa, was obtained. The enzyme was found to be active in the pH range of 9.0-9.5 and its optimum temperature was $40^{\circ}C$. The protease activity retained more than 96% at the temperature of $50^{\circ}C$ for 60 min. Phenylmethylsulfonyl fluoride (PMSF) inhibited the enzyme activity, thus confirming that this protease isolated from Bacillus sp. QDV-3 is an alkaline serine protease. Metal ions, $Mn^{2+}$ and $Mg^{2+}$, were determined to enhance the protease activity, whereas $Ba^{2+}$, $Zn^{2+}$, and $Cu^{2+}$ were found to inactivate the enzyme.

• Journal of Aquaculture
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• v.21 no.1
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• pp.47-53
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• 2008

The purpose of this study was to suggest that selecting method of rotifer with high activity of digestive enzymes for the enrichment effect of rotifer and the increasing of digestive enzymes of fish larvae. the populations assayed the activities of the digestive enzymes were randomly selected out of several population communities cultured with freshwater condensed Chlorella. The relationship with the population density and the growth rate of selected populations was shown to RD=5865 SGR-350.08(P<0.001). The relationships with fecundity of the growth rate and the population density were shown to F=-36.147 SGR+61.652(P<0.05) and F=-0.0085 RD+66.38(P<0.001), respectively. The relationships of the growth rate and the individual activities of digestive enzymes in rotifer were assayed to Amyl=-1.6482 SGR+3.2498(P<0.05), TAP=-0.8115 SGR+1.1361(P<0.001) and TGL+0.0055 SGR+0.0079(P=0.239), respectively. But in TG-lipase was not related significantly with the growth rate. Also the relationships of the fecundity and the individual activities of digestive enzymes in rotifer were shown to Amyl=0.0296 F+1.0981(P<0.001), TAP=0.0252 F+0.0975(P<0.001) and TGL=-6E-06 F+0.0113(P=0.915), respectively. But in TG-lipase was not related significantly with the fecundity. And the relationships with the specific activity of TG-lipase of the fecundity, the growth rate and the population density were TGL=-0.024 F+0.2332(P=0.132), TGL=0.1267 SGR+0.005(P<0.01) and TGL=0.0002 F-0.0594(P<0.001), respectively. In this case, specific activity of TG-lipase was shown the significant relationship with the population density and the growth rate, but it was not related significantly with fecundity. Therefore, Because a population shown the high activity of digestive enzymes for increasing a lipid enrichment effect of a rotifers and receiving the many exogenous digestive enzymes to fish larvae was the population of high fecundity than the population of high rotifer density, to select the population of a high fecundity was suggested to benefit than a high growth rate for fish larvae.

• Journal of Aquaculture
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• v.16 no.3
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• pp.196-201
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• 2003

Sessile organisms such as the oyster Crassostrea gigas have been given much attention as a potential biomonitoring indicator to assess the impact of toxicants on aquatic organism. In this study, we exposed cells isolated from gill of oyster (Crassostrea gigas) to hydrogen peroxide in vitro. In addition oysters were in vivo exposed to pyrene and benzo(a)pyrene at various concentrations for 2 weeks. Comet assay was used to detect DNA single strand breaks and to investigate the application of this technique as a tool for aquatic biomonitoring. Hydrogen peroxide increased DNA single strand break with increasing concentration after 30 minutes exposure in vitro. Pyrene and benzo(a)pyrene caused DNA damage only at very high concentration (100 $\mu\textrm{g}$/L or 1000 $\mu\textrm{g}$/L) at two week exposure in vivo. DNA damage was relatively higher at hemocyte than at gill. It suggested that metabolized PAHs are transferred to hemolymph from digestive gland which have a relatively high enzyme activity, and attacked the DNA of hemocyte, while gill accumulated PAHs without degrading them to their metabolites due to low enzyme activity at gill. Both in vitro and in vivo exposure experiments showed that the comet assay is an effective tool on screening whether the organism are exposed to genotoxic contaminants.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.16 no.2
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• pp.147-153
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• 1983

To check the differences of the digestive enzymes by the bait habits and the proteolytic activities of the fissile extracts from the fish, omnivorous filefish (Navodon modestus), carnivorous cat shark (Scilliorhinus tarazame) and bloodsucking hag fish (Eptatretus burgeri) were sampled for this experiment. The activity of crude alkaline protease extracted from the muscle and the internal organs of the samples was determined with casein as substrate. The activity of the proteolytic enzymes showed remarkable differences by the organs of the fish. The optimum condition of the pretenses from the muscle revealed in range of pH 7.8-8.3, at $60-65^{\circ}C$, while those of the enzymes from the internal organs were at about pH 8.2, $45-55^{\circ}C$, but those of hag fish were at about pH 6.7, $45-55^{\circ}C$. The proteolytic activity of the enzyme of alimentary canal in filefish and in hag fish was 57 and 11 times stronger than that of muscle, respectively. The crude enzyme from the alimentary canal of file fish showed the strongest proteolytic activity in samples submitted and that of cat shark was the lowest. The activity of pancreatic alkaline protease in cat shark was 50 fold higher than that of muscle alkaline protease in the fish.

