• KSBB Journal
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• v.25 no.6
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• pp.513-519
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• 2010

Marine bacterium producing pigment was isolated from the solar saltern of Mijo-myeon, Namhae, Korea. Based on phenotypic characteristics and 16S rRNA sequence analysis, the strain was identified as Kocuria sp., which produced a yellow pigment. The pigment showed UV absorption maximum at 469nm. The bacterial strain grew well on Marine broth 2216 culture medium. Productivity of the pigment reached the maximum value after 44 hours at $30^{\circ}C$, 2% NaCl and pH 6.0. The pigment was produced best when supplied by 1% lactose as a carbon source and 1% beef extract as a nitrogen source. The result of the color stability study showed that pigment extracted from the strain by ethanol was stable at $-20-25^{\circ}C$ and also showed higher stability over 70% for 14 days in light conditions at $25^{\circ}C$. The pigment extract was also stable for all metal ions tested, except for $FeCl_2$.

• Microbiology and Biotechnology Letters
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• v.45 no.3
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• pp.226-235
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• 2017

Carotenoids such as phytoene, lycopene, and ${\beta}-carotene$ are used as food colorants, animal feed supplements, and for human nutrition and cosmetic purposes. Previously, we reported the isolation of a novel marine bacterium, Kocuria gwangalliensis, which produces a pink-orange pigment. Phytoene desaturase (CrtI), encoded by the gene crtI, catalyzes lycopene formation from phytoene and is an essential enzyme in the early steps of carotenoid biosynthesis. CrtI is one of the key enzymes regulating carotenoid biosynthesis and has been implicated as a rate-limiting enzyme of the pathway in various carotenoid synthesizing organisms. Here, we report the cloning of the crtI gene responsible for lycopene biosynthesis from K. gwangalliensis. The gene consisted of 1,584 bases encoding 527 amino acid residues. The nucleotide sequence of the crtI gene was compared with that of other species, including Kocuria rhizophila and Myxococcus xanthus, and was found to be well conserved during evolution. An expression plasmid containing the crtI gene was constructed (pCcrt1), and Escherichia coli cells were transformed with this plasmid to produce a recombinant protein of approximately 57 kDa, corresponding to the molecular weight of phytoene desaturase. Lycopene biosynthesis was confirmed when the plasmid pCcrtI was co-transformed into E. coli containing the plasmid pRScrtEB carrying the crtE and crtB genes required for lycopene biosynthesis. The results from this study will provide valuable information on the primary structure of K. gwangalliensis CrtI at the molecular level.

• Journal of Digital Convergence
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• v.15 no.6
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• pp.339-344
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• 2017

In this study, microorganisms were isolated from nosocomial environment and are identified by biochemical analysis as the part of biochemical and technical convergence. Microorganisms were collected at intense care unit of general hospital located in Pyeongtak (2014.11.28. - 2014. 11. 30). Using a VITEK2 equipment of biochemical approaches, eleven microorganisms e.g., Micrococcus luteus (or M. lylae), Granulicatella adiacens (M. luteus or M. lylae), Staphylococcus caprae, Sphingomonas paucimobilis, Kocuria kristinae, G elegans, Aerococcus viridans (or Staphylococcus arlettae), Methylobacterium spp., Dermacoccus nishinomiyaensis (or Kytococcus sedentarius), Kocuria kristinae (or M. luteus, M. lylae), Pseudomonas oryzihabitans were identified. And then identified bacteria plates were applied with a plastic stick, so called with "FarmeTok (medistick/Puristic) to produce ClO2. ClO2-releasing plastic stick showed the very strong inhibition of bacterial growth with about 99.9%. There were no bacterial colonies on the ClO2-incubated plate. Taken together, it is suggested that chlorine dioxide should be very strong inhibitor to microorganisms of nosocomial infections.

