• Journal of Microbiology and Biotechnology
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• v.33 no.10
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• pp.1257-1267
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• 2023

Although rational genetic engineering is nowadays the favored method for microbial strain improvement, building up mutant libraries based on directed evolution for improvement is still in many cases the better option. In this regard, the demand for precise and efficient screening methods for mutants with high performance has stimulated the development of biosensor-based high-throughput screening strategies. Genetically encoded biosensors provide powerful tools to couple the desired phenotype to a detectable signal, such as fluorescence and growth rate. Herein, we review recent advances in engineering several classes of biosensors and their applications in directed evolution. Furthermore, we compare and discuss the screening advantages and limitations of two-component biosensors, transcription-factor-based biosensors, and RNA-based biosensors. Engineering these biosensors has focused mainly on modifying the expression level or structure of the biosensor components to optimize the dynamic range, specificity, and detection range. Finally, the applications of biosensors in the evolution of proteins, metabolic pathways, and genome-scale metabolic networks are described. This review provides potential guidance in the design of biosensors and their applications in improving the bioproduction of microbial cell factories through directed evolution.

• Microbiology and Biotechnology Letters
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• v.4 no.3
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• pp.105-110
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• 1976

In the studies of microbiological utilization of cellulosic wastes, cellulolytic fungi were isolated and screened out. At the first stage, 221 cellulolytic fungi were isolated from different sources such as soils, humus, composts and rotten wood debris by enrichment culture techniques. In the second stage, 36 strains of fungi out of those previously isolated were selected for their cellulase activities estimated by means of filter paper degradation, carboxy methyl cellulose liquefaction and cup method. Activities of C$_1$-cellulase, C$\sub$x/-cellulase and filter paper activity were adopted on the final screening stage and five different strains which are tentatively identified as Aspergillus sp.(strain No. AS-9), Penicillium sp. (strain No. KNI-1-2), Trichoderma, sp. (strain No. KI-7-2, KI-7-5, KI-4-1-1B) were selected for their high potency of C$_1$ and C$\sub$x/-cellulase activities. When rice straw milled and treated with NH$_4$OH was hydrolyzed with the crude enzyme Prepared from the culture broth of Trichoderma sp. (strain No. KI-4-1-1B), saccharification rate was obtained up to 26％.

• Journal of Microbiology and Biotechnology
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• v.20 no.3
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• pp.602-608
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• 2010

An atrazine-degrading bacterium, strain KU001, was obtained from a sugarcane field at the Cane and Sugar Research and Development Center at the Kasetsart University, Kamphaeng Saen Campus, Thailand. Strain KU001 had a rod-to-coccus morphological cycle during growth. Biolog carbon source analysis indicated that the isolated bacterium was Arthrobacter histidinolovorans. Sequence analysis of the PCR product indicated that the 16S rRNA gene in strain KU001 was 99% identical to the same region in Arthrobacter sp. The atrazine degradation pathway in strain KU001 consisted of the catabolic genes trzN, atzB, and atzC. Strain KU001 was able to use atrazine as a sole nitrogen source for growth, and surprisingly, atrazine degradation was not inhibited in cells grown on ammonium, nitrate, or urea, as compared with cells cultivated on growth-limiting nitrogen sources. During the atrazine degradation process, the supplementation of nitrate completely inhibited atrazine degradation activity in strain KU001, whereas ammonium and urea had no effect on atrazine degradation activity. The addition of strain KU001 to sterile or nonsterile soils resulted in the disappearance of atrazine at a rate that was 4- to 5-fold more than that achieved by the indigenous microbial community. The addition of citrate to soils resulted in enhanced atrazine degradation, where 80% of atrazine disappeared within one day following nutrient supplementation.

• Journal of Microbiology and Biotechnology
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• v.27 no.6
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• pp.1171-1179
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• 2017

Butanol is a promising alternative to ethanol and is desirable for use in transportation fuels and additives to gasoline and diesel fuels. Microbial production of butanol is challenging primarily because of its toxicity and low titer of production. Herein, we compared the transcriptome and phenome of wild-type Escherichia coli and its butanol-tolerant evolved strain to understand the global cellular physiology and metabolism responsible for butanol tolerance. When the ancestral butanol-sensitive E. coli was exposed to butanol, gene activities involved in respiratory mechanisms and oxidative stress were highly perturbed. Intriguingly, the evolved butanol-tolerant strain behaved similarly in both the absence and presence of butanol. Among the mutations occurring in the evolved strain, cis-regulatory mutations may be the cause of butanol tolerance. This study provides a foundation for the rational design of the metabolic and regulatory pathways for enhanced biofuel production.

