• Title/Summary/Keyword: microbial transformation

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Microbial Transformation of Aniline to Acetaminophen

  • Lee, Sang-Sup;Jin, Hyung-Jong;Son, Mi-Won
    • Archives of Pharmacal Research
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    • v.15 no.1
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    • pp.30-34
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    • 1992
  • In order to obtain acetaminophen, a popular analgesic-antipyretic, through microbial p-hydroxylation and N-acetylation of aniline, various fungi and bacteria were secreened. Among them, Streptomyces species were chosen for strain improvement by the use of interspecific protoplast fusion technique. Two interspecific fused strains were developed between S. rimosus (N-cetylation function) and S. aureofaciens (p-hydroxylation function) and also between S. lividans and S. globisporus. For efficient protoplast fusion and cell wall regeneration, various conditions were examined. In a typical experiment of mixed S rimosus ($pro^- \;his^-$) and S. aureofaciens ($ilv^-$) protoplasts with 40% (w/v) polythylene glycol 3350 (PEG) for 3 min gave $8.3\times10^{-7}$ of fusion frequency. Treatment of mixed S. lividans (pant-) and S. globisporus (leu-) protoplasts with 50% (w/v) PEG for 3 min at $30^\circ{C}$ gave $1.2\times10^{-6}$ of frequency. Among the fused strains, up to 40-50% increase in p-hydroxylation power was observed. To investigate the possibility of plasmid involvement in p-hydroxylation power was observed. To investigate the possibility of plasmid involvement in p-hydroxylation of acetanilide, plasmid curing was attempted. We found that cells treated with acriflavine (at the frequency of 100%) and cells regenerated from protoplsts of S. auroefaciens (2% frequency) lost their p-hydroxylation function.

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Microbial $9{\alpha}$-Hydroxylase:Epoxidation of 9(11)-dehydro-$17{\alpha}$-methyl-testosterone

  • Kang, Hee-Kyoung;Lee, Sang-Sup
    • Archives of Pharmacal Research
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    • v.20 no.6
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    • pp.525-528
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    • 1997
  • Steroid $9{\alpha}$.-hydroxylase is a key enzyme system in steroid nucleus degradation in company with ${\Delta}$-dehydrogenase. To examine $9{\alpha}$-hydroxylase activity during microbial transformation of steroids, 9(11)-dehydro-$17{\alpha}$-methyl-testosterone was adopted as a stable substrate for preventing the rupture of steroid nucleus. Using Nocardia restrictus ATCC 14887 capable of introducing a $9{\alpha}$-hydroxyl group into steroids, $9{\alpha}$,$11{\alpha}$-oxido-$17{\beta}$-hydroxy-$17{\alpha}$-methyl-4-androstene-3-one and $9{\alpha}$-hydroxyl group into steroids,$9{\alpha}$,$11{\alpha}$-oxido-$17{\beta}$-hydroxy-$17{\alpha}$-methyl-1,4-androstadiene-3- one were obtained. These microbiologically transformed products could be used as reference compounds in the enzyme assay.

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Gut Microbial Metabolites on Host Immune Responses in Health and Disease

  • Jong-Hwi Yoon;Jun-Soo Do;Priyanka Velankanni;Choong-Gu Lee;Ho-Keun Kwon
    • IMMUNE NETWORK
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    • v.23 no.1
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    • pp.6.1-6.24
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    • 2023
  • Intestinal microorganisms interact with various immune cells and are involved in gut homeostasis and immune regulation. Although many studies have discussed the roles of the microorganisms themselves, interest in the effector function of their metabolites is increasing. The metabolic processes of these molecules provide important clues to the existence and function of gut microbes. The interrelationship between metabolites and T lymphocytes in particular plays a significant role in adaptive immune functions. Our current review focuses on 3 groups of metabolites: short-chain fatty acids, bile acids metabolites, and polyamines. We collated the findings of several studies on the transformation and production of these metabolites by gut microbes and explained their immunological roles. Specifically, we summarized the reports on changes in mucosal immune homeostasis represented by the Tregs and Th17 cells balance. The relationship between specific metabolites and diseases was also analyzed through latest studies. Thus, this review highlights microbial metabolites as the hidden treasure having potential diagnostic markers and therapeutic targets through a comprehensive understanding of the gut-immune interaction.

