Enterococcus bacteria are studied in various sectors including fermentation, food and dairy industries,as well as studied for their probiotic properties but have limited use due to their possible pathogenic behavior. The present report talks about the metabolites produced, by the previously isolated Enterococcus strain, E.villorum SB2 (accession number KX830968), from the vaginal source. The growth of the bacteria in three types of media (M17, MRS and LAPTg) was compared, where the M17 media gave better bacterial colonies, also maximum growth rate was observed in M17 media (T_d = 1.6 h & k = 0.4 h^-1), and thus was selected as the metabolite production media. Further, the studied bacteria did not show any hemolytic activity, making it safe for industrial applications. The HR-LCMS results showed the production of various amino acids, organic acids, peptides, and other metabolites like flavonoids (Quercetin 3-O-Manoglucoside), terpenoids (7',8',Dihydro-8'-hydroxycitraniaxanthin, O-Methylganoderic acid O, Thalicsessine, Austinol, Valdiate), indole derivatives produced by tryptophan metabolism (5-hydroxykynurenamine, 2S,4R)-4-(9H-Pyrido[3,4-b]indol-1-yl)-1,2,4-butanetriol, Indoleacrylic acid), antimicrobial compounds (Fortimicin A) and fatty acids (Stearic acid, Myristic acid), which were earlier unreported form Enterococcus species opening new scope for discovering new industrial applications of the strain. As the studied bacteria has been reported to be a potential probiotic, the detection of these industrially important metabolites can be studied further in future studies to reveal the potential industrial applications of the strain.

28 Mar 2019·Journal of Microbiology and BiotechnologyQ4 · ENGINEERING & TECHNOLOGY

In Vivo Characterization of Phosphotransferase-Encoding Genes istP and forP as Interchangeable Launchers of the C3��,4��-Dideoxygenation Biosynthetic Pathway of 1,4-Diaminocyclitol Antibiotics

Q4 · ENGINEERING & TECHNOLOGY

Article

Author: Hoang, Nguyen ; Park, Je Won ; Kim, Hye Ji ; Huong, Nguyen ; Hwang, Hyun Ha ; Lee, Na Joon ; Seo, Eun Gyo ; Son, Hye Bin

Deactivation of aminoglycosides by their modifying enzymes, including a number of aminoglycoside O-phosphotransferases, is the most ubiquitous resistance mechanism in aminoglycoside-resistant pathogens. Nonetheless, in a couple of biosynthetic pathways for gentamicins, fortimicins, and istamycins, phosphorylation of aminoglycosides seems to be a unique and initial step for the creation of a natural defensive structural feature such as a 3',4'- dideoxy scaffold. Our aim was to elucidate the biochemical details on the beginning of these C3',4'-dideoxygenation biosynthetic steps for aminoglycosides. The biosynthesis of istamycins must surely involve these 3',4'-didehydroxylation steps, but much less has been reported in terms of characterization of istamycin biosynthetic genes, especially about the phosphotransferase-encoding gene. In the disruption and complementation experiments pointing to a putative gene, istP, in the genome of wild-type Streptomyces tenjimariensis, the function of the istP gene was proved here to be a phosphotransferase. Next, an in-frame deletion of a known phosphotransferase-encoding gene forP from the genome of wild-type Micromonospora olivasterospora resulted in the appearance of a hitherto unidentified fortimicin shunt product, namely 3-O-methyl-FOR-KK1, whereas complementation of forP restored the natural fortimicin metabolite profiles. The bilateral complementation of an istP gene (or forP) in the ΔforP mutant ( or ΔistP mutant strain) successfully restored the biosynthesis of 3',4'- dideoxy fortimicins and istamycins , thus clearly indicating that they are interchangeable launchers of the biosynthesis of 3',4'-dideoxy types of 1,4-diaminocyclitol antibiotics.

01 Feb 2016·Analytical and Bioanalytical ChemistryQ2 · CHEMISTRY

Characterization of fortimicin aminoglycoside profiles produced from Micromonospora olivasterospora DSM 43868 by high-performance liquid chromatography-electrospray ionization-ion trap-mass spectrometry

Q2 · CHEMISTRY

Article

Author: Sohng, Jae Kyung ; Yoon, Yeo Joon ; Huong, Nguyen Lan ; Hoang, Nguyen Huu ; Park, Je Won ; Hong, Sung-Yong

In this study, an efficient high-performance liquid chromatography (HPLC)-electrospray ionization (ESI)-ion trap-tandem mass spectrometry (MS/MS) was developed for the identification of the biosynthetic congeners involved in the aminocyclitol aminoglycosidic fortimicin pathway from Micromonospora olivasterospora fermentation. The usage of both acid extraction (pH ∼2.5) followed by an cationic-exchanging SPE cleanup and pentafluoropropionic acid mediated ion-pairing chromatography with ESI-ion trap-MS/MS detection was determined to be sufficiently practical to profile the fortimicin (FOR) congeners produced in a culture broth. The limit of the quantification for the fortimicin A (FOR-A) standard spiked in the culture broth was ∼1.6 ng mL(-1). The average recovery rate was 93.6%, and the intra- and inter-day precisions were <5% with accuracy in the range from 87.1 to 94.2%. Moreover, the epimeric mixtures including FOR-KH, FOR-KR, and FOR-B were separately resolved through a macrocyclic glycopeptide (teicoplanin)-bonded chiral column. As a result, ten natural FOR pseudodisaccharide analogs were identified and semi-quantified in descending order as follows: FOR-A, FOR-B, DCM, FOR-KH plus FOR-KR, FOR-KK1, FOR-AP, FOR-KL1, FOR-AO, and FOR-FU-10. This is the first report on both the simultaneous characterization of diverse structurally closely related FORs derived from bacterial fermentation using HPLC-ESI-ion trap-MS/MS analysis and the chromatographic separation of the three FOR epimers.

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