Enzymatic synthesis of SFAEs using the commercial ionic liquid CYPHOS 104

Sugar fatty acid esters (SFAEs) are compounds formed by a carbohydrate linked to one or more fatty acid (FA) chains. These molecules present interesting technological properties and are commonly used as biodegradable surfactants and emulsifiers in the food, pharmaceutical and cosmetics industries [1]. Recently, the synthesis of SFAEs using enzymes as biocatalysts proved to be a greener alternative. The main problem associated with the synthesis of SFAEs is the different chemical nature of the substrates, which can significantly affect the effectiveness of the enzymatic synthesis and greatly compromise the yield of the process. Lipases are quite stable biocatalysts and have been successfully used in traditional media and alternative media containing ionic liquids (ILs) [2,3]. ILs present an interesting strategy to overcome the process limitations since they generally increase enzyme stability and can be tailored to improve the solubility of a wide range of substrates. Moreover, ILs are non-volatile, non-flammable, chemically and thermally stable and biocompatible being greener alternatives to the hazardous and volatile organic solvents commonly applied [4]. In the present work, the enzymatic synthesis of SFAEs in the commercial IL CYPHOS 104 was followed and qualitatively evaluated by thin layer chromatography. The esterification between a disaccharide and vinyl laurate was performed by immobilized Lipase B from Candida antarctica, at 60 °C, using two agitation strategies (rotatory and orbital). The results showed that SFAEs were successfully biosynthesized in the IL CYPHOS 104 using a rotatory agitation.We thank the Fundação de Amparo à Pesquisa do estado de São Paulo (process 2018/07522-6; 2014/50884-5), and Conselho Nacional de Dsenvolvimento Científico (process 301963/2017-7; 465319/2014-9).info:eu-repo/semantics/publishedVersio

Biocompatible and biodegradable functional coatings with natural occurring materials for the corrosion protection of Mg alloys

Magnesium alloys are amidst the most innovative materials for biomedical applications, as they show a set of unique properties, namely appropriate mechanical properties and biodegradability, when compared to other alloys. Although these properties make them suitable for medical implants, the main challenge is the uncontrolled corrosion. Mg degradation is fast, inhomogeneous, localized and often accompanied by hydrogen formation which can lead to complications in vivo. Here, we propose the development of a functional coating, containing natural-based capsules for the controlled release of biocompatible corrosion inhibitors and well known pharmaceutical agents. Empty and loaded capsules toxicity tests were performed as a first step for materials selection. Subsequently, they were incorporated into polyetherimide (PEI) coatings and tested using electrochemical impedance spectroscopy (EIS) under aggressive conditions. The obtained results showed a successful synthesis of natural-based microcapsules, constituting a fast, simple and environmentally friendly method. Additionally, the high cell proliferation observed in the presence of the aforementioned materials demonstrates their low toxicity. Preliminary results carried out with capsule-modified coatings show that the incorporation of Ca2+-loaded gelatin capsules in PEI coatings leads to barrier and active corrosion protection properties improvement and that anti-inflammatory agent ibuprofen may have a role in active corrosion protection as well.publishe

Identification of candidate genes for yeast engineering to improve bioethanol production in very high gravity and lignocellulosic biomass industrial fermentations

<p>Abstract</p> <p>Background</p> <p>The optimization of industrial bioethanol production will depend on the rational design and manipulation of industrial strains to improve their robustness against the many stress factors affecting their performance during very high gravity (VHG) or lignocellulosic fermentations. In this study, a set of <it>Saccharomyces cerevisiae </it>genes found, through genome-wide screenings, to confer resistance to the simultaneous presence of different relevant stresses were identified as required for maximal fermentation performance under industrial conditions.</p> <p>Results</p> <p>Chemogenomics data were used to identify eight genes whose expression confers simultaneous resistance to high concentrations of glucose, acetic acid and ethanol, chemical stresses relevant for VHG fermentations; and eleven genes conferring simultaneous resistance to stresses relevant during lignocellulosic fermentations. These eleven genes were identified based on two different sets: one with five genes granting simultaneous resistance to ethanol, acetic acid and furfural, and the other with six genes providing simultaneous resistance to ethanol, acetic acid and vanillin. The expression of <it>Bud31 </it>and <it>Hpr1 </it>was found to lead to the increase of both ethanol yield and fermentation rate, while <it>Pho85</it>, <it>Vrp1 </it>and <it>Ygl024w </it>expression is required for maximal ethanol production in VHG fermentations. Five genes, <it>Erg2</it>, <it>Prs3</it>, <it>Rav1</it>, <it>Rpb4 </it>and <it>Vma8</it>, were found to contribute to the maintenance of cell viability in wheat straw hydrolysate and/or the maximal fermentation rate of this substrate.</p> <p>Conclusions</p> <p>The identified genes stand as preferential targets for genetic engineering manipulation in order to generate more robust industrial strains, able to cope with the most significant fermentation stresses and, thus, to increase ethanol production rate and final ethanol titers.</p

