• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.101-101
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• 2002

GH-transgenic coho salmon (Oncorhynchus kitsutch) juveniles in tGH＊T$_3$and tGH*PTU were fed with the diets containing 1 ug/g fish of 3,5,3'-triiodo-L-thyronine (T$_3$) and 30 ug/g fish of 6-n-propyl-2- thiouracil (PTU), respectively, to assess the effect of these drugs on the change of physiological activity, growth and survival rate in comparison with normal transgenic (tGH＊C) and nontransgenic coho salmon (Wild) for 90 days. Although the daily food intakes of all transgenic (tGH)-groups were higher than Wild, the amount was reduced by exogenous PTU supply. The fred efficiencies of tGH-groups were lower than Wild, but the efficiency was reduced both by T$_3$and PTU. The survival rate of tGH-group was significantly higher than that of Wild, but there was no significant difference among tGH-groups. Although the growth of tGH-coho salmon was faster than Wild. the growth rate of transgenic salmon was increased by exogenous T$_3$, but was reduced by PTU Plasma TT$_4$levels of tGH-groups was approximately 2-fold higher relative to Wild, but there were no difference of plasma TT$_4$levels among tGH-groups. plasma TT$_3$level or tGH-coho salmon was increased by exogenous T$_3$administration, but was reduced by exogenous PTU. In addition, although plasma GH levels of all tGH-groups were higher than that of Wild, the GH level in plasma of transgenic coho salmon was increased by exogenous T$_3$and reduced by exogenous PTU. In the meantime, the transgenic fishes also displayed head, jaw and opercular abnormalities typical of the offsets of this gene construct in coho salmon, indicating that some imbalance in growth processes has been induced. However, the abnormalities of transgenic coho salmon was reduced following exogenous PTU administration.

• Fisheries and Aquatic Sciences
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• v.25 no.1
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• pp.1-11
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• 2022

Catfish is one of the most important freshwater fish farming commodities in Indonesia. Higher catfish production can be achieved by cultivating transgenic catfish carrying the growth hormone (GH) gene of African catfish (Clarias gariepinus GH, CgGH). This research focuses on analysis of the presence of the CgGH gene in transgenic G₁, G₂, and G₃ mutiara catfish broodstock, as an indication of stable CgGH inheritance. CgGH gene was isolated using the RNeasy mini kit and RT-PCR. RT-PCR revealed amplicons measuring approximately 600 bp in transgenic G₀, G₁, G₂, and G₃ mutiara catfish. The CgGH consensus sequence similarities ranged from 93.76% to 97.06%, with four functional domain sites (somatotropin-1, somatotropin-2, four α-helix, N-glycosylation, four cysteine residues) of fish GH proteins. The functional domains of fish GH proteins are conserved in G₁, G₂, and G₃ and indicate stable exogenous GH inheritance to produce transgenic catfish strains in each generation.

• Journal of the Korean earth science society
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• v.29 no.3
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• pp.255-262
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• 2008

Late Cretaceous to early Tertiary paleostress was evaluated by analyzing the healed microcracks in the Cretaceous granite of the Goheung area, south Korea. Healed microcracks in five granite samples (GH-1, GH-3, GH-4, GH-5, GH-8) were investigated and measured according to direction. The directions of maximum horizontal principal stress in GH-1, GH-3, and GH-4 are dominantly $N60^{\circ}W\;and\;N70^{\circ}E,\;N20^{\circ}W\;and\;N50^{\circ}W$, while minor directions are N-S and $N30^{\circ}E$. In GH-5 and GH-8, $N40^{\circ}E\;and\;N10^{\circ}E$ are the most dominant directions, while $N40^{\circ}W$ is the minor direction. Thus overall, the most dominant directions of healed microcracks in the study area are oriented $N60^{\circ}W$, while minor directions are oriented $N20^{\circ}W,\;N20^{\circ}E\;and\;N70^{\circ}E$, essentially NE. Combining the paleostress results of this study with other studies, the direction of the maximum horizontal principal stress in the study area during the late Cretaceous to the early Tertiary should perhaps be changed WNW to NE. The reason for this is thought to be the complex tectonic movements which occurred in northeast Asia at that time.

