• Title/Summary/Keyword: diacetyl

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Characterization of Two Metagenome-Derived Esterases That Reactivate Chloramphenicol by Counteracting Chloramphenicol Acetyltransferase

  • Tao, Weixin;Lee, Myung-Hwan;Yoon, Mi-Young;Kim, Jin-Cheol;Malhotra, Shweta;Wu, Jing;Hwang, Eul-Chul;Lee, Seon-Woo
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1203-1210
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    • 2011
  • Function-driven metagenomic analysis is a powerful approach to screening for novel biocatalysts. In this study, we investigated lipolytic enzymes selected from an alluvial soil metagenomic library, and identified two novel esterases, EstDL26 and EstDL136. EstDL26 and EstDL136 reactivated chloramphenicol from its acetyl derivates by counteracting the chloramphenicol acetyltransferase (CAT) activity in Escherichia coli. These two enzymes showed only 27% identity in amino acid sequence to each other; however both preferentially hydrolyzed short-chain p-nitrophenyl esters (${\leq}C_5$) and showed mesophilic properties. In vitro, EstDL136 catalyzed the deacetylation of 1- and 3-acetyl and 1,3-diacetyl derivates; in contrast, EstDL26 was not capable of the deacetylation at $C_1$, indicating a potential regioselectivity. EstDL26 and EstDL136 were similar to microbial hormone-sensitive lipase (HSL), and since chloramphenicol acetate esterase (CAE) activity was detected from two other soil esterases in the HSL family, this suggests a distribution of CAE among the soil microorganisms. The isolation and characterization of EstDL26 and EstDL136 in this study may be helpful in understanding the diversity of CAE enzymes and their potential role in releasing active chloramphenicol in the producing bacteria.

Photopodegradation efficiency of visible light cured dental resin composites with novel photosensitizers (가시광선 중합형 복합수지용 광증감제의 분해율 비교)

  • Sun, Gum-Ju
    • Journal of Technologic Dentistry
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    • v.33 no.4
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    • pp.291-297
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    • 2011
  • Purpose: The purpose of this study was to know the availability of three photosensitizers, CQ, PD, DA, as a photosensitizer of dental resin composite. We want to know abortion band around visible light region for the using potential possibility as a photosensitizer for visible light cured dental composite resin. And I studied to know the relative photodecomposition ratio of three photosensitizers with or without photoinitiator, DAEM. Methods: The photodecomposition of three photosensitizers were studied by UV absorption spectroscopy in ethanol and determined by same instrument with irradiation time for relative photodecomposition. In order to study the effect of amine on photodecomposition was added the DAEM in the photosensitizer solution and the relative rate was measured by the same procedure with aove mentioned. Results: The all of three photosensitizers are absorbed around visible light region. The relative rate of decrease in absorbance incereased in the order: CQ < BD < PD. The effect of DAEM on the photodecomposition of the photosensitizers was appeared different results without DAEM. The photodecomposition rate of PD and DA decreased somewhat with the addition of amine, while that of CQ increased. The rtealtive photodecomposition rate increased in the oprder: BD ${\leq}$ CQ < PD with the addition of amine, but the differnce was not significant. Conclusion: PD and DA like CQ gives to the possibility of use as a photosensitizer for visible light cured dental composite resin by absorption around visible light region and photodecomposition in the maximum absorption wavelength. And it is showed that PD and DA are not effective decomposed with amine initiator, DAEM but CQ decomposed with DAEM effectively. This result may be due to a different mechanism operating for the decomposition of photosensitizers in the presence of amine.

Purification and Characterization of Phocaecin PI80: An Anti-Listerial Bacteriocin Produced by Streptococcus phocae PI80 Isolated from the Gut of Peneaus indicus (Indian White Shrimp)

