• 한국미생물·생명공학회지
• /
• 제38권3호
• /
• pp.227-234
• /
• 2010

Cytochrome P450 enzymes which require the supply of electrons from NAD(P)H have a great biotechnological impact as they catalyze valuable reactions on a vast variety of substrates. However, very limited biotechnological application has been reported so far due to their functional complexity, limited stability (instability) and, in most cases, low catalytic activity. In this present review, we introduce some possibilities for improving their defect by exploring electron transport system and substrate flexibility in field of Streptomyces cytochrome P450.

• 환경생물
• /
• 제23권1호
• /
• pp.52-56
• /
• 2005

8일간 생장한 보리 유식물에 72시간 동안 dimethipin을 처리하면서 엽록소 함량의 변화와 광합성적 전자전달 활성을 측정하였다. 10/sup -3/M dimethipin을 72시간 처리한 경우 엽록소 함량이 33% 감소하였다. 이에 비해 7일간 암소에서 생장시킨 후 dimethipin을 처리한 보리 유식물은 녹화 48시간에 10/sup -4/M에서 대조구에 비해 43% 엽록소 함량이 감소하였으며, 엽록소 a/b 비율이 증가하였다. 24시간의 dimethipin처리는 PSⅡ+PSⅠ활성에 거의 영향을 미치지 않았다. 10/sup -4/M dimethipin처리는 48시간 배양시 10% 그리고 72시간에는 25%의 PSⅡ+PSⅠ 활성을 억제하였다. Water splitting system을 제외한 PSⅡ+PSⅠ 활성은 10/sup -4/M dimethipin을 48시간 처리할 경우 16% 그리고 72시간에는 27% 억제되었다. 10/sup -4/M dimethipin을 24시간 처리할 경우 PSⅡ활성은 8%, 48시간에서는 13% 그리고 72시간에서는 18% 억제되었다. 10/sup -4/M dimethipin을 24시간 처리할 경우 PSⅠ활성은 4%, 48시간에서는 8% 그리고 72시간에서는 10% 억제되었다. 7일간 암소에서 자란 황백화된 보리 유식물을 녹화시간 6, 12, 24와 48시간에 dimethipin을 처리할 경우 PSⅡ+PSⅠ 활성은 5, 10, 10 그리고 11% 억제되었다. 반면에 water splitting system을 제외한 PSⅡ+PSⅠ 활성은 모든 처리시간에서 그 효과가 없었다. 또한 PSⅡ와 PSⅠ의 활성도 대조구와 큰 차이가 없었다. 이와 같이 dimethipin에 의해 야기된 광합성적 전자전달활성의 억제는 엽록체 합성과정에서의 영향보다는 엽록체의 기능에 영향을 주는 것으로 사료되며, 이러한 엽록체 기능의 감소는 dimethipin에 의해 야기되는 30∼40%의 엽록소 함량감소에 의한 것으로 사료된다.

• Journal of Plant Biology
• /
• 제31권3호
• /
• pp.217-226
• /
• 1988

Effects of simazine [2-chloro-4,6-bis(methylamino)-s-triazine] on the photochemical reactions of isolaed spinach chloroplasts and crude thylakoids of Anabaena inequalis UTEX B-381 were compared. Simazine inhibited photosynthetic O2 evolution and increased the chlorophyll fluorescence in whole cells of Anabaena. The electron transfer from diphenylcarbazide to 2,6-dichlorophenolindophenol was inhibited by simazine treatment in spinach chloroplasts, but not in crude thylakoids of Anabaena. In spinach chloroplasts, the chlorophyll fluorescence was increased by simazine treatment in the presence of diphenylcarbazide and ferricyanide, but not in the presence of diphenylcarbazide and silicomolybdate. In crude thylakoids of Anabaena, simazine treatment did not increase the chlorophyll fluorescence in the presence of either diphenylcarbazide and silicomolybdate, or diphenylcarbazide and ferricyanide. There results suggest that the inhibitory site of simazine on photosynthetic electron transport chain of anabaena is different from that of spinach chloroplasts. And there may be a possiblity that the inhibition site of simazine in Anabaena lies on the donor side of photosystem II, before the site of electron donation by diphenylcarbazide.