Microbial Tansformatin of $\gamma$-Butyrobetaine into L-Carnitine by Achromobacter cylcoclast (Achromobacter cycloclast에 의한 $\gamma$-Butyrobetaine의 L-Carnitine에로의 생물전환)

  • 이은구;이인영;박영훈
    • KSBB Journal
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    • v.14 no.2
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    • pp.205-211
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    • 1999
  • We investigated optimal conditions for the microbial transformation of $\gamma$-butyrobetaine into L-carnitine by using Achromobacter cycloclast ATCC 21921. When the cells were cultivated in the medium containing $\gamma$-butyrobetaine as the sole carbon source for both cell growth and L-carnitine production, the maximum L-carnitine production was 2.9 g/L and the conversion yield from $\gamma$-butyrobetaine to L-carnitine was as low as 30.9 mol%. In order to enhance the L-carnitine production and the conversion yield, various carbon sources were added to the $\gamma$-butyronetaine containing basal medium. In the medium supplemented with glycerol, L-carnitine production was as high as 4.6 g/L and the conversion yield was 88.2 mol%, showing a significant improvement in L-carnitine synthesis compared to those in the medium without glycerol. We also examined the additional effect of quaternary ammonium compounds such as betaine and choline, which are similar in structure to $\gamma$-butyrobetaine and L-carnitien. It was observed that in the presence of those quaternary ammonium compounds, both the L-carnitine production rate and the conversion yield increased. In addition, we found that cell growth was inhibited by a $\gamma$-butyrobetaine concentration of more than 3%, while L-carnitine production was efficient at the $\gamma$-butyrobetaine concentration of 2-3%. By cultivating the cells in the optimal medium containing glycerol and choline, we obtained an L-carnitine concentration of 7.2 g/L with the conversion yield of 98.7 mol% in 4 days.

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Study of Functional Verification to Abiotic Stress through Antioxidant Gene Transformation of Pyropia yezoensis (Bangiales, Rhodophyta) APX and MnSOD in Chlamydomonas

  • Lee, Hak-Jyung;Yang, Ho yeon;Choi, Jong-il
    • Journal of Microbiology and Biotechnology
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    • v.28 no.7
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    • pp.1217-1224
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    • 2018
  • Seaweeds produce antioxidants to counteract environmental stresses, and these antioxidant genes are regarded as important defense strategies for marine algae. In this study, the expression of Pyropia yezoensis (Bangiales, Rhodophyta) ascorbate peroxidase (PyAPX) and manganese-superoxide dismutase (PyMnSOD) was examined by qRT-PCR in P. yezoensis blades under abiotic stress conditions. Furthermore, the functional relevance of these genes was explored by overexpressing them in Chlamydomonas. A comparison of the different expression levels of PyAPX and PyMnSOD after exposure to each stress revealed that both genes were induced by high salt and UVB exposure, being increased approximately 3-fold after 12 h. The expression of the PyAPX and PyMnSOD genes also increased following exposure to $H_2O_2$. When these two genes were overexpressed in Chlamydomonas, the cells had a higher growth rate than control cells under conditions of hydrogen peroxide-induced oxidative stress, increased salinity, and UV exposure. These data suggest that Chlamydomonas is a suitable model for studying the function of stress genes, and that PyAPX and PyMnSOD genes are involved in the adaptation and defense against stresses that alter metabolism.

Biosequestration, Transformation, and Volatilization of Mercury by Lysinibacillus fusiformis Isolated from Industrial Effluent