• Journal of Aquaculture
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• v.19 no.2
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• pp.125-132
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• 2006

Black rockfish larvae and juveniles were reared for 95 days after parturition (DAP) in order to determine four enzyme activities (trypsin, pepsine-like enzyme, lipase, amylase) during ontogeny. Larvae were fed rotifers Brachionus plicatilis from 1 to 25 DAP, Artemia nauplii from 10 to 78 DAP and then gradually changed to pelleted feed from 30 DAP. Temperature was kept between $13.5{\sim}14.9^{\circ}C$. Trypsin and lipase activities were found in 2 DAP larvae ($7.0{\pm}1.5$ unit and $4.5{\pm}1.4$ unit, $mean{\pm}SD$, respectively). The evolution of both enzymes activities showed a profile marked by drastic increases between postflexion and juvenile stage. There is an increment on specific trypsin activity at 10 DAP, corresponding with the beginning of Artemia feeding. Pepsin-like enzyme activity was found at 11 DAP and increased drastically from 56 DAP, cor-responding with the initiation of juvenile stage. Amylase activity was also found at 11 DAP and maintained at a low level up to 38 DAP followed by a drastic increase from 39 DAP to 50 DAP. Considering our results of both trypsin and pepsin-like enzyme activities, it might be concluded that higher somatic growth of Sebastes inermis could be possible with the initiation juvenile of stage and the early juvenile stage is a suit-able period for feeding an artificial diet for fish.

• Journal of Life Science
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• v.30 no.1
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• pp.82-87
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• 2020

The gene expression of amylase, trypsin, and lipase in the digestive organs of the two-spotted cricket (Gryllus bimaculatus) was tested to understand how it overcomes starvation. Amylase gene expression in the foregut was reduced by digesting no food until starvation-3 days. Although that expression persisted to starvation-6 days, it returned to normal at refeeding-2 days. The expression of trypsin peaked at around 8 times as starvation started and at around 4 times at starvation-3 days. After refeeding, trypsin expression rose up to 14 times and then fell back to normal as feeding continued. Lipase gene expression remained elevated at 1.5-2 times when starvation started and returned to normal at refeeding-2 days. In the midgut, amylase expression decreased until starvation-3 days, increasing to about 2 times at starvation-6 days; it did not rise again by refeeding. Trypsin was constantly expressed regardless of starvation and refeeding, while lipase expression was reduced by 0.6-0.7 times by starvation and refeeding. Amylase gene expression in the hindgut was 0.2-0.3 times lower than starvation-6 days, and it increased by 0.5 times on refeeding-1 day and more than 1.5 times on refeeding-3 days. The gene expression of trypsin was almost identical to amylase.

• Korean journal of applied entomology
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• v.53 no.1
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• pp.15-26
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• 2014

The black soldier fly, Hermetia illucens, larvae may depend on indigenous bacteria in the intestine to feed and digest diverse food sources. To prove this hypothesis, we isolated and identified the intestinal bacteria of the black soldier fly for their digestive and antimicrobial abilities. The last instar larvae had long digestive tracts, which were about seven times longer than its body length. An individual of H. illucens larvae possessed a total of $5.0{\pm}10^6$ bacteria in the whole intestine, of which more than 98% bacteria were located in the hindgut. Three different bacterial isolates cultured on nutrient agar (NA) medium were detected in the intestine and identified as Morganella morganii, Providencia rettgeri and Bacillus halodurans by Biolog microbial identification system. Analysis of 16S rDNA sequences of the intestinal bacteria detected the additional bacteria of Proteus mirabilis, Providencia alcalifaciens, and Providencia sp. These intestinal bacteria cultured on NA medium exhibited high resistance to 4 antibiotics and inhibited growth of other microbes which are mainly plant pathogens. Also, these bacteria exhibited catalytic activities to degrade cellulose, lipid, proteins, and carbohydrates. These results suggest that H. illucens larvae possess intestinal bacteria that may play crucial roles in their digestive physiology.

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

• Korean Journal of Poultry Science
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• v.48 no.1
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• pp.31-39
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• 2021

We aimed to investigate the age-dependent development of digestive organs, intestinal enzymes, and hepatic antioxidant defense system in White Leghorn chicks aged 0, 3, 7, 14, and 21 days. Body weight (BW) did not significantly change between days 0 and 7 but significantly increased (P<0.05) after day 7. The relative liver weight (g/100 g of BW) was significantly lower at day 0 than at the other ages but markedly increased at days 3 and 7 (P<0.05). The relative pancreatic weight changed similar to the change in liver weight, with the maximum development at 7 days (P<0.05). The relative intestinal and mucosal tissue weights increased rapidly after hatching (P<0.05), with the maximum growth at 7 days. Furthermore, maltase and sucrase activities were significantly higher at day 3 than at day 0 (P<0.05). Leucine aminopeptidase activity was high at day 0 and remained constant as age increased. Superoxide dismutase and glutathione S-transferase activities in the liver were the lowest at day 0 but significantly increased after 7 days (P<0.05). Glutathione peroxidase activity increased significantly after day 14 compared with that at days 0 and 7 (P<0.05). Lipid peroxidation was not affected by age. In conclusion, the digestive organ weights and hydrolase activity of chicks increased rapidly during the first 3 or 7 days post-hatching. Hepatic antioxidant enzyme activity increased simultaneously with the increase in digestive organ weights, after 7 days.