  • Satish Kumar, Ramraj;Arul, Venkatesan
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1393-1400
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    • 2009
  • A bacteriocin-producing strain PI80 was isolated from the gut of Penaeus indicus (Indian white shrimp) and identified as Streptococcus phocae PI80. The bacteriocin was purified from a culture supernatant to homogeneity as confirmed by Tricine SDS-PAGE. Reverse-phase HPLC analysis revealed a single active fraction eluted at 12.94 min, and MALDI-TOF mass spectrometry analysis showed the molecular mass to be 9.244 kDa. This molecular mass does not correspond to previously described streptococcal bacteriocins. The purified bacteriocin was named phocaecin PI80 from its producer strain, as this is the first report of bacteriocin production by Streptococcus phocae. The bacteriocin exhibited a broad spectrum of activity and inhibited important pathogens: Listeria monocytogenes, Vibrio parahaemolyticus, and V. fischeri. The antibacterial substance was also sensitive to proteolytic enzymes: trypsin, protease, pepsin, and chymotrypsin, yet insensitive to catalase, peroxidase, and diastase, confirming that the inhibition was due to a proteinaceous molecule (i.e., the bacteriocin), and not due to hydrogen peroxide or diacetyl. Phocaecin PI80 moderately tolerated heat treatment (up to $70^{\circ}C$ for 10 min) and resisted certain solvents (acetone, ethanol, and butanol). A massive leakage of $K^+$ ions from E. coli $DH5\alpha$, L. monocytogenes, and V. parahaemolyticus was induced by phocaecin PI80, as measured by Inductively Coupled Plasma Optical Emission Spectrometry (ICPOES). Therefore, the results of this study show that phocaecin PI80 may be a useful tool for inhibiting L. monocytogenes in seafood products that do not usually undergo adequate heat treatment, whereas the cells of Streptococcus phocae PI80 could be used to control vibriosis in shrimp farming.

Construction of Amylolytic Industrial Brewing Yeast Strain with High Glutathione Content for Manufacturing Beer with Improved Anti-Staling Capability and Flavor

  • Wang, Jin-Jing;Wang, Zhao-Yue;He, Xiu-Ping;Zhang, Bo-Run
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1539-1545
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    • 2010
  • In beer, glutathione works as the main antioxidant compound, which also correlates with the stability of the beer flavor. In addition, high residual sugars in beer contribute to major nonvolatile components, which are reflected in a high caloric content. Therefore, in this study, the Saccharomyces cerevisiae GSH1 gene encoding glutamylcysteine synthetase and the Saccharomycopsis fibuligera ALP1 gene encoding ${\alpha}$-amylase were coexpressed in industrial brewing yeast strain Y31 targeting the ${\alpha}$-acetolactate synthase (AHAS) gene (ILV2) and alcohol dehydrogenase gene (ADH2), resulting in the new recombinant strain TY3. The glutathione content in the fermentation broth of TY3 increased to 43.83 mg/l as compared with 33.34 mg/l in the fermentation broth of Y31. The recombinant strain showed a high ${\alpha}$-amylase activity and utilized more than 46% of the starch as the sole carbon source after 5 days. European Brewery Convention tube fermentation tests comparing the fermentation broths of TY3 and Y31 showed that the flavor stability index for TY3 was 1.3-fold higher, whereas its residual sugar concentration was 76.8% lower. Owing to the interruption of the ILV2 gene and ADH2 gene, the contents of diacetyl and acetaldehyde as off-flavor compounds were reduced by 56.93% and 31.25%, respectively, when compared with the contents in the Y31 fermentation broth. In addition, since no drug-resistant genes were introduced to the new recombinant strain, it should be more suitable for use in the beer industry, owing to its better flavor stability and other beneficial characteristics.

Application of Saccharomyces rouxii for the Production of Non-alcoholic Beer

  • Sohrabvandi, Sarah;Razavi, Seyed Hadi;Mousavi, Seyed Mohammad;Mortazavian, Amir;Rezaei, Karamathollah
    • Food Science and Biotechnology
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    • v.18 no.5
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    • pp.1132-1137
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    • 2009
  • Successive application of Saccharomyces cerevisiae DSM 70424 and Saccharomyces rouxii DSM 2535 or DSM 2531 in the production of non-alcoholic beer was investigated. The aim of the study was to consider the impact of the 2 mentioned strains of S. rouxii on the reduction of alcohol content in wort fermented at 12 or $24^{\circ}C$ for 96 hr, applying periodic aeration. The 2 S. rouxii strains were added at the $48^{th}$ hr of fermentation after thermal inactivation of S. cerevisiae cells. The greatest alcohol decrease rate was observed for the treatment containing S. rouxii DSM 2535-fermented at $24^{\circ}C$ (from 1.56 to 0.36%). The concentration of acetaldehyde, diacetyl, and 2,3-pentandione, that have a key role in appearance of 'wort' and 'buttery' off flavors, were significantly lower in S. rouxii-containing treatments fermented at $24^{\circ}C$. S. rouxii-containing treatment fermented at $24^{\circ}C$ showed slightly lower overall flavor acceptability compared to S. cerevisiae-containing treatment fermented at the same temperature. Such score was improved for the products obtained at $12^{\circ}C$.

