• Journal of Microbiology and Biotechnology
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• 제34권2호
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• pp.436-456
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• 2024

Several thermostable proteases have been identified, yet only a handful have undergone the processes of cloning, comprehensive characterization, and full exploitation in various industrial applications. Our primary aim in this study was to clone a thermostable alkaline protease from a thermophilic bacterium and assess its potential for use in various industries. The research involved the amplification of the SpSKF4 protease gene, a thermostable alkaline serine protease obtained from the Geobacillus thermoglucosidasius SKF4 bacterium through polymerase chain reaction (PCR). The purified recombinant SpSKF4 protease was characterized, followed by evaluation of its possible industrial applications. The analysis of the gene sequence revealed an open reading frame (ORF) consisting of 1,206 bp, coding for a protein containing 401 amino acids. The cloned gene was expressed in Escherichia coli. The molecular weight of the enzyme was measured at 28 kDa using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The partially purified enzyme has its highest activity at a pH of 10 and a temperature of 80℃. In addition, the enzyme showed a half-life of 15 h at 80℃, and there was a 60% increase in its activity at 10 mM Ca²⁺ concentration. The activity of the protease was completely inhibited (100%) by phenylmethylsulfonyl fluoride (PMSF); however, the addition of sodium dodecyl sulfate (SDS) resulted in a 20% increase in activity. The enzyme was also stable in various organic solvents and in certain commercial detergents. Furthermore, the enzyme exhibited strong potential for industrial use, particularly as a detergent additive and for facilitating the recovery of silver from X-ray film.

• Journal of Microbiology and Biotechnology
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• 제25권5호
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• pp.559-568
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• 2015

Kimchi is a traditional Korean vegetable product that is naturally fermented by various microorganisms present in the raw materials. Among these microorganisms, lactic acid bacteria dominate the fermentation process. Natural fermentation with unsterilized raw materials leads to the growth of various lactic acid bacteria, resulting in variations in the taste and quality of kimchi, which may make it difficult to produce industrial-scale kimchi with consistent quality. The use of starter cultures has been considered as an alternative for the industrial production of standardized kimchi, and recent trends suggest that the demand for starter cultures is on the rise. However, several factors should be carefully considered for the successful application of starter cultures for kimchi fermentation. In this review, we summarize recent studies on kimchi starter cultures, describe practical problems in the application of industrial-scale kimchi production, and discuss the directions for further studies.

• Journal of Microbiology and Biotechnology
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• 제10권6호
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• pp.817-822
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• 2000

A proper flashing light is expected to enhance microalgal biomass productivity and photosynthetic efficiency. The effect of flashing light on high-density Chlorella kessleri (UTEX 398) cultures was studied using light-emitting diodes. A frequency modulator was designed to flash LEDs, and the device successfully provided wide range of frequencies and various duty cycles of flashing. A relatively high frequencies of 10, 20 and 50 kHz were used in this study. These frequencies have very short flashing time ($2-50{\mu}s$), which corresponded to the time constant of the light reaction of photosynthesis. The specific oxygen production rates of photosynthesis under flashing light were compared with those under an equivalent continuous light in specially designed illumination cuvette. The specific oxygen production rates under flashing light were 5-25% higher than those under the continuous light. A range of cell concentration was discovered, where the benefit of flashing light was maximized. The photosynthetic efficiency was also higher under flashing light with frequencies of over 1 kHz, which was a clear indication of flashing light effect and the degree of mutual shading could by overcome by flashing lights, particularly at high-density algal cultures.

• Journal of Microbiology and Biotechnology
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• 제28권3호
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• pp.409-417
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• 2018

Biologically produced 2,3-butanediol (2,3-BDO) has diverse industrial applications. In this study, schematic isolation and screening procedures were designed to obtain generally regarded as safe (GRAS) and efficient 2,3-BDO producers. Over 4,000 candidate strains were isolated by pretreatment and enrichment, and the isolated Bacillus strains were further screened by morphological, biochemical, and genomic analyses. The screened strains were then used to test the utilization of the most common carbon (glucose, xylose, fructose, sucrose) and nitrogen (yeast extract, corn steep liquor) sources for the economical production of 2,3-BDO. Two-stage fed-batch fermentation was finally carried out to enhance 2,3-BDO production. In consequence, a newly isolated Bacillus licheniformis GSC3102 strain produced 92.0 g/l of total 2,3-BDO with an overall productivity and yield of 1.40 g/l/h and 0.423 g/g glucose, respectively, using a cheap and abundant nitrogen source. These results strongly suggest that B. licheniformis, which is found widely in nature, can be used as a host strain for the industrial fermentative production of 2,3-BDO.

• Journal of Microbiology and Biotechnology
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• 제13권5호
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• pp.745-750
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• 2003

Zoogloea ramigera produces an extracellular polysaccharide called zooglan, which adsorbs heavy metals. In the current study, Zoogloea ramigera was cultured in media containing various carbon sources. When different carbon sources were included in the cultivation medium, there was a change in the composition of zooglan that is mainly composed of glucose, galactose, and pyruvic acid. The various zooglan compositions were analyzed by HPLC, and changes in the heavy metal (lead (II) and cadmium) adsorption characteristics relative to a change in the composition were examined using an atomic absorption spectrophotometer. A high adsorption capacity was observed at a pH higher than 3.0. The adsorption of metal ions was the highest at $35^{\circ}C$, and a higher adsorption was obtained with a lower flow rate. Changes in the zooglan composition did result in changes in the heavy metal adsorption characteristics. Furthermore, it was also found that the pyruvic acid content was more important than the glucose or galactose content for heavy metal adsorption.

