Advances on the production of organic acids by yeast / 生物工程学报

Organic acids are organic compounds that can be synthesized using biological systems. They often contain one or more low molecular weight acidic groups, such as carboxyl group and sulphonic group. Organic acids are widely used in food, agriculture, medicine, bio-based materials industry and other fields. Yeast has unique advantages of biosafety, strong stress resistance, wide substrate spectrum, convenient genetic transformation, and mature large-scale culture technology. Therefore, it is appealing to produce organic acids by yeast. However, challenges such as low concentration, many by-products and low fermentation efficiency still exist. With the development of yeast metabolic engineering and synthetic biology technology, rapid progress has been made in this field recently. Here we summarize the progress of biosynthesis of 11 organic acids by yeast. These organic acids include bulk carboxylic acids and high-value organic acids that can be produced naturally or heterologously. Finally, future prospects in this field were proposed.

Thermo-resistant enzyme-producing microorganisms isolated from composting / Microrganismos produtores de enzimas termo-resistentes isolados de compostagem

Organo-mineral fertilizers supplemented with biological additives are an alternative to chemical fertilizers. In this study, thermoresistant microorganisms from composting mass were isolated by two-step procedures. First, samples taken at different time points and temperatures (33 days at 52 ºC, 60 days at 63 ºC, and over 365 days at 26 ºC) were pre-incubated at 80 oC for 30 minutes. Second, the microbial selection by in vitro culture-based methods and heat shock at 60 oC and 100 oC for 2h and 4h. Forty-one isolates were able to grow at 60 °C for 4h; twenty-seven at 100 °C for 2h, and two at 100 °C for 4h. The molecular identification by partial sequencing of the 16S ribosomal gene using universal primers revealed that thirty-five isolates were from eight Bacillus species, one Brevibacillus borstelensis, three Streptomyces thermogriseus, and two fungi (Thermomyces lanuginosus and T. dupontii). Data from amylase, phytase, and cellulase activity assays and the enzymatic index (EI) showed that 38 of 41 thermo-resistant isolates produce at least one enzyme. For amylase activity, the highest EI value was observed in Bacillus licheniformis (isolate 21C2, EI= 4.11), followed by Brevibacillus borstelensis (isolate 6C2, EI= 3.66), Bacillus cereus (isolate 18C2, EI= 3.52), and Bacillus paralicheniformis (isolate 20C2, EI= 3.34). For phytase, the highest EI values were observed for Bacillus cereus (isolate 18C2, EI= 2.30) and Bacillus licheniformis (isolate 3C1, EI= 2.15). Concerning cellulose production, B. altitudinis (isolate 6C1) was the most efficient (EI= 6.40), followed by three Bacillus subtilis (isolates 9C1, 16C2, and 19C2) with EI values of 5.66, 5.84, and 5.88, respectively, and one B. pumilus (isolate 27C2, EI= 5.78). The selected microorganisms are potentially useful as a biological additive in organo-mineral fertilizers and other biotechnological processes.

Os fertilizantes organo-minerais suplementados com aditivos biológicos são uma alternativa aos adubos químicos. Neste estudo, microrganismos termoresistentes foram isolados de compostagem por procedimentos de duas etapas. Inicialmente, as amostras tomadas em diferentes períodos e temperaturas (33 dias a 52 ºC, 60 dias a 63 ºC e mais de 365 dias a 26 ºC) foram pré-incubadas a 80 oC por 30 minutos. Posteriormente, a seleção microbiana foi conduzida por métodos baseados em cultura in vitro e choque térmico a 60 oC e 100 oC por 2h e 4h. Quarenta e um isolados foram capazes de crescer a 60 °C por 4h; vinte e sete a 100 °C por 2h e dois a 100 °C por 4h. A identificação molecular por sequenciamento parcial do gene ribossomal 16S usando primers universais revelou que trinta e cinco isolados eram de oito espécies de Bacillus, um Brevibacillus borstelensis, três Streptomyces thermogriseus e dois fungos (Thermomyces lanuginosus e T. dupontii). Os dados dos ensaios de atividade de amilase, fitase e celulase e o índice enzimático (IE) mostraram que 38 dos 41 isolados termorresistentes produziram pelo menos uma enzima. Para a atividade da amilase, o maior valor de IE foi observado em Bacillus licheniformis (isolado 21C2, IE = 4,11), seguido por Brevibacillus borstelensis (isolado 6C2, IE = 3,66), Bacillus cereus (isolado 18C2, IE = 3,52) e Bacillus paralicheniformis (isolado 20C2, IE = 3,34). Para a fitase, os maiores valores de IE foram observados para B. cereus (isolado 18C2, IE = 2,30) e B. licheniformis (isolado 3C1, IE = 2,15). Em relação à produção de celulose, B. altitudinis (isolado 6C1) foi o mais eficiente (IE = 6,40), seguido por três Bacillus subtilis (isolados 9C1, 16C2 e 19C2) com valores de IE de 5,66, 5,84 e 5,88, respectivamente, e um B. pumilus (isolado 27C2, IE = 5,78). Pode-se inferir que os microrganismos selecionados são potencialmente úteis como aditivos biológicos em fertilizantes organo-minerais e outros processos biotecnológicos.

