Progress in the molecular mechanism of KaiA regulating cyanobacterial circadian clock / 生物工程学报

The cyanobacterial circadian clock has three relatively independent parts: the input path, the core oscillator, and the output path. The core oscillator is composed of three clock proteins: KaiA, KaiB, and KaiC. The interactions among these three proteins generate a rhythmic signal and convey the input signals to the output signals to maintain the accuracy and stability of the oscillation of downstream signals. Based on the cyanobacterial circadian clock and the structure, function, and interaction of the clock proteins of the core oscillator, combining the recent results from our laboratory, this review summarized the recent progresses of the molecular mechanism of KaiA in regulating KaiC's enzymatic activity, mediating phase reset of the oscillator, and competing with CikA for the binding site of KaiB.

Efecto de ácidos húmicos sobre el crecimiento y la composición bioquímica de Arthrospira platensis / Effect of humic acids on the growth and the biochemical composition of Arthrospira platensis

RESUMEN Con el propósito de evaluar el efecto de tres concentraciones de ácidos húmicos (AH) 1, 10, 100 mg/L contra una concentración de ácido indol acético (AIA), sobre la producción de biomasa, pigmentos, proteínas, carbohidratos y lípidos de A. platensis; se realizaron cultivos en discontinuo en medio Zarrouk 25% suplementados con AH y AIA, bajo condiciones de aireación constante y fotoperiodos de 12:12 horas, durante 30 días. Se encontró que la producción máxima de biomasa fue mayor en el cultivo suplementado con 10 mg/L de AH, comparado con el control y los cultivos suplementados con AIA. De la misma manera fue observado en este tratamiento la producción más alta de pigmentos, proteínas y carbohidratos. Por tanto, se logró incrementar el crecimiento y la producción de metabolitos de A. platensis, lo cual permite observar el uso potencial de estas sustancias como estimulantes biológicos orgánicos.

ABSTRACT The purpose of this research was to evaluate the effect of three concentrations of humic acids (HA) 1, 10, 100 mg/L against a concentration of indole acetic acid (IAA) on production of biomass, pigments, proteins, carbohydrates and lipids of A. platensis. Cultures discontinuous in Zarrouk 25% medium supplemented with HA and IAA under conditions of aeration constant and photoperiod from 12:12 hours, during 30 days were made. It was found that the maximum biomass production was higher in the culture supplemented with 10 mg / L of AH, compared to the control and cultures supplemented with AIA. In the same way the highest production of pigments, proteins and carbohydrates were observed in this treatment. Therefore, the growth and production of metabolites of A. platensis was increased, which makes it possible to observe the potential use of these substances as organic biological stimulants.

Toxic concentrations of metronidazole to Microcystis protocystis / Concentrações tóxicas de metronidazol para Microcystis protocystis

Antimicrobials are among the most commonly used drugs and have become a class of contaminants with great environmental importance. Metronidazole is an antimicrobial used for the therapeutic management of several human diseases. The toxicity of antimicrobials on aquatic species may affect sensitive microorganisms and reduce metabolic processes. Cyanobacteria is a group of organisms that are of great ecological importance in aquatic environments. Studies indicate that cyanobacteria are very sensitive to some antimicrobials. Therefore, it is necessary to evaluate the effects of metronidazole contamination on phytoplankton. The aim of this study was to investigate the effects of metronidazole on the growth of the cyanobacterium Microcystis protocystis and to evaluate the stability of this antimicrobial agent in the culture medium over a period of 96 hours. M. protocystis was resistant to growth inhibition by metronidazole. The EC50 of this antimicrobial for M. protocystis was 117.3 mg L–1. Under the growth inhibition test conditions, neither a significant change in the MNZ concentration nor the presence of drug metabolites or degradation products was observed. These results indicate low cellular uptake of the antimicrobial agent and its persistence in the culture medium.

