• Journal of Microbiology and Biotechnology
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• v.34 no.2
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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.

• Korean Journal of Environmental Agriculture
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• v.12 no.2
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• pp.144-152
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• 1993

A bacterium which degrades efficiently synthetic detergents was isolated from the polluted waters, activated sludge of wastewater treatment plants or polluted soil. This bacterium showed considerably higher growth rate in the agar plate containing $2,000{\mu}g/ml$ of synthetic detergents than any other isolated strains, was identified as a Pseudomonas fluorescens or strains similar to it. The strain was named as a Pseudomonas fluorescens S1. Optimum pH and temperature for the growth of the Pseudomonas fluorescens S1 were pH 7.0 and $30^{\circ}C$, respectively. The strain was resistant to streptomycin and gentamycin, but sensitive to kanamycin. The strain was greatly resistant to zinc chloride, lead nitrate and copper sulfate, but unable to grow in the presence of relatively low concentrations of mercury chloride and silver nitrate. This strain utilized benzene, catechol, cyclohexane and xylene as a sole carbon source. The strain was well grown in the medium containing ABS 10,000${\mu}g$/ml. Degradation of ABS was 55% and 60% at 20${\mu}g$/ml and 100${\mu}g$/ml of ABS, respectively. Benzene ring was degraded 45% in 100${\mu}g$/ml of ABS. During the incubation of the strain in the medium containing ABS 100${\mu}g$/ml and COD 10,000${\mu}g$/ml for 4 days, degradation of ABS and COD were reduced to 40${\mu}g$/ml and 3,200${\mu}g$/ml, respectively. Total amino acid content of the Pseudomonas fluorescens S1 grown with 1,000${\mu}g$/ml of ABS was 115mg/g cell, whereas its content was decreased in the bacterium grown without synthetic detergent by 9.4%.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.19 no.1
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• pp.331-336
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• 2018

Oral health is an essential requirement for maintaining and promoting health and quality of life while aging. It is known that there is a deep connection with hygienic management of dentures. However, despite the emphasis on the importance of clean management of dentures, there is a lack of quantitative research on denture-related detergents. Therefore, the purpose of this study is to evaluate food stain removal performance of three denture cleansers containing proteolytic enzymes. In addition, this study provides basic data on self-cleaning methods using detergents and suggests ways to improve effective treatment for oral health of elders in domiciliary care. In order to obtain quantitative data concerning food stain removal performance of each proteinase denture cleaner, the test evaluation method proposed by the Korean Consumer Agency, ISO 20795, was used. A total of 18 specimens were discolored for 48 hours in water dissolved with turmeric, and then washed with each denture cleaner for 24 hours. The cleaned denture specimens were measured quantitatively using a spectrophotometer to measure the color values of the denture specimens before the discoloration as well as the color values of the discolored specimens after cleaning with the denture cleanser. The experimental values were analyzed through one-way ANOVA with post-hoc Tukey's test, and statistically significant differences were found among the experimental groups. It was found that C had statistically significant stain removal performance compared with A and B(p<.05). All experiments were conducted for a total of 21 days, from July 3- 23, 2017. This study confirms that denture cleansers is a beneficial self-denture management method for domiciliary seniors. Therefore, it is necessary to extend accessibility for denture cleansers in order to to optimize domiciliary care for elderly oral hygiene and health.

• Korean Journal of Microbiology
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• v.54 no.1
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• pp.60-68
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• 2018

A cold-active and alkaline serine protease (Pro21717) was partially purified from the Antarctic marine bacterium Pseudoalteromonas arctica PAMC 21717. On a zymogram gel containing skim milk, Pro21717 produced two distinct clear-zones of approximately 37 kDa (low intensity) and 74 kDa (high intensity). These were found to have identical N-terminal sequences, suggesting they arose from an identical precursor and that the 37 kDa protease might homodimerize to the more active 74 kDa form of the protein. Pro21717 displayed proteolytic activity at $0-40^{\circ}C$ (optimal temperature of $40^{\circ}C$) and maintained this activity at pH 5.0-10.0 (optimal pH of 9.0). Notably, relative activities of 30% and 45% were observed at $0^{\circ}C$ and $10^{\circ}C$, respectively, in comparison to the 100% activity observed at $40^{\circ}C$, and this enzyme showed a broad substrate range against synthetic peptides with a preference for proline in the cleavage reaction. Pro21717 activity was enhanced by $Cu^{2+}$ and remained stable in the presence of detergent surfactants (linear alkylbenzene sulfonate and sodium dodecyl sulfate) and other chemical components ($Na_2SO_4$ and metal ions, such as $Ba^{2+}$, $Mg^{2+}$, $Ca^{2+}$, $Zn^{2+}$, $Fe^{2+}$, $K^+$, and $Na^{2+}$), which are often included in commercial detergent formulations. These data indicate that the psychrophilic Pro21717 has properties comparable to the well-characterized mesophilic subtilisin Carlsberg, which is commercially produced by Novozymes as the trademark Alcalase. Thus it has the potential to be used as a new additive enzyme in laundry detergents that must work well in cold tap water below $15^{\circ}C$.

• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.349-354
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• 2000

Abstract A facultative alkalophilic gram-negative Vibrio metschnikovii strain RH530, isolated from the wastewater, produced several alkaline proteases (VAP) including six alkaline serine proteases and a metalloprotease. From this strain, high yielding YAP mutants were isolated by NTG treatment. The isolated mutant KS1 showed nine times more activity than the wild-type after optimization of the culture media. The production was regulated by catabolite repression when glucose was added to the medium. The effects of several organic nitrogen sources on the production of the YAP were investigated to avoid catabolite repression. The combination of 4% wheat gluten meal (WGM), 1.5% cotton seed flour (eSF), and 5% soybean meal (SBM) resulted in the best production when supplemented with 1% NaCl. The YAP showed a resistance to surfactants such as $sodium-{\alpha}-olefin$ sulfonate (AOS), polyoxy ethylene oxide (POE), and sodium dodecyl sulfate (SDS), yet not to linear alkylbenzene sulfonate (LAS). However, the activity of the YAP was restored completely when incubated with LAS in the presence of POE or $Na_2SO_4$. The YAP was stable in a liquid laundry detergent containing 6.6% SLES (sodium lauryl ether sulfate), 6.6% LAS, 19.8% POE, and stabilizing agents for more than two weeks at $40^{\circ}C$, but the stability was sharply decreased even after 1 day when incubated at $60^{\circ}C$. A washing performance test with the YAP exhibited it to be a good washing power by showing 51 % and 60% activity at $25^{\circ}C{\;}and{\;}40^{\circ}C$, respectively, thereby indicating that the YAP also has a good detergency at a low temperature. All the results suggest that the YAP produced from the mutant strain KSI has suitable properties for use in laundry detergents.rgents.

• Journal of Distribution Science
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• v.17 no.10
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• pp.125-134
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• 2019

Purpose - Environment-friendly behaviors are diverse and can be motivated by different factors. For example, magnitude of factors affecting recycling behavior could be different from those affecting purchasing of environmentally-friendly products. In this study, the focus is on usage satisfaction of consumers who have purchased environmentally-friendly products. Research design, data, and methodology - For this purpose, data were collected from a total of 345 users of environmentally-friendly products, consisting of 177 users of electric vehicles and 168 users of environmentally-friendly detergents. The study analyzed the relationships among consumer's innovation propensity, consumer's environmental concern, and usage satisfaction. In addition, the moderating effect of the product's innovativeness was examined. Results - The result shows that there is a relationship between consumer's innovation propensity and consumer's environmental concern. In particular, it is shown that both consumer's innovation propensity and consumer's environmental concern have positive effects on usage satisfaction. Further, there was a moderating effect of the product's innovativeness. Namely, products rated higher in innovativeness showed a greater effect of consumer's innovation propensity on usage satisfaction. The degree of the product's innovativeness also affected the directional paths of the relationships. In the electric vehicle (i.e., high product innovativeness) case, consumer's innovation propensity had a direct effect on usage satisfaction, as well as an indirect effect through consumer's environmental concern. In the environmentally-friendly detergent (i.e., low product innovativeness) case, consumer's innovation propensity only had an indirect effect on usage satisfaction, through consumer's environmental concern. Conclusions - Theoretical contributions of this study are as follows. Foremost, this is the first study that suggests an existence of a relationship between consumer's innovation propensity and consumer's environmental concern. Second, this study showed the positive effect of consumer's innovation propensity on usage satisfaction of environmentally-friendly products. Third, this study expanded the scope of previous studies by showing the interaction between consumer's innovation propensity and consumer's environmental concern. Lastly, this study showed that the effect of these variables on usage satisfaction can differ by the degree of innovativeness of the environmentally-friendly product. The results of the study will have implications for marketers and policy makers in formulating marketing activities and policies.

