• Title/Summary/Keyword: Purified salt

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Simple Purification of Bromelain from Pineapple

  • Ko, Bo-Sung;Hwang, Yong-Il;Lee, Seung-Cheol
    • Preventive Nutrition and Food Science
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    • v.1 no.1
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    • pp.106-110
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    • 1996
  • Bromelain(EC 3.4.22.4), the collective name for the proteolytic enzymes found in tissues of the plant family Bromeliaceae(pineapple), has been used as a tenderizing agent in food processing, and as an antiinflammatory agent in pharmaceuticals. In this paper, we describe the simple purification method of bromelain using Korean pineapple fruit. After removing contaminants at 30% saturation of ammonium sulfate, the supernatant obtained was treated again with ammonium sulfate to 80% saturation. Wit the above salt fractionation, partially purified bromelain could be obtained. To get highly purified bromelain, the previous 30% to 80% ammonium sulfate treated precipitate was dialyzed against 25mM sodium acetate buffer(pH 5.0) followed by passing through a CM- cellulose cation exchange column. Fruit bromelain was eluted as a major peak at 0.5~0.8M NaCI gradient. The present method is simpler with high wield than the traditional purification method-acetone treatment and several consecutive chromatographic processes.

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Simple Purification of the Human Antimicrobial Peptide Dermcidin (MDCD-1L) by Intein-Mediated Expression in E. coli

  • Hong, In-Pyo;Kim, Yong-Seok;Choi, Shin-Geon
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.350-355
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    • 2010
  • Among human antimicrobial peptides (hAMPs), DCD-1L has a broad spectrum of antimicrobial activity over a wide pH range and in high salt concentrations. It offers a promising alternative to conventional antibiotics. The 458-bp-long dermcidin cDNA was amplified by PCR using a human fetal cDNA library as a template. The 147-bp fragment of the MDCD-1L gene encoding an additional methionine residue was subcloned into the pTYB11 vector. Recombinant MDCD-1L was expressed as an intein fusion protein in E. coli, and then purified by affinity chromatography using chitin beads. A small peptide with a molecular mass of about 5 kDa was detected by tricine gel electrophoresis. The recombinant MDCD-1L peptide was purified from the gel and its amino acid sequence was determined by nanoLC-ESI-MS/MS analysis. The initiating amino acid, methionine, remained attached to the N-terminal region of recombinant MDCD-1L. Purified MDCD-1L showed antimicrobial activity against a Micrococcus luteus test strain.

Purification and Characterization of Recombinant Human Interferon Alpha 2a Produced from Saccharomyces cerevisiae

  • Rae, Tae-Ok;Chang, Ho-Jin;Kim, Jung-Ho;Park, Soon-Jae
    • BMB Reports
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    • v.28 no.6
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    • pp.477-483
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    • 1995
  • The recombinant human interferon alpha 2a ($rhIFN-{\alpha}2a$), expressed in Saccharomyces cerevtsiae, was purified from insoluble aggregates. The inclusion body of $rhIFN-{\alpha}$ was solubilized by guanidine salt in the presence of disulfide reducing agent. The refolding of denatured $rhIFN-{\alpha}2a$ was achieved by simple dilution. The authentic interferon alpha, which has two correctly matched disulfide bonds, was seperated from incompletely oxidized $IFN-{\alpha}$ and dimeric $IFN-{\alpha}$ by use of a CM-Sepharose column, followed by size exclusion columns at two different pH conditions. The purified protein has been subjected to detailed physicochemical characterization including sequence determination. Unlike other $rhIFN-{\alpha}2a$ from E. coli reported, the $rhIFN-{\alpha}2a$ from S. cerevisiae has no methionine residue at its N-terminus originating from the start codon, ATG. The pI of the protein was determined to be 6.05 with a single band in the pI gel, which demonstrated that the purified $rhIFN-{\alpha}$ was homogeneous. The structural study using circular dichroism showed that the protein retains its three dimensional structure in the wide range of pH conditions between pH 3 and 9, and only minor strucural deformation was observed at pH 1.0.

