• Title/Summary/Keyword: Eptatretus burgeri

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Cloning, Expression Analysis and Enzymatic Characterization of Cathepsin L from the Inshore Hagfish (Eptatretus burgeri) (먹장어 Cathepsin L의 분자생물학적 클로닝, 발현 및 효소학적 특성 분석)

  • JANG, Jin-Hyeon;SON, So-Hee;JO, Hyeon-Kyeong;CHUNG, Joon-Ki;LEE, Hyung-Ho
    • Journal of Fisheries and Marine Sciences Education
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    • v.28 no.4
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    • pp.903-912
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    • 2016
  • Hagfish which belongs to the chordate contact cyclostomata, is important phylogenetic relationship between vertebrate and invertebrate. Cathepsins of the cysteine protease family have traditionally been thought to play a major role in intracellular protein degradation and turnover in lysosomes. In this study, Catepsin L was cloned from Inshore hagfish (Eptatretus burgeri), the cDNA encoding ORF of the Eptatretus burgeri Cathepsin L (EbCtL) is 978 bp. The cDNA encoding proEbCtL was expressed in Escherichia coli strain BL21(DE3) using the pGEX-4T-1 expression vector system. The recombinant proEbCtL protein was overexpressed as a approximately 55 kDa fusion protein. The overproduced soluble GST-fusion protein was then applied to glutathione-Sepharose 4B column chromatography; the sample harboring the fusion protein evidenced a high degree of purity when analyzed via SDS-PAGE and Western blot analysis. Its activity was quantied by cleaving the synthetic peptide Z-FR-AMC, Z-LLE-AMC, and Suc-AAF-AMC, and the optimal pH for the protease activity was 8, 9.5, and 9, respectively.

Reproduction characteristics of hagfish Eptatretus burgeri in the South Sea of Korea (한국 남해에 출현하는 먹장어 Eptatretus burgeri의 재생산 특성)

  • KIM, Doo-Nam;HWANG, Kang-Seok;CHA, Hyung-Kee;PARK, Jun-Su;KIM, Jung-Nyun;MOON, Seong-Yong;LEE, Jeong-Hoon
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.54 no.2
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    • pp.188-192
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    • 2018
  • The reproduction characteristics of hagfish Eptatretus burgeri were examined using individuals caught in the South Sea of Korea. The spawning season and size at minimum sexual maturity of this species were characterized based on a gonad-somatic index (GSI) and monthly variation egg size (long axis). From monthly variation of GSI, the spawning season was estimated to be from August to September. Developing eggs larger than 10 mm were found in March, and the largest egg size was found in July. The first spawning length was 34.2 cm TL. Batch fecundity ranged from 13 to 117 eggs for hagfish sized from 34.2 cm TL to 77.0 cm TL, respectively, and increased linearly with total length.

Purification of a Bradykinin-Related Peptide from the Skin of Hagfish. Eptatretus burgeri (먹장어 (Eptatretus burgeri)의 피부로부터 Bradykinin-Related Peptide의 정제)

  • SHIN Mi Jung;KIM Eun Jung;KIM Chan-Hee;GO Hye-Jin;KIM In Hae;RYU Hong-Soo;Huh Min-Do;CHUNG Joon-Ki;PARK Nam Gyu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.36 no.1
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    • pp.30-34
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    • 2003
  • A hagfish bradykinin(BK)-related peptide was isolated and characterized from the skin of hagfish, Eptatretus burgeri. The hagfish BK with a molecular mass of 875.8 Da was purified to a homogeneity using $C_{18}$ reverse-phase and cation-exchange high performance liquid chromatography. The primary structure of the hagfish BK was determined as Gly-Thr-Ala-Gly-Ile-Gly-Pro-Phe-Arg by a combination of an automated amino acid sequencing and MALDI-TOF mass spectrometry. This amino acid sequence contains five substitutions $(Arg^1{\rightarrow}Gly,\;Pro^2{\rightarrow}Thr,\;Pro^3{\rightarrow}Ala,\;Phe^5{\rightarrow}Ile,\;Ser^6{\rightarrow}Gly)$ compared with that of mammalian BK. The hagfish BK showed a contractile action on the intestine of hagfish, Eptatretus burgeri. The threshold concentration of hagfish BK was around $10^{-11}\;M.$

