• Molecules and Cells
• /
• 제19권2호
• /
• pp.256-261
• /
• 2005

In silico analysis of genome of the cyanobacterium Synechocystis sp. PCC 6803 identified two genes, slr0329 and sll0593, that might participate in glucose (Glc) phosphorylation (www.kazusa.or.jp/cyano). In order to determine the functions of these two genes, we generated deletion mutants, and analyzed their phenotypes and enzymatic activities. In the presence of 10 mM Glc, wild-type (WT) and slr0329 defective strain (M1) grew fast with increased respiratory activity and NADPH production, whereas the sll0593 deletion mutant (M2) failed to show any of the Glc responses. WT and M1 were not significantly different in their glucokinase activity, but M2 had 90% less activity. Therefore, we propose that Sll0593 plays a major role in the phosphorylation of glucose in Synechocystis cells.

• BMB Reports
• /
• 제39권4호
• /
• pp.394-399
• /
• 2006

The transport of putrescine into a moderately salt tolerant cyanobacterium Synechocystis sp. PCC 6803 was characterized by measuring the uptake of radioactively-labeled putrescine. Putrescine transport showed saturation kinetics with an apparent $K_m$ of $92{\pm}10\;{\mu}M$ and $V_{max}$ of $0.33{\pm}0.05\;nmol/min/mg$ protein. The transport of putrescine was pH-dependent with highest activity at pH 7.0. Strong inhibition of putrescine transport was caused by spermine and spermidine whereas only slight inhibition was observed by the addition of various amino acids. These results suggest that the transport system in Synechocystis sp. PCC 6803 is highly specific for polyamines. Putrescine transport is energy-dependent as evidenced by the inhibition by various metabolic inhibitors and ionophores. Slow growth was observed in cells grown under salt stress. Addition of low concentration of putrescine could restore growth almost to the level observed in the absence of salt stress. Upshift of the external osmolality generated by either NaCl or sorbitol caused an increased putrescine transport with an optimum 2-fold increase at 20 mosmol/kg. The stimulation of putrescine transport mediated by osmotic upshift was abolished in chloramphenicol-treated cells, suggesting possible involvement of an inducible transport system.

• 아시안잔디학회지
• /
• 제11권1호
• /
• pp.45-58
• /
• 1997

Pigments and thylakoid membrane proteins were investigated in wild type and PS I- mutants from Synechocystis sp. PCC6803 Comparing morphological features, B2 was less fluorescent than the other strains. The contents of chlorophyll a were propotional to the FNR activity in thylakoid membrane. The FNR activity of mutants was lower than that of wild type. In the result of pigments analysis, mutants had smaller cholophyll a than that of wild type. The major carotenoid was found to he $\beta$-caroene, but aeaxanthin was barely detected in thylakoid membrane of mutants. The polypeptide, 14.8kD was detected by electrophoresis in mutants. It was considered to be the modification of 15.4kD in wild type. Membrane polypeptides of 17.6 and 19.7kD were not detected in mutants. In the result of western blotting, subunit I was detected in all strains, but subunit II was barely detected in mutants. Subunit II was not detected in B2 at all. In view of the results so far achieved, the changes of contents of chlorophyll and zeaxanthin were affected by the defficiency or modification of functional domain in subunit I. Also the modification in subunit I affected the subunit II- binding site in PS I. As the result, efficiency of photosynthesis was decreased. Key words: Synechoystis sp. PCC6803, PS I - mutant, Photosynthetic efficiency, Pigment,Thylakoid membrane proteins, Subunit I, II.