• Journal of Plant Biology
• /
• 제26권2호
• /
• pp.91-99
• /
• 1983

The formation of chlorophyll-protein complexes (CP-complexes) during the greening of rape cotyledons (Brassica napus cv. Yongdang) was investigated by the SDS-polyacrylamide gel electrophoresis. The total chlorophyll content and Chl a/b ratio were also determined. In addition, the effects of dark treatment on the CP-complex patterns during greening have been examined with respect to their photosynthetic electron transport activity. Greening has brought about the increasein total chlorophyll content and the decrease in Chl a/b ratio, but there have been no changes in Chl a/b ratio after 24 hrs of greening. The light-harvesting chlorophyll a/b-protein complex (LHCP-complex0 was predominant during the initial greening period. Thereafter, the amout of chlorophyll a-protein complex (CP I-complex) was gradually increased. Twenty-four-hr dark treatment immediately after illumination for 6 hrs and 12 hrs resulted in the increase of the Chl a/b ration and the CP I complex, otherwise the decrease of the LHCP-complex. The LHCP/CP I ratio was gradually decreased with further greening, and appeared no change after 48 hrs illumination. The investigation of the photosynthetic electron transport activity indicated that photosystem (PS) II activity (H2Olongrightarrowp-PD*＋FeCy**) did not change, but the activity of PS I was increased suddenly due to the dark treatment. The data suggests that the increase of CP I-complex may result in that of P-700, that is, the increase of PS I activity.

• BMB Reports
• /
• 제38권4호
• /
• pp.481-485
• /
• 2005

The response of Spirulina (Arthrospira) platensis to high salt stress was investigated by incubating the cells in light of moderate intensity in the presence of 0.8 M NaCl. NaCl caused a decrease in photosystem II (PSII) mediated oxygen evolution activity and increase in photosystem I (PSI) activity and the amount of P700. Similarly maximal efficiency of PSII (Fv/Fm) and variable fluorescence (Fv/Fo) were also declined in salt-stressed cells. Western blot analysis reveal that the inhibition in PSII activity is due to a 40% loss of a thylakoid membrane protein, known as D1, which is located in PSII reaction center. NaCl treatment of cells also resulted in the alterations of other thylakoid membrane proteins: most prominently, a dramatic diminishment of the 47-kDa chlorophyll protein (CP) and 94-kDa protein, and accumulation of a 17-kDa protein band were observed in SDS-PAGE. The changes in 47-kDa and 94-kDa proteins lead to the decreased energy transfer from light harvesting antenna to PSII, which was accompanied by alterations in the chlorophyll fluorescence emission spectra of whole cells and isolated thylakoids. Therefore we conclude that salt stress has various effects on photosynthetic electron transport activities due to the marked alterations in the composition of thylakoid membrane proteins.

• 약학회지
• /
• 제31권6호
• /
• pp.361-369
• /
• 1987

The four azo dyes such as Amaranth (FD & C Red No. 2), Tartrazine (FD & C Yellow No. 4), sunset Yellow (FD & C Yellow No. 5) and Allura red (FD & C Red No. 40) are currently employed as a food additives in Korea. In this study, the effects of these azo dyes on the hepatic microsomal mixed function oxidase systems in Rats. (i.e., Cyt. P-450, Cyt. b$_5$, NADPH cyt. c-reductase and azo reductase) were investigated. Furthermore, to determine the relationship among the electron transport systems, each level of azo reductase, Cyt. P-450 and NADPH cyt. c-reductase was measured upon the administration of phenobarbital (known as an inducer of Cyt. P-450), 3-methylcholanthrene (Known as an inducer of Cyt. P-448), CoCl$_2$ (inhibitor on Cyt. P-450) or $CCl_4$ (inhibitor on Cyt. P-450). The results of these studies are as follows; (1) The levels of Cyt. P-450 and Cyt. b$_5$ were decreased upon the administration of these azo dyes. (2) When the level of Cyt. P-450 was decreased, the azo reductase activity was also decreased. (3) These azo dyes did not show any significant effect on the level of NADPH cyt. c-reductase. (4) The administration of 3-methylcholanthrene resulted in the elevation of azo reductase activity. The 3-methylcholanthrene may be responsible for the induction of CO-insensitive electron transport system.

