• Title/Summary/Keyword: peptide mass

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Molecular Cloning and Characterization of Chitosanase Gene from Bacillus amyloliquefaciene MJ-1 (Bacillus amyloliquefaciens MJ-1 유래의 chitosanase 유전자의 클로닝 및 특성)

  • Park Chan-Soo;Oh Hae-Geun;Hong Soon-Kwang;Park Byung-Chul;Hyun Young;Kang Dae-Kyung
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.142-148
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    • 2006
  • In order to develop chitosanase for the production of chitosan oligosaccharides, a chitosanase-producing bacterium was isolated from the traditional fermented soybean, Meju, and identified as Bacillus amyloliquefaciene MJ-1. The cloned chitosanase gene, 825 bp in size, encoded a single peptide of 274 amino acids with a estimated molecular mass of 30.9 kDa. The deduced amino acid sequence showed significant homology with microbial chitosanases. The recombinant chitosanase was expressed in Escherichia coli upon induction with isopropyl-D-thiogalactopyranoside, and purified using $Ni^{2+}-NTA$ agarose column chromatography. The maximal activity of the recombinant chitosanase is at pH 5.0 and $60^{\circ}C$. The recombinant chitosanase is stable between pH 5.0 and pH 7.0 at $37^{\circ}C$ for 30 min, and more than 75% of the activity still remain at $80^{\circ}C$ for 30 min incubation.

Proteomic Analyses of Chinese Cabbage(Brassica campestris L. pekinensis) Affected by High Temperature Stresses in Highland Cultivation During Summer in Korea (Proteomics를 이용한 고랭지 배추의 고온장해 해석)

  • Shin, Pyung-Gyun;Hong, Sung-Chang;Chang, An-Cheol;Kim, Sang-Hyo;Lee, Ki-Sang
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1649-1653
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    • 2007
  • High temperature stresses have caused growth inhibition and delayed heading in highland cultivation Chinese cabbage during summer in Korea. We have studied high temperature stress responses in the terms of changes of inorganic components and proteins by proteomic analyses. Insufficiencies of nitrogen and phosphorus have affected growth rate and calcium deficiency has caused blunted heading. Proteins extracted from Brassica seedling grown at the altitude of 600m and 900m in the Mount Jilun were extracted and analysed by 2-dimentional polyacrylamide gel electrophoresis. Profiles of protein expression was then analyzed by 2-dimentional gel analyses. Protein spots showing different expression level were picked using the spot handling workstation and subjected to MALDI-TOF MS. Total 48 protein spots were analyzed by MALDI-TOF MS and 30 proteins spots out of 48 were identified by peptide mass fingerprinting analyses. Fourteen proteins were up-regulated in extracts from the altitude of 900m and they were identified as oxygen-evolving proteins, rubisco activase and ATPase etc. Sixteen proteins were up-regulated in extracts from the altitude of 600m and they were identified as glutathione S-transferase(1, 28kD cold induced- and 24 kD auxin-binding proteins) and salt-stress induced protein etc. These stress-induced proteins were related to the mediated protective mechanism against oxidative damage during various stresses. The results indicated that physiological phenomenon in response to high temperature stresses might be resulted by complex and multiple array of responses with drought, heat, oxidative, salt, and cold by high temperature.

Identification of Brassinosteroid-Related Protein, BAK1 from Nutrition Deficient Tomato Cultivated by Soilless Cultivation System (수경재배 영양결핍토마토에서 브레시노스테로이드관련 신호전달 단백질 BAK1의 동정)

