• Title/Summary/Keyword: 단백질 가수분해 효소

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Expression of Membrane-Type Matrix Metalloproteinase 1 and 2 in Mouse Oocytes, Embryos, Ovary and Oviduct (생쥐 난자와 배아 및 난소와 수란관의 Membrane-Type Matrix Metalloproteinase 1 및 2의 유전자 발현)

  • 김지영;이희진;김소라;김해권;강성구;이승재;조동제
    • Development and Reproduction
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    • v.4 no.1
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    • pp.45-52
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    • 2000
  • Membrane type matrix metalloproteinases(MT-MMPs) have been suggested to play an important role during structural remodeling of various tissue. Expression patterns of MT1-MMP and MT2-MMP mRNAs were investigated in oocytes, embryos, ovary and oviduct of mouse during their differentiation or periovulatory period using RT-PCR technique. Both cDNA products of MT1- and MT2-MMP of immature oocytes were barely discernable with a minimum amount but the expressions were distinct in mature oocytes regardless that they were matured in vivo or in vitro. MT2-MMP was not expressed by 2-cell embryos but was expressed by 4-cell stage embryos. From the morula stage untill hatched blastocyst stage, embyos showed intesnse expression of MT2-MMP with a sudden increase at blastocyst stage. While mouse ovarian tissues showed both expression of MT1- and MT2-MMP, there was no stage-specific difference throughout the estrous cycle. Mouse oviducts also exhibit constant amount of both MT1- and MT2-MMP expressions throughout periovulatory period, i.e., before or after ovulation. These observations lead to suggest that the differential expressions of maternal MT1- and MT2-MMP during meiotic resumption of mouse oocytes and embryonic expression of MT2-MMP particularly at blastocyst stage might play a role in the differentiation of mouse oocytes and/or embryos. The precise function of MT1- and MT2-MMP with regards to their participation in the remodeling of ovarian and oviductal tissues remains in a question.

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Draft genome sequence of Senegalimassilia sp. KGMB 04484 isolated from healthy Korean human feces (건강한 한국인 분변으로부터 분리된 Senegalimassilia sp. KGMB 04484 균주의 유전체 염기서열 초안)

  • Han, Kook-Il;Kang, Se Won;Kim, Ji-Sun;Lee, Keun Chul;Eom, Mi Kyung;Suh, Min Kuk;Kim, Han Sol;Park, Seung-Hwan;Lee, Ju Huck;Park, Jam-Eon;Oh, Byeong Seob;Yu, Seung Yeob;Choi, Seung-Hyeon;Lee, Dong Ho;Yoon, Hyuk;Kim, Byung-Yong;Lee, Je Hee;Lee, Jung-Sook
    • Korean Journal of Microbiology
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    • v.55 no.2
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    • pp.160-163
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    • 2019
  • Senegalimassilia sp. KGMB 04484 was isolated from fecal samples obtained from a healthy Korean. The whole-genome sequence of Senegalimassilia sp. KGMB 04484 was analyzed using the PacBio Sequel platform. The genome comprises a 2,748,041 bp chromosome with a G+C content of 61.18%, 2,300 total genes, 2,139 protein-coding gene, 21 rRNA genes, and 51 tRNA genes. Also, we found that strain KGMB 04484 had some genes for hydrolysis enzyme, fatty acid biosynthesis and metabolism in its genome based on the result of genome analysis. Those genes of KGMB 04484 may be related to regulation of human health and digest.

Development and Research on a Functional Hydrolyzed Whey Protein Powder Product with Sialic Acid as a Marker Compound - II. Repeated 90-day Oral Administration Toxicity Test using Rats Administered Whey Protein Powder containing Highly Concentrated Sialic Acid (23%) produced by Enzyme Separation and Solvent Enrichment Method - (Sialic Acid를 지표성분으로 하는 유청가수분해단백분말의 기능성식품 개발연구 - II. 효소분리 용매정제로 고농도 Sialic Acid가 함유된 유청가수분해단백분말(23%)의 랫드를 이용한 90일 반복경구투여 독성시험 평가 연구 -)