• 한국수소및신에너지학회논문집
• /
• 제19권6호
• /
• pp.529-536
• /
• 2008

Cyanobacteria Synechocystis PCC 6803 or the extracted thylakoid membrane from this strain was appled to photosynthetic bio-electrochemical fuel cell(PBEFC) for the production of hydrogen under the illumination of 48Klux using halogen lamp. PBEFC was composed of anode, cathode and membrane between them. Electrode material was carbon paper while electron mediator and receptor were added phenazine methosulfate(PMS) and potassium ferricyanide respectively. When water and 50 mM tricine buffer and $300{\mu}M$ PMS were added to the anode under the light condition, PBEFC produced the current density $4.4{\times}10^{-5}\;mA/cm^2$, $1.4{\times}10^{-4}\;mA/cm^2$ and $2.4{\times}10^{-4}\;mA/cm^2$, respectively. And the addition of the thylakoid membrane to the system increased current density to $1.3{\times}10^{-3}\;mA/cm^2$. Two times increase of the thylakoid membrane into the anode doubled the current density to $2.6{\times}10^{-3}\;mA/cm^2$. But the current density was not increased proportionally to the amount of thylakoid membrane increased. The system was unstable to measure the electricity output due to the foam production in the anode. Addition of triton X-100 and tween 80 stabilized the system to measure the electricity output but the current density was not increased higher than $8.4{\times}10^{-4}\;mA/cm^2$ and $2.3{\times}10^{-3}\;mA/cm^2$. When the thylakoid membrane was substituted to Synechocystis PCC 6803 cells of four-day culture which has chlorophyll contents $20.5{\mu}g/m{\ell}$, maximum current density was $1.3{\times}10^{-3}\;mA/cm^2$ with $1\;k{\Omega}$ resistance.

• KSBB Journal
• /
• 제13권1호
• /
• pp.101-107
• /
• 1998

광합성 미생물의 고농도 배양에 의한 이산화탄소 고정능에 대한 기초 연구로써 관형 광생물반응기를 이용하여 이산화탄소 조성 및 초기균체농도에 따른 성장 경향을 보았다 배지의 pH가 지어되고 있는 조건하에서 20% 이산화탄소 혼합공기가 공급되는 조건에서도 성장이 이루어졌다 $45.5{\mu}E/m^2{\cdot}s$의 광강도에서 5% 이산화탄소 혼합공기 조성과 0.45 g/L의 초기균체농도에서 성장속도가 가장 우수하였으며, 비성장속도는 0.0258 $h^{-1}$를 나타냈고, 단위 시간당 균체생성량은 0.278 g/L . day 이다. 관형 반응기에서 최대균체농도는 2.03 g/L 까지 배양되었다. 배양된 균체의 원소성분분석을 통하여 Synechocystis PCC 6803의 분자식은 $C_{1.0}H_{2.022}N_{0.194}O_{0.443}S_{0.002}$로 계산되었고, 이산화탄소 고정화속도는 0.482g-$C0_2/L$ . day의 결과를 얻었다.

• 한국응용곤충학회지
• /
• 제35권1호
• /
• pp.66-73
• /
• 1996

Bacillus thuringiensis subsp. morrisoni PG-14의 cryIVD 유전자를 포함하고 있는 발현벡터 pCYASK5-1을 제작하여 Synechocystis PCCC6803에 형질전환시킨 후, 학질모기(Anopheles sinensis)유충에 대한 독성을 검정하였다. Kanamycin이 포함된 BG-11배지에서 선발된 형질전환체의 cryIVD 유전자 발현은 SDS-PAGE와 West-ern blot분석으로 확인하였다. 형질전환체는 A. sinensis 유충에 대해 높은 독성을 나타내었으며, 성장은 야생주인 Synechocystis Pccc6803과 유사하였다. 또 수심에 따른 형질전환체의 분포도 조사에서 전체적으로 살충농도의 세포수로 분포함을 확인하였다. 이러한 결과들은 B. thuringiensis subsp. morrisoni PG-14의 cryIVD 유전자로 형질전환된 Synechocystis PCCC6803이 모기유충 방제에 효율적으로 이용될 수 있음을 나타내었다.

• 한국응용곤충학회지
• /
• 제43권1호
• /
• pp.35-41
• /
• 2004

Bacillus thuringiensis는 포자형성기 동안에 위생해충이나 농업해충에 독성을 보이는 내독소 단백질을 생성한다. 내독소 단백질의 모기 유충 방제효과를 높이기 위해, 광합성에 관여하는 psbA promoter로 모기 살충성 cry11Aa유전자를 발현하는 pSyn4D벡터를 제작하고, 모기 유충이 먹이로 이용하는 Synechocystis PCC6803에 형질 전환시켰다. 형질 전환체들은 kanamycin이 포함된 배지에서 선발되었으며, 정상적인 생물검정을 통해 형질 전환체 Tr2C를 선발하였다. cry11Aa 유전자는 형질전환체의 genomic DNA에 안정적으로 결합되어 있는 것을 PCR을 이용하여 확인하였다. 형질전환체 Tr2C는 약 72-kDa크기의 Cry11Aa 단백질을 발현하였으며, 작은빨간집모기(Culex tritaeniorhynchus) 3령 유충과 중국얼룩날개모기(Anopheles sinensis) 3령 유충에 75%가 넘는 살충력을 보였다. 모기 유충에 대한 형질전환체의 반수치사시간(LT$_{50}$)은 작은빨간집모기 유충과 중국얼룩날개모기 유충에 대해 각각 2.1일과 0.7일이었다. 이상의 결과들은 형질전환체 Tr2C가 모기 유충방제에 유용하게 이용될 수 있음을 보여준다.