• Korean Journal of Environmental Biology
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• v.36 no.2
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• pp.190-198
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• 2018

The petroleum industry is an important part of the world economy. However, the massive exposure of petroleum in nature is a major cause of environmental pollution. Therefore, the microbial mediated biodegradation of petroleum residues is an emerging scientific approach used to resolve these problem. Through the screening of diesel contaminated soil we isolated a rapid phenanthrene and a diesel degrading bacterium identified as Enterobacter cancerogenus DA1 strain through 16S rRNA gene sequence analysis. The strain was registered in NCBI with an accession number MG270576. The optimal growth condition of the DA1 strain was determined at pH 8 and $35^{\circ}C$, and the highest degradation rate of the diesel was achieved at this condition. At the optimal condition, growth of the strain on the medium containing 0.05% phenanthrene and 0.1% of diesel-fuel was highest at 45 h and 60 h respectively after the incubation period. Biofilm formation was found significantly higher at $35^{\circ}C$ as compared to $30^{\circ}C$ and $40^{\circ}C$. Likewise, the lipase activity was found significantly higher at 48 h after the incubation compared to 24 h and 72 h. These results suggest that the Enterobacter cancerogenus DA1 could be an efficient candidate, for application through ecofriendly scientific approach, for the biodegradation of petroleum products like diesel.

• Journal of Life Science
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• v.11 no.1
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• pp.11-13
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• 2001

Several culture conditions affecting cellulose production by a newly isolated Acetobacter sp. A9 were examined by cultivating cells under shaking cultures. The inoculum size in the range of 1-10% (v/v) did not influence cellulose production. Maximum cellulose production was obtained with 200 rpm of agitation speed. The cells grown in the 75 ml of medium in a 250-ml conical flask produced the highest level of cellulose. The strain was able to produce cellulose at 25-3$0^{\circ}C$ with a maximum at 3$0^{\circ}C$. Cellulose production occurred at pH 4.5-7.5 with a maximum at pH6.5.

• Journal of Microbiology and Biotechnology
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• v.10 no.2
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• pp.187-194
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• 2000

An actinomycetes strain, T-2, which produces transglutaminase (EC 2.3.2.13), was isolated from soil and identified as belonging to the Actinomadura sp., based on taxonomc studies. The conditions for the transglutaminase production and its enzymatic properties were investigated. The optimum components for the transglutaminase production were 2% glucose, 1% polypeptone and soytone, and 0.1% MnCl2. The optimum pH and temperature of the enzyme reaction were pH 8.0 and $45^{\circ}C$, respectively. The enzyme was stable within the pH range of 5.0-9.0 and $30^{\circ}C-45^{\circ}C$. The novel enzyme required no calcium ions for its activity. This enzyme polymerized various proteins such as casien, soy protein, hemoglobin, egg white, gelatin, and soybean milk.

• Journal of Ginseng Research
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• v.8 no.1
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• pp.1-7
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• 1984

In order to confirm the antimutagenicity of ginseng extracts, mutagenicity of ginseng and several medicinal herbs of which extracts were used as drinks, was examined through the method of Ames test. The obtained results were as follows. 1. Strong mutagenicities for Salmonella tyhimurium TA98, TA100, TA1535 were observed in all sample herbs Paenonia aalbiflora, Rehmannia glutinosa, Astragalus membranaceus, Angelica acutiloba, Cnidium officinale, Laurus nobilis and Panax ginseng without S-9 mix metabolic activation. 2. In the case of S-9 mix metabolic activation, even in a low concentraction, the extracts of Angelica autilobu, Cnidium officinale and Paenonla albtilora showed also a high mutagenicities for the strain TA98 and TA1535. 3. Even in high concentration of ginseng extracts, mutagenicity was almost neglectable through the metabolic activation of S-9 mix, compared with other extracts of medicinal herbs.

• The Korean Journal of Mycology
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• v.3 no.2
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• pp.25-29
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• 1975

Twelve strains of wood decaying Basidiomycetes were tested for ability to decomposelignin in saw dust of Pine, Quelcus, Rawan and Zelkowa, wood and to transform inorganic nitrogen in the medium into organic crude protein. In this test, one strain of Pleurotus ostreatus showed to be 49% of its ability of lignin decomposition. The increasing rate of the crude protein went up to 26%. It is considered, therefore, saw dust can he a useful animal food by microbial treatment.

• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.563-566
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• 2012

To investigate the possibility of horizontal gene transfer between agricultural microorganisms and soil microorganisms in the environment, Bacillus subtilis KB producing iturin and the PGPR recombinant strain Pseudomonas fluorescens MX1 were used as model microorganisms. The soil samples of cucumber or tomato plants cultivated in pots and the greenhouse for a six month period were investigated by PCR, real-time PCR, Southern hybridization, and terminal restriction fragment length polymorphism (T-RFLP) fingerprinting. Our data from Southern blotting and T-RFLP patterns suggest that the model bacteria do not give significant impacts on the other bacteria in the pots and greenhouse during cultivation.