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Purification and Characterization of Polyphenol Oxidase from Oyster Mushroom (Pleurotus ostreatus) (느타리버섯(Pleurotus ostreatus)의 Polyphenol Oxidase 분리 정제 및 특성 조사)

  • Choi, Ju-Hee;Kim, Hyun-Jin;Park, Sun-Young;Ham, Kyung-Sik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.10
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    • pp.1447-1452
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    • 2011
  • Polyphenol oxidase (PPO) isoforms were partially purified from oyster mushroom (Pleurotus ostreatus) using various chromatography techniques, and their characteristics of heat stability, substrate affinity, optimum pH, and optimum temperature were investigated. Three PPO isoforms named PO-I, PO-II-1, and PO-II-2 were partially purified from oyster mushroom. The molecular weight of PO-II-1 was 70 kDa and PO-I and PO-II-2 were less than 6 kDa each. Characterization was carried out using a PPO isoform partially purified by hydrophobic interaction chromatography. Optimum temperature was $55^{\circ}C$ and optimum pH 5.0. However, the PPO was inactivated at neutral pH or by heating at $80^{\circ}C$ for 30 min, while the 40% PPO still remained active after heating at $60^{\circ}C$ for 45 min. The PPO isoform showed the highest substrate affinity to chlorogenic acid and pyrogallol, in which KM values were 1.01 and 2.06 mM, respectively. Therefore, these results suggested that the mushrooms should be stored at a pH higher than 7.0 and at a low temperature to prevent enzymatic browning.

Korean solar salts reduce obesity and alter its related markers in diet-induced obese mice

  • Ju, Jaehyun;Song, Jia-Le;Park, Eui-Seong;Do, Myoung-Sool;Park, Kun-Young
    • Nutrition Research and Practice
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    • v.10 no.6
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    • pp.629-634
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    • 2016
  • BACKGROUND/OBJECTIVE: The aim of this experiments was to show anti-obesity effects of Korean solar salt from different salt fields in diet-induced obese mice. MATERIALS/METHODS: Diet-induced obesity (DIO) was induced by a high-fat diet (HFD; 45% cal from fat) in C57BL/6J mice for eight weeks. The mice were fed with the designated diets (chow diet for Normal, HFD for Control, 0.47%-salt-mixed HFD for purified salt (PS), Guerande solar salt from France (SS-G), solar salt from Y salt field (SS-Y), solar salts from T salt field (SS-T) and S salt field (SS-S)) for another eight weeks. We checked body weight, food efficiency ratio (FER) and tissue weights (liver and epididymal adipose tissue (EAT)), and observed serum concentrations of triacylglycerol (TG), total cholesterol (TC), leptin and insulin. We also evaluated gene expressions of adipogenic / lipogenic mRNAs of $C/EBP{\alpha}$, $PPAR{\gamma}$ and FAS and beta-oxidation-related factors ($PPAR{\alpha}$ and CPT-1) in liver and EAT. The mineral composition of salt samples were analyzed using inductively coupled plasma optical emission spectrometry (ICP-OES). RESULTS: SS-T and SS-S significantly reduced body weight gain, FER, and weight of EAT compared to control and other samples (P < 0.05). SS-T and SS-S also significantly decreased serum levels of TG, TC, leptin and insulin (P < 0.05). SS-T and SS-S suppressed expressions of adipogenic / lipogenic mRNAs in liver and EAT, while promoting expression of beta-oxidation-related factors. The lowest sodium concentration was observed in SS-T ($30.30{\pm}0.59%$), and the lowest sodium-to-potassium (Na/K) ratio was found in SS-S (17.81). CONCLUSIONS: Our study shows that well-processed Korean solar salt may have anti-obesity effects in vivo, probably owing to its differences in mineral composition and other components, presumably resulting from the manufacturing processes. Further research is needed into the mechanism and to explore optimal manufacturing processes.

Effects of Different Kinds of Salt on the Quality of Wet Noodles (소금종류별에 따른 첨가가 생면의 품질에 미치는 효과)

  • Lee, Jeong-Mi;Kim, So-Young;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.12
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    • pp.1776-1780
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    • 2011
  • The effects of different kinds of salt on the qualities of weight, volume, water absorption rate, turbidity, pH, textural characteristics, and sensory evaluation of wet noodles were studied. The salts employed in this study were purified salt (PS), solar salt (SS), solar salt without bittern (SSWB), roasted salt (RS) and bamboo salt (BS, $1{\times}$). The weight, volume, and water absorption rate of the noodles decreased with the 2% addition of different kinds of salts compared to the noodles without any added salt. The turbidity of the noodles decreased when BS (0.484), RS (0.489), or SSWB (0.489) were added to the noodles much more than when PS (0.508) was added. The pH of both wet noodles and cooked noodles was higher when BS, RS, or SS were added than when the others were added. The springiness and cohesiveness of the noodles were both increased with the addition of the salts. BS, RS and SS especially increased the springiness and cohesiveness of the noodles. In sensory evaluations of appearance, taste, texture and overall acceptance, BS received the highest scores. From the results, adding salt to wet noodle preparations is important to increase the quality of the noodles. BS showed the best overall results among the salts employed in this study.

