• 제목/요약/키워드: Substrate preference

검색결과 59건 처리시간 0.02초

Impact of Lactic Acid and Hydrogen Ion on the Simultaneous Fermentation of Glucose and Xylose by the Carbon Catabolite Derepressed Lactobacillus brevis ATCC 14869

  • Jeong, Kyung Hun;Israr, Beenish;Shoemaker, Sharon P.;Mills, David A.;Kim, Jaehan
    • Journal of Microbiology and Biotechnology
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    • 제26권7호
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    • pp.1182-1189
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    • 2016
  • Lactobacillus brevis ATCC 14869 exhibited a carbon catabolite derepressed phenotype that has ability to consume fermentable sugars simultaneously with glucose. To evaluate this unusual phenotype under harsh conditions during fermentation, the effects of lactic acid and hydrogen ion concentrations on L. brevis ATCC 14869 were examined. Kinetic equations describing the relationship between specific cell growth rate and lactic acid or hydrogen ion concentration were deduced empirically. The change of substrate utilization and product formation according to lactic acid and hydrogen ion concentration in the media were quantitatively described. Although the simultaneous utilization has been observed regardless of hydrogen ion or lactic acid concentration, the preference of substrates and the formation of two-carbon products were changed significantly. In particular, acetic acid present in the medium as sodium acetate was consumed by L. brevis ATCC 14869 under extreme pH of both acid and alkaline conditions.

Bacillus subtilis의 mannanase에 의한 갈락토만난과 만노올리고당의 가수분해 (Hydrolysis of Galactomannan and Manno-oligosaccharides by A Bacillus subtiis Mannanase)

  • 권민아;윤기홍
    • 한국미생물·생명공학회지
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    • 제32권4호
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    • pp.347-351
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    • 2004
  • Hydrolysis of manno-oligosaccharides and galactomannan was studied with the purified Bacillus subtilis WL-7 mannanase from recombinant Eschericoli. The predominant products of hydrolysis were mannose, mannobiose and mannotriose. The enzyme could hydrolyze $\beta$-1 A-linked manno-oligosaccharides larger than mannobiose, but was not active on mannobiose. When the mannanase hydrolyzed manno-oligo saccharides of degree of polymerization(DP) 4-6, it was more active on the substrate of higher DP. Based on analysis of transient reaction products by TLC, the enzyme was found to have a preference for internal $\beta$-IA-mannosidic linkages, which are the central mannosidic bond of mannotetraose and the two middle mannosidic bonds of mannopentaose. The $\beta$-l A-mannosidic bonds situated at the second and fourth positions from the nonreducing end of mannohexaose were preferenhydrolyzed by the mannanase. Locust bean gum(LBG) was enzymatically hydrolyzed with higher efficiency than guar gum, resulting that amount of reducing sugars was liberated more efficiently from LBG than guar gum with same activity of mannanase.

Purification and Characterization of an α-D-Galactosidase from Grape Berry

  • Kang, Han-Chul;Kim, Tae-Su
    • Journal of Applied Biological Chemistry
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    • 제43권3호
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    • pp.141-146
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    • 2000
  • Glycosidase activities were tested from the grape berries, Vitis labruscana B. Takasumi. Among various glycosidases, $\alpha$-D-galactosidase was found to be the most active in the flesh and other glycosidases were considerably active in the order of the following: $\alpha$-D-mannosidase>$\alpha$-D-glucosidase>$\beta$-D-glucosidase>$\beta$-D-galactosidase. In the seeds, $\alpha$-D-glucosidase activity was the highest and other glycosidases such as $\alpha$-D-galactosidase, $\beta$-D-glucosidase, and $\beta$-D-galactosidase were still significantly active. The $\alpha$-D-galactosidase in the grape flesh was purified over 83-folds through salting-out with $(NH_4)_2SO_4$ and a series of chromatographies employing Sephadex G-50, Octyl-Sepharose, Q-Sepha- rose, and Biogel P-100. The enzyme was a monomer of 45 kDs as determined through SDS-PAGE and Sephacryl S-200 chromatography. The purified enzyme showed a preference of $\alpha$-D-galactose to $\beta$-D-galactose as a substrate about 5.4 times. Sulfhydryl specific reagents such as N-ethylmaleimide and iodoacetamide significantly inhibited the enzyme activity to the extents of 48 and 52% of its initial activity, respectively. The optimumpH range of $\alpha$-D-galactosidase was around 6.5-7.0. The enzyme activity increased by 46% in the presence of 1mM $Fe^{2+}$.

