• Journal of Life Science
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• v.25 no.12
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• pp.1458-1469
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• 2015

Many organic solvent-tolerant bacteria have been isolated from all environments such as soil, waste-water, even deep sea after first isolation report of organic solvent-tolerant bacterium. Most organic solvent- tolerant isolates have been determined to be Gram-negative bacteria, because Gram-negative bacteria have inherent tolerance property toward hostile organic solvents more than Gram-positive bacteria. The mechanisms of organic solvent tolerance have been elucidated extensively using mainly organic solvent-tolerant Gram-negative bacteria. The solvent-tolerance mechanisms in Gram-positive bacteria can be found in comparatively recent research. Organic solvents exhibited different toxicity depending on the solvent, and the tolerance levels of organic solvent-tolerant bacteria toward organic solvents were also highly changeable among species and strains. Therefore, organic solvent-tolerant bacteria could coped with solvent toxicity and adapted to solvent stress through the multifactorial and multigenic adaptative strategies. They could be survived even in the hyper concentrations of organic solvents by mechanisms which include: changes in cell morphology and cell behaviour, cell surface modifications, cell membrane adaptations, solvent excretion pumps, chaperones and anti-oxidative response. The aim of this work is to review the representative solvent tolerant bacteria and the adaptative and tolerance strategies toward organic solvents in organic solvent-tolerant bacteria, and their potential industrial and environmental impact.

• Journal of Life Science
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• v.27 no.11
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• pp.1381-1392
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• 2017

This review described solvent-tolerant lipases and their potential industrial, biotechnological and environmental impacts. Although organic solvent-tolerant lipase was first reported in organic solvent-tolerant bacterium, many organic solvent-tolerant lipases are in not only solvent-tolerant bacteria but also solvent-intolerant bacterial and fungal strains, such as the well-known Bacillus, Pseudomonas, Streptomyces and Aspergillus strains. As these lipases are not easily inactivated in organic solvents, there is no need to immobilize them in order to prevent an enzyme inactivation by solvents. Therefore, the solvent-tolerant lipases have the potential to be used in many biotechnological and biotransformation processes. With the solvent-tolerant lipases, a large number insoluble substrates become soluble, various chemical reactions that are initially impossible in water systems become practical, synthesis reactions (instead of hydrolysis) are possible, side reactions caused by water are suppressed, and the possibility of chemoselective, regioselective and enantioselective transformations in solvent and non-aqueous systems is increased. Furthermore, the recovery and reuse of enzymes is possible without immobilization, and the stabilities of the lipases improve in solvent and non-aqueous systems. Therefore, lipases with organic-solvent tolerances have attracted much attention in regards to applying them as biocatalysts to biotransformation processes using solvent and non-aqueous systems.

• Journal of Life Science
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• v.29 no.12
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• pp.1401-1407
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• 2019

To elucidate whether or not solvent-tolerant bacteria use anti-oxidative defense systems to defend themselves against toxic solvents, oxidative enzyme activity and total anti-oxidative capacity (T-AOC) were investigated in two tolerant strains of Pseudomonas sp. under toluene stress. The superoxide dismutase (SOD) activities of solvent tolerant BCNU 106 exhibited relatively increased levels at a toluene concentration of 100 mg/l, where those of solvent tolerant BCNU 171 increased at 200 mg/l. A greater than three-fold increase in catalase (CAT) levels was observed at concentrations of 200 and 300 mg/l in BCNU 106, and a two-fold increase was monitored at the same concentrations in BCNU 171. High glutathione S-transferase (GST) levels were also observed in the solvent tolerant bacteria. Higher levels of T-AOC was expressed in the solvent tolerant strains than in the ordinary non-tolerant KACC 10266. The highest plateau of SOD in BCNU 171 was observed at 1 hr of toluene exposure. CAT levels plateaued at 1 hr and 14 hr in BCNU 106 and reached the highest plateau at 3 hr in BCNU 171. The highest peak of T-AOC occurred at 9 hr in BCNU 106, and two high peaks occurred in BCNU 171, at 1 hr and at 9 hr of toluene exposure. The solvent-tolerant bacteria showed active antioxidant responses and could survive under harsh environments, including the presence of solvents, through means of antioxidant defense systems.

• KSBB Journal
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• v.27 no.1
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• pp.61-66
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• 2012

In the screening process of organic solvent tolerant bacteria showing good growth in media containing several kinds of organic solvents, one strain was isolated and identified as Bacillus sp. BCNU 5006. The strain was able to tolerate many organic solvents including benzene, toluene, xylene, octane, dodecane, butanol and ethylbenzene. Likewise, it could also utilize these solvents as the sole source of carbon with significant enzyme production. The lipolytic enzyme stability of Bacillus sp. BCNU 5006 was studied in the presence of several kinds of solvents at a 25% (v/v) concentration. The highest enzyme stability was observed in the presence of octane (107%), followed by ethylbenzene (88%), decane (86%), and chloroform (85%). Especially, BCNU 5006 lipase was determined to be more stable than immobilized enzyme (Novozyme 435) in the presence of octane, chloroform and xylene. This organic solvent tolerant Bacillus sp. BCNU 5006 could be expected as a potential bioremediation agent and biocatalyst for biodegradation and provide on organic-solvent-based enzymatic synthetic method in industrial chemical processes.