  • Noh, Hye-Ji;Cho, Hyang-Hyun;Kim, Hee-Kyong;Koh, Hong-Bum
    • Journal of Dairy Science and Biotechnology
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    • v.34 no.2
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    • pp.117-135
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    • 2016
  • The present study was performed to develop a functional raw food material from hydrolyzed whey protein powder (23%-GNANA) medication containing sialic acid as a marker compound that is naturally occurring at 7% concentration in GMP (glycomacropeptide). GMP is used worldwide in foodstuffs for babies and infants and is obtained from the milk protein as safe food. While the purpose of our detailed evaluation was aimed to assess preliminary NOAEL values for and above 2,000 mg/kg/day, a clinical dose allowance for 23%-GNANA (as per characteristic of a functional health product, a highly refined test substance of 23% (v/v) sialic acid combined in GMP), at the same time we also wanted to assess the safety of GMP hydrolyzate lacking sialic acid but with identical properties as GMP. Animal safety evaluation was conducted using 23%-GNANA as the test substance, produced from hydrolyzed whey protein powder (product name: HELICOBACTROL-23; provided by Medinutrol Inc. [Korea]; composed of 23% sialic acid and GMP protein) after isolating the sialic acid using enzymes approved as food additives, with GMP as a raw material, and subsequently increasing the content of xx up to 23% through 80% (v/v) ethanol soaking and concentrating, in accordance with GLP Guideline. The animal safety evaluation mentioned above was made on the basis of toxicity in SPF Sprague-Dawley female and male rats dosed with 10 mL of the test substance diluted to 0, 1,250, 2,500, and 5,000 mg/kg directly into their stomachs for 90 d. This was determined in terms of the general symptoms and animal viability, weight and amount of feed intake, eye examination, uracrasia tests, hematological and blood biochemical disorder tests, blood coagulation test, abnormal intestine weight, abnormalities during postmortem and histopathological examinations. Statistical significance was set at P<0.05. Based on the toxicity determination, a certain minor effect associated with the test substance was observed in male rats with no major effects of the tested substance, in comparison with the control group dosed with sterilized water. Nevertheless, the NOAEL value, evaluated as per toxicity criteria, was verified as 5,000 mg/kg/day (P<0.05). Similarly, for female rats, a certain minor effect associated with the test substance was observed in 5,000 mg/kg/day dosed group, with no major effect, yet the NOAEL value (as assessed as per toxicity criteria) was determined to be 5,000 mg/kg/day (P<0.05), which was the same as for male rats. Accordingly, the NOAEL values of the test substances for all female and male rats were finally verified as 5,000 mg/kg/day (P<0.05). In conclusion, it was determined that the 23%-GNANA test substance exceeds 2,000 mg/kg/day, the clinical allowance characteristic for functional health food, and was finally evaluated to cause no safety concerns when used as a raw material in functional health food production, which was the ultimate goal of the present study.

Biochemical Characterization of Phospholipase C$\delta$from liver of Mud loach (Misgurnus mizolepis) (미꾸라지 간으로부터 포스포리파아제 C델타 단백질의 생화학적 특성)

  • Seo, Jung-Soo;Lim, Sang-Uk;Kim, Na-Young;Lee, Sang-Hwan;Oh, Hyun-Suk;Lee, Hyung-Ho;Chung, Joon-Ki
    • Journal of fish pathology
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    • v.18 no.1
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    • pp.67-80
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    • 2005
  • Phosphoinositide-specific phospholipase $C\delta$ $PLC\delta$) plays an important role in many cellular responses and is involved in the production of second messenger. The present study was conducted to obtain the biochemical characteristics of the expressed recombinant $PLC\delta$ in E. coli cloned from Misgurnus mizolepis and partially purified $PLC\delta$ enzymes from liver tissues of M. mizolepis (wild ML-$PLC\delta$). The ML $PLC\delta$ gene was cloned and expressed under the previous report (Kim et al., 2004), and purified the recombinant protein by successive chromatography using $Ni^{2+}$-NTA affinity column and gel iltration FPLC column. The wild ML-$PLC\delta$ protein was solublized with 2 M KCI and purified by successive chromatography on open heparin-Sephagel and analytical TSKgel heparin-5PW. Both the recombinant and wild ML-$PLC\delta$ form of protein showed a concentration-dependent PLC activity to phosphatidylinositol 4,5-bis-phosphate (PIP$_2$) or phosphatidylinositol (PI). Its activity was absolutely $Ca^{2+}$- dependant, which was similar to mammalian $PLC\delta$ isozymes. Maximal PI-hydrolytic activations of recombinant and wild ML- TEX>$PLC\delta$ was at pH 7.0 and pH 7.5, respectively. In addition, the enzymatic activities of recombinant and wild ML-$PLC\delta$ were increased in concentration-dependent manner by detergent, such as sodium deoxycholate SDC), phosphatidylethanolamine (PE) and phosphatidylcholine (PC). The activities decreased in contrast by a polyamine, such as spermine. Western blotting showed that several types of $PLC\delta$ isozymes exist in various organs. Taken together our results, it suggested that the biochemical characteristics of ML-$PLC\delta$ are similar with those of mammalian $PLC\delta1$ and ${\delta}3$ isozymes.

