• Title/Summary/Keyword: Substrate preference

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Isolation and Biochemical Characterization of Bacillus pumilus Lipases from the Antarctic

  • Arifin, Arild Ranlym;Kim, Soon-Ja;Yim, Joung Han;Suwanto, Antonius;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.5
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    • pp.661-667
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    • 2013
  • Lipase-producing bacterial strains were isolated from Antarctic soil samples using the tricaprylin agar plate method. Seven strains with relatively strong lipase activities were selected. All of them turned out to be Bacillus pumilus strains by the 16S rRNA gene sequence analysis. Their corresponding lipase genes were cloned, sequenced, and compared. Finally, three different Bacillus pumilus lipases (BPL1, BPL2, and BPL3) were chosen. Their amino acid sequence identities were in the range of 92-98% with the previous Bacillus pumilus lipases. Their optimum temperatures and pHs were measured to be $40^{\circ}C$ and pH 9. Lipase BPL1 and lipase BPL2 were stable up to $30^{\circ}C$, whereas lipase BPL3 was stable up to $20^{\circ}C$. Lipase BPL2 was stable within a pH range of 6-10, whereas lipase BPL1 and lipase BPL3 were stable within a pH range of 5-11, showing strong alkaline tolerance. All these lipases exhibited high hydrolytic activity toward p-nitrophenyl caprylate ($C_8$). In addition, lipase BPL1 showed high hydrolytic activity toward tributyrin, whereas lipase BPL2 and lipase BPL3 hydrolyzed tricaprylin and castor oil preferentially. These results demonstrated that the three Antarctic Bacillus lipases were alkaliphilic and had a substrate preference toward short- and medium-chain triglycerides. These Antarctic Bacillus lipases might be used in detergent and food industries.

Cloning, Expression, and Characterization of a Cold-Adapted Lipase Gene from an Antarctic Deep-Sea Psychrotrophic Bacterium, Psychrobacter sp. 7195

  • Zhang, Jinwei;Lin, Shu;Zeng, Runying
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.604-610
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    • 2007
  • A psychrotrophic strain 7195 showing extracellular lipolytic activity towards tributyrin was isolated from deep-sea sediment of Prydz Bay and identified as a Psychrobacter species. By screening a genomic DNA library of Psychrobacter sp. 7195, an open reading frame of 954 bp coding for a lipase gene, lipA1, was identified, cloned, and sequenced. The deduced LipA1 consisted of 317 amino acids with a molecular mass of 35,210 kDa. It had one consensus motif, G-N-S-M-G (GXSXG), containing the putative active-site serine, which was conserved in other cold-adapted lipolytic enzymes. The recombinant LipA1 was purified by column chromatography with DEAE Sepharose CL-4B, and Sephadex G-75, and preparative polyacrylamide gel electrophoresis, in sequence. The purified enzyme showed highest activity at $30^{\circ}C$, and was unstable at temperatures higher than $30^{\circ}C$, indicating that it was a typical cold-adapted enzyme. The optimal pH for activity was 9.0, and the enzyme was stable between pH 7.0-10.0 after 24h incubation at $4^{\circ}C$. The addition of $Ca^{2+}\;and\;Mg^{2+}$ enhanced the enzyme activity of LipA1, whereas the $Cd^{2+},\;Zn^{2+},\;CO^{2+},\;Fe^{3+},\;Hg^{2+},\;Fe^{2+},\;Rb^{2+}$, and EDTA strongly inhibited the activity. The LipA1 was activated by various detergents, such as Triton X-100, Tween 80, Tween 40, Span 60, Span 40, CHAPS, and SDS, and showed better resistance towards them. Substrate specificity analysis showed that there was a preference for trimyristin and p-nitrophenyl myristate $(C_{14}\;acyl\; groups)$.

Amorphous Cr-Ti Texture-inducing Layer Underlying (002) Textured bcc-Cr alloy Seed Layer for FePt-C Based Heat-assisted Magnetic Recording Media

  • Jeon, Seong-Jae;Hinata, Shintaro;Saito, Shin
    • Journal of Magnetics
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    • v.21 no.1
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    • pp.35-39
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    • 2016
  • $Cr_{100-x}Ti_x$ amorphous texture-inducing layers (TIL) were investigated to realize highly (002) oriented $L1_0$ FePt-C granular films through hetero-epitaxial growth on the (002) textured bcc-$Cr_{80}Mn_{20}$ seed layer (bcc-SL). As-deposited TILs showed the amorphous phase in Ti content of $30{\leq}x(at%){\leq}75$. Particularly, films with $40{\leq}x{\leq}60$ kept the amorphous phase against the heat treatment over $600^{\circ}C$. It was found that preference of the crystallographic texture for bcc-SLs is directly affected by the structural phase of TILs. (002) crystallographic texture was realized in bcc-SLs deposited on the amorphous TILs ($40{\leq}x{\leq}70$), whereas (110) texture was formed in bcc-SLs overlying on crystalline TILs (x < 30 and x > 70). Correlation between the angular distribution of (002) crystal orientation of bcc-SL evaluated by full width at half maximum of (002) diffraction (FWHM) and a grain diameter of bcc-SL indicated that while the development of the lateral growth for bcc-SL grain reduces FWHM, crystallization of amorphous TILs hinders FWHM. $L1_0$ FePt-C granular films were fabricated under the substrate heating process over $600^{\circ}C$ with having different FWHM of bcc-SL. Hysteresis loops showed that squareness ($M_r/M_s$) of the films increased from 0.87 to 0.95 when FWHM of bcc-SL decreased from $13.7^{\circ}$ to $3.8^{\circ}$. It is suggested that the reduction of (002) FWHM affects to the overlying MgO film as well as FePt-C granular film by means of the hetero-epitaxial growth.