Antimicrobianos estão entre os fármacos mais usados e tem se tornado uma classe emergente de contaminantes com grande importância ambiental. A toxicidade dos antimicrobianos sobre as espécies aquáticas podem afetar microrganismos sensíveis e reduzir seus processos metabólicos. O Metronidazol é um antimicrobiano usado para o manejo terapêutico de várias doenças humanas. Cyanobacteria é um grupo de organismos de grande importância ecológica no ambiente aquático. O estudo visou investigar os efeitos do metronidazol- MNZ sobre o crescimento da cianobactéria Microcystis protocystis e avaliar a persistência do antimicrobiano no meio de cultivo durante 96h. A M. protocystis mostrou-se resistente à inibição de crescimento pelo MNZ. A EC50% do MNZ para a espécie foi 117,3 mg L–1. Nas condições do teste de inibição de crescimento não foi observada variação significativa da concentração de MNZ nem a presença de metabólitos ou produtos de degradação do fármaco, indicando baixa captação celular do antimicrobiano e sua persistência no meio de cultivo.

Toxic concentrations of metronidazole to Microcystis protocystis

Antimicrobials are among the most commonly used drugs and have become a class of contaminants with great environmental importance. Metronidazole is an antimicrobial used for the therapeutic management of several human diseases. The toxicity of antimicrobials on aquatic species may affect sensitive microorganisms and reduce metabolic processes. Cyanobacteria is a group of organisms that are of great ecological importance in aquatic environments. Studies indicate that cyanobacteria are very sensitive to some antimicrobials. Therefore, it is necessary to evaluate the effects of metronidazole contamination on phytoplankton. The aim of this study was to investigate the effects of metronidazole on the growth of the cyanobacterium Microcystis protocystis and to evaluate the stability of this antimicrobial agent in the culture medium over a period of 96 hours. M. protocystis was resistant to growth inhibition by metronidazole. The EC50 of this antimicrobial for M. protocystis was 117.3 mg L1. Under the growth inhibition test conditions, neither a significant change in the MNZ concentration nor the presence of drug metabolites or degradation products was observed. These results indicate low cellular uptake of the antimicrobial agent and its persistence in the culture medium.

Antimicrobianos estão entre os fármacos mais usados e tem se tornado uma classe emergente de contaminantes com grande importância ambiental. A toxicidade dos antimicrobianos sobre as espécies aquáticas podem afetar microrganismos sensíveis e reduzir seus processos metabólicos. O Metronidazol é um antimicrobiano usado para o manejo terapêutico de várias doenças humanas. Cyanobacteria é um grupo de organismos de grande importância ecológica no ambiente aquático. O estudo visou investigar os efeitos do metronidazol- MNZ sobre o crescimento da cianobactéria Microcystis protocystis e avaliar a persistência do antimicrobiano no meio de cultivo durante 96h. A M. protocystis mostrou-se resistente à inibição de crescimento pelo MNZ. A EC50% do MNZ para a espécie foi 117,3 mg L1. Nas condições do teste de inibição de crescimento não foi observada variação significativa da concentração de MNZ nem a presença de metabólitos ou produtos de degradação do fármaco, indicando baixa captação celular do antimicrobiano e sua persistência no meio de cultivo.

Cultivos discontinuos alimentados con urea de la cianobacteria Phormidium sp. en función de la salinidad y edad del cultivo / Urea fed-batch cultures of the cyanobacterium Phormidium sp. as a function of the salinity and age of cultures

Se comparó la eficiencia de sistemas de cultivos discontinuos alimentados versus cultivos discontinuos convencionales, en cuanto a concentración de nitrógeno, adicionando 0,2 mM de urea cada tres días al final de la fase exponencial, durante 21 días. Se realizaron cultivos con un volumen de 1500 mL a 15 y 35 UPS de salinidad, enriquecidos con medio ALGAL 8mM NaNO3, a 238 µmol q m-2 s-1, aireación constante, fotoperiodo 12:12 horas y temperatura de 29 ±3°C. Phormidium sp. posee la capacidad de hidrolizar la urea; mostrando una asimilación de 65±7,07% de la misma, con la mayor producción (p<0,05) de clorofila a, ficocianina y proteínas de 20,26±1,24; 203,47±12,83 y 707,87±28,47 µg mL-1en los cultivos alimentados. La producción de pigmentos vario en el tiempo, independientemente a la salinidad y sistema de cultivo, mientras que la producción de proteínas y carbohidratos totales fue directamente proporcional a la edad del cultivo, con valores máximos de 612,74 ± 5,41 µg mL-1 y 8,96±0,08 mg mL-1 respectivamente a los 31 días. La síntesis de lípidos y EPS fueron influenciadas (p<0,05) por la salinidad, presentando los máximos de lípidos a 15 UPS con 12,22±2,91µg mL-1, y los EPS se incrementaron a 35 UPS con 2,00 ± 0,26 y 2,03 ± 0,15 mg mL-1. Estos resultados determinan que los cultivos de Phormidium sp. alimentados con urea y a salinidades de 15 y 35 UPS, representan una alternativa económica para la producción de clorofila a, ficocianina y proteínas, incrementándose un 31,04; 40,72 y 31,94 % respectivamente en comparación con cultivos no alimentados.