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The Purification and Characterization of a Bacillus stearothermophilus Methionine Aminopeptidase (MetAP)

  • Chung, Jae-Min;Chung, Il-Yup;Lee, Young-Seek
    • BMB Reports
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    • 제35권2호
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    • pp.228-235
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    • 2002
  • Methionine aminopeptidase (MetAP) catalyzes the removal of an amino-terminal methionine from a newly synthesized polypeptide. The enzyme was purified to homogeneity from Bacillus stearothermophilus (KCTC 1752) by a procedure that involves heat precipitation and four sequential chromatographs (including DEAE-Sepharose ion exchange, hydroxylapatite, Ultrogel AcA 54 gel filtration, and Reactive red 120 dye affinity chromatography). The apparent molecular masses of the enzyme were 81,300 Da and 41,000 Da, as determined by gel filtration chromatography and sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), respectively. This indicates that the enzyme is comprised of two identical subunits. The MetAP specifically hydrolyzed the N-terminal residue of Met-Ala-Ser that was used as a substrate, and exhibited a strong preference for Met-Ala-Ser over Leu-Gly-Gly, Leu-Ser-Phe, and Leu-Leu-Tyr. The enzyme has an optimal pH at 8.0, an optimal temperature at $80^{\circ}C$, and pI at 4.1. The enzyme was heat-stable, as its activity remained unaltered when incubated at $80^{\circ}C$ for 45 min. The Km and Vmax values of the enzyme were 3.0mM and 1.7 mmol/min/mg, respectively. The B. stearothernmophilus MetAP was completely inactivated by EDTA and required $Co^{2+}$ ion(s) for activation, suggesting the metal dependence of this enzyme.

Preference for Heated Substrate in Captive River Cooters (Pseudemys concinna): A Potential Use for the Control of Invasive Populations

  • Kang, Hakyung;Borzee, Amael;Chuang, Ming-Feng;Jang, Yikweon
    • Animal Systematics, Evolution and Diversity
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    • 제37권1호
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    • pp.9-14
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    • 2021
  • Invasive species threaten global biodiversity as well as human livelihood and much of the global lands are vulnerable to these threats. Numerous freshwater turtles from the northern hemisphere have been introduced in East Asian countries, including the Republic of Korea. Knowing turtle's behavioral ecology is valuable to manage introduced populations and a distinctive behavior is basking for behavioral thermoregulation. To understand the possibility of using basking to enhance trapping, we tested thermotaxis in the river cooter (Pseudemys concinna). Turtles were placed in an aquarium containing heated and non-heated mats under controlled water and air temperature, air humidity and light. We found that P. concinna stayed significantly longer on heated mats than on unheated control mats in 11 out of 18 trials, demonstrating that heat source is a potential attractant for P. concinna. We recommend the use of heat source to bait traps used for population control of invasive freshwater turtles.

Mortierella sp. 유래 ${\alpha}$-Galactosidase의 기질특이성 (Substrate Specificities of ${\alpha}$-Galactosidase from Mortierella sp.)