Advances in bacterial adsorption and transport of aromatic compounds / 生物工程学报

Aromatic compounds are a class of organic compounds with benzene ring(s). Aromatic compounds are hardly decomposed due to its stable structure and can be accumulated in the food cycle, posing a great threat to the ecological environment and human health. Bacteria have a strong catabolic ability to degrade various refractory organic contaminants (e.g., polycyclic aromatic hydrocarbons, PAHs). The adsorption and transportation are prerequisites for the catabolism of aromatic compounds by bacteria. While remarkable progress has been made in understanding the metabolism of aromatic compounds in bacterial degraders, the systems responsible for the uptake and transport of aromatic compounds are poorly understood. Here we summarize the effect of cell-surface hydrophobicity, biofilm formation, and bacterial chemotaxis on the bacterial adsorption of aromatic compounds. Besides, the effects of outer membrane transport systems (such as FadL family, TonB-dependent receptors, and OmpW family), and inner membrane transport systems (such as major facilitator superfamily (MFS) transporter and ATP-binding cassette (ABC) transporter) involved in the membrane transport of these compounds are summarized. Moreover, the mechanism of transmembrane transport is also discussed. This review may serve as a reference for the prevention and remediation of aromatic pollutants.

Correlation of Lipin gene expression with hepatic fat content in rats with intrauterine growth retardation / 中国当代儿科杂志

OBJECTIVES@#To study the correlation of the expression of Lipin1 in visceral adipose tissue and Lipin2 in liver tissue with hepatic fat content in rats with intrauterine growth retardation (IUGR).@*METHODS@#Pregnant rats were given a low-protein (10% protein) diet during pregnancy to establish a model of IUGR in neonatal rats. The pregnant rats in the control group were given a normal-protein (21% protein) diet during pregnancy. The neonatal rats were weighed and liver tissue was collected on day 1 and at weeks 3, 8, and 12 after birth, and visceral adipose tissue was collected at weeks 3, 8, and 12 after birth. The 3.0T 1H-magnetic resonance spectroscopy was used to measure hepatic fat content at weeks 3, 8, and 12 after birth. Real-time PCR was used to measure mRNA expression levels of Lipin2 in liver tissue and Lipin1 in visceral adipose tissue. Western blot was used to measure protein levels of Lipin2 in liver tissue and Lipin1 in visceral adipose tissue. A Pearson correlation analysis was performed to investigate the correlation of mRNA and protein expression of Lipin with hepatic fat content.@*RESULTS@#The IUGR group had significantly higher mRNA and protein expression levels of Lipin1 in visceral adipose tissue than the control group at weeks 3, 8, and 12 after birth (P<0.05). Compared with the control group, the IUGR group had significantly lower mRNA and protein expression levels of Lipin2 in liver tissue on day 1 after birth and significantly higher mRNA and protein expression levels of Lipin2 at weeks 1, 3, 8, and 12 after birth (P<0.05). At week 3 after birth, there was no significant difference in hepatic fat content between the IUGR and control groups (P>0.05), while at weeks 8 and 12 after birth, the IUGR group had a significantly higher hepatic fat content than the control group (P<0.05). The protein and mRNA expression levels of Lipin1 were positively correlated with hepatic fat content (r=0.628 and 0.521 respectively; P<0.05), and the protein and mRNA expression levels of Lipin2 were also positively correlated with hepatic fat content (r=0.601 and 0.524 respectively; P<0.05).@*CONCLUSIONS@#Upregulation of the mRNA and protein expression levels of Lipin1 in visceral adipose tissue and Lipin2 in liver tissue can increase hepatic fat content in rats with IUGR and may be associated with obesity in adulthood.

Advances in the metabolic engineering for the production of tetracarbon organic acids / 生物工程学报

Tetracarbon organic acids are important platform chemicals that are widely used in the food, chemical, medicine, material industries and agriculture. Compared with the traditional petrochemical process, the production of tetracarbon organic acids by microbial fermentation is more promising due to milder reaction conditions, greener process and better environmental compatibility. This review summarizes the biosynthetic pathways and metabolic mechanisms for the production of tetracarbon organic acids, and illustrates recent advances, challenges, and future perspectives in the production of tetracarbon organic acids by naturally selected or purposefully engineered strains.

Co-fermentation of kitchen waste and excess sludge for organic acid production: a review / 生物工程学报

Resource utilization is an effective way to cope with the rapid increase of kitchen waste and excess sludge, and volatile fatty acids produced by anaerobic fermentation is an important way of recycling organic waste. However, the single substrate limits the efficient production of volatile fatty acids. In recent years, volatile fatty acids produced by anaerobic co-fermentation using different substrates has been widely studied and applied. In this paper, we analyze the characteristics of fermentation to produce acid using kitchen waste and excess sludge alone or mixture. Influences of environmental factors and microbial community structure on the type and yield of volatile fatty acids in the anaerobic fermentation system are discussed in detail. Moreover, we propose future research directions, to provide a reference for recycling kitchen waste and excess sludge.