Physical habitat and chemical water quality characteristics on the distribution patterns of ecologically disturbing fish (Largemouth bass and Bluegill) in Dongjin-River Watershed (동진강 수계에서 생태 교란어종(큰입배스, 불루길)의 분포에 대한 물리적 서식지 및 화학적 수질 특성)

  • Kang, Yu-Jin;Lee, Sang-Jae;An, Kwang-Guk
    • Korean Journal of Environmental Biology
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    • v.37 no.2
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    • pp.177-188
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    • 2019
  • The objective of this study was to analyze the distribution patterns of two exotic disturbing fish species (largemouth bass, Micropterus salmoides and bluegill, Lepomis macrochirus), their physical habitat features, and chemical water qualities from 13 sites of Dongjin River. The relationships between the fish distributions and water chemistry and physical habitat characteristics (i.e., bottom substrate) were evaluated by comparing dominant vs. absent regions of largemouth bass and bluegill. The relative abundance of the two species decreased sharply with decreased in the water quality in terms of BOD, PO4-P and pH, but not with NO3-N and conductivity. The bluegill showed no difference in habitat preference between the two regions with regard to the bottom structure, while the largemouth bass had significant differences in the bottom structures of silt, coarse gravels and boulders. The dominant species were Zacco platypus (14.6%), Lepomis macrochirus (14.0%), and Micropterus salmoides (9.8%) in the order of relative abundance, indicating that exotic species (24% in total) dominated the fish community, perhaps ensuing large ecological disturbances on the ecosystem. In conclusion, the results of this study suggest that the distribution of the two exotic species may be largely influenced by water quality of organic matter and nutrient pollutions.

Acyltransferases for production of industrial oils in transgenic plants (식물의 산업용 지방산 생산을 위한 오일합성 유전자의 기능과 이용 전망)

  • Kim, Hyun-Uk;Lee, Kyeong-Ryeol;Park, Jong-Sug;Roh, Kyung-Hee;Kim, Sun-Hee;Kim, Jong-Bum
    • Journal of Plant Biotechnology
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    • v.37 no.2
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    • pp.220-227
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    • 2010
  • Fatty acids in seed oil from plants are essential for human nutrients and have been used for industrial purpose. The growing demands of seed oil as food resources and feedstocks for industrial uses have attempted to modify fatty acid composition and to increase oil content in transgenic plants. However, production of unusual fatty acids in transgenic plants are limited, which is not synthesized the level same as original plants. This bottleneck was common for production of several unusual fatty acids in transgenic plants and suggests that there is different for substrate preference in oil metabolic pathway enzymes between host oil plants and original wild plants. Review of acyltransferases involved in acyl-editing and seed oil accumulation of oil plant and wild-plant producing unusual fatty acids will design strategies to maximize the production of unusual fatty acids in transgenic plants. In here, we identified eleven acyltransferase genes in castor based on sequence homology, which will be useful to increase hydroxy unusual fatty acids in transgenic plants.

Zooplankton Community Distribution in Aquatic Plants Zone: Influence of Epiphytic Rotifers and Cladocerans in Accordance with Aquatic Plants Cover and Types (수생식물이 발달된 습지에서 동물플랑크톤 군집 분포: 수생식물의 밀도 및 종류가 부착성 윤충류와 지각류에게 미치는 영향)

  • Choi, Jong-Yun;La, Geung-Hwan;Kim, Seong-Ki;Jeong, Kwang-Seuk;Joo, Gea-Jae
    • Korean Journal of Ecology and Environment
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    • v.46 no.1
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    • pp.86-93
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    • 2013
  • We monitored 32 wetlands in order to investigate the influence of aquatic plants on zooplankton density and diversity in the littoral zone in Gyeongsangnam-do from May to June in 2011. A total of 65 zooplankton species were identified in the study sites. Among them, the diversity of epiphytic zooplankton were higher (40 species) than planktonic zooplankton. Littoral zones of all wetlands were covered by various aquatic plants, and influenced the epiphytic zooplankton assemblages. Based on the data from $1{\times}1$ (m) quadrat sampling, epiphytic and planktonic rotifer density showed no significant relationships with macrophyte cover. However, the epiphytic cladocerans density significantly increased under high aquatic plant cover ($r^2=0.39$, p<0.05, n=32). Types of aquatic plants strongly influenced epiphytic zooplankton density. Upo and Jangcheok are locations which have well developed Phragmites communis and Ceratophyllum demersum communities in the littoral zone, and a higher density of epiphytic zooplankton was recorded on the surface of C. demersum. Especially, rotifers such as Lepadella, Monostyla and Testudinella showed obvious differences (One-way ANOVA, p<0.05 for all three species). This result suggests that epiphytic zooplankton have a substrate preference for larger surface areas, likely for adherence, on C. dimersum. In conclusion, the complex structure of the littoral plant community is expected to provide diverse refuge and microhabitats to epiphytic zooplankton.