Genetic analysis of enteropathogenic Escherichia coli (EPEC) adherence factor (EAF) plasmid reveals a new deletion within the EAF probe sequence among O119 typical EPEC strains

Enteropathogenic Escherichia coli (EPEC) are classified into typical and atypical strains based on the presence of the E. coli adherence factor (EAF) plasmid. The EAF plasmid contains the bfp (bundle-forming pilus) operon and the perABC (plasmid encoded regulator) gene cluster. A 1-kb cryptic region of EAF plasmid has been widely used as a genetic probe for EPEC detection. However, some EPEC strains may harbor an EAF plasmid lacking the EAF probe sequence, which makes the differentiation between typical and atypical a complex task. In this study, we report the genetic analysis of the EAF plasmid-encoded genes in a collection of EPEC clinical isolates. A total of 222 EPEC clinical isolates, which were previously classified as typical (n = 70) or atypical (n = 152) by EAF probe reactivity, were screened for the presence of different EAF sequences by PCR and DNA hybridization. All typical strains possessed intact bfpA and perA genes, and most of them were positive in the PCR for EAF probe sequence. However, a subset of 30 typical strains, 22 of which belonged to O119 serogroup, presented a 1652 pb deletion in the region between 1093-bp downstream perC and 616-bp of the EAF fragment. The bfpA, bfpG, and per genes were found in all typical strains. In addition, 32 (21 %) atypical strains presented the perA gene, and 20 (13.2 %) also presented the bfpA gene. Among the 32 strains, 16 belonged to the O119:H2, O119:HND, and ONT:HND serotypes. All 32 atypical strains contained perA mutation frameshifts and possessed an IS1294 element upstream of the per operon as detected by PCR followed by restriction fragment length polymorphism (RFLP) typing and multiplex PCR. Among the 20 bfpA probe-positive strains, eight O119 strains possessed deletion in the bfp operon at the 3'end of bfpA due to an IS66 element. Our data show that typical O119 strains may contain a deletion within the EAF probe sequence not previously reported. This new finding suggests that care should be taken when using the previously described EAF PCR assay in epidemiological studies for the detection of typical O119 strains. In addition, we were able to confirm that some atypical strains carry vestiges of the EAF plasmid.1520

Design of a lipid nanovesicle system encapsulating bacteriophages integrated in a multiple emulsion formulation: A proof-of-concept

Development of a biotechnological process for the inhalational administration of a bacteriophage was pursued, using strategies of nanoencapsulation within lipid nanovesicles. As a proof-of-concept for the nanoencapsulation strategy, a bacteriophage with broad lytic spectrum was entrapped within W/O/W multiple nanoemulsions. Physicochemical characterization of the optimized bacteriophage-encasing nanovesicles encompassed determination of particle hydrodynamic size, size distribution and particle charge via DLS, surface morphology via CRYO-SEM, and thermal analysis via DSC, whereas antimicrobial activity of the nanoemulsions produced was assessed in vitro using several bacterial strains. The optimized nanosystems showed no phase separation and encompassed nanovesicles with an average size of ca. 114 nm and an average Zeta Potential of ca. -13 mV, which were maintained stable over a storage timeframe of ca. 3 months.info:eu-repo/semantics/publishedVersio

Et Blad af Sønderskov Mølles Historie

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Background: Enteropathogenic Escherichia coli (EPEC) are classified into typical and atypical strains based on the presence of the E. coli adherence factor (EAF) plasmid. The EAF plasmid contains the bfp (bundle-forming pilus) operon and the perABC (plasmid encoded regulator) gene cluster. A 1-kb cryptic region of EAF plasmid has been widely used as a genetic probe for EPEC detection. However, some EPEC strains may harbor an EAF plasmid lacking the EAF probe sequence, which makes the differentiation between typical and atypical a complex task. In this study, we report the genetic analysis of the EAF plasmid-encoded genes in a collection of EPEC clinical isolates. Methods: A total of 222 EPEC clinical isolates, which were previously classified as typical (n = 70) or atypical (n = 152) by EAF probe reactivity, were screened for the presence of different EAF sequences by PCR and DNA hybridization. Results: All typical strains possessed intact bfpA and perA genes, and most of them were positive in the PCR for EAF probe sequence. However, a subset of 30 typical strains, 22 of which belonged to O119 serogroup, presented a 1652 pb deletion in the region between 1093-bp downstream perC and 616-bp of the EAF fragment. The bfpA, bfpG, and per genes were found in all typical strains. In addition, 32 (21 %) atypical strains presented the perA gene, and 20 (13.2 %) also presented the bfpA gene. Among the 32 strains, 16 belonged to the O119: H2, O119: HND, and ONT: HND serotypes. All 32 atypical strains contained perA mutation frameshifts and possessed an IS1294 element upstream of the per operon as detected by PCR followed by restriction fragment length polymorphism (RFLP) typing and multiplex PCR. Among the 20 bfpA probe-positive strains, eight O119 strains possessed deletion in the bfp operon at the 3' end of bfpA due to an IS66 element. Conclusion: Our data show that typical O119 strains may contain a deletion within the EAF probe sequence not previously reported. This new finding suggests that care should be taken when using the previously described EAF PCR assay in epidemiological studies for the detection of typical O119 strains. In addition, we were able to confirm that some atypical strains carry vestiges of the EAF plasmid.15Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Girişimciliğin ilk aşamasında finansman temin yolları ve finansman temininde karşılaşılan sorunlar: Gaziantep ilinde bir uygulama