Identification and control of wood-deteriorating fungi / Identificação e controle de fungos deterioradores de madeira

Wood is rich in organic compounds; thus, it is susceptible to attacks by several deteriorating agents because they acknowledge such attacks as a necessary energy supply. Fungi stand out among these agents since they can attack the wood in trees (before felling) and in several post-felling stages. Fungi are biological agents that use organic compounds as food sources, and that is the reason why they degrade most chemical components found in wood. Wood attacked by fungi suffers significant value loss, since these degrading agents affect their mechanical and aesthetic properties. Fungi ­ which are classified as staining, molding and rotting ­ are one of the main responsible for the biggest losses in the timber industry. Wood can be virtually used in a whole range of environments, a fact that makes it susceptible to attacks by different fungal species, since the ideal conditions for fungal development change from species to species. In other words, the fact that one or more fungal species are capable of deteriorating wood is directly related to conditions wood is subjected to. Thus, it is essential knowing how to evaluate fungal attacks, symptoms to be taken into consideration at the time to identify the type of attacking organism and the attack stage, as well as the control and prevention measures to be applied to these organisms.(AU)

Rica em compostos orgânicos, a madeira é suscetível ao ataque de diversos agentes deterioradores por ser reconhecida como suprimento necessário de energia. Entre esses agentes, destacam-se os fungos, cujo início de ataque pode se dar ainda na árvore, antes do abate e nas diversas fases posteriores. Os fungos são agentes biológicos que necessitam de compostos orgânicos como fontes de alimento e, por essa razão, degradam grande parte dos componentes químicos da madeira. As madeiras, quando atacadas por fungos, sofrem grandes perdas de valor, afetando suas propriedades mecânicas e estéticas. Os fungos são considerados um dos principais responsáveis pelos maiores prejuízos na indústria madeireira e são classificados como manchadores, emboloradores e apodrecedores. Praticamente, a madeira pode estar em uso em uma variedade de ambientes; sabendo-se que as condições ideais de desenvolvimento de um fungo são diferentes para cada espécie, a madeira ficará suscetível ao ataque por diferentes espécies de fungos. Ou seja, o fato de uma ou mais espécies de fungos serem encontradas deteriorando a madeira está diretamente relacionado às condições em que ela se encontra. Dessa forma, torna-se importante saber como avaliar os ataques feitos por fungos, os sintomas a serem considerados para identificar o tipo de organismo e o estágio do ataque, bem como as medidas de controle e prevenção desses organismos.(AU)

Mitochondrial Toxins and Healthy Lifestyle Meet at the Crossroad of Hormesis

Mitochondrial function is crucial for the maintenance of cellular homeostasis under physiological and stress conditions. Thus, chronic exposure to environmental chemicals that affect mitochondrial function can have harmful effects on humans. We argue that the concept of hormesis should be revisited to explain the non-linear responses to mitochondrial toxins at a low-dose range and develop practical methods to protect humans from the negative effects of mitochondrial toxins. Of the most concern to humans are lipophilic chemical mixtures and heavy metals, owing to their physical properties. Even though these chemicals tend to demonstrate no safe level in humans, a non-linear dose-response has been also observed. Stress response activation, i.e., hormesis, can explain this non-linearity. Recently, hormesis has reemerged as a unifying concept because diverse stressors can induce similar stress responses. Besides potentially harmful environmental chemicals, healthy lifestyle interventions such as exercise, calorie restriction (especially glucose), cognitive stimulation, and phytochemical intake also activate stress responses. This conceptual link can lead to the development of practical methods that counterbalance the harm of mitochondrial toxins. Unlike chemical hormesis with its safety issues, the activation of stress responses via lifestyle modification can be safely used to combat the negative effects of mitochondrial toxins.

Actividad antimicrobiana de cuatro variedades de plantas frente a patógenos de importancia clínica en Colombia / Antimicrobial activity of four varieties of plants against pathogens clinical significance in Colombia

Resumen Objetivo. Evaluar la actividad antimicrobiana de los extractos de las plantas Bauhinia sp., Sambucus nigra, Eichhornia crassipes y Taraxacum officinale frente a patógenos de importancia clínica. Método. La metodología incluyó la adquisición, secado, maceración, molienda, preparación de los extractos crudos etanólicos y concentración por rotaevaporación, análisis fitoquimico y se separaron las fracciones por cromatografía en capa fina. Las pruebas antimicrobianas se realizaron con diferentes concentraciones de los extractos según las indicaciones de Clinical and Laboratory Standars Institute. Los microorganismos utilizados fueron Enterococcus faecium resistente a vancomicina, Streptococcus pneumoniae, Klebsiella pneumoniae con presencia de KPC, Providencia rettgeri con presencia de ESBLs, Pseudomonas aeruginosa, Enterobacter cloacae, Escherichia coli, Staphylococcus aureus (3-lisina y Candida albicans. Resultados. Las cromatografías permitieron comprobar la presencia de flavonoides, terpenos, saponinas, fenoles, quinonas y alcaloides que han sido reportados con actividad antimicrobiana. En los ensayos de susceptibilidad antimicrobiana se encontró que los extractos presentaban diversos grados de inhibición frente a los microrganismos de estudio, siendo el más eficaz los tallos de T. officinale. Conclusión. Se puede concluir que los extractos vegetales podrían ser una alternativa de tratamiento para infecciones nosocomiales.