• Food Science of Animal Resources
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• v.42 no.2
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• pp.252-265
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• 2022

Kocuria salsicia can survive in extreme environments and cause infections, including catheter-related bacteremia, in humans. Here, we investigated and evaluated the characteristics of nine K. salsicia strains (KS1-KS9) isolated from cheese brine from a farmstead cheese-manufacturing plant in Korea from June to December, 2020. Staphylococcus aureus American Type Culture Collection (ATCC) 29213 was used as a positive control in the growth curve analysis and biofilm-formation assays. All K. salsicia isolates showed growth at 15% salt concentration and temperatures of 15℃, 25℃, 30℃, 37℃, and 42℃. KS6 and KS8 showed growth at 5℃, suggesting that they are potential psychrotrophs. In the biofilm-formation analysis via crystal violet staining, KS6 exhibited the highest biofilm-forming ability at various temperatures and media [phosphate buffered saline, nutrient broth (NB), and NB containing 15% sodium chloride]. At 25℃ and 30℃, KS3, KS6, and KS8 showed higher biofilm-forming ability than S. aureus ATCC 29213. The antimicrobial resistance of the isolates was evaluated using the VITEK^® 2 system; most isolates were resistant to marbofloxacin and nitrofurantoin (both 9/9, 100%), followed by enrofloxacin (7/9, 77.8%). Five of the nine isolates (5/9, 55.6%) showed multidrug resistance. Our study reports the abilities of K. salsicia to grow in the presence of high salt concentrations and at relatively low temperatures, along with its multidrug resistance and tendency to form biofilms.

• Microbiology and Biotechnology Letters
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• v.44 no.3
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• pp.293-302
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• 2016

To determine the dominant bacterial species during the Saeu-jeotgal ripening process, the cultivable bacterial population was examined over a 135-day period using six different growth media. The greatest numbers of bacteria were identified when marine agar was used for culture, with maximum cell density identified at day 65 (2.51 × 10⁷ colony forming units/g). Over the course of 135 days, the bacterial diversity was analyzed eight times. A total of 467 isolates, comprising 87 species from 42 genera, as well as 16 isolates belonging to previously unknown species, were identified. The number of species detected decreased from 39 at day 1 to 13 at day 135. The order of dominance at the genus level was as follows: Staphylococcus, Salimicrobium, Kocuria, and Psychrobacter. Staphylococcus and Salimicrobium accounted for 2% of the diversity at day 1, and then increased to 39% and 36%, respectively, at day 135. The dominant species Staphylococcus equorum, Salimicrobium salexigens, and Kocuria palustris accounted for 23.6%, 16.1%, and 10.9% of all isolates, respectively. Importantly, both St. equorum and Sm. salexigens remained viable at a NaCl concentration of 21% (w/v), which indicates their strong involvement in the ripening of Saeu-jeotgal.

• Journal of Life Science
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• v.22 no.8
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• pp.1024-1033
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• 2012

A gene encoding a novel geranylgeranyl pyrophosphate (GGPP) synthase from Kocuria gwangalliensis has been cloned and expressed in Escherichia coli. The deduced amino acid sequence showed 59.6% identity with a putative GGPP synthase (CrtE) from K. rhizophila. An expression plasmid containing the crtE gene was constructed, and E. coli cells containing this plasmid produced a recombinant protein with a theoretical molecular mass of 41 kDa, corresponding to the molecular weight of GGPP synthase. Due to the lack of crtE, crtB, and crtI in E. coli, the biosynthesis of lycopene was only obtained when the plasmid pCcrtE was co-transformed into E. coli expressing the pRScrtBI-carrying carotenoid biosynthesis crtB and crtI genes, which were sub-cloned from Paracoccus haeundaensis. The biochemical studies on the expressed proteins were performed via HPLC. The results obtained from this study will provide a wider base of knowledge regarding the primary structure of CrtE cloned from K. gwangalliensis at the molecular level.

• Journal of Microbiology and Biotechnology
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• v.22 no.2
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• pp.244-247
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• 2012

To remediate lead (Pb)-contaminated soils, it is proposed that microbially induced calcite precipitation (MICP) would provide the best alternative to other remediation technologies. In this study, Pb bioremediation in soils was investigated using the calcite-precipitating bacterium Kocuria flava. Results indicate that the Pb is primarily associated with the carbonate fraction in bioremediated soil samples. The bioavailability of Pb in contaminated soil was reduced so that the potential stress of Pb was alleviated. This research provides insight into the geochemistry occurring in the MICP-based Pb-remediated soils, which will help in remediation decisions.

