• Journal of Microbiology and Biotechnology
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• v.17 no.10
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• pp.1638-1644
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• 2007

The effect of carbon sources on tacrolimus production by a mutant strain of Streptomyces clavuligerus CKD 1119, an isolate from soil, was examined. Among the carbohydrates and oils tested in this work, a mixed carbon source of soluble starch and com oil was the best. An analysis of the culture kinetics also showed that, in contrast to the carbohydrates, the com oil was consumed later in the antibiotic production phase, implying that the oil substrate was the principal carbon source for the biosynthesis of tacrolimus, and this was directly proven by experiments using $^{14}C$-glucose and $^{14}C$-oleate substrates. Furthermore, com oil induced the formation of lipase by the mutant strain, whereas the addition of glucose significantly repressed lipase activity. The lipase activity exhibited by the FK-506-overproducing mutants was also observed to be directly proportional to their tacrolimus yield, indicating that a high lipase activity is itself a crucial factor for tacrolimus production. A feasibility study with a 200-1 pilot-scale fermentor and the best strain (Tc-XII-15322) identified in this work revealed a high volumetric and specific productivity of about 495 mg/l and 0.34 mg/mg dry mycelium, respectively.

• Journal of Microbiology and Biotechnology
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• v.26 no.3
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• pp.477-482
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• 2016

A uridine diphosphate-glucose:sterol glycosyltransferase-encoding gene was isolated and cloned from the established fosmid library of Micromonospora rhodorangea ATCC 27932 that usually produces the aminoglycoside antibiotic geneticin. The gene consists of 1,185 base pairs and encodes a 41.4 kDa protein, which was heterologously expressed in Escherichia coli BL21(DE3). In silico analyses of the deduced gene product suggested that it is a member of the family 1 glycosyltransferases. The recombinant protein MrSGT was able to catalyze the transfer of a glucosyl moiety onto the C-3 hydroxy function in sterols (β-sitosterol, campesterol, and cholesterol), resulting in the corresponding steryl glucosides (β-sitosterol-3-O-β-ᴅ-glucoside, campesterol-3-O-β-ᴅ-glucoside, and cholesterol-3-O-β-ᴅ-glucoside). This enzyme prefers phytosterols to cholesterol, and also shows substrate flexibility to some extent, in that it could recognize a number of acceptor substrates.

• Journal of Microbiology and Biotechnology
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• v.19 no.2
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• pp.121-127
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• 2009

Sequencing analysis of a 5-kb DNA fragment from Streptomyces venezuelae ATCC 15439 revealed the presence of one 3.1-kb open reading frame(ORF), designated as afsR-sv. The deduced product of afsR-sv(1,056 aa) was found to have high homology with the global regulatory protein AfsR. Homology-based analysis showed that aftR-sv represents a transcriptional activator belonging to the Streptomyces antibiotic regulatory protein(SARP) family that includes an N-terminal SARP domain containing a bacterial transcriptional activation domain(BTAD), an NB-ARC domain, and a C-terminal tetratricopeptide repeat domain. Gene expression analysis by reverse transcriptase PCR(RT-PCR) demonstrated the activation of transcription of genes belonging to pikromycin production, when aftR-sv was overexpressed in S. venezuelae. Heterologous expression of the aftR-sv in different Streptomyces strains resulted in increased production of the respective antibiotics, suggesting that afsR-sv is a positive regulator of antibiotics biosynthesis.

• Journal of the Korean Magnetic Resonance Society
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• v.6 no.1
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• pp.69-77
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• 2002

During the screening of material which has the antimicrobial activity against aminoglycoside-resistant bacteria, A new material $\varepsilon$-(L-$\beta$-Iysine) polypeptide from a culture medium of Streptomyces sp.(DWGS2) was isolated, and the structure and the physicochemical properties of the new material were elucidated. The new material was separated by column chromatography of the culture medium using Dowex1$\times$2, Silica gel, and Sephadex LH20 etc. The chemical structure and molecular weight were determined with the data of various NMR experiments, MALDI mass, and ESI mass experiments. The antimicrobial activity of $\varepsilon$-(L-$\beta$-Iysine) polypeptide is not only better than equal to the activity of known aminoglycoside type of antibiotics(MIC=3.125 - 6.25ug/mL) but also effective against aminoglycoside-resistant bacteria and fungi. If the mechanism of antimicrobial activity against aminoglycoside- resistant bacteria is figured out, the $\varepsilon$-(L-$\beta$-Iysine) polypeptide can be utilized for the treatment of diseases caused by aminoglycoside-resistant bacteria.