• Journal of Life Science
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• v.31 no.3
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• pp.356-365
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• 2021

Recently, we sequenced the entire genome of a freshwater agar-degrading bacterium Cellvibrio sp. KY-GH-1 (KCTC13629BP) to explore genetic information encoding agarases that hydrolyze agarose into monomers 3,6-anhydro-L-galactose (L-AHG) and D-galactose. The KY-GH-1 strain appeared to possess nine β-agarase genes and two α-neoagarobiose hydrolase (α-NABH) genes in a 77-kb agarase gene cluster. Based on these genetic information, the KY-GH-1 strain-caused agarose degradation into L-AHG and D-galactose was predicted to be initiated by both endolytic GH16 and GH86 β-agarases to generate NAOS (NA4/NA6/NA8), and further processed by exolytic GH50 β-agarases to generate NA2, and then terminated by GH117 α-NABHs which degrade NA2 into L-AHG and D-galactose. More recently, by employing E. coli expression system with pET-30a vector we obtained three recombinant His-tagged GH50 family β-agarases (GH50A, GH50B, and GH50C) derived from Cellvibrio sp. KY-GH-1 to compare their enzymatic properties. GH50A β-agarase turned out to have the highest exolytic β-agarase activity among the three GH50 isozymes, catalyzing efficient NA2 production from the substrate (agarose, NAOS or AOS). Additionally, we determined that GH117A α-NABH, but not GH117B α-NABH, could potently degrade NA2 into L-AHG and D-galactose. Sequentially, we examined the enzymatic characteristics of GH50A β-agarase and GH117A α-NABH, and assessed their efficiency for NA2 production from agarose and for production of L-AHG and D-galactose from NA2, respectively. In this review, we describe the benefits of recombinant GH50A β-agarase and GH117A α-NABH originated from Cellvibrio sp. KY-GH-1, which may be useful for the enzymatic hydrolysis of agarose for mass production of L-AHG and D-galactose.

• Journal of Life Science
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• v.18 no.11
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• pp.1617-1624
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• 2008

Chitin and chitosan, which is deacetylated form of chitin, are one of the most abundant biomass on the earth. They showed various biological activities including antimicrobial activity, heavy metal chelating, immune system activation, and have very diverse applications in food, pharmaceutical, medicinal, and environmental industry. There have been reported many chitin/chitosan-hydrolyzing enzymes, their structures and genes from three domains, archaea, bacteria, and eukarya. Carbohydrate hydrolyzing enzymes are classified in CAZy (Carbohydrate Active Enzymes) database according to their amino acid sequence similarity. Interestingly, chitinases and chitosanases are classified in various glycosyl hydrolase(GH) families, GH2, GH5, GH7, GH8, GH18, GH19, GH20, GH46, GH48, GH73, GH75, GH80, GH84, and GH85. Here, we review characteristics and structures of chitin/chitosan hydrolyzing enzymes according to glycosyl hydrolase families in order to provide information about gene mining.

• KSBB Journal
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• v.10 no.3
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• pp.271-282
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• 1995

The authentic hGH converted from met-hGH by immobilized ApM was purified by successive chromatographic processes based on the differences in isoelectric points, hydrophobicities and charges. The final recovery yield was about 14.1% and the specific activity of the purified hGH was 2.75IU per mg when assayed by enzyme immunoassay. The purified hGH was verified to be authentic hGH through the analysis of amino acid composition, amino-terminal amino acid sequence, carboxy-terminal amino acid and tryptic peptide map. The purity of purified hGH was higher than that of commercial hGH when assessed by SDS-PAGE, PAGE, IEF and HSGF. In weight-gain assay and tibia test with hypophysectomized rats, the hGH produced in this study showed the same growth effect as the commercial hGH.

• Animal Bioscience
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• v.34 no.5
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• pp.867-879
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• 2021

Objective: Fibronectin 3 (FN3) and immunoglobulin like modules (Ig) are usually collocated beside modular cellulase catalytic domains. However, very few researches have investigated the role of these modules. In a previous study, we have sequenced and analyzed bacterial metagenomic DNA in Vietnamese goats' rumen and found that cellulase-producing bacteria and cellulase families were dominant. In this study, the properties of modular cellulases and the role of a FN3 in unique endoglucanase belonging to glycosyl hydorlase (GH) family 5 were determined. Methods: Based on Pfam analysis, the cellulases sequences containing FN3, Ig modules were extracted from 297 complete open reading frames (ORFs). The alkaline, thermostability, tertiary structure of deduced enzymes were predicted by AcalPred, TBI software, Phyre2 and Swiss models. Then, whole and truncated forms of a selected gene were expressed in Escherichia coli and purified by His-tag affinity column for assessment of FN3 ability to enhance enzyme activity, solubility and conformation. Results: From 297 complete ORFs coding for cellulases, 148 sequences containing FN3, Ig were identified. Mostly FN3 appeared in 90.9% beta-glucosidases belonging to glycosyl hydrolase family 3 (GH3) and situated downstream of catalytic domains. The Ig was found upstream of 100% endoglucanase GH9. Rarely FN3 was seen to be situated downstream of X domain and upstream of catalytic domain endoglucanase GH5. Whole enzyme (called XFN3GH5 based on modular structure) and truncate forms FN3, XFN3, FN3GH5, GH5 were cloned in pET22b (+) and pET22SUMO to be expressed in single and fusion forms with a small ubiquitin-related modifier partner (S). The FN3, SFN3 increased GH5 solubility in FN3GH5, SFN3GH5. The SFN3 partly served for GH5 conformation in SFN3GH5, increased modules interaction and enzyme-soluble substrate affinity to enhance SXFN3GH5, SFN3GH5 activities in mixtures. Both SFN3 and SXFN3 did not anchor enzyme on filter paper but exfoliate and separate cellulose chains on filter paper for enzyme hydrolysis. Conclusion: Based on these findings, the presence of FN3 module in certain cellulases was confirmed and it assisted for enzyme conformation and activity in both soluble and insoluble substrate.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.541-545
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• 2000