  • 박귀근
    • 한국미생물·생명공학회지
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    • 제39권3호
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    • pp.245-251
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    • 2011
  • Mortierella sp. 유래 효소 정제는 CM-sephadex C-50 column chromatography와 Sephadex G-100 column에 의해 수행하여 SDS-전기영동에서 단일밴드를 확인하였고 분자량은 57kDa로 결정되었다. melibiose, raffinose 및 stachyose의 세 종류의 기질에 대한 특이성에서는 Mortierella sp. 유래 정제 ${\alpha}$-galactosidase는 세 종류 기질의 비환원말단에 위치하고 있는 galactose를 모두 유리하는 특이성이 있음을 확인하였다. Bacillus sp. 유래의 $Gal^3Man_4$에 대해서 반응초기 3시간부터 가수분해가 진행되어 반응말기에서는 galactose, mannotetraose 그리고 분해되지 않고 일부 남아있는 $Gal^3Man_4$의 spot이 출현된 반면, 중합도 7의 $Gal^{2,3}Man_5$에 대해서는 반응초기부터 말기까지 전혀 특이성이 없음을 시사하였다. Trichoderma harzianum 유래의 $Gal^2Man_3$에 대해서는 반응초기 3시간부터 가수분해가 진행되어 일부 galactose와 mannotriose spot이 출현되고 있는 반면, 중합도 7의 $Gal^2Man_6$에 대해서는 Bacillus sp. 유래의 중합도 7의 $Gal^{2,3}Man_5$와 동일하게 galactose를 절단하는 능력이 없는 특이성을 보이고 있다. Xylogone sphaerospora 유래의 $Gal^2Man_3$는 Trichoderma harzianum 유래의 중합도 4와 동일한 구조로서 가수분해 시간 경과에 따른 반응말기에서 역시 galactose, mannotriose 및 잔존하는 $Gal^2Man_3$ spot이 출현하고 있는 반면 중합도 6인 $Gal^2Man_5$에 대해서는 mannopentaose의 환원말단부터 2번 mannose에 결합하고 있는 galactose에 대해서는 역시 특이성을 나타내지 않아 반응 초기부터 말기까지 가수분해 pattern에 대한 변화를 보이지 않고 있다.

바위털갯지렁이 Marphysa sanguinea의 인공종묘생산에 미치는 사육환경의 영향 (Effects of Rearing Conditions on the Artificial Seed Production of a Polychaete Marphysa sanguinea)

  • 김창훈;장성욱
    • 한국양식학회지
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    • 제21권1호
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    • pp.34-40
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    • 2008
  • 갯지렁이류 중 산업적으로 가치가 높은 바위털갯지렁이 Marphysa sanguinea의 인공종묘생산 기술 개발을 위하여 산란습성, 유생발달 및 치충의 성장에 미치는 사육환경의 영향을 조사하였다. 바위털갯지렁이는 수온 $18^{\circ}C{\sim}22^{\circ}C$에 서식공 주변으로 담륜자(trochophore) 시기의 유생을 방출하였고, 방출된 유생은 서식공 주변에 잠시 머무른 후 수조 전체로 퍼져나가는 것이 관찰되었다. 서식공에서 방출된 유생은 8일이 지나면 개구하면서 점액질이 분비되어 몸 전체를 둘러싸는 것이 관찰되었고, 방출 후 20일이 지나면 10체절을 형성하였다. 시판용 모래의 입도별 선택성을 조사한 결과 생존율은 ${\phi}1{\sim}2mm$인 실험구에서 가장 높았고, ${\phi}2{\sim}3mm$ 실험구에서 가장 낮게 나타났다. 수온에 따른 치충의 성장은 $24^{\circ}C$ 실험구의 성장이 가장 좋았고, 50일간의 실험기간동안 이 실험구의 일간성장률은 1.1%을 나타내어 $21^{\circ}C,\;18^{\circ}C$$15^{\circ}C$구보다 2-3배가 높은 체중과 일간 성장률을 보였다. 염분 또한 치충의 성장과 생존율에 큰 영향을 주었으며, 30 psu 조건에서 가장 높은 증중 및 성장률을 보였다.