Objective. To evaluate the antimicrobial activity of extracts of plants Bauhinia sp., Sambucus nigra, Taraxacum officinale and Eichhornia crassipes against clinically important pathogens. Method. The methodology included the acquisition, drying, soaking, grinding, preparing ethanolic crude extracts and concentration by rotary evaporation; phytochemical analysis, visualization by thin layer chromatography. Antimicrobial tests were performed with different concentrations of extracts as indicated by the Clinical and Laboratory Standards Institute. The microorganisms used were Enterococcus faecium vancomycin-resistant, Streptococcus pneumoniae, Klebsiella pneumoniae with KPC, Providencia rettgeri presence of ESBLs, Pseudomonas aeruginosa, Enterobacter cloacae, Escherichia coli, Staphylococcus aureus (3-lysine and Candida albicans. Results. Chromatographies allowed checking the presence of flavonoids, terpenes, saponins, phenols, quinones and alkaloids have been reported having antimicrobial activity. In antimicrobial susceptibility tests it found that extracts showed varying degrees of inhibition against microorganisms study, the most effective stems T. officinale. Conclusion. It can be concluded that plant extracts may be alternative in the treatment of nosocomial infections.

Toxicidade equivalente da atmosfera por dioxinas, furanos e bifenilas policloradas,com uso de duas técnicas de coleta, passiva e ativa / Equivalent toxicity of the atmosphere for dioxins, furans and polychlorinated biphenyls, using two air sampling techniques, passive and active

Introdução: Dioxinas, furanos e bifenilas policloradas são poluentes tóxicos para a saúde humana incluindo riscos de incidência de cânceres, efeitos de neurodesenvolvimento, lesões dérmicas, cloroacne. Estes compostos são poluentes orgânicos persistentes (POPs) que podem ser transportados de longas distâncias da fonte de emissão e se bioacumular em ecossistemas. A atmosfera poluída foi recentemente classificada como carcinogênica para os seres humanos pela Organização Mundial da Saúde, mostrando a importância de sua caracterização, principalmente para compostos tóxicos. Entretanto, técnica de coleta ativa tem custo elevado para POPs, e existem poucos estudos de calibração que validem a substituição. Objetivos: Avaliar a toxicidade equivalente da atmosfera por dioxinas, furanos e bifenilas cloradas, utilizando técnicas de coleta ativa e passiva, e verificar gradiente de concentração nos ambientes urbano, urbano/industrial e de background. Método: Amostras de ar foram coletadas, utilizando coletores ativos e passivos, durante dois períodos consecutivos de quatro meses: de setembro a dezembro de 2014 (período 1) e de maio a agosto de 2015 (período 2) em três cidades de São Paulo, SP, em ambientes urbano, urbano/industrial e de background. Todas as amostras foram extraídas com solução de tolueno:acetona (9:1) em Soxhlet por 24 h e padrões marcados (13C12-PCDD/Fs e 13C12-PCBs) foram adicionados em cada amostra antes do processo de extração. Os extratos foram purificados em coluna de sílica mista (40 por cento H2SO4 e 10 por cento AgNO3) seguida por coluna de alumina. O procedimento analítico foi realizado utilizando HRGC/HRMS (High Resolution Gas Chromatograph/High Resolution Mass Spectrometer) operando em ionização de impacto de elétrons com energia de 35 eV no modo SIM (Select Ion Monitoring) e resolução de 10.000. Resultados mostraram que: (1) existe variação sazonal para concentrações de PCDD/Fs no ar entre os períodos 1 e 2 (p=0,03), enquanto as concentrações de dl-PCBs não foram estatisticamente diferentes nestes períodos (p=0,52); (2) existe gradiente de concentração de PCDD/Fs e dl- PCBs que aumenta na seguinte ordem: background

Introduction: Dioxins, furans and polychlorinated biphenyls are toxic pollutants for human health including risks of cancer incidence, neurodevelopmental effects, dermal lesions, chloracne. These compounds are persistent organic pollutants (POPs) that can be transported to long distances from the emission source and they are bioaccumulated in ecosystems. Recently, the outdoor air pollution were classified as carcinogenic to humans by the World Health Organization, showing the importance of its characterization for toxic compounds. However, active air monitoring has a high cost for POPs, and there is a few calibration studies which support that substitution. Objective: To assess the equivalent toxicity of the atmosphere regarding the measurement of dioxins, furans and polychlorinated biphenyls, using active and passive air samplers, and to evatuate the contrasting concentrations at urban, urban/industrial and background sites. Method: Air samples were collected, using active and passive samplers, over two consecutive periods of four months: from September to December 2014 (period 1) and from May to August 2015 (period 2) at three cities in São Paulo, SP, covering urban, urban/industrial and background sites. All samples were extracted with toluene:acetone (9:1) in a Soxhlet apparatus for 24 hours and surrogate standards (13C12-PCDD/F and 13C12-PCBs) were spiked on each sample media prior to extraction procedure. The extracts were purified on an silica column (40 per cent H2SO4 and 10 per cent AgNO3) followed by an alumina column. The analytical procedure was carried out using HRGC/HRMS (High Resolution Gas Chromatograph/High Resolution Mass Spectrometer) operating in electron impact ionization with an energy of 35 eV in SIM (selected ion monitoring) mode and 10.000 resolution power. Results show that (1) there are seasonal variations for PCDD/F concentrations in air between period 1 and 2 (p=0.03), whereas dl-PCB levels were not statistically different (p=0.52) in those periods. (2) PCDD/F and dl-PCB air levels are in the following order: background