For the sustained release formulation of recombinant human growth hormone (rhGH), dissociable rhGH aggregates were microencapsulated within poly(D,L-lactic-co-glycolic acid) [PLGA] microparticles. rhGH aggregates with 2 - 3 m Particle diameter were first produced by adding a small volume of aqueous rhGH solution into a partially water miscible organic solvent phase(ethyl acetate) containing PLGA. These rhGH aggregates were then microencapsulated within PLGA polymer phase by extracting ethyl acetate into an aqueous phase pre-saturated with ethyl acetate. The resultant microparticles were 2 - 3 m in diameter similar to the size of rhGH aggregates, suggesting that PLGA polymer was coated around the protein aggregates. Release profiles of rhGH from these microparticles were greatly affected by changing the volume of the incubation medium. The release rhGH species consisted of mostly monomeric form with having a correct conformation. This study reveals that sustained rhGH release could be achieved by microencapsulating reversibly dissociable protein aggregates within biodegradable polymers.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.8
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• pp.1193-1198
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• 2003

An increase in frequency of administration of exogenous growth hormone (GH) or GH-releasing hormone was reported to be a model to increase blood circulating insulin-like growth factor-1 (IGF-1) and to improve growth performance in animals. We have investigated the effect of twice daily administration of GH-releasing peptide-2 (GHRP-2) on growth performance, GH responsiveness and plasma insulin-like growth factor IGF-1 in swine. We administered to eight swine, 3 control and 5 treatment, a twice daily s.c. injections of GHRP-2 ($30{\mu}g/kg\;BW$) for a period of 10 days. Every day blood samples immediately taken before injections of GHRP-2 or saline, at 08:00 h and 16:00 h, were measured for IGF-1 concentrations. Blood samples for GH assay were collected every 20 min on days 1, 6 and 10, from 1 hour before and 3 h after GHRP-2 or saline injections at 08:00 h. GH peak concentrations and GH area under curve (GH AUC) on day 1, 6 and 10 in treatment group of swine were higher than those in control swine (p<0.05). Twice daily administration of GHRP-2 caused a significantly attenuation (p<0.05) of GH peak concentrations ($80.25{\pm}13.87$, $39.73{\pm}5.72$ and $27.57{\pm}6.06ng/ml$ for day 1, 6 and 10, respectively) and GH AUCs ($3,536.15{\pm}738.35$, $1,310.31{\pm}203.55$ and $934.37{\pm}208.99ng/ml$ for day 1, 6 and 10, respectively). However, there was no significant difference in GH peak concentration and GH AUC between day 6 and 10. Plasma IGF-1 concentration levels were higher in treatment than control group of swine (p<0.05) after 3 days of the treatment, and the levels reached a plateau from day 3 to 10 of experiment. Growth performance did not alter by GHRP-2 administration, even though a numerical increase of body weight gain and feed efficiency was observed. These results indicate that twice daily administration of GHRP-2 for 10 days in swine did not significantly influence on growth performance, caused an overall attenuation of GH response, and that elevation of plasma GH concentrations caused by GHRP-2 administration increased plasma IGF-1 concentrations, even though an attenuation of GH response was observed.

• Fisheries and Aquatic Sciences
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• v.15 no.1
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• pp.43-47
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• 2012

Growth hormone (GH) transgenesis in fish has the potential to improve aquaculture efficiency and capacity. However, many fast-growing transgenic fish have experienced side effects caused by excess GH expression. To overcome this unwanted issue associated with several GH transgenic mud loach Misgurnus mizolepis lines carrying GH construct driven by a strong ${\beta}$-actin regulator ($pml{\beta}$-actGH), we performed an alternative version of GH autotransgenesis using a weaker but more stable regulator, the mud loach lectin promoter. GH transgenesis with a pmlectGH construct consisting of the mud loach GH gene driven by the 2.3-kb lectin promoter exhibited significant growth stimulation. However, the extent of the growth acceleration in pmlectGH transgenics (six times maximum when assessed 2 months post hatching) was much less than that in transgenic individuals carrying the $pml{\beta}$-actGH construct. Additionally, the extraordinary gigantism that was common in $pml{\beta}$-actGH-transgenic mud loaches was diminished in transgenic loaches harboring the pmlectGH construct. Transgenic founders (pmlectGH) successfully transmitted their transgene into the next generation with up to 41% frequency. Growth stimulation also persisted in the transgenic F1 strains, with a seven-fold increase in maximum body weight at 6 months of age.