• Applied Chemistry for Engineering
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• v.9 no.3
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• pp.413-418
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• 1998

Methoxy polyoxyethylene dodecanoates are a kind of nonionic surfactants obtainable from reaction of fatty acid methyl ester with ethylene oxide utilizing a solid catalyst. Methoxy polyoxyethylene dodecanoates have economical advantage compared with polyoxyethylene dodecyl ethers using fatty alcohol. In this work, the solubilizing capacity concerned with phase behavior of ternary systems composed of nonionic surfactant/water/oil, interfacial tension and detergency at the phase inversion temperature(PIT) were investigated and compared with those of polyoxyethylene dodecyl ethers in order to confirm the applicability of methoxy polyoxyethylene dodecanoates in the detergents. Methoxy polyoxyethylene dodecanoates showed the solubilizing capacity of 10~18% for hexadecane which were about 6% higher than polyoxyethylene dodecyl ethers. At the PIT condition, methoxy polyoxyethylene dodecanoates' interfacial tension were 0.0124~0.0176 dyne/cm while polyoxyethylene dodecyl ethers have the value of 0.013~0.0163 dyne/cm and methoxy polyoxyethylene dodecanoates showed higher detergency of 82.1~83.2% than polyoxyethylene dodecyl ethers of 76.5~77.3%. The good detergency performance of methoxy polyoxyethylene dodecanoates would be due to the higher oil solubilizing power and lower interfacial tension than polyoxyethylene dodecyl ethers at the PIT condition.

• Applied Biological Chemistry
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• v.38 no.4
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• pp.320-324
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• 1995

Solubilization of microsomal ${\Delta}^{5}-3{\beta}$-hydroxy steroid acyl transfearse(${\Delta}^{5}-3{\beta}$-OH-SAT) of rat brain and its reconstitution into liposomes were investigated. Among the detergents utilized for the solubilization, deoxycholic acid was superior to Tween 80 or Triton X-100 for the reconstituted activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. The enzyme activity was shown to be affected by the nature of phospholipids used for the preparation of the liposome. Phosphatidylcholines from egg yolk and soybean showed the highest activity of ${\Delta}^{5}-3{\beta}$-OH-SAT and phosphatidylethanolamine came next. However phosphatidylserine and phosphatidic acid showed a lower activity than those obtained before the reconstitution. This study suggests that the presence of quaternary ammonium salt or amine group in the phospholipids stimulates the activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. However the presence of a carboxylic group or the absence of the amine group may have an inhibitory effect on the ${\Delta}^{5}-3{\beta}$-OH SAT.

• Applied Biological Chemistry
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• v.42 no.4
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• pp.288-292
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• 1999

It was previously reported that cobalt(II) effectively suppresses the postharvest gravitropic response of snapdragon (Antirrhinum majus L.) (Kim et al. (1997) Agric. Chem. and Biotechnol. 40, 567-571). In this study, several factors affecting the suppression of the gravitropic response by cobalt ion were examined. When flower stalks of snapdragon were pretreated with several different cobalt salts, cobalt nitrate turned out to be the most effective not only in reducing the response but also in preserving the flower quality. We also tested the effects of various detergents which were added to cobalt(II) solution, finding that Tween-40 was the best among the tested with respect to the effectiveness as well as the flower quality. Based on these results, we optimized a protocol for the chemical treatment; that is, a suppressor solution containing 10 mM $CO(NO_3)_2$ and 0.05% Tween-40 was directly sprayed on the gravitropically sensitive region of cut flowers of snapdragon. The suppressor treatment gave rise to a significantly improved results when the flower stalks were stored at a lower temperature after the chemical treatment.

• Journal of Applied Biological Chemistry
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• v.54 no.2
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• pp.94-100
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• 2011

A gene encoding a putative phospholipase D was isolated from Bacillus licheniformis and cloned into pGEM-T easy vector. The gene was expressed in E. coli BL21 (DE3) using a pET-21(a) vector containing His6 tag. Affinity purification of the recombinant phospholipase D with nickel-nitrilotriacetic acid (Ni-NTA) resin resulted major one-band by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The purified enzyme showed a molecular weight of 44 kDa. The optimum activity of enzyme was around pH 7.0 and the enzyme was also the most stable around this condition. The optimum temperature was about $40-45^{\circ}C$ and the enzyme still showed considerable activities at wide range of temperature. Among various detergents, Triton X-100 significantly increased the enzyme activity, resulting in 181% activity of control at 0.6 mM of the detergent. Calcium ion did not significantly affect the enzyme activity, suggesting that the enzyme might be classified into $Ca^{2+}$-independent PLD.