• Journal of Microbiology and Biotechnology
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• v.25 no.11
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• pp.1801-1809
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• 2015

A phytoene synthase gene, crtB, was isolated from Kocuria gwangalliensis. The crtB with 1,092 bp full-length has a coding sequence of 948 bp and encodes a 316-amino-acids protein. The deduced amino acid sequence showed a 70.9% identity with a putative phytoene synthase from K. rhizophila. An expression plasmid, pCcrtB, containing the crtB gene was constructed, and E. coli cells containing this plasmid produced the recombinant protein of approximately 34kDa , corresponding to the molecular mass of phytoene synthase. Biosynthesis of lycopene was confirmed when the plasmid pCcrtB was co-transformed into E. coli containing pRScrtEI carrying the crtE and crtI genes encoding lycopene biosynthetic pathway enzymes. The results obtained from this study will provide a base of knowledge about the phytoene synthase of K. gwangalliensis and can be applied to the production of carotenoids in a non-carotenoidproducing host.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.873-880
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• 2009

A novel xylanase gene, Kxyn, was cloned from Kocuria sp. Mn22, a bacteria isolated from the deep sea of the east Pacific. Kxyn consists of 1,170 bp and encodes a protein of 390 amino acids that shows the highest identity (63%) with a xylanase from Thermohifida fusca YX. The mature protein with a molecular mass of approximately 40 kDa was expressed in Escherichia coli BL21 (DE3). The recombinant Kxyn displayed its maximum activity at $55^{\circ}C$ and at pH 8.5. The $K_m,\;V_{max}$, and $k_{cat}$ values of Kxyn for birchwood xylan were 5.4 mg/ml, $272{\mu}mol/min{\cdot}mg$, and 185.1/s, respectively. Kxyn hydrolyzed birchwood xylan to produce xylobiose and xylotriose as the predominant products. The activity of Kxyn was not affected by $Ca^{2+},\;Mg^{2+},\;Na^+,\;K^+$, ${\beta}$-mercaptoethanol, DTT, or SDS, but was strongly inhibited by $Hg^{2+},\;Cu^{2+},Zn^{2+}$, and $Pb^{2+}$. It was stable over a wide pH range, retaining more than 80% activity after overnight incubation at pH 7.5-12. Kxyn is a cellulase-free xylanase. Therefore, these properties make it a candidate for various industrial applications.

• Korean Journal of Soil Science and Fertilizer
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• v.42 no.4
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• pp.257-265
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• 2009

The long-term effects of soil management history on microbial communities are still poorly understood. Our objectives were to determine the impact of long-term application of soil amendments on microbial communities in rice paddy fields. The treatments selected were control where crops were grown without any nutrient application (CON); nitrogen-phosphorus-potassium (NPK); NPK plus compost (CNPK); NPK plus lime (LNPK); and NPK plus silicate (WNPK). The long-term addition of organic and inorganic amendments significantly changed soil chemical properties. The amount of organic carbon increased in the treatments with fertilizer and amendments over that in the soil without inputs. However, we could not observe the differences of bacterial population among the treatments, but the number of aerobic bacteria increased by the addition of amendments. Isolates from the rice paddy soils before irrigation were Dactylosporangium, Ewingella, Geobacillus, Kocuria, Kurthia, Kytococcus, Lechevalieria, Micrococcus, Micromonospora, Paenibacillus, Pedobacter, Pseudomonas, Pseudoxanthomonas, Rhodococcus, Rothia, Sphingopyxis, Stenotrophomonas, and Variovorax. Dominant genera were Arthrobacter, Kocuria, Kurthia, and Bacillus in the long-term field. Microbial biomass was the highest in the compost treatment (CNPK), and was the lowest in the CON. Dehydrogenase activity in soils treated with rice compost straw was the highest and the activity showed an increasing trend according to treatment as follows: CON < WNPK < NPK = LNPK < CNPK. These results demonstrate that soil management practice, such as optimal application of fertilizer and amendment, that result in accumulations of organic carbon may increase microbial biomass and dehydrogenase activity in long-term rice paddy soils.