  • Shin, Pyung-Gyun;Chang, An-Cheol;Hong, Sung-Chang;Lee, Ki-Sang
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1729-1733
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    • 2007
  • Brassinolide insensitive associated receptor kinase 1(BAK1) is a critical component that play an important roles in signaling of brassinosteroid biosynthesis. Brassinosteroid-deficient and -insensitive mutants showed the characteristic of dwarf symptom. The nutrient deficient tomato showing stunt phenomenon was selected from soiless cultivation system using modified Sonneveld hydroponic solution. Twenty eight protein spots showing different expression levels compared to the control were isolated from extracts of stunted tomato leaves by 2D PAGE analyses. Significantly down-regulated 6 protein spots out of 28 protein spots were analyzed and sequenced by MALDI-TOF mass spectrometry. The protein spot having pI=4.5 and MW=24 kDa was identified as a signal protein, BAK1, which is directly related to brassinosteroid biosynthesis. In addition, five other protein spots were identified as BCK1, cystein proteinase, sulfutase, peroxidase and zinc finger factor respectively, and they were also signal proteins related to brassinosteroid biosynthesis. Furthermore, amplification of 500bp of BAK1 mRNA by RT-PCR using a primer set of peptide matched regions was inhibited conpared to that of the wild type. The results sugested that the BAK1 might be regulated at the transcription level in response to nutrition applications.

Analysis of Pine Nut Oil Composition and Its Effects on Obesity (잣기름 성분분석 및 비만 예방효과 연구)

  • Kim, Kyoung Kon;Kang, Yun Hwan;Kim, Dae Jung;Kim, Tae Woo;Choe, Myeon
    • Korean Journal of Food Science and Technology
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    • v.46 no.5
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    • pp.630-635
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    • 2014
  • Pine nut oil (PNO) is well known to impart beneficial effects in overweight individuals, but the mechanisms underlying PNO-mediated weight loss remain unclear. To investigate how PNO promotes weight loss, its composition was determined by gas chromatography coupled with mass spectrometry (GC-MS). In addition, the effects of PNO on cytotoxicity, lipid accumulation, expression of lipid metabolism-related biomarkers, and leptin secretion were assessed in 3T3-L1 cells. GC-MS analyses revealed that PNO contains several components, including linoleic acid, oleic acid, palmitic acid, and stearic acid. Moreover, PNO did not have a cytotoxic effect on 3T3-L1 cells. However, it inhibited the expression of peroxisome proliferator-activated receptor (PPAR) and adipocyte protein 2 (aP2). Finally, PNO significantly increased leptin secretion in a dose-dependent manner. Taken together, these results support the notion that PNO is useful for weight management in overweight individuals.

Effects of Green Tea, Cocoon Hydrolysates and Capsulated Components Mixture on Body Fat and Serum Lipid Composition in College Women in Terms of Supplemented Periods (녹차와 실크 펩타이드 및 캡슐 소재 함유 조성물이 여대생들의 복용 기간별 체지방 및 혈청 지질 성상 변화에 미치는 영향)

  • Lee, Sung-Hee;Cho, Byung-Nam;Hong, I-Jin;Lee, Min-Sook;Jew, Sang-Sup;Kim, Dong-Myung;Jin, Dong-Kyu;Rho, Sook-Nyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.5
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    • pp.715-722
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    • 2003
  • This study is to investigate effects of green tea and capsulated components mixture on body fat and serum lipid composition and fat distribution in college women students in terms of supplemented periods. During 3 months of this research, 34 college women students (average age 20.3 yr) were selected as subjects. Nutrient intake was investigated by questionnaire and 24-hr recall method. Anthropolatric assessments of the subjects were investigated by SBIA method (Segmental Bioimpedance Assay, Inbody 3.0). The results were as follows: mean body height was 162.5cm and mean body weight and mean BMI (Body Mass Index, kg/$m^2$) were 57.9kg and 21.9, respectively. Status of carbohydrate intake decreased significantly (p<0.05) and dietary fiber intake increased significantly (p<0.001) after supplementation. WHR (waist-hip ratio) decreased significantly (p<0.05) and body fat and percent body fat decreased significantly after supplementation (p<0.05). Total cholesterol and triglyceride decreased significantly after supplementation (p<0.001). Above results of this study show that green tea, silk peptide and dietary fiber supplementation-added routine diet improves body fat distribution, total cholesterol, triglyceride. Especially, declination of abdominal fat and WHR was notable, since that means diminution of risk factors for obesity and chronic degenerative diseases.