Busca por oxidantes quirais para a transformação enantiosseletiva de compostos orgânicos de boro / Search for chiral oxidants for the enantioselective transformation of organic boron compounds

Neste trabalho, avaliou-se o potencial do uso de oxidantes quirais em oxidações enantiosseletivas de compostos orgânicos de boro. É de conhecimento geral que compostos orgânicos de boro, especialmente ésteres e ácidos borônicos são facilmente oxidados por hidroperóxidos em meio básico. No entanto, são escassos na literatura exemplos destas reações de modo enantiosseletivo. A fim de realizar as reações mencionadas, sintetizou-se os hidroperóxidos quirais TADOOH ({(4R,5R)-5-[(hidroperoxidifenil)metil]-2,2-dimetil-1,3-dioxolan-4il}difenilmetanol) e o hidroperóxido quiral derivado de carboidrato, 2,3-dideoxi1-O-oxidanil-4,6-di-O-pivaloil-α-D-eritro-hex-2-enopiranose (di-O-PivOOH). Estes compostos apresentaram resultados interessantes na literatura em oxidações enantiosseletiva de sulfetos orgânicos, em epoxidações de alcenos e em oxidações de Baeyer-Villiger. Inicialmente o potencial oxidativo de ambos hidroperóxidos, bem como a seletividade destes, foi avaliado frente a diversos ésteres borônicos, sendo que somente o TADOOH apresentou resultados promissores. (Ver esquema no PDF) Observou-se uma melhor seletividade do TADOOH frente a ésteres borônicos que possuíam grupos carbonílicos em sua estrutura. Ao submeter o ß-boronil-éster, 3-fenil-3-(4,4,5,5-tetrametil-1,3,2-dioxaborolan-2-il)propanoato de etila, à oxidação com o TADOOH em THF utilizando NaOH como base, a -30°C por 1 hora, obteve-se o respectivo álcool com 40% de e.e. Cálculos de DFT para o estado de transição na oxidação dos ésteres borônicos com o TADOOH foram realizados em colaboração com o grupo do Prof. Dr. Ataualpa Albert Carmo Braga. Estes cálculos demonstraram que o estado de transição é estabilizado por uma ligação de hidrogênio não clássica entre o oxigênio da carbonila e umas das ligações C-H dos grupos fenila do TADOOH. Além dos estudos relatados, a reconhecida metodologia de Sharpless na epoxidação assimétrica de alcoóis alílicos foi adaptada para a oxidação enantiosseletiva de ésteres borônicos. Ao trocar o ligante derivado de éster tártarico, normalmente utilizado nas epoxidações de Sharpless, por (-)-efedrina observou-se uma moderada seletividade deste sistema frente ao pinacol l-fenietilboronato. Investigações mais detalhadas demonstraram que a presença do Ti(IV) não era necessária, sendo que a (-)efedrina era a responsável pela ativação e indução quiral nesta reação.

In this work, it was investigated the potential use of chiral oxidants in organic boron compound oxidation. It is known in the literature, that organic boron compounds can be easily oxidized by hydroperoxides. However, an enantioselective approach in literature is scarce. In order to perform these reactions, hydroperoxide TADOOH ({(4R,5R)-5[(hydroperoxydiphenyl)methyl]-2,2-dimethyl-l,3-dioxolan-4-yl}diphenylmethanol) and carbohydrate derived hydroperoxide, 2,3-dideoxy-1-O-oxidanyl-4,6-di-O-pivaloyl-α-D-erythro-hex-2-enopyranose (di-O-PivOOH), have been synthesized. These compounds showed interesting results in several enantioselective oxidations, as like, organic sulfides oxidation, alkenes epoxidation and Baeyer-Villiger oxidations. The oxidative potential of both hydroperoxides, as well as their selectivity, were evaluated against several boronic esters. Only TADOOH has shown promissing results for further studies. (See Scheme on PDF). Boronic esters containing a carbonyl moiety showed better selectivities with TADOOH, for example, the reaction of ß-boronyl-ester, ethyl 3-phenyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)propanoate, gave the correponding alcohol with 40% e.e. DFT calculations for the transition state in the oxidation of the boronic esters with TADOOH were carried out in collaboration with the group of Prof. Dr. Ataualpa Albert Carmo Braga. These calculations have shown that the transition state is stabilized by a non-classical hydrogen bond between the carbonyl oxygen and one of the C-H bonds of the TADOOH phenyl groups. In addition to the studies, the well-known Sharpless protocol for asymmetric epoxidation of allylic alcohols was adapted in the enantioselective oxidation of boronic esters. By replacing the tartaric ester-derived, commonly used in the Sharpless experiments, for (-)-ephedrine moderate selectivity was observed with pinacol 1-phenylethyl boronate. Further investigations showed that the presence of Ti (IV) was not necessary, and (-)-ephedrine was responsible for the activation and chiral induction in this reaction.