Purification and Characterization of Novel Antimicrobial Peptide from the Skin of the Hagfish , Eptatretus burgeri

  • Hwang, Eun-Young;Seo, Jung-Kil;Kim, Chan-Hee;Go, Hye-Jin;Kim, Eun-jung;Chung, Joon-Ki;Rye, Hong-Soo;Park, Nam-Gyu
    • Preventive Nutrition and Food Science
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    • v.4 no.1
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    • pp.28-32
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    • 1999
  • A novel antimicrbial peptide , named HFS-I, was isolated and characterized from the skin of the hagfish, Eptatretus bugeri. The decapeptide with a molecular mass of 1279.5 Da was purified to homogeneity using a gel-filtration column, ion-exchange and C18 reverse-phase high performance liquid chromatograpy . The complete amino acid sequence of HFS-I, which was determined by a combination of an automated amino acid sequencing and FAB-MS, was F-P-W-W-L-S-G-K-Y-P-NH2. Comparison of the amino acid sequence with those of other known antimicrobial peptides revealed that HFS-I was a novel antimicrobial peptide. HFS-I showed a weak antimicrobial activity in vitro aganinst a broad spectrum of microorganism without hemolytic acitivity.

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Effects of Water Temperature and Salinity on Blood Properties and Oxygen Consumption in Hagfish (Eptatretus burgeri) (먹장어(Eptatretus burgeri)의 혈액성상과 산소소비에 미치는 수온 및 염분의 영향)

  • Do, Yong-Hyun;Min, Byung-Hwa;Myeong, Jeong-In;Jee, Young-Ju;Chang, Young-Jin
    • Journal of Fisheries and Marine Sciences Education
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    • v.26 no.1
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    • pp.214-222
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    • 2014
  • Hagfish Eptatretus burgeri is classified as a agnathans and has many different physiological properties compared with vertebrates. In this study, we examined effects of water temperature and salinity on blood properties and oxygen consumption in hagfish. In the experiment of water temperature change, hematocrit (Ht), red blood cell (RBC) and glucose of hagfish blood revealed the lowest values at $15^{\circ}C$. Oxygen consumption of hagfish had significantly increased with rising water temperature, and the increasing rate was twice as much when the temperature was manipulated every $5^{\circ}C$. Also, oxygen consumption during the night time (a short photoperiod) was significantly higher than that of the daytime. Q10 level was 3.50 in the light period and 3.92 in the dark period. No significant change in plasma glucose level was showned in changing salinity from 30 psu to 22 psu, while it had rapidly increased at 20 psu ($13.7{\pm}4.0mg/dL$) and thereafter all hagfish were dead at 18 psu. However, osmolarity, $Na^+$, $K^+$ and $Cl^-$ levels had significantly decreased when salinity decreased. This results are expected to develop the artificial rearing techniques of natural hagfish.

Seasonal and Interannual Variation in Species Composition of Fish Assemblages Collected by Pots in the Southern Coast of East Sea, Korea (동해 남부 연안해역 통발에서 채집된 어류 군집의 계절변동과 연간변동)

  • Park, Joo Myun;Huh, Sung-Hoi
    • Korean Journal of Ichthyology
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    • v.27 no.4
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    • pp.310-316
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    • 2015
  • The seasonal and interannual variation in the species composition and abundance of the fish assemblage were studied. Fish samples were collected seasonally over 4 year (2005 to 2008) using pots in the southern coast of East Sea. During the study period, a total of 32 fish species belonging to 24 families were collected. The dominant species were Conger myriaster, Paracentropogon rubripinnis, Liparis tanakae, Parapercis sexfasciata, Eptatretus burgeri and Clupea pallasii pallasii, which accounted for 91.9% of the total number of individuals collected. The number of species, number of individuals, biomass, and diversity fluctuated with season. The peak number of species and number of individuals occurred were in August 2008, whereas the biomass was the highest in August 2005 and diversity was in May 2008. The abundance of dominant species also showed some seasonal and interannual changes. A one-way analysis of similarity (ANOSIM) showed that the fish assemblage in 2008 was significantly different from those of 2005, 2006 and 2007, whereas there was no significant difference among seasons over 4 years.