• 한국미생물·생명공학회지
• /
• 제31권2호
• /
• pp.103-110
• /
• 2003

그람 양성세균에서 PyrR단백질에 의하여 피리미딘의 생합성이 조절된다는 발견을 바탕으로 하여, Synechocystis sp.PCC6803과 Haemophilus influenzae의 PyrR orthologue 유전자를 Bacillus subtilis에서 형질전환 시켜 피리미딘 생합성의 조절 유무를 조사하였다. Synechocystis sp.PCC6803과 H. influenzae의 PyrR orthologue유전자를 pUC19과 T-vector에 클로닝 한후 pKH1, pKH2, pHPSK1, pHPSK2으로 각각 명명하였다. 이것을 다시 Escherichia. coli와 B. subtiius용 shuttle vector인 pHPS9에 클로닝 하여 pKH3, pKH4, pHPSK3, pHPSK4로 각각 명명하였다. B. subtilis DB104Δ PyrR에 pKH3, pKH4, pHPSK3, pHPSK4을 형질전환후 ATCase 활성을 측정결과 pHPSK3을 가진 균주만 피리미딘에 의한 조절작용이 일어난다는 사실을 통하여, H. influenzae의 PyrR orthologue 유전자의 선도 부분에 조절에 관여하는 미지의 부분이 있음을 예측할 수 있었다. 서로 다른 유래의 PyrR orthologue단백질을 정제하기 위하여 pET14b에 클로닝후 pKH5, pHPSK5으로 각각 명명하였다. SDS-PAGE로 분석한 결과 각각 약 18 kDa과 21 kDa의 분자량을 나타내었다. 정제된 PyrR orthologue 단백질의 UPRTase 활성을 측정한 결과 H. infuenzae의 PyrR orthologue 단백질은 UPRTase 활성을 나타내었으며 다양한 pH에서 측정한 결과 pH 5에서 가장 높은 활성을 나타내었다. 반면에, Synechocystis sp. PCC6803의 PyrR orhologue 단백질은 UPRTase 활성을 나타내지 않았다. 여러 가지 균주의 PyrR 아미노산 서열을 비교한 계통수 분석은 PyrR 단백질의 조절기작과 어느 정도 연관됨을 시사해 주었다.

• 미생물과산업
• /
• 제17권3호
• /
• pp.2-14
• /
• 1991

• Journal of Microbiology and Biotechnology
• /
• 제24권8호
• /
• pp.1015-1025
• /
• 2014

Microorganisms that live in the rhizosphere play a pivotal role in the functioning and maintenance of soil ecosystems. The study of rhizospheric cyanobacteria has been hampered by the difficulty to culture and maintain them in the laboratory. The present work investigated the production of the plant hormone indole-3-acetic acid (IAA) and the potential of biofilm formation on the rhizoplane of pea plants by two cyanobacterial strains, isolated from rice rhizosphere. The unicellular cyanobacteria Chroococcidiopsis sp. MMG-5 and Synechocystis sp. MMG-8 that were isolated from a rice rhizosphere, were investigated. Production of IAA by Chroococcidiopsis sp. MMG-5 and Synechocystis sp. MMG-8 was measured under experimental conditions (pH and light). The bioactivity of the cyanobacterial auxin was demonstrated through the alteration of the rooting pattern of Pisum sativum seedlings. The increase in the concentration of L-tryptophan and the time that this amino acid was present in the medium resulted in a significant enhancement of the synthesis of IAA (r > 0.900 at p = 0.01). There was also a significant correlation between the concentration of IAA in the supernatant of the cyanobacteria cultures and the root length and number of the pea seedlings. Observations made by confocal laser scanning microscopy revealed the presence of cyanobacteria on the surface of the roots and also provided evidence for the penetration of the cyanobacteria in the endorhizosphere. We show that the synthesis of IAA by Chroococcidiopsis sp. MMG-5 and Synechocystis sp. MMG-8 occurs under different environmental conditions and that the auxin is important for the development of the seedling roots and for establishing an intimate symbiosis between cyanobacteria and host plants.