Chemical and physical properties of an anthropogenic dark earth soil from Bragança, Para, Eastern Amazon / Propriedades físicas e químicas de Terra Preta Antropogênica em Bragança, Pará, Amazônia Oriental

Although anthropogenic dark earth (ADE) is generally found in non-floodable land, it also occurs on floodplains but, there is no information about the chemical and physical characteristics of ADE in this environment. In this study, we propose to check the hypothesis that a Gleysol, classified as ADE, presents improved chemical and physical conditions than an adjacent soil, no anthropogenic. Thus, the objective of this study was to characterize the chemical and physical properties of the top layer of two ADE profiles in a Gleysol and compare them with an adjacent soil. Samples were taken from two areas classified as ADE in Bragança, Pará State, Brazil, at the "Jabuti" archaeological site, and from an adjacent non-anthropogenic site. Disturbed and undisturbed soil samples were collected at the soil depth of 0.05-0.10 m for chemical (pH, potential acidity, exchangeable cations, and soil organic carbon) and physical (soil particle size distribution, particles density, water retention curve, total porosity, microporosity, macroporosity, and bulk density) analysis. The two areas of ADE in a Gleysol, showed improved soil chemical properties compared to the adjacent soil, particularly in relation to phosphorus and calcium levels that contributed to higher cation exchange capacity which, in turn, was positively related to organic carbon content. Changes in soil physical properties were less noticeable but both areas of ADE presented higher water retention capacity, particularly at low tension. The improved conditions of the ADE soil under Gleysols shows that these areas are adequate for soil cultivation, especially with plants adapted to floodplain.

Embora as terras pretas antropogênicas (TPA) geralmente são encontradas em áreas de terra firme, elas também ocorrem em áreas e várzeas, mas, não há informações sobre as características químicas e físicas de TPA neste ambiente. Neste estudo, nós propomos verificar a hipótese de que um Gleissolo, classificado como TPA, apresenta melhores condições químicas e físicas do que o solo adjacente, não antropogênico. Assim, o objetivo do estudo foi caracterizar as propriedades químicas e físicas da camada superficial de dois perfis de TPA em um Gleissolo e compará-los com um solo adjacente. Amostras de solo foram coletadas em duas áreas classificadas como TPA em Bragança, Estado do Pará, Brasil, no sítio arqueológico "Jabuti" e em uma área adjacente não antropogênica. Amostras de solo com estrutura deformada e indeformada foram coletadas na profundidade de 0,05-0,10 m para análises químicas (pH, acidez potencial, cátions trocáveis, e carbono orgânico do solo) e físicas (distribuição do tamanho das partículas do solo, densidade de partículas, curva de retenção de água, porosidade total, microporosidade, macroporosidade e densidade do solo). As duas áreas de TPA em Gleissolo, apresentaram melhores propriedades químicas comparadas com o solo adjacente, principalmente em relação aos níveis de fósforo e cálcio que contribuíram para a maior capacidade de troca catiônica que, por sua vez, foi positivamente relacionada com o conteúdo de carbono orgânico. Mudanças nas propriedades físicas do solo foram menos pronunciadas, mas as duas áreas de TPA exibiram maior capacidade de retenção de água, sobretudo em baixa tensão.

Tomato yield and quality under various combinations of organic compost / Produção e qualidade de tomate cultivado sob oito tipos de composto orgânico

The number of growers and planted area dedicated to organic farming have increased considerably. Consequently, demand has increased for alternative organic composts in terms of quantity and new options for direct use with crops. Making use of local industrial byproducts is also important throughout Brazil and the rest of the world. Therefore, it was evaluated the effect of eight types of organic composts, produced with different percentages of poultry litter and crushed sugarcane on tomato yield and quality. The experiment was carried out during the 2013 growing season at the experimental farm of the Agricultural Research Company of Minas Gerais state (Empresa de Pesquisa Agropecuária do Estado de Minas Gerais) located in Prudente de Morais, MG, Brazil. It was evaluated different formulations of poultry litter and crushed sugarcane (0, 10, 20, 30, 40, 50, 50, 60 and 70%) yielding eight types of alternative Bokashi used as organic compost in the production of two tomato cultivars (Santa Clara and the Verano hybrid). It was found that the various formulations of alternative Bokashi, except the alternative Bokashi B1, strengthened components of tomato yield and quality. We found that composts of poultry litter and crushed sugarcane increase tomato yield and quality and can be and excellent alternative, especially for organic farmers.

O cenário atual da agricultura orgânica demonstra aumento considerável do número de produtores e área plantada. Consequentemente, há maior demanda por novas alternativas de compostos orgânicos, não só em volume, mas também disponibilizar novas opções para uso direto no sistema de cultivo. A busca pelo aproveitamento de subprodutos gerados pelas indústrias locais é pauta no Brasil e no mundo. O objetivo do trabalho foi avaliar o efeito de oito tipos de compostos orgânicos produzidos com diferentes porcentagens de cama de frango e cana de açúcar triturada na produção e qualidade do tomate.O experimento foi instalado na fazenda experimental da Empresa de Pesquisa Agropecuária do Estado de Minas Gerais localizada no município de Prudente de Morais, MG no ano agrícola de 2013. Foram avaliadas diferentes formulações entre cama de frango e cana de açúcar triturada(0, 10, 20, 30, 40, 50, 50, 60 e 70%) resultando em oito tipos de Bokashi alternativo utilizado como composto orgânico na produção de duas cultivares de tomate (Santa Clara e Híbrido Verano). Pode-se verificar que as diferentes formulações de Bokashi Alternativo, com exceção da B1, potencializaram os componentes de produção e qualidade do tomate. O uso de cama de frango associado com cana de açúcar triturada promove incrementos na produção e qualidade de tomate demonstrando ser uma excelente alternativa, especialmente para produtores orgânicos.