Effect of Mineral-rich Salt Intake on Diabetic Goto-Kakizaki Rats (미네랄이 풍부한 천일염이 Goto-Kakizaki Rat에 미치는 영향)

  • Jin, Yong-Xie;Kim, Haeng-Ryan;Kim, So-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.3
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    • pp.355-359
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    • 2014
  • The objective of this study was to determine the hyperglycemic effects of mineral-rich salt in type 2 diabetic Goto-Kakizaki (GK) rats and normal Wistar rats. Animals were divided into five groups, including a normal group, fed three different experimental salts [purified salt (PS), mineral-rich salt (WS1 and WS2), and bamboo salt (BS)] in the form of 1% salt solution for 12 weeks. Liver, kidney, and spleen weights were significantly increased in GK rats of salt groups as compared to Wistar normal group without salt. However, there was no difference among the salt groups. For serum lipids, total cholesterol level in the BS group and triglyceride level in the WS group were significantly reduced compared to those of the PS group. The concentration of blood glucose in the GK-PS group increased continuously during the experimental period, whereas that in the GK-WS group was significantly reduced at 12 weeks. In GK rats, glucose levels among the salt groups in OGTT by glucose were not significantly different compared to normal rats. Insulin and glucagon levels in blood were not significantly different among the groups, and no such association was observed for insulin. Pancreatic lslets of Langerhans in the PS group showed irregular formation compared to those of the normal, WS, and BS groups.

Characteristics of salt-tolerant pretense purified from the fermented anchovy sauce

  • Kim, Woo-Jae;Kim, Sang-Moo
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2001.10a
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    • pp.91-92
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    • 2001
  • Enzymes have been used as processing aids in the manufacture of food products to improve their quality, solubility and stability for centuries. About 50% of the enzymes used as industrial processing aids are protein hydrolases which have been used in a number of industrial application including laundry detergents, feed, leather treatment, silk degumming, cheese making, chill proofing, meat tenderzing, fermented sauces, and pharmaceuticals. (omitted)

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Thin Layer Chromatogram by an Extracellular ${\beta}$-Amylase of Bacillus sp. KYJ 963 and its Amino Acid Composition

  • Kim, Young-Jae
    • Journal of Life Science
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    • v.11 no.2
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    • pp.92-93
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    • 2001
  • Bacillus sp. KYJ 963, which was isolated from Korean salt-fermented anchovy (anchovy-jeot), produces an extracellular ${\beta}$-amylase. The analysis of the digestion products of substrates by thin layer chromatography from the purified protein revealed that the enzyme could not hydrolyze maltose or ${\alpha}$-cyclodextrin. In the amino acid composition analysis, the major characteristic of the ${\beta}$-amylase was the high proportion of amino acids that possess short side chain such as glycine and alanine.

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Cloning and Characterization of a Bile Salt Hydrolase from Enterococcus faecalis Strain Isolated from Healthy Elderly Volunteers (사람 분변에서 분리한 Enterococcusfaecalis가 생성하는 BileSaltHydrolase의 특징)

  • Eom, Seok-Jin;Kim, Geun-Bae
    • Journal of Dairy Science and Biotechnology
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    • v.29 no.1
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    • pp.49-54
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    • 2011
  • Bile salt hydrolase (BSH, EC 3.5.1.24) activity, which cleaves amide bond between carboxyl group (bile acid) and amino group (glycine or taurine), is commonly detected in gut-associated species of human and animal. During the screening of BSH active strains from the fecal samples of elderly human volunteers, strain CU30-2 was isolated on the basis of the highly active BSH producing activity. A bsh gene of the isolate was cloned into the pET22b expression vector and overexpressed in Escherichia coli BL21 (DE3) Gold by induction with 1mM IPTG. The overexpressed BSH enzyme with 6x His-tag was purified with apparent homogeneity using a $Ni^+$-NTA agarose column and characterized. The BSH enzyme of E. faecalis CU30-2 exhibited approximately 50 times higher activity against glycol-conjugated bile salts than tauro-conjugated bile salts having the highest activity against glycocholic acid. Considering the prevalence of E. faecalis strains in the human GI tract and glycol-conjugates dominated bile acid composition of human bile, further study is needed to investigate the impact of the BSH activity exerted by E. faecalis strains to the host as well as to the BSH producing strains.

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