남한강 보 구간 유수성 저서성 대형무척추동물의 시·공간적 분포 특성 (Spatio-temporal Distribution Patterns of Lotic Benthic Macroinvertebrate Communities in Namhan-River Weir Section)

  • 김진영;이승현;이경락;노성유;신유나;이수웅;이재관;원두희;임성호;권용주;공동수
    • 생태와환경
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    • 제51권4호
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    • pp.331-344
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    • 2018
  • 한강수계 남한강의 보(강천보, 여주보, 이포보) 공사 후 수변부 여울 구간의 물리적 환경의 변화에 따른 유수성 저서동물의 군집 변화를 확인한 결과 다음과 같은 결론을 얻었다. 첫째, 보에 의한 여울구간의 소실은 유속, 하상 등 물리적 환경의 변화를 야기하여 수질요인과는 독립적으로 수생태계에 영향을 미칠 수 있다. 둘째, 남한강 보 구간의 수변부 물리적 서식환경의 복합적인 변화는 유수성 분류군의 군집구성에 영향을 미친 것으로 판단된다. 셋째, 저서성 대형무척추동물 유수성 지표 후보종 157 분류군은 수생태계 변화의 원인 분석을 비롯한 하천 수생태계 복원 및 재자연화 등에 유용한 지표로 활용될 수 있을 것으로 기대된다. 넷째, 향후 유속변화에 대한 수생태계 영향을 면밀하게 파악하기 위해서는 분류군별 유속에 대한 저항성 또는 선호도를 바탕으로 한 지표연구가 필요할 것으로 사료된다.

Janthinobacterium sp. 유래 저온활성 lipase의 발현, 정제 및 효소 특성 연구 (Expression, Purification, and Characterization of a Cold-adapted Lipase from Janthinobacterium sp.)

  • 박성호;박성주;최종일
    • 한국미생물·생명공학회지
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    • 제46권1호
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    • pp.51-58
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    • 2018
  • 본 연구에서는 극지에서 유래한 Janthinobacterium sp. PAMC25641로부터 분리한 리파아제 유전자를 클로닝 하고 과발현시켜 정제하였으며, 이 분리한 재조합 리파아제 효소의 생화학적 특성에 대해 분석하였다. 이 효소는 $15^{\circ}C$ 이하의 온도에서 장시간 활성을 유지하는 효소로서 산업적으로 활용될 가능성이 높을 것으로 기대된다.

Recombinant Expression and Characterization of Thermoanaerobacter tengcongensis Thermostable $\alpha$-Glucosidase with Regioselectivity for High-Yield Isomaltooligosaccharides Synthesis

  • Zhou, Cheng;Xue, Yanfen;Zhang, Yueling;Zeng, Yan;Ma, Yanhe
    • Journal of Microbiology and Biotechnology
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    • 제19권12호
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    • pp.1547-1556
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    • 2009
  • A novel thermostable $\alpha$-glucosidase (TtGluA) from Thermoanaerobacter tengcongensis MB4 was successfully expressed in E. coli and characterized. The TtgluA gene contained 2,253 bp, which encodes 750 amino acids. The native TtGluA was a trimer with monomer molecular mass of 89 kDa shown by SDS-PAGE. The purified recombinant enzyme showed hydrolytic activity on maltooligosaccharides, p-nitrophenyl-$\alpha$-D-glucopyranide, and dextrin with an exotype cleavage manner. TtGluA showed preference for short-chain maltooligosaccharides and the highest specific activity for maltose of 3.26 units/mg. Maximal activity was observed at $60^{\circ}C$ and pH 5.5. The half-life was 2 h at $60^{\circ}C$. The enzyme showed good tolerance to urea and SDS but was inhibited by Tris. When maltose with the concentration over 50 mM was used as substrate, TtGluA was also capable of catalyzing transglycosylation to produce $\alpha$-1,4-linked maltotriose and $\alpha$-1,6-linked isomaltooligosaccharides. More importantly, TtGluA showed exclusive regiospecificity with high yield to produce $\alpha$-1,6-linked isomaltooligosaccharides when the reaction time extended to more than 10 h.