Process optimization for production and purification of a thermostable, organic solvent tolerant lipase from Acinetobacter sp. AU07

ABSTRACT The purpose of this study was to isolate, purify and optimize the production conditions of an organic solvent tolerant and thermostable lipase from Acinetobacter sp. AU07 isolated from distillery waste. The lipase production was optimized by response surface methodology, and a maximum production of 14.5 U/mL was observed at 30 ºC and pH 7, using a 0.5% (v/v) inoculum, 2% (v/v) castor oil (inducer), and agitation 150 rpm. The optimized conditions from the shake flask experiments were validated in a 3 L lab scale bioreactor, and the lipase production increased to 48 U/mL. The enzyme was purified by ammonium sulfate precipitation and ion exchange chromatography and the overall yield was 36%. SDS-PAGE indicated a molecular weight of 45 kDa for the purified protein, and Matrix assisted laser desorption/ionization time of flight analysis of the purified lipase showed sequence similarity with GDSL family of lipases. The optimum temperature and pH for activity of the enzyme was found to be 50 ºC and 8.0, respectively. The lipase was completely inhibited by phenylmethylsulfonyl fluoride but minimal inhibition was observed when incubated with ethylenediaminetetraacetic acid and dithiothreitol. The enzyme was stable in the presence of non-polar hydrophobic solvents. Detergents like SDS inhibited enzyme activity; however, there was minimal loss of enzyme activity when incubated with hydrogen peroxide, Tween 80 and Triton X-100. The kinetic constants (Km and Vmax) revealed that the hydrolytic activity of the lipase was specific to moderate chain fatty acid esters. The Vmax, Km and Vmax/Km ratio of the enzyme were 16.98 U/mg, 0.51 mM, and 33.29, respectively when 4-nitrophenyl palmitate was used as a substrate.

Evaluation and optimization of SYBR Green real-time reverse transcription polymerase chain reaction as a tool for diagnosis of the Flavivirus genus in Brazil

Abstract: INTRODUCTION: The genus Flavivirus includes several pathogenic species that cause severe illness in humans. Therefore, a rapid and accurate molecular method for diagnosis and surveillance of these viruses would be of great importance. Here, we evaluate and optimize a quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) method for the diagnosis of the Flavivirus genus. METHODS: We evaluated different commercial kits that use the SYBR Green system for real-time RT-PCR with a primer set that amplifies a fragment of the NS5 flavivirus gene. The specificity and sensitivity of the assay were tested using twelve flaviviruses and ribonucleic acid (RNA) transcribed from the yellow fever virus. Additionally, this assay was evaluated using the sera of 410 patients from different regions of Brazil with acute febrile illness and a negative diagnosis for the dengue virus. RESULTS: The real-time RT-PCR amplified all flaviviruses tested at a melting temperature of 79.92 to 83.49°C. A detection limit of 100 copies per ml was determined for this assay. Surprisingly, we detected dengue virus in 4.1% (17/410) of samples from patients with febrile illness and a supposedly negative dengue infection diagnosis. The viral load in patients ranged from 2.1×107to 3.4×103copies per ml. CONCLUSIONS: The real-time RT-PCR method may be very useful for preliminary diagnoses in screenings, outbreaks, and other surveillance studies. Moreover, this assay can be easily applied to monitor viral activity and to measure viral load in pathogenesis studies.

Effect of organo and inorganic lithium salt on human blood plasma glutathione- a comparative study

Investigation of toxicological effect of various metals is the field of interest for toxicological scientists since four to five decades and especially the toxicological effect of those drugs containing metals and there use is common because there is no other choice except to use these metal containing drugs. Inorganic as well as organic salts of lithium are commonly used in prophylaxis and treatments of many psychiatric disorders. The aim of the present study was to see the difference between the effect of organic and inorganic salt of lithium commonly used in psychiatric disorders on the GSH of human blood plasma. It is the scientific fact that ionic dissociation of organic and inorganic salts of any metal is always quite different hence to prove this fact, the effect of lithium citrate [organic salt of lithium] and lithium carbonate [inorganic salt of lithium] was investigated on human blood plasma GSH to find the difference between the effect of two. Ellman's method was used for the quantification of glutathione contents in plasma. It was found that lithium citrate decrease plasma GSH contents less than lithium carbonate indicating that organic salts of lithium are safe than inorganic salts of lithium when are used in psychiatric disorders. Further to analyze the effect of organic and inorganic salt of lithium on blood plasma GSH with the increase in incubation time was also evaluated and was found that both concentration and time dependent effect of organic salt of lithium shows that this salt has decreased plasma GSH contents of human blood less than inorganic salt of lithium either by promoting oxidation of GSH into GSSG or by lithium glutathione complex formation. These results suggest the physicians that the use of organic lithium salts is much safer than inorganic salts of lithium in terms of depletion of blood plasma GSH contents