Fed-batch system efficiency versus batch cultures was compared in relation to nitrogen concentration, adding 0,2mM urea at the end of the exponential phase, during 21 days. Cultures were carried out in 1500 mL to 1.5 and 3.5 UPS of salinity, enriched with Algal medium 8mM NaNO3, 238 mol q m-2 s-1, constant aeration, photoperiod 12:12 h. and 29 ±3°C. Phormidium sp. is able to hydrolyze urea; showing a total assimilation of 65±7.07%, with the highest (p< 0.05) chlorophyll a, phycocyanin and protein production of 20.26 ± 1.24, 203.47 ± 12.83 and 707.87 ± 28.47 µg mL-1 in the fed-batch cultures. On the other hand, pigment production varies in time, regardless salinity and culture system. Proteins and total carbohydrate production were directly proportional to the age of cultures, with maximum values of 612.74 ± 5.41 µg mL-1 and 8.96 ± 0.08 mg mL-1, respectively. Lipid and EPS were influenced (p< 0.05) by salinity, showing maximum of lipids at 15 UPS with 12.22±2.91 µg mL-1, and EPS at 15 and 35 UPS with 2.00 ± 0.26 and 2.03 ± 0.15 mg mL-1. These results determine that Phormidium sp. cultures fed with urea, to salinities of 15-35 UPS, represent an economic alternative for chlorophyll a, phycocyanin and protein production, with an increase of 31.04, 40.72 and 31.94% respectively in comparison with non-fed cultures.

Adaptive responses of cyanobacterium plectonema boryanum to herbicide butachlor.

The Present paper deals with the herbicide Butachlor (5,10 ,20,40 and 80ppm)-induced changes in physiological and biochemical parameters related to photosynthesis and defense systems in paddy field cyanobacterium Plectonema boryanum grown under laboratory conditions. Growth and photosynthetic pigments, i.e., chlorophyll a and carotenoids were adversely affected by Butachlor treatment and the inhibition was found to be dose dependent. The toxic effect of Butachlor was more pronounced protein; however, a considerable reduction in chlorophyll a, and carotenoids was also noticed. Furthermore, Butachlor with increasing doses accelerated the formation of active oxygen species, i.e., O2- and H2O2, in cells progressively. As a consequence of active oxygen species (AOS) generation in Butachlor -treated cells, the activity of superoxide dismutase (SOD) and peroxidase (POD) was enhanced considerably. Besides the accelerated action of enzymatic defense systems, Protein damage also showed an increasing trend with the rising concentration of Butachlor (5, 10, 20, 40 and 80 ppm).

Uso da Spirulina platensis na recuperação de ratos submetidos à dieta de restrição proteica / Use of Spirulina platensis for recovering rats under protein restriction diet feeding