Purification of Neuropeptide with the Contractile Activity on the Smooth Muscle from the Skin of Conger Eel Conger myriaster (붕장어(Conger myriaster)의 피부로부터 평활근 수축작용을 지닌 신경성 펩타이드의 정제)

  • Go, Hye-Jin;Park, Nam-Gyu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.45 no.4
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    • pp.358-366
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    • 2012
  • A novel neuropeptide was isolated from the skin of the conger eel Conger myriaster using hagfish Eptatretus burgeri intestine as a bioassay system. The sequence of the purified peptide was analyzed using automated amino acid sequencing and MALDI-TOF mass spectrophotometry. The molecular ion peak in the MALDI-TOF mass spectrum of the peptide was at m/z 962.89 $(M+H)^+$. The sequence of the peptide was determined to be L-P-M-L-E-T-Q-M, and was tentatively named comyrin. To investigate the complete primary structure of comyrin, comyrin-OH and comyrin-$NH_2$ were synthesized and the chemical and pharmacological properties of the synthetic peptides were compared with those of the native peptide. However, the elution time of synthetic peptides did not match that of the native peptide on the reverse-phase HPLC chromatogram. In addition, the synthetic peptides did not cause contractile activity in the intestinal smooth muscle of the hagfish. Based on these results, one possible reason for this disagreement may be the presence of a D-amino acid in comyrin.

Alkaline Pretense Activity of the Tissue Extracts from Some Different Kinds of Fish (어류의 조직중에 분포하는 알카리성 단백질분해효소의 활성조건)

  • NAM Taek Jeung;PYEUN Jae Hyeung
    • 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.

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Protein Quality Evaluation of Cooked Hagfish (Eptatretus burgeri) Meats

  • Hwang, Eun-Young;Lee, Jin-Hwa;Ryu, Hong-Soo;Park, Nam-Gyu;Chun, Soon-Sil
    • Preventive Nutrition and Food Science
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    • v.7 no.3
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    • pp.287-292
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    • 2002
  • The effect of cooking methods on in vivo and in vivo indices of the protein quality of hagfish meat were investigated. In vivo protein digestibilities of cooked meats (81.3~83.5 %) were not significant different (p<0.05) from those of van meat (82.9%), with the exception of steamed (11$0^{\circ}C$, 15 min) meat (86.3 %). Convection oven cooking (22$0^{\circ}C$, IS min) resulted in a higher trypsin indigestible substrate (TIS, 49.2 mg/g solid) compared with that of raw meat (38.9 mg/g solid). free amino acid content of raw meat was decreased after boiling (10$0^{\circ}C$, 10min). Both convection oven and microwave cooking (2,450 MHz, 3 min) decreased available lysine from 4.9g/16g N to 3.8~4.1g/16g N. In vivo apparent protein digestibilites (AD) of hagfish meat were similar fur raw (92.4%) and cooked meats, but were somewhat lower than ANRC (Animal Nutrition Research Council) casein (945%). The PERs (3.7~4.1) and NPRs (3.7~4.9) of cooked meats were significantly higher (p<0.05) than those of raw meat (PER 3.3, NPR 3.6 and ANRC casein (PER 2.5, NPR 2.6), despite their lower in vivo protein digestibilities. These results demonstrate that cooking at optimal conditions resulted in remarkably positive effects on in vivo and in vivo protein qualities of hagfish meats. Therefore, steamed hagfish meat is an excellent source of high quality protein from seafood products.