Nutritional Effects and Antimicrobial Activity of Kefir (Grains)

  • Shen, Ying;Kim, Dong-Hyeon;Chon, Jung-Whan;Kim, Hyunsook;Song, Kwang-Young;Seo, Kun-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.36 no.1
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    • pp.1-13
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    • 2018
  • Kefir exhibits antimicrobial activity in vitro against gram-positive and gram negative bacteria, as well as some fungi. The ability of LAB to inhibit the growth of closely related bacteria is well known. This inhibition of pathogenic and spoilage microbes may be due to the production of organic acids, hydrogen peroxide, acetaldehyde, diacetyl, carbon dioxide, or bacteriocins. Lactobacilli are the major contributors to acid production and, hence, a determining factor in the flavor development in kefir. Lactic acid, proteolytic activity, and acetaldehyde are the essential flavor compounds in kefir. Both acid and bacteriocins contribute to the antimicrobial activity of kefir and kefir grains. Kefir is rich in proteins, calcium, vitamin $B_{12}$, niacin, and folic acid. Many studies have investigated the benefits of consuming kefir and have shown that it is a natural probiotic, which when consumed regularly, can help relieve intestinal disorders, promote bowel movement, reduce flatulence, and improve the overall health of the digestive system. Tibetan kefir, which is different from traditional kefir, is produced in China. It has been reported to exhibit antimicrobial activity that is nearly identical to that of traditional kefir. Kefir production is considered a rapidly growing food industry in China.

Analytical Method of L-Ascorbic Acid Content in Green Tea (녹차중(綠茶中)의 L-Ascorbic Acid의 정량법(定量法)에 관한 연구(硏究))

  • Shin, Mee-Gyung;Nam, Chang-Woo
    • Korean Journal of Food Science and Technology
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    • v.11 no.2
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    • pp.77-80
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    • 1979
  • Effects of interfering substances on the determination of L-ascorbic acid (AsA) in green tea and it's extracts by 2,4-dinitrophenylhydrazine (DNP) method was studied. and the removal of these interfering substances was also investigated. Under the condition prescribed for DNP method, AsA content of green tea are effected by some sugar, reductones, dicarbonyl compounds, organic acids, amino acids and others. All interfering substances except amino acids were eliminated by the chloroform extraction after adding o-phenylendiamine to sample solution. and remaining amino acids were eliminated almost completely by the treatment with ion exchange resin$(Amberlite{\;}IR-120H^{+})$. After removing the interfering substances by the above mentioned procedure, total AsA in green tea was determined by DNP method. The values obtained by this method were in good agreement with those by thin layer chromatography (TLC) method. and the method was more rapid and simple than TLC method.

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Photopolymerization Efficiency of Bis-GMA Dental Resin Composites with New Photosensitizers (새로운 광증감제의 양에 따른 치과용 Bis-GMA 복합수지의 중합효율)

  • Sun, Gum-Ju;Jung, Jong-Hyun
    • Journal of dental hygiene science
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    • v.9 no.2
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    • pp.189-195
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    • 2009
  • Two diketones, 1,2-phenylpropanedione (PD) and diacetyl (DA) were investigated as new visible light photosensitizers for dental composite resin of bis-GMA in order to improve the photopolymerization effect. The photopolymerization efficiency of bis-GMA composite resin containing PD and DA was studied by IR absorption spectroscopy. And the results were compared with that of camphorquinone (CQ). Relative photopolymerization efficiency of the photosensitizers increased in the order of DA < CQ < PD. Thus. PO is a new visible light photosensitizer for dental composite resin with higher photopolymerization efficiency than that of CQ.