Bugün işletmeler gelişen dünya pazarlarında rekabette bulunarak ekonomik ve finansal açıdan avantaj sağlamaktadırlar. İşletmeler, girişimcilerin temel özellikleri ile teknolojiyi kullanan ve geliştiren işletmeler olarak ülkelerin kalkınmasında önemli rol oynamaktadır. Teknolojik yeniliklerin yanında inovasyona da yatırım yaparak gelişimlerini tamamlarlar. Ancak her alanda olduğu gibi, girişimciler finansal açıdan da sorunlarla karşılaşmaktadırlar. Dolayısıyla işletme sayısının artarak ülkeye katkıda bulunabilmesi için finansman teminindeki sorunları farklı olan girişimcilerin desteklenmeleri ve yönlendirilmeleri gerekmektedir. Bu noktada, girişimcileri desteklemek amacıyla kamu kurum ve kuruluşları ve diğer kuruluşlar faaliyetlerini sürdürmektedir. Bu araştırmanın temel amacı, girişimciliğin ilk aşamasında finans temin yollarının belirlenerek finansman temin yönteminde karşılaşılan sorunları ortaya koymaktır. Bununla birlikte, bu çalışmada Türkiye’de girişimciliğin temel özellikleri, girişimcilik finansman türleri ve finansman temininde karşılaşılan sorunlara yönelik önerilerin sunulması amaçlanmaktadır. Araştırma kapsamında nitel ve nicel araştırma yöntemleri uygulanmıştır. Nicel araştırma kapsamında, 408 girişimciye anket uygulanmış, nitel araştırma kapsamında ise 180 girişimciyle yüz yüze görüşmeler yapılmıştır. Araştırma yöntemlerinden elde edilen veriler, çeşitli istatistiksel analizlere tabi tutulmuştur. Girişimciliğin ilk aşamasında, girişimcilerin finansman temininde karşılaştıkları sorunlar belirlenerek, bu sorunlara yönelik çözüm önerilerinde bulunulmuştur. Bu araştırmanın sonucunda, girişimciliğin ilk aşamasında, girişimcilerin finansman temin yöntemi konusunda yeterli derecede bilgi sahibi olmadıkları ve kendi kaynaklarını daha çok kullandıkları tespit edilmiştir

Identification of a DNA-binding site for the transcription factor Haa1, required for Saccharomyces cerevisiae response to acetic acid stress

The transcription factor Haa1 is the main player in reprogramming yeast genomic expression in response to acetic acid stress. Mapping of the promoter region of one of the Haa1-activated genes, TPO3, allowed the identification of an acetic acid responsive element (ACRE) to which Haa1 binds in vivo. The in silico analysis of the promoter regions of the genes of the Haa1-regulon led to the identification of an Haa1-responsive element (HRE) 5′-GNN(G/C)(A/C)(A/G)G(A/G/C)G-3′. Using surface plasmon resonance experiments and electrophoretic mobility shift assays it is demonstrated that Haa1 interacts with high affinity (KD of 2 nM) with the HRE motif present in the ACRE region of TPO3 promoter. No significant interaction was found between Haa1 and HRE motifs having adenine nucleotides at positions 6 and 8 (KD of 396 and 6780 nM, respectively) suggesting that Haa1p does not recognize these motifs in vivo. A lower affinity of Haa1 toward HRE motifs having mutations in the guanine nucleotides at position 7 and 9 (KD of 21 and 119 nM, respectively) was also observed. Altogether, the results obtained indicate that the minimal functional binding site of Haa1 is 5′-(G/C)(A/C)GG(G/C)G-3′. The Haa1-dependent transcriptional regulatory network active in yeast response to acetic acid stress is proposed

Electrospinning polypropylene with an amino acid as a strategy to bind the antimicrobial peptide Cys-LC-LL-37

Hospital isolation gowns are increasingly competitive, with brands and manufacturers contesting consumer preferences. The textile materials in contact with the skin can acquire secretions and multiresistant microorganisms, causing discomfort and health risks, respectively. A new nanofibrous substrate---polypropylene grafted with l-Cys---was developed with an increased crystallinity, providing its surface with --SH hooks necessary to efficiently cross-link the antimicrobial peptide Cys-LC-LL-37 in order to protect against nosocomial pathogens and their spread to community. Furthermore, this application does not require environmental control of humidity, and it is not susceptible to enzyme and microorganism degradation.The authors acknowledge the Fundação para a Ciência e Tecnologia (FCT) for the PhD Grant SFRH/ BD/91444/2012 and Programa Operacional Capital Humano (POCH) and European Union for co-funding the work.info:eu-repo/semantics/publishedVersio