• Journal of Microbiology and Biotechnology
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• v.11 no.5
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• pp.804-811
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• 2001

In many Gram-negative bacteria, autoinducers, such as N-acyl-L-homoserine lactone(acyl-HSL) and its derivative molecules, mediate the cell-density-dependnet expression of certain operons. The current study identified the autoinducers produced by Burkholderia cepacia G4, a trichloroethylene-degrading lagoon isolate, using TLC bioassays with Agrobacterium tumefaciens NT1(pDCI141E33) and Chromobacterium violaceum CVO26, and a GC-MS analysis. The ${R_f}\;and\;{R_t}$ values and mass spectra were compared with those of synthetic compounds. Based on the analyses, it was confirmed that G4 produces N-hexanoyl (C6)-, N-octanoyl (C8)-, N-decanoyl (C10)-, N-dodecanoyl (C12)-HSL, and an unknown active species. The integration of the GC peak areas exhibited a ratio of C8-HSL:C10-HSL:C12-HSL at 3:17:1 with C6-HSL and C10-HSL production at trace and micromolar levels, respectively, in the culture supernatants. Nutants partially defective in producing acyl-HSLs were also partially defective in the biosynthesis of an antibiotic substance. These results indicate that the autoinducer-dependent gene regulation in G4 is dissimilar to the clinical B. cepacia strains isolated from patients with cystic fibrosis.

• Journal of Microbiology and Biotechnology
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• v.15 no.3
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• pp.491-496
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• 2005

The biosynthetic gene cluster for the aminoglycoside antibiotic kasugamycin was isolated and characterized from the kasugamycin producing strain, Streptomyces kasugaensis KACC 20262. By screening a fosmid library using kasA, the gene encoding aminotransferase, we isolated a 22 kb DNA fragment. The fragment contained seventeen complete open reading frames (ORFs); one of these ORFs, kasD, was identified as the gene for dNDP-glucose 4,6-dehydratase, which catalyzes the conversion of dNDP-glucose to 4-keto-6-deoxy-dNDP-glucose. The enzyme showed a broad spectrum of substrate specificity. In addition, ksR was overexpressed in E. coli BL21 and proved to be a self-resistance gene against kasugamycin. These findings suggest that the isolated gene cluster is highly likely responsible for the biosynthesis of kasugamycin.

• Journal of Microbiology
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• v.35 no.1
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• pp.40-46
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• 1997

The pWHM220 cosmid with a 24-kb insert cloned from Streptomyces albus ATCC 21838 induces the biosynthesis of a polysther antibiotic similar to salinomycin in Streptomyces invidans. We have analyzed this region by DNA sequencing as well as Southern blot hybridization with type I and type II polyketide synthase (PKS) probes. Surprisingly, we found another set of type II SKS genes only 10-kb from the original PKS genes, salABCDE. The DNA sequence revealed two complete open reading frames (ORFs) named salB2 and salC2, and one partial ORF that does not resemble any known DNA or deduced protein sequence. The salC2 should code for chain length determining factor while the deduced amino acid sequence encoded by salB2 exhibits high similarity to .betha.-ketoacyl synthase from different PKS gene clusters. The highest identity was found for .betha.-keetoacyl synthases from S. argillaceus (MtmP. 59.1% identity), the mithramycin producer and from S. venezuelae ISP5230 (JadA, 52.3% identity), the jadomycin producer. The SalC2 protein clearly resembles its counterparts in order aromatic PKS gene clusters that are believed to influence the length of the polyketide chain. The highest identities observed were to that of S. argillaceus (MtmK, 62.3%) and S. venezuelae ISP 5230 (JadB, 55.1%) proteins, Moreover, the deduced amino acid sequences of the salB2 and salC2 products were 29.0% identical.