Study on Consequent Body Fat and Serum Lipid Metabolism after Cocoon Hydrolysate, Green Tea Leaves and Dietary Fiber Supplementation (누에고치 유래 실크 펩타이드와 녹차잎 및 식이 섬유소 보충이 체지방 및 혈청 지질 대사에 미치는 영향에 관한 연구)

  • Lee, Min-Sook;Kim, Dong-Myung;Cho, Byung-Nam;Koo, Seung-Ja;Jew, Sang-Sup;Jin, Dong-Kyu;Lee, Sung-Hee
    • Applied Biological Chemistry
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    • v.46 no.2
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    • pp.123-129
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    • 2003
  • This study is to investigate consequent nutrient intake status, Influences of body mass index (BMI) and serum lipid composition, and fat distribution on the cocoon hydrolysate, green tea leaves and dietary fiber supplementation. During 2 months of this research (April to May, 2002), 47 women aged 20 yr-30 yr (average age 26.2 yr) were selected as subjects. Nutrient intake was investigated by questionnaire and 24-hr recall method. Antropometric assessments of the subjects were investigated by SBIA method (Segmental bioimpedance assay, Inbody 3.0). The results were as follows: mean body weight was 60.7 kg, mean body height 161.7 cm and mean BMI 23.4. Status of energy intakes significantly decreased (p<0.01) and dietary fiber intakes significantly increased (p<0.001) after supplementation. BMI and WHR (waist-hip ratio) significantly decreased (p<0.01) and body fat significantly decreased after supplementation (p<0.001). Total cholesterol and LDL-cholesterol significantly decreased after supplementation (p<0.05). HDL-cholesterol was negatively correlated with BMI and WHR (p<0.01). LDL/HDL ratio was positively correlated with BMI (p<0.01) and WHR (p<0.05). Above results of this study show that low-molecule peptide, green tea leaves and dietary fiber supplementation-added routine diet improves lessening body fat distribution, total cholesterol, LDL-cholesterol. Especially, decrease of abdominal fat and WHR were notable. That meant decrease of risk factors.

Characterization of Bacteriocin Produced from Isolated Strain of Bacillus sp. (Bacillus 속 분리주가 생산하는 박테리오신의 특성 조사)

  • Ham, Seung-Hee;Choi, Nack-Shick;Moon, Ja-Young;Baek, Sun-Hwa;Lee, Song-Min;Kang, Dae-Ook
    • Journal of Life Science
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    • v.27 no.2
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    • pp.202-210
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    • 2017
  • As an effort to find a potential biopreservative, we isolated bacterial strains producing bacteriocin from fermented foods. A strain was finally selected and characteristics of the bacteriocin were investigated. The selected strain was identified as Bacillus subtilis E9-1 based on the 16S rRNA gene analysis. The culture supernatant of B. subtilis E9-1 showed antimicrobial activity against Gram-positive bacteria. Subtilisin A, ${\alpha}$-chymotrypsin, trypsin and proteinase K inactivated the antimicrobial activity, which means its proteinaceous nature, a bacteriocin. The bacteriocin activity was fully retained at the pH range from 2.0 to 8.0 and stable at up to $100^{\circ}C$ for 60 min. Solvents such as ethanol, isopropanol and methanol had no effect on the antimicrobial activity at the concentration of 100% but acetone and acetonitrile reduced the activity at up to 100% concentration. Cell growth of four indicator strains was dramatically decreased in dose-dependent manner. Listeria monocytogenes was the most sensitive, but Enterococcus faecium was the most resistant. Bacillus cereus and Staphylococcus aureus showed the medium sensitivity. The bacteriocin showed its antimicrobial activity against B. cereus and L. monocytogenes via bactericidal action. The number of viable cells of L. monocytogenes started to reduce after addition of bacteriocin to the minced beef. The bacteriocin was purified through acetone concentration, gel filtration chromatography and RP-HPLC. The whole purification step led to a 6.82 fold increase in the specific activity and 6% yield of bacteriocin activity. The molecular weight of the purified bacteriocin was determined to be 3.3 kDa by MALDI-TOF/TOF mass spectrometry.