  • Gupta, Saurabh;Goyal, Richa;Nirwan, Jashan;Cameotra, Swaranjit Singh;Tejoprakash, Nagaraja
    • Journal of Microbiology and Biotechnology
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    • v.22 no.5
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    • pp.684-689
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    • 2012
  • In the present study, an efficient mercury-tolerant bacterial strain (RS-5) was isolated from heavy-metalcontaminated industrial effluent. Under shake flask conditions, 97% of the supplemented mercuric chloride was sequestered by the biomass of RS-5 grown in a tryptone soy broth. The sequestered mercuric ions were transformed inside the bacterial cells, as an XRD analysis of the biomass confirmed the formation of mercurous chloride, which is only feasible following the reaction of the elemental mercury and the residual mercuric chloride present within the cells. Besides the sequestration and intracellular transformation, a significant fraction of the mercury (63%) was also volatilized. The 16S rRNA gene sequence of RS-5 revealed its phylogenetic relationship with the family Bacillaceae, and a 98% homology with Lysinibacillus fusiformis, a Gram-positive bacterium with swollen sporangia. This is the first observation of the sequestration and volatilization of mercuric ions by Lysinibacillus sp.

Fusaric Acid Production in Fusarium oxysporum Transformants Generated by Restriction Enzyme-Mediated Integration Procedure (Restriction Enzyme-Mediated Integration 방법으로 확보한 Fusarium oxysporum 형질전환체의 후자리산 생성능 분석)

  • Lee, Theresa;Shin, Jean Young;Son, Seung Wan;Lee, Soohyung;Ryu, Jae-Gee
    • Research in Plant Disease
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    • v.19 no.4
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    • pp.254-258
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    • 2013
  • Fusaric acid (FA) is a mycotoxin produced by Fusarium species. Its toxicity is relatively low but often associated with other mycotoxins, thus enhancing total toxicity. To date, biosynthetic genes or enzymes for FA have not been identified in F. oxysporum. In order to explore the genetic element(s) for FA biosynthesis, restriction enzyme mediated integration (REMI) procedure as an insertional mutagenesis was employed using FA producing-F. oxysporum strains. Genetic transformation of two F. oxysporum strains by REMI yielded more than 7,100 transformants with efficiency of average 3.2 transformants/${\mu}g$ DNA. To develop a screening system using phytotoxicity of FA, eleven various grains and vegetable seeds were tested for germination in cultures containing FA: Kimchi cabbage seed was selected as the most sensitive host. Screening for FA non-producer of F. oxysporum was done by growing each fungal REMI transformant in Czapek-Dox broth for 3 weeks at $25^{\circ}C$ then observing if the Kimchi cabbage seeds germinated in the culture filtrate. Of more than 5,000 REMI transformants screened, fifty-three made the seeds germinated, indicating that they produced little or fewer FA. Among them, twenty-six were analyzed for FA production by HPLC and two turned out to produce less than 1% of FA produced by a wild type strain. Sequencing of genomic DNA regions (252 bp) flanking the vector insertion site revealed an uncharacterized genomic region homologous (93%) to the F. fujikuroi genome. Further study is necessary to determine if the vector insertion sites in FA-deficient mutants are associated with FA production.

Production of cis, cis-Muconic Acid from Benzoic Acid via Microbial Transformation

  • SangGu Bang;Won
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.1 no.1
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    • pp.36-40
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    • 1996
  • For the production of cis, cis-muconic acid via biocatalytic conversion reactions from a toxic cosubstrate, benzoic acid, a fed-batch process using computer-controlled DO-stat feeding was developed. The mutant strain of Pseudomonas putida BM014 produced cis, cis-muconic acid from benzoic acid with high conversion yield. More than 32 g/L of cis, cis-muconic acid was accumulated in 42h and a productivity of 1.4g/(L.h)was achieved.

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Microbial Transformation of a Monoterpene, Geraniol, by the Marine-derived Fungus Hypocrea sp.

  • Leutou, Alain S.;Yang, Guohua;Nenkep, Viviane N.;Siwe, Xavier N.;Feng, Zhile;Khong, Thang T.;Choi, Hong-Dae;Kang, Jung-Sook;Son, Byeng-Wha
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1150-1152
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    • 2009
  • Geraniol (1) is the biogenetic precursor of a number of monoterpenes. We tested various marine-derived microorganisms to determine their ability to biotransform 1. Only Hypocrea sp. was capable of transforming 1 into its oxidized derivative, 1,7-dihydroxy-3,7-dimethyl-(E)-oct-2-ene (2). The structure of the metabolite obtained was assigned on the basis of detailed spectroscopic data analyses.