• Journal of Microbiology and Biotechnology
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• v.33 no.3
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• pp.410-418
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• 2023

Bacilysin is a dipeptide antibiotic composed of L-alanine and L-anticapsin produced by certain strains of Bacillus subtilis. Bacilysin is gaining increasing attention in industrial agriculture and pharmaceutical industries due to its potent antagonistic effects on various bacterial, fungal, and algal pathogens. However, its use in industrial applications is hindered by its low production in the native producer. The biosynthesis of bacilysin is mainly based on the bacABCDEF operon. Examination of the sequence surrounding the upstream of the bac operon did not reveal a clear, strong ribosome binding site (RBS). Therefore, in this study, we aimed to investigate the impact of RBS as a potential route to improve bacilysin production. For this, the 5' untranslated region (5'UTR) of the bac operon was edited using the CRISPR/Cas9 approach by introducing a strong ribosome binding sequence carrying the canonical Shine-Dalgarno sequence (TAAGGAGG) with an 8 nt spacing from the AUG start codon. Strong RBS substitution resulted in a 2.87-fold increase in bacilysin production without affecting growth. Strong RBS substitution also improved the mRNA stability of the bac operon. All these data revealed that extensive RBS engineering is a promising key option for enhancing bacilysin production in its native producers.

• Journal of Life Science
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• v.33 no.10
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• pp.842-850
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• 2023

Pyrrolnitrin, pyrrolomycin, and pyoluteorin are functional halogenated phenylpyrrole derivatives (HPDs) derived from microorganisms with diverse antimicrobial activities. Pyrrolnitrin is a secondary metabolite produced from L-tryptophan through four-step reactions in Pseudomonas fluorescens, Burkholderia cepacia, Serratia plymuthica, etc. It is currently used for the treatment of superficial dermatophytic fungal infections, has high antagonistic activities against soil-borne and foliar fungal infections, and has many industrial applications. Since pyrrolnitrin is easily decomposed by light, it is difficult to widely use it outdoors. As an alternative, fludioxonil, a synthetically produced non-systemic surface fungicide that is structurally similar and has excellent light stability, has been commercialized for seed and foliar treatment of plants. However, due to its high toxicity to aquatic organisms and adverse effects in human cell lines, many countries have established maximum residue levels and strictly control its levels. Pyrrolomycin and pyoluteorin, which have antibiotic/antibiofilm activity against Gram-positive bacteria and high anti-oomycete activity against the plant pathogen Pythium ultimum, respectively, were isolated and identified from microorganisms. This review summarizes the biosynthesis and production of natural pyrrolnitrin derived from bacteria and the characteristics of synthetic fludioxonil and other natural phenylpyrrole derivatives among the HPDs. We expect that a plethora of highly effective, novel HPDs that are safe for humans and environments will be developed through the generation of an HPD library by microbial biosynthesis and chemical synthesis.

• Journal of Aquaculture
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• v.19 no.4
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• pp.225-230
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• 2006

A long-term (26 weeks) feeding experiment was conducted to examine the essentiality and requirement of inositol in diets for olive flounder because no information is available in the species. Five casein-gelatin based semi-purified diets were formulated to contain four different levels of myo-inositol (0, 0+antibiotic, 400, 800, and 1600 mg/kg, designated as M0, M0+, M400, M800, and M1600, respectively). One (M0+) of the control diets contained tetracycline hydrochloride (0.4%, wt/wt) as an antibiotic to inhibit biosynthesis of inositol by micro-organism in intestine of fish. Olive flounder at the early juvenile stage (initial body weight 1.22 g) were randomly distributed into fifteen 35 L tanks (48 fish/tank) and fed with one of the experimental diets (3 replicates per diet). At the end of the feeding trial, the weight gain, feed intake, specific growth rate, and protein efficiency ratio of fish fed diets containing higher levels of myo-inositol (M800 & M1600) were significantly higher than those of fish fed the other diets (P<0.05). Feed conversion ratio, survival, hematocrits, and hemoglobin of fish fed experimental diets were not significantly different among all the fish groups. Whole body compositions of fish were not different except for lipid content. The lipid content was significantly different between M0 and M400 diet groups. These results indicate that juvenile olive flounder requires dietary supplementation of myo-inositol in diets for normal growth and its optimum level seems to be approximately 800 mg myo-inositol/kg diet.