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Isolation and Characterization of Constituent Compounds from Leaves and Stems of Chrysanthemum boreale Makino (산국 잎과 줄기의 유효성분 분리 및 특성 연구)

  • Park, Sook Jahr;Park, Moon Ki;Lee, Jong Rok
    • Journal of Environmental Science International
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    • v.28 no.11
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    • pp.993-1004
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    • 2019
  • Chrysanthemum boreale Makino (C. boreale) is widely distributed in Asian countries, and has traditionally been used to treat various inflammatory diseases including bronchitis. In this study, we aimed to isolate biologically active compounds from leaves and stems of C. boreale. Chemical components were purified by column chromatograpy and recyclic HPLC, and characterized from their spectral data (IR, MS, NMR). Biological activity experiments were conducted for Farnesyl-protein transferase (FPTase) activity, apoptosis and nitirc oxide (NO) release. As a results, three sesquiterpene lactones were isolated. Compound 1 (4-methoxy-8-O-acetyl-10-hydroxy-2,11(13)-guaiadiene-12,6-olide) showed strong cytotoxic activities having an average growth inhibition of 50% ($GI_{50}$) value of $1.89{\mu}g/m{\ell}$ against human colon adenocarcinoma cells. Compound 1 also showed a low half maximal inhibitory concentration ($IC_{50}$) value of $10{\mu}g/m{\ell}$ for NO release. In the caspase 3 activity, compound 1 and compound 2 (8-O-(2-carbonyl-2-butyl)-3,10-dihydroxy-4,11(13) -guaiadiene-12,6-olide) exhibited 94% and 90% apoptosis inhibition activity, respectively. Compound 3 (4,8-O-diacetyl -10-hydroxy-2(3),11(13)-guaiadiene-12,6-olide) showed a strong inhibitory effect on FPTase activity with 90% inhibitory activity at a concentration of $100{\mu}g/m{\ell}$. These results clearly show the presence of lactone compounds in the leaves and stems, which may partially contribute to the pharmacological activity of C. boreale.

The Manufactures and Characteristics of Chestnut Yoghurt (The Volatile Flavour Compounds and the Sensory Properties of Chestnut Yoghurt) (밤 Yoghurt의 제조와 특성 (밤 Yoghurt의 휘발성 향미성분 및 관능적 특성))

  • Shin, Hyeon-Su;Kim, Sang-Beom;Kim, Gi-Yeong;Ryu, Jin-Su;Lim, Jong-U
    • Journal of Dairy Science and Biotechnology
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    • v.19 no.1
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    • pp.4-12
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    • 2001
  • The effects of the addition of raw and dried chestnut with levels of 1%($T_1$),2%($T_2$),3%($T_3$) and 4%($T_4$) in skim milk substrate on the volatile flavour compounds and sensory properties of yoghurt fermented with the mixed cultures of YC-380 and ABT-4 during fermentation and storage period were investigated. In all treatments, the contents of acetaldehyde and acetone were detected at 2 hrs. and 1 hr. of fermentation, respectively and decreased with the storage period. The contents of ethanol and diacetyl were detected in all treatments at 3 hrs. and 4 hrs. of fermentation, respectively and increased significantly(p<0.05) with the storage period. The contents of volatile flavour compounds of treatments were increased gradually with decreasing the level of addition of raw and dried chestnut, and increased in order of fermented with YC-380 and ABT-4. The contents of volatile flavour compounds of raw chestnut yoghurt were slightly high compared to those of dried chestnut yoghurt. The taste, flavour and texture of $T_1$ were slightly higher than those of all treatments immediately after fermentation and during the storage period. The scores of sensory evaluation of treatments except $T_1$ were lowered significantly(p<0.05) with increasing the level of addition of raw and dried chestnut. The quality of both of flavour and texture, and taste were superior to chestnut yoghurt fermented with YC-380 and ABT-4, respectively. Generally the scores of sensory evaluation of dried chestnut yoghurt were slightly high compared to those of raw chestnut yoghurt. From the results mentioned above, the addition of raw and dried chestnut at 1%(w/v) level in skim milk substrate were suitable for volatile flavour compounds and sensory property of raw and dried chestnut yoghurt.

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