• Journal of Microbiology and Biotechnology
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• 제15권2호
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• pp.337-345
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• 2005

The degradation of phenanthrene, a model PAH compound, by microorganisms either in the mixed culture or individual strain, isolated from oil-contaminated soil in oil refmery vicinity sites, was examined. The effects of pH, temperature, initial concentration of phenanthrene, and the addition of carbon sources on biodegradation potential were also investigated. Results showed that soil samples collected from four oil refinery sites in Korea had different degrees of PAH contamination and different indigenous phenanthrene-degrading microorganisms. The optimal conditions for phenanthrene biodegradation were determined to be 30$^{circ}C$ and pH 7.0. A significantly positive relationship was observed between the microbial growth and the rate of phenanthrene degradation. However, the phenanthrene biodegradation capability of the mixed culture was not related to the degree of PAH contamination in soil. In low phenanthrene concentration, the growth and biodegradation rates of the mixed cultures did not increase over those of the individual strain, especially IC10. High concentration of phenanthrene inhibited the growth of microbial strains and biodegradation of phenanthrene, but was less inhibitory on the mixed culture. Finally, when non-ionic surfactants such as Brij 30 and Brij 35 were present at the level above critical micelle concentrations (CMCs), phenanthrene degradation was completely inhibited and delayed by the addition of Triton X100 and Triton N101.

• Journal of Microbiology and Biotechnology
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• 제12권4호
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• pp.695-698
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• 2002

The biodegradation of 2,4,6-trinitrotoluene (TNT) was performed on a laboratory scale using P. putida originally isolated from explosive-contaminated soil. One hundred mg/1 of TNT was completely degraded within 20 h under optimum conditions. Various supplemental energy sources (carbon sources, nitrogen sources, and surfactant) were tested, with the main objective of identifying an inexpensive source and enhancing the degradation rate for large-scale biodegradation. Based on the degradation rate, molasses was selected as a possible supplemental carbon source, along with NH$_4$Cl and Tween 80 as a nitrogen source and surfactant, respectively. The degradation rate increased about 3.3 fo1d when supplemental energy sources were added and the degradation rate constant increased from 0.068 h$\^$-1/ to 0.224 h$\^$-1/. These results appear to be promising in application of the process to TNT-contaminated soil applications.

• Journal of Microbiology and Biotechnology
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• 제23권7호
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• pp.885-896
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• 2013

2,3-Butanediol (2,3-BDO) has immense industrial applications. Recently, microbial fermentation has emerged as an alternative way to produce this industrially important chemical. Although 2,3-BDO is produced by several microorganisms, the Klebsiella genera has an excellent production compared with other 2,3-BDO-producing microorganisms. In order to produce 2,3-BDO on a large scale, the challenges of removing pathogenic factors from Klebsiella pneumoniae need to be addressed. K. pneumoniae produces a number of virulence factors that contribute to its pathogenesis, including lipopolysaccharides, capsules, fimbrial adhesins, etc. Removal of these pathogenic factors from 2,3-BDO-producing Klebsiella strains will result in avirulent strains for the safe, economic, and efficient production of 2,3-BDO. In this review, we summarize the current trends in 2,3-BDO production using K. pneumoniae and insights into the removal of its virulence factors for industrial applications.

• Journal of Microbiology and Biotechnology
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• 제29권3호
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• pp.419-428
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• 2019

Phytases are enzymes that can hydrolyze phytate and its salts into inositol and phosphoric acid, and have been utilized to increase the availability of nutrients in animal feed and mitigate environmental pollution. However, the enzymes' low thermostability has limited their application during the feed palletization process. In this study, a combination of B-value calculation and protein surface engineering was applied to rationally evolve the heat stability of Escherichia coli phytase. After systematic alignment and mining for homologs of the original phytase from the histidine acid phosphatase family, the two models 1DKL and 1DKQ were chosen and used to identify the B-values and spatial distribution of key amino acid residues. Consequently, thirteen potential amino acid mutation sites were obtained and categorized into six domains to construct mutant libraries. After five rounds of iterative mutation screening, the thermophilic phytase mutant P56214 was finally yielded. Compared with the wild-type, the residual enzyme activity of the mutant increased from 20% to 75% after incubation at $90^{\circ}C$ for 5 min. Compared with traditional methods, the rational engineering approach used in this study reduces the screening workload and provides a reference for future applications of phytases as green catalysts.

• Journal of Microbiology and Biotechnology
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• 제18권12호
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• pp.1919-1926
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• 2008

Statistical experimental designs; involving (i) a fractional factorial design (FFD) and (ii) a central composite design (CCD) were applied to optimize the culture medium constituents for production of a unique antifreeze protein by the Antartic micro algae Chaetoceros neogracile. The results of the FFD suggested that NaCl, KCl, $MgCl_2$, and ${Na}_{2}{SiO}_{3}$ were significant variables that highly influenced the growth rate and biomass production. The optimum culture medium for the production of an antifreeze protein from C. neogracile was found to be Kalle's artificial seawater, pH of $7.0{\pm}0.5$, consisting of 28.566 g/l of NaCl, 3.887 g/l of $MgCl_2$, 1.787 g/l of $MgSO_4$, 1.308 g/l of $CaSO_4$, 0.832 g/l of ${K_2}{SO_4}$, 0.124 g/l of $CaCO_3$, 0.103 g/l of KBr, 0.0288 g/l of $SrSO_4$, and 0.0282 g/l of ${H_3}{BO_3}$. The antifreeze activity significantly increased after cells were treated with cold shock (at $-5^{\circ}C$) for 14 h. To the best of our knowledge, this is the first report demonstrating an antifreeze-like protein of C. neogracile.