Preface for special issue on bio-based materials (2016) / 生物工程学报

Bio-based materials are new materials or chemicals with renewable biomass as raw materials such as grain, legume, straw, bamboo and wood powder. This class of materials includes bio-based polymer, biobased fiber, glycotechnology products, biobased rubber and plastics produced by biomass thermoplastic processing and basic biobased chemicals, for instance, bio-alcohols, organic acids, alkanes, and alkenes, obtained by bio-synthesis, bio-processing and bio-refinery. Owing to its environmental friendly and resource conservation, bio-based materials are becoming a new dominant industry taking the lead in the world scientific and technological innovation and economic development. An overview of bio-based materials development is reported in this special issue, and the industrial status and research progress of the following aspects, including biobased fiber, polyhydroxyalkanoates, biodegradable mulching film, bio-based polyamide, protein based biomedical materials, bio-based polyurethane, and modification and processing of poly(lactic acid), are introduced.

Effects of Leukemia Inhibitory Factor Combined with Basic Fibroblast Growth Factor on Self-maintenance and Self-renewal of Human Umbilical Cord Mesenchymal Stem Cells In Vitro / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the effects of LIF combined with bFGF on the proliferation, stemness and senescence of hUC-MSC.</p><p><b>METHODS</b>Experiments were divided into 4 groups: control group, in which the cells were treated with complete medium (α-MEM containing 10% FBS); group LIF, in which the cells were treated with complete medium containing 10 ng/ml LIF; group bFGF, in which the cells were treated with complete medium containing 10 ng/ml bFGF; combination group, in which the cells were treated with complete medium containing 10 ng/ml LIF and 10 ng/ml bFGF. The growth curves of hUC-MSC at passage 4 in different groups were assayed by cell counting kit 8. Cellular morphologic changes were observed under inverted phase contrast microscope; hUC-MSC senescence in different groups was detected by β-galactosidase staining. The expression of PCNA, P16, P21, P53, OCT4 and NANOG genes was detected by RT-PCR.</p><p><b>RESULTS</b>The cell growth curves of each group were similar to the S-shape; the cell proliferation rate from high to low as follows: that in the combination group > group bFGF > group LIF > control group. Senescence and declining of proliferation were observed at hUC-MSC very early in control group; the cells in group LIF maintained good cellular morphology at early stage, but cell proliferation was slow and late senescence was observed; a few cells in group bFGF presented signs of senescence, but with quick proliferation; the cells in combination group grew quickly and maintained cellular morphology of hUC-MSC for long time. The LIF and bFGF up-regulated the expression of PCNA, OCT4 and NANOG, while they down-regulated the expression of P16, P21, P53, and their combinative effects were more significant.</p><p><b>CONCLUSION</b>LIF combined with bFGF not only can promote the proliferation and maintenance of stemness of hUC-MSC, but also can delay the senescence of hUC-MSC.</p>

Fluorescent membrane markers elucidate the association of Borrelia burgdorferi with tick cell lines

This study aimed to describe the association of Borrelia burgdorferi s.s. with ixodid tick cell lines by flow cytometry and fluorescence and confocal microscopy. Spirochetes were stained with a fluorescent membrane marker (PKH67 or PKH26), inoculated into 8 different tick cell lines and incubated at 30°C for 24 h. PKH efficiently stained B. burgdorferi without affecting bacterial viability or motility. Among the tick cell lines tested, the Rhipicephalus appendiculatus cell line RA243 achieved the highest percentage of association/internalization, with both high (90%) and low (10%) concentrations of BSK-H medium in tick cell culture medium. Treatment with cytochalasin D dramatically reduced the average percentage of cells with internalized spirochetes, which passed through a dramatic morphological change during their internalization by the host cell as observed in time-lapse photography. Almost all of the fluorescent bacteria were seen to be inside the tick cells. PKH labeling of borreliae proved to be a reliable and valuable tool to analyze the association of spirochetes with host cells by flow cytometry, confocal and fluorescence microscopy.

Enhanced production and organic solvent stability of a protease fromBrevibacillus laterosporus strain PAP04

A bacterial strain (PAP04) isolated from cattle farm soil was shown to produce an extracellular, solvent-stable protease. Sequence analysis using 16S rRNA showed that this strain was highly homologous (99%) to Brevibacillus laterosporus. Growth conditions that optimize protease production in this strain were determined as maltose (carbon source), skim milk (nitrogen source), pH 7.0, 40°C temperature, and 48 h incubation. Overall, conditions were optimized to yield a 5.91-fold higher production of protease compared to standard conditions. Furthermore, the stability of the enzyme in organic solvents was assessed by incubation for 2 weeks in solutions containing 50% concentration of various organic solvents. The enzyme retained activity in all tested solvents except ethanol; however, the protease activity was stimulated in benzene (74%) followed by acetone (63%) and chloroform (54.8%). In addition, the plate assay and zymography results also confirmed the stability of the PAP04 protease in various organic solvents. The organic solvent stability of this protease at high (50%) concentrations of solvents makes it an alternative catalyst for peptide synthesis in non-aqueous media.