Future Development of Genetics and the Broiler (BROILER 육종기술의 전망)

  • 오봉국
    • Korean Journal of Poultry Science
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    • v.8 no.1
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    • pp.1-5
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    • 1981
  • In trying to predict the effect of genetics on the broiler in the year 2000, this is a relatively short period of time as far as broiler genetics in concerned. Modern broiler genetics started around 1945 and tremendous gains when made in past 35 years. Futher improvements on broiler will depend on the evolution and revolution: 1. Evolution: (1) Growth rate has been made 4-5% per year. (2) Feed conversion has improved approximately 1% per year. (3) Abdominal fat is becoming a major complaint in broiler. (4) Because of the changing life-style, broiler meat sales in the future will be more and more in cut-up form. (5) Breeding for stress resistance and selection for docile temperament can be important in order to funker improve fled efficiency. (6) In female parent stock, reproduction characteristics are in many can negatively correlated with the desired broiler traits. (7) Egg production and hatchability in moot commercial parent nod m at a fairly high level. (8) In male parent stock, the heavier and mon super-meat-type male lines are desired to Product better broilers. 2. Revolution: Trying to forecast revolutionary change in broiler genetics is highly speculative, as sudden change are aften unpredictable. (1) Species hybridization, such as a turkey-chicken cross (2) Biochemical tools, such as blood typing. (3) Mutation breeding by radiation or chemical mutagentia. (4) Broiler breeding would be to change the phenotypic appearance by single gene, such as naked, wingless. (5) Changes in production techniques. such as growing in cage or growing in filtered air positive pressure houses.

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Production of Organic Rice (Oryza sativa L.) using Organic Cultivation Manual (유기재배매뉴얼을 활용한 유기쌀 생산)

  • Cha, Kwang-Hong;Oh, Hwan-Jung;Seo, Dong-Jun;Song, Yong-Su;Ahn, Joon-Seob;An, Kyu-Nam;Jung, Woo-Jin
    • Korean Journal of Organic Agriculture
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    • v.22 no.1
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    • pp.97-113
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    • 2014
  • An actual proof experiment was carried out to establish the production of high quality organic rice at farm household in Gwangjin, Junnam, Korea 2011. Experimental plot was designed with conventional organic cultivation and manual organic cultivation in both Hopeongbyeo and Onnuri varities. Experimental field was consisted with suitable drainage loam soil, anryong series, and normal paddy field. Chemical characteristics of the soil were investigated from initial soil and soil after harvest in experimental sites. Seed disinfection was treated with hot water dipping method at $60^{\circ}C$ for 10 min. Rice transplanting was carried out at 17 June with 25 days seedling using machine transplanting. Golden apple snail was used to prevent weed growth as middle size (1.5~1.7 kg/10a) at one week after rice transplanting. Amount of applied fertilizer in experimental plot was supplied with organic fertilizer for the shortage after harvest of hairy vetch (1,100 kg/10a). Bordeaux mixture was used to control of sheath blight and neck blast disease in all experimental plot at 19 Aug. Application of chitin culture solution was treated in manual organic cultivation at 2 and 15 Aug. At late growth stage, incidence rate of neck blast was the highest level at Hopyeongbyeo in conventional organic cultivation. Incidence rate of sheath blight in both Hopyeongbyeo and Onnuri was higher in conventional organic cultivation than in manual organic cultivation. Incidence of rice leaf folder was the highest level at Onnuri in conventional organic cultivation. Yield index in manual organic cultivation was increased by 13% and 38% at Onnuri and Hopyeongbyeo, respectively, compared with conventional organic cultivation. Head rice percentage in both Hopyeongbyeo and Onnuri was slightly increased in manual organic cultivation. These results indicate that high quality production of rice organic cultivation could be depending on management of safe fertilization with nutrient content in soil.