Characterization of an Antarctic alkaline protease, a cold-active enzyme for laundry detergents (세탁세제 첨가용 효소 개발을 위한 남극 해양세균 유래 저온성 단백질분해효소의 특성 연구)

  • Park, Ha Ju;Han, Se Jong;Yim, Joung Han;Kim, Dockyu
    • 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 $15^{\circ}C$.

In vitro mass propagation and acclimatization of Haworthia truncata (하월시아 옥선(Haworthia truncata)의 기내 대량 증식 및 순화 조건 구명)

  • Kim, Youn Hee;Lee, Gee Young;Kim, Hye Hyeong;Lee, Jae Hong;Jung, Jae Hong;Lee, Sang Deok
    • Journal of Plant Biotechnology
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    • v.46 no.2
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    • pp.127-135
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    • 2019
  • The purpose of this study was to investigate suitable parts for callus induction and optimal concentrations of growth regulators, contained in the medium affecting shoot and rooting for in vitro mass production of Haworthia truncata. Leaves and flower bud showed 100% callus formation rate at NAA $1{\sim}2mgL^{-1}$ treatment, and NAA $1mgL^{-1}$ + TDZ $2mgL^{-1}$ treatment. The flower stalk showed 75% callus formation rate, at NAA $2mgL^{-1}$ + TDZ $2mgL^{-1}$ treatment in H. truncata. While the rate of callus formation was high in leaves and flower bud, leaves were the most efficient in obtaining most culture parts. Shoot induction rate from callus was highest, at NAA $0.1mgL^{-1}$ treatment in H. truncata. Additionally, the number of shoots formation was 66.3 shoots high, in NAA $1mgL^{-1}$ + BA $0.1mgL^{-1}$ treatment in H. truncata. In the case of acclimatization of regenerated plant, growth characteristics did not show significant difference (95%) shading with respect to the different ratio of substrate mixture, and it was determined that would be appropriate, considering plant height and appearance preference of H. truncata. It was established that optimization of culture condition, was responsible for mass propagation in vitro cultures of H. truncata.

Establishment of tissue culture and acclimatization method for in vitro mass propagation of Echeveria laui and Echeveria elegans (에케베리아 라우이(Echeveria laui)와 엘레강스(Echeveria elegans)의 대량증식을 위한 조직배양 및 순화 조건 확립)

  • Kim, Youn Hee;Lee, Gee Young;Kim, Hye Hyeong;Lee, Jae Hong;Jung, Jae Hong;Lee, Sang Deok
    • Journal of Plant Biotechnology
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    • v.46 no.1
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    • pp.22-31
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    • 2019
  • The objective of this study was to investigate the suitable parts for callus induction and optimal concentrations of growth regulators contained in the medium affecting shooting and rooting Echeveria laui and Echeveria elegans for in vitro mass production. To determine the suitable plant parts for callus induction, the leaves were divided into upper, medium and bottom parts and cultured on MS medium at different concentrations with $0{\sim}2mgL^{-1}\;NAA$ and $0{\sim}4 mgL^{-1}BA$. The upper and middle parts of leaves both showed 100% callus formation rate with $NAA\;1\;mgL^{-1}$ and $BA\;1\;mgL^{-1}$ treatment in E. laui. The middle parts of leaves showed 83.3% callus formation rate at $NAA\;2\;mgL^{-1}$ and BA 4 mgL-1 treatment in E. elegans. The shoot induction rate from callus was highest at $NAA\;0.1\;mgL^{-1}$ and $BA\;3\;mgL^{-1}$ treatment in E. laui and $NAA\;0.3\;mgL^{-1}$ in E. elegans. In addition, the number of shoots formation was 10.4 shoots high in $NAA\;1\;mgL^{-1}$ and $BA\;1\;mgL^{-1}$ treatment in E. laui and 12.0 shoots in most effective $NAA\;1\;mgL^{-1}$ and $BA\;0.1\;mgL^{-1}$ treatment in E. elegans. In the case of acclimatization of regenerated plant, growth characteristics did not show any significant difference (35 ~ 55%) shading with respect to the different ratio of substrate mixture, and it was determined that would be appropriate considered plant height and appearance preference of E. laui and E. elegans. It was established that the optimization of culture condition was responsible for the mass propagation in vitro cultures of E. laui and E. elegans.