• Journal of Life Science
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• v.21 no.5
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• pp.700-705
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• 2011

Using enrichment procedures, we isolated organic solvent-tolerant Bacillus sp. BCNU 5005 from waste water and soil in the Ulsan industrial plant region. BCNU 5005 had a maximum similarity of 98% with B. subtilis and was designated as B. subtilis based on phylogenetic analyses using 16S rDNA sequences. Generally, most bacteria and their enzymes are destroyed or inactivated in the presence of high concentrations of organic solvents. However, the lipase activity of B. subtilis BCNU 5005 was very stable in the presence of various kinds of solvents (25%, v/v) except chloroform, ethylbenzene and decane. Furthermore, BCNU 5005 was determined to have a degradative ability towards organic solvents. This organic solvent tolerant Bacillus sp. BCNU 5005 could be used as a new potential resource for biotransformation and bioremediation.

• Journal of Microbiology
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• v.44 no.6
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• pp.583-590
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• 2006

In this study, an organic solvent tolerant bacterial strain was isolated. This strain was identified as Pseudomonas sp. strain S5, and was shown to degrade BTEX (Benzene, Toluene, Ethyl-Benzene, and Xylene). Strain S5 generates an organic solvent-tolerant lipase in the late logarithmic phase of growth. Maximum lipase production was exhibited when peptone was utilized as the sole nitrogen source. Addition of any of the selected carbon sources to the medium resulted in a significant reduction of enzyme production. Lower lipase generation was noted when an inorganic nitrogen source was used as the sole nitrogen source. This bacterium hydrolyzed all tested triglycerides and the highest levels of pro-duction were observed when olive oil was used as a natural triglyceride. Basal medium containing Tween 60 enhanced lipase production to the most significant degree. The absence of magnesium ions ($Mg^{2+}$) in the basal medium was also shown to stimulate lipase production. Meanwhile, an alkaline earth metal ion, $Na^+$, was found to stimulate the production of S5 lipase.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.474-477
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• 2000

An organic solvent-tolerant Pseudomonas savastanoi strain BCNU 106 was isolated.. This bacterium was gram negative rod, and showed oxidase positive, catalase positive, and decaboxylase positive reactions. The Pseudomonas strain oxidized cholesterol on the medium supplemented with an organic solvent. The strain BCNU 106 is able to grow in presence of organic solvents of which log $P_{ow}$ is between 1.5 to 7.0. The strain was well grown in a medium supplemented with a 10% volume of a organic solvent (toluene, p-xylene, cyclohexane, mesitlylene) containing $cholesterol(20mg/m{\ell)$. Major conversion compound were 7-dehydrocholesteryl dimethylphosphate, cholesta-4,7-dien-3-one, and cholesta-3,5-dieone.

• Journal of Life Science
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• v.23 no.10
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• pp.1246-1251
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• 2013

An organic solvent-tolerant lipase of Pseudomonas sp. BCNU 154 that was isolated from wastewater in the industrial complex region had optimal activity at $37^{\circ}C$ and pH 8. This crude extracellular lipase from BCNU 154 exhibited maximum stability in toluene, retaining about 6.01 U/ml (117.53%) activity for 2 h. $Ca^{2+}$, $Mg^{2+}$, $NH_4{^+}$, and $Na^+$ ions and triton X-100 activated the enzymes, whereas $Ba^{2+}$, $Hg^{2+}$, and $Zn^{2+}$ ions inhibited their activity. Pseudomonas sp. BCNU 154 lipase revealed stable activity comparable to that of the commercial immobilized Novozym 435. Thus, this organic solvent-tolerant lipase could have potential as a whole cell biocatalyst in industrial chemical processes without the use of immobilization.

• Journal of Microbiology and Biotechnology
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• v.20 no.5
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• pp.881-888
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• 2010

solvent-tolerant bacterium strain, MH6, was isolated by hydrophilic organic solvent DMSO enrichment in the medium and identified as Serratia marcescens. The extracellular protease with novel organic-solvent-stable properties from strain MH6 was purified and characterized. The molecular mass of the purified protease was estimated to be 52 kDa on SDS-PAGE. The open reading frame (ORF) of the MH6 protease encoded 504 amino acids with 471 amino acid residues in the mature protease. Based on the inhibitory effects of EDTA and 1,10-phenathroline, the MH6 protease was characterized as a metalloproteinase. The enzyme activity was increased in the presence of $Ni^{2+}$, $Mg^{2+}$, and $Ca^{2+}$. The protease could also be activated by the nonionic surfactants Tween 80 (1.0%) and Triton X-100 (1.0%). The protease showed remarkable solvent stability in the presence of 50% (v/v) solutions of long-chain alkanes and long-chain alcohols. It was also fairly stable in the presence of 25% solutions of hydrophilic organic solvents. Owing to its high stability in solvents and surfactants, the MH6 protease is an ideal candidate for applications in organic catalysis and other related fields.

• Journal of Microbiology and Biotechnology
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• v.28 no.9
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• pp.1502-1510
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• 2018

Organic solvent-tolerant (OST) enzymes are widely applied in various industries for their activity and stability in organic solvents, for their higher substrate solubility, and for their greater stero-selectivity. However, the criteria for identifying OST enzymes largely remain undefined. In this study, we compared the amino acid composition of 19 OST esterases with that of 19 non OST esterases. OST esterases have increased the ratio of Ala and Arg residues and decreased the ratio of Asn, Ile, Tyr, Lys, and Phe residues. Based on our amino acid composition analysis, we cloned a carboxylesterase (EstSP2) from a psychrophilic bacterium, Sphingomonas glacialis PAMC 26605, and characterized its recombinant protein. EstSP2 is a substrate specific to p-nitrophenyl acetate and hydrolyzed aspirin, with optimal activity at $40^{\circ}C$; at $4^{\circ}C$, the activity is approximately 50% of its maximum. As expected, EstSP2 showed tolerance in up to 40% concentration of polar organic solvents, including dimethyl sulfoxide, methanol, and ethanol. The results of this study suggest that selecting OST esterases based on their amino acid composition could be a novel approach to identifying OST esterases produced from bacterial genomes.