Studies on the Mutation of Aspergillus niger (흑국균(黑麴菌)의 인공변이(人工變異)에 관(關)한 연구(硏究))

  • Park, Yoon-Joong;Sohn, Cheon-Bae
    • Korean Journal of Food Science and Technology
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    • v.14 no.1
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    • pp.72-79
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    • 1982
  • Several mutants were isolated from the parent strain of Aspergillus niger CF: the first mutant strain CF-11 was obtained by UV irradiation, and the second mutant strain CF-21 and CF-22 were from NTG (N-methyl-N'-nitroso-N-nitroso-guanidine) treatment on the CF-11. These mutants were characterized, and their enzyme and acid production on wheat bran Koji and wheat flour Koji were studied. Asp. niger CF-22 mutant appeared to be tan type which conidial heads were discolored. It's glucoamylase activity was inreased approximately two times and its ${\alpha}-amylase$ about 50 percent as compared with that of the parent strain of Asp. niger CF, when grown on wheat bran Koji under the optimal conditions. Asp. niger CF-21 mutant showed slower growth rate and poor sporulation than the wild type, although its conidial heads were not discolored. Approximately 4-fold increment in its acid production was observed as compared with the weld type. The activities of glucoamylase and ${\alpha}-amylase$ of the Asp. niger CF-22 and CF-21 mutants were higher than those of the wild type, but their protease activity was rather lower. The maximum production of glucoamylase by the Asp. niger CF-22 mutant was obtained after 2 to 3 days incubation on wheat bran at 30 to $35^{\circ}C;$ ${\alpha}-amylase$2 days incubation at 30 to $35^{\circ}C$. The maximal levels of acid production by the mutant CF-21 was appeared after 2 days incubation on wheat bran Koji, and after 3 days on wheat flour Koji at $30^{\circ}C$. Little differences in the levels of acid production were observed between on wheat bran and flour Koji.

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Food Component Characteristics of Seafood Cooking Drips (수산 자숙액의 식품성분 특성)

  • Oh, Hyeun-Seok;Kang, Kyung-Tae;Kim, Hye-Suk;Lee, Jae-Hyoung;Jee, Seung-Joon;Ha, Jin-Hwan;Kim, Jin-Soo;Heu, Min-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.5
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    • pp.595-602
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    • 2007
  • This study was conducted to investigate on food component characteristics of seafood cooking drips (skipjack tuna cooking drip, octopus cooking drip and oyster cooking drip) as a source of functional seasoning. Heavy metal contents of seafood cooking drips were below food safety level. Among seafood cooking drips concentrated to 5 folds, the crude protein content was the highest (18.1%) in skipjack tuna cooking drip (SCD). The free amino acid content and taste value were higher in SCD than in other seafood cooking drips, and the major free amino acids were glutamic acid and aspartic acid. Total amino acid content of SCD was 16.2 g/100 mL and the major amino acids were glutamic acid (11.9%), proline (9.2%), glycine (9.1%) and histidine (11.5%). SCD in comparison with other seafood cooking drips showed the highest angiotensin I converting enzyme (ACE) inhibitory activity at $IC_{50}$ of 14.1 mg/mL. These results suggested that SCD could be used as a source of functional seasoning.

Development and Research into Functional Foods from Hydrolyzed Whey Protein Powder with Sialic Acid as Its Index Component - I. Repeated 90-day Oral Administration Toxicity Test using Rats Administered Hydrolyzed Whey Protein Powder containing Normal Concentration of Sialic Acid (7%) with Enzyme Separation Method - (Sialic Acid를 지표성분으로 하는 유청가수분해단백분말의 기능성식품 개발연구 - I. 효소분리로 7% Siailc Acid가 표준적으로 함유된 유청가수분해단백분말(7%)의 랫드를 이용한 90일 반복경구투여 독성시험 평가 연구 -)