O presente estudo avaliou a ação da espirulina como fonte protéica na recuperação do estado nutricional de ratos machos adultos. Foram estudados 48 animais, divididos em quatro grupos de acordo com a dieta: grupo controle de caseína (CC) e grupo controle de espirulina (CE), que receberam dieta equilibrada durante todo o experimento; grupo recuperado de caseína (RC) e grupo recuperado de espirulina (RE), que consumiram dieta com restrição protéica durante 60 dias e, a seguir, receberam a dieta equilibrada à base de caseína e espirulina, respectivamente, durante 90 dias. Foram analisadas a composição centesimal e a microbiológica da Spirulina platensis, bem como o ganho em peso dos animais. Após o sacrifício dos animais, foram realizadas as análises para determinação do peso do fígado e do aspecto histopatológico, da gordura corporal e dos constituintes sanguíneos. A espirulina utilizada continha 59,65% de proteína, 3,72% de lipídios, 17,53% de resíduo mineral fixo e 7,81 de umidade, e não apresentou contaminação por microrganismos. Não houve diferenças significativas quanto à gordura corporal e constituintes protéicos séricos entre os quatro grupos de animais. Em relação ao ganho de peso não foi constatada diferença entre os grupos CC e CE, nas diferentes semanas de números 1, 5, 6, 7 e de 10 a 18, contudo esta ocorrência não foi observada nos grupos RC e RE. O peso relativo do fígado diminuiu nos animais com restrição protéica, principalmente no grupo RC, porém sem alterações histopatológicas, e os constituintes lipídios tiveram valores diminuídos. Conclui-se que o uso da Spirulina platensis na recuperação de ratos com dieta de restrição protéica reduz a lipemia.

This investigation aims at evaluating the effects of spirulin as a protein source for recovering the nutritional status of adult male rats. Forty-eight animals were studied and divided by four groups according to the feed diet: casein control (CC) group and spirulin control (SC) group, which received balanced diet during the whole experiment; recovered casein (RC) group and recovered spirulin (RS) group, which received a protein-restriction diet for 60 days and, afterwards, they were fed with balanced diet based on casein and spirulin, respectively, for 90 days. The centesimal and microbiologicalcomposition of Spirulina platensis and animals weight gains could be evaluated. After being euthanized the animal’s liver weight and its histopathologic aspects were analyzed, besides the body fat and blood components. The spirulinused contained 59.65% of protein, 3.72% of lipids, 17.53 of fixed mineral residue and 7.81% of humidity, and showed no microorganism contamination. Significant differences on body fat and serum protein contents could not be observedamong the four animal groups. Regarding the weight gains, there was no difference among the animals of groups CC and SC in different weeks of experiments numbers 1, 5, 6, 7 and from 10 to 18, but these findings were not observed among the RC and RS groups. The relative liver weight decreased in animals under protein restriction, mainly in the group RC, but no histopathological alterations could be observed, and lipid contents decreased. In conclusion, the useof Spirulina platensis for recovering rat on protein restriction diet induces a reduction of lipemia.

Responses of cyanobacterium Anabaena doliolum during nickel stress.

Growth and biochemical responses of heterocystous nitrogen fixing cyanobacteria Anabaena doliolum were studied upon exposure to various concentrations of nickel (0.1 to 100 mM) for duration of upto 4 days, in view of its tolerance. The growth of the cyanobacterium was increased in terms of cell density (O.D), heterocyst frequency, chlorophyll-a, carotenoid and c-phycocynin upto moderate exposure (10 mM for 96 hr of Ni treatment). Protein and total non protein thiol were measured as stress responsive metabolites and frequency of heterocyst and spores were observed in responses to Ni. The present study demonstrated that the tested cyanobacterium exhibited dose specific responses of metal towards studied different morphological, physiological and biochemical parameters.

Control of sporulation in the filamentous cyanobacterium A nabaena torulosa.

In the cyanobacterium Anabaena torulosa, sporulation occurred even during the logarithmic growth phase. Sporulation was initiated by differentiation of the vegetative cell on one side, adjoining the heterocyst followed by differentiation of the vegetative cell on the other side. Subsequently, spores were differentiated alternately on either side to form spore strings. The sequence of sporulation supports the previous notion that a gradient of spore maturation exists in cyanobacteria and also indicates that the gradient is manifested unequally on either side of heterocysts. Sporulation was absent or negligible in a minerally enriched medium but ocurred readily in a minimal medium. The extent of sporulation was inversely related to phosphate concentration. Sporulation was enhanced at higher temperature. Incandescent light, but not fluorescent light, greatly stimulated sporulation suggesting possible involvement of red light in spore differentiation. Addition of filtrate, from 5 to 8 day old cultures, to freshly inoculated A. torulosa greatly enhanced sporulation indicating the influence of extracellular products in spore formation.