• BMB Reports
• /
• 제29권1호
• /
• pp.58-62
• /
• 1996

The structural changes in the electron donor side of the PSII reaction center have been monitored since heat treatment ($45^{\circ}C$ for 5 min) of thylakoids is known to decrease the oxygen evolving activity. In heat-treated spinach chloroplast thylakoids, the inhibitory effect of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) on the electron transport activity of the PSII reaction center from diphenyl carbazide to dichlorophenolindophenol became reduced approximately 3.8 times and [$^{14}C$]-labeled DCMU binding on the D1 polypeptide decreased to 25~30% that of intact thylakoid membranes, implying that the conformational changes of the DCMU binding pocket, residing on the D1 polypeptide, occur by heat treatment. The accessibility of trypsin to the $NH_2$-terminus of the cytochrome b-559 ${\alpha}$-subunit, assayed with Western blot using an antibody generated against the synthetic peptide (Arg-68 to Arg-80) of the COOH-terminal domain, was also increased, indicating that heat-treatment caused changes in the structural environments near the stromal side of the cytochrome b-559 ${\alpha}$-subunit, allowing trypsin more easily to cleave the $NH_2$-terminal domain. Therefore, the structural changes in the electron donor side of the PSII reaction center complexes could be one of the reasons why the oxygen evolving activity of the heat-treated thylakoid membranes decreased.

• Journal of Plant Biology
• /
• 제36권3호
• /
• pp.195-201
• /
• 1993

The room temperature fluorescence induction of chloroplasts was utilized as a probe to locate the site of inhibition by mercury, copper and zinc on PS II by mercury. Inhibitory effect of Hg2+ on electron transport activity was notable as compared with Cu2+ and Zn2+. At concentrations of HgCl2 over 50 $\mu$M, activities of PS II and whole-chain electron transport decreased more than 70%, while that of PS I decreased about 10~30%. This suggests that PS II is more susceptible to Hg2+ than PS I is. In the presence of diphenylcarbazide (DPC), 50 $\mu$M HgCl2 inhibited the reduction of dichlorophenolindophenol (DCPIP) about 50%. Addition of heavy metals induced marked decrease in maximal variable fluorescence/initial fluorescence [(Fv)m/Fo], but no changes in Fo. With various concentrations of heavy metals, changes of chlorophyll a fluorescence emitted by PS II showed gradual decrease in photochemical quenching (qQ), which indicates an increase in reduced state of electron acceptor, QA. Especially, the addition of HgCl2 caused a notable decrease of qQ. In the presence of 50 $\mu$M CuCl2, energy-depended quenching (qE) was completely reduced, whereas in the presence of 50 $\mu$M CuCl2 and ZnCl2 it was still remained. The above results are discussed on the effects of mercury in relation to water-splitting system and plastoquinone (PQ) shuttle system.

• BMB Reports
• /
• 제43권9호
• /
• pp.614-621
• /
• 2010

Propamidine, an aromatic diamidine compound, is widely used as an antimicrobial agent. To uncover its mechanism on pathogenetic fungi, Botrytis cinerea as an object was used to investigate effects of propamidine in this paper. The transmission electron microscope results showed that the mitochondrial membranes were collapsed after propamidine treatment, followed that mitochondria were disrupted. Inhibition of whole-cell and mitochondrial respiration by propamidine suggested that Propamidine is most likely an inhibitor of electron transport within Botrytis cinerea mitochondria. Furthermore, the mitochondrial complex III activity were inhibited by propamidine.

• 대한약리학회지
• /
• 제23권2호
• /
• pp.113-121
• /
• 1987

국소마취제가 mitochondria에서의 전자이동 및 superoxide라디칼의 생성 그리고 지질의 과산화에 따른 malondialdehyde생성에 미치는 영향을 관찰하였다. 국소마취제는 전자이동계 의 효소활성도에 영향을 나타내었다. NADH dehydrogenase, NADH oxidase와 NADH-ubiquinone oxidoreductase의 활성도는 lidocaine, procaine과 dibucaine에 의하여 효과적으로 억제되었고 cocaine에 의하여 약간 억제되었다. Succinate dehydrogenase, succinate cytochrome c oxidoreductase와 succinate-ubiquinone oxidoreductase 활성도는 lidocaine 과 dibucaine에 의하여 억제되었으나 succinate oxidase는 국소마취제에 의하여 활성화되었다. 국소마취제는 dihydroubiquinone-cytochrome c oxidoreducatse와 cytochrome c oxidase의 활성도를 억제하였다. 이와 같은 반응에서 국소마취제에 대한 complex I segment의 반응이 다른 complex segment보다 크게 나타났다. 국소마취제는 succinate 또는 NADH에 의한 superoxide 생성과 이에 대한 antimycin의 자극효과를 억제하였다. 또한 국소마취제는 산소라디칼에 의한 지질의 과산화를 억제하였다. 이상의 결과로부터 국소마취제는 mitochondria의 전자전달 과정 중 Complex I segment때 또는 인접한 부위에 작용하여 전자이동을 억제함으로써 superoxide 생성과 지질의 과산화를 억제할 것으로 시사되었다.