  • Noh, Hye-Ji;Cho, Hyang-Hyun;Kim, Hee-Kyong
    • Journal of Dairy Science and Biotechnology
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    • v.34 no.2
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    • pp.99-116
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    • 2016
  • We herein performed animal safety assessment in accordance with Good Laboratory Practice (GLP) regulations with the aim of developing sialic acid from glycomacropeptide (hereafter referred to as "GMP") as an index ingredient and functional component in functional foods. GMP is a type of whey protein derived from milk and a safe food, with multiple functions, such as antiviral activity. A test substance was produced containing 7% (w/w) sialic acid and mostly-hydrolyzed whey protein (hereafter referred to as "7%-GNANA") by enzymatic treatment of substrate GMP. The maximum intake test dose level was selected based on 5,000 mg/kg/day dose set for male NOEL (no-observed-effect-level) and female NOAEL (no-observed-adverse-effect-level) determined by a dose-range finding (DRF) test (GLP Center of Catholic University of Daegu, Report No. 15-NREO-001) that was previously conducted with the same test substance. To evaluate the toxicity of a repeated oral dose of the test substance in connection with the previous DRF study, 1,250, 2,500, and 5,000 mg/kg of the substance were administered by a probe into the stomachs of 6-week-old SPF Sprague-Dawley male and female rats for 90 d. Each test group consisted of 10 male and 10 female rats. To determine the toxicity index, all parameters, such as observation of common signs; measurements of body weight and food consumption; ophthalmic examination; urinalysis, electrolyte, hematological, and serum biochemical examination; measurement of organ weights during autopsy; and visual and histopathological examinations were conducted according to GLP standards. After evaluating the results based on the test toxicity assessment criteria, it was determined that NOAEL of the test substance, 7%-GNANA, was 5,000 mg/kg/day, for both male and female rats. No animal death was noted in any of the test groups, including the control group, during the study period, and there was no significant difference associated with test substance, as compared with the control group, with respect to general symptoms, body weight changes, food consumption, ophthalmic examination, urinalysis, hematological and serum biochemical examination, and electrolyte and blood coagulation tests during the administration period (P<0.05). As assessed by the effects of the test substance on organ weights, food consumption, autopsy, and histopathological safety, change in kidney weight as an indicator of male NOAEL revealed up to 20% kidney weight increase in the high-dose group (5,000 mg/kg/day) compared with the change in the control group. However, it was concluded that this effect of the test substance was minor. In the case of female rats, reduction of food consumption, increase of kidney weight, and decrease of thymus weight were observed in the high-dose group. The kidney weight increased by 10.2% (left) and 8.9% (right) in the high-dose group, with a slight dose-dependency compared with that of the control group. It was observed that the thymus weight decreased by 25.3% in the high-dose group, but it was a minor test substance-associated effect. During the autopsy, botryoid tumor was detected on the ribs of one subject in the high-dose group, but we concluded that the tumor has been caused by a naturally occurring (non-test) substance. Histopathological examination revealed lesions on the kidney, liver, spleen, and other organs in the low-dose test group. Since these lesions were considered a separate phenomenon, or naturally occurring and associated with aging, it was checked whether any target organ showed clear symptoms caused by the test substance. In conclusion, different concentrations of the test substance were fed to rats and, consequently, it was verified that only a minor effect was associated with the test substance in the high-dose (5,000 mg/kg/day) group of both male and female rats, without any other significant effects associated with the test substance. Therefore, it was concluded that NOAEL of 7%-GNANA (product name: Helicobactrol) with male and female rats as test animals was 5,000 mg/kg/day, and it thus was determined that the substance is safe for the ultimate use as an ingredient of health functional foods.

Draft genome sequence of Ruminococcus sp. KGMB03662 isolated from healthy Korean human feces (건강한 한국인 분변으로부터 분리된 Ruminococcus sp. KGMB03662 균주의 유전체 염기서열 초안)

  • Han, Kook-Il;Kang, Se Won;Eom, Mi Kyung;Kim, Ji-Sun;Lee, Keun Chul;Suh, Min Kuk;Kim, Han Sol;Park, Seung-Hwan;Lee, Ju Huck;Park, Jam-Eon;Oh, Byeong Seob;Ryu, Seoung Woo;Yu, Seung Yeob;Choi, Seung-Hyeon;Lee, Dong Ho;Yoon, Hyuk;Kim, Byung-Yong;Lee, Je Hee;Lee, Jung-Sook
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.274-277
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    • 2019
  • Ruminococcus sp. KGMB03662 was isolated from fecal samples obtained from a healthy Korean. The whole-genome sequence of Ruminococcus sp. KGMB03662 was analyzed using the PacBio Sequel platform. The genome comprises a 2,707,502 bp chromosome with a G + C content of 43.09%, 2,484 total genes, 2,367 protein-coding gene, 14 rRNA genes, and 53 tRNA genes. In the draft genome, genes involved in the hydrolysis enzyme, fatty acid biosynthesis, fatty acid metabolite, antibiotic biosynthesis, and antibiotic resistance have been identified. Those genes of KGMB03662 may be related to the regulation of human health and disease.