Industrial Biotechnology

Background: Tannase is an enzyme that catalyses the breakdown of ester bonds in gallotannins such as tannic acid. In recent years, the interest on bacterial tannases has increased because of its wide applications. The lactic acid bacteria (LAB) plays an important role in food tannin biotransformation, it has the ability of hydrolyse tannins in ruminants intestine. The finding of tannin hydrolysis by LAB has sparked their use as tannase producer. The bacterial strains used in the present work were identified as Bacillus subtilis AM1 and Lactobacillus plantarum CIR1. The maximal tannase production levels were 1400 and 1239 U/L after 32 and 36 h of fermentation respectively, for B. subtilis AM1 and L. plantarum CIR1. Maximum gallic acid release was 24.16 g/L for B. subtilis AM1 and 23.73 g/L for L. plantarum CIR1. HPLC analysis showed the formation of another peaks in the retention time range of 9-14 min, which could be attributed to the formation of di or tri-galloyl glucose. Conclusions: According to database, the strains were identified as Bacillus subtilis AM1 and Lactobacillus plantarum CIR1. In conclusion, both strains had the capability to produce good titres of extracellular tannase and release gallic acid.

One of the species of mushrooms authorized for sale in some countries-Tricholoma equestre-can be harmful, and its excessive consumption can lead to serious health problems. Many authors have pointed to the relationship between the consumption of the T. equestre and the occurrence of rhabdomyolysis. The aim of this study was to analyze the composition of biologically active substances in the T. equestre fruiting bodies, and determine their biological activity. The main aim of the analyses performed in the present study was to determine whether the T. equestre is safe or dangerous for consumption by humans. The obtained results, in addition to the existing reports on poisoning caused by T. equestre consumption, are another reason to not take the risk of consuming the fruiting bodies of this species, as the natural environment is rich in other species of edible mushrooms that are not only tasty, but also exhibit healing properties.

Background: A consolidated bioprocessing (CBP), where lignocellulose is converted into the desired product(s) in a single fermentative step without the addition of expensive degradative enzymes, represents the ideal solution of renewable routes to chemicals and fuels. Members of the genus Geobacillus are able to grow at elevated temperatures and are able to utilise a wide range of oligosaccharides derived from lignocellulose. This makes them ideally suited to the development of CBP. In this study, we engineered Geobacillus thermoglucosidasius NCIMB 11955 to utilise lignocellulosic biomass, in the form of nitric acid/ammonia treated wheat straw to which expensive hydrolytic enzymes had not been added. Two different strains, BZ9 and BZ10, were generated by integrating the cglT (β-1,4-glucosidase) gene from Thermoanaerobacter brockii into the genome, and localising genes encoding different cellulolytic enzymes on autonomous plasmids. The plasmid of strain BZ10 carried a synthetic cellulosomal operon comprising the celA (Endoglucanase A) gene from Clostridium thermocellum and cel6B (Exoglucanase) from Thermobifida fusca; whereas, strain BZ9 contained a plasmid encoding the celA (multidomain cellulase) gene from Caldicellulosiruptor bescii. All of the genes were successfully expressed, and their encoded products secreted in a functionally active form, as evidenced by their detection in culture supernatants by Western blotting and enzymatic assay. In the case of the C. bescii CelA enzyme, this is one of the first times that the heterologous production of this multi-functional enzyme has been achieved in a heterologous host. Both strains (BZ9 and BZ10) exhibited improved growth on pre-treated wheat straw, achieving a higher final OD600 and producing greater numbers of viable cells. To demonstrate that cellulosic ethanol can be produced directly from lignocellulosic biomass by a single organism, we established our consortium of hydrolytic enzymes in a previously engineered ethanologenic G. thermoglucosidasius strain, LS242. We observed approximately twofold and 1.6-fold increase in ethanol production in the recombinant G. thermoglucosidasius equivalent to BZ9 and BZ10, respectively, compared to G. thermoglucosidasius LS242 strain at 24 h of growth. We engineered G. thermoglucosidasius to utilise a real-world lignocellulosic biomass substrate and demonstrated that cellulosic ethanol can be produced directly from lignocellulosic biomass in one step. Direct conversion of biomass into desired products represents a new paradigm for CBP, offering the potential for carbon neutral, costeffective production of sustainable chemicals and fuels.

A study of the presence of hepatitis A virus (HAV) and enterovirus (EV) in shellfish from the northwestern coast of Spain, one of the most important mussel producers in the world, was carried out employing dot-blot hybridization and RT-PCR techniques. In addition, bacterial contamination of the samples was evaluated by Escherichia coli (EC) counts, according to the European Union (EU) standards of shellfish microbiological quality. Shellfish samples included raft-cultured and wild mussels, as well as wild clams and cockles. Bacterial counts showed that the majority of samples (40.8%) could be classified as moderately polluted following the EU standards, and therefore should undergo depuration processes. However, differences in bacterial contamination were observed between cultured mussel and wild shellfish. Thus, percentage of clean samples ( < 230 EC/100 g shellfish) was clearly higher in cultured mussels (49.1%) than in wild mussels (22.8%) or clams and cockles (10.7%). HAV was detected in 27.4% and EV in 43.9% of the samples that were analyzed. Simultaneous detection of both viral types occurred in 14.1% of the samples. Statistical tests of dependence (chi-square test) showed no relationship either between viral and bacterial contamination, or between the presence of HAV and EV. Comparative analysis of hybridization and RT-PCR for viral detection yielded different results depending on the virus type that was studied, RT-PCR being effective for HAV but not for EV detection. The obtained results reinforce once again the inadequacy of bacteriological standards to assess viral contamination and suggest that although virological analysis of shellfish is possible by molecular techniques, interlaboratory standardization and validation studies are needed before the routine use in monitoring shellfish microbiological safety.

Background: The recent CRISPR-Cas coupled with λ recombinase mediated genome recombineering has become a common laboratory practice to modify bacterial genomes. It requires supplying a template DNA with homology arms for precise genome editing. However, generation of homology arms is a time-consuming, costly and inefficient process that is often overlooked. In this study, we first optimized a CRISPR-Cas genome engineering protocol in the Escherichia coli (E. coli) BL21 strain and successfully deleted 10 kb of DNA from the genome in one round of editing. To further simplify the protocol, asymmetric homology arms were produced by PCR in a single step with two primers and then purified using a desalting column. Unlike conventional homology arms that are prepared through overlapping PCR, cloning into a plasmid or annealing synthetic DNA fragments, our method significantly both shortened the time taken and reduced the cost of homology arm preparation. To test the robustness of the optimized workflow, we successfully deleted 26 / 27 genes across the BL21 genome. Noteworthy, gRNA design is important for the CRISPR-Cas system and a general heuristic gRNA design has been proposed in this study. To apply our established protocol, we targeted 16 genes and iteratively deleted 7 genes from BL21 genome. The resulting strain increased lycopene yield by ~ threefold. Our work has optimized the homology arms design for gene deletion in BL21. The protocol efficiently edited BL21 to improve lycopene production. The same workflow is applicable to any E. coli strain in which genome engineering would be useful to further increase metabolite production.

The recent CRISPR-Cas coupled with λ recombinase mediated genome recombineering has become a common laboratory practise to modify bacterial genomes. It requires supplying a template DNA or homolog arms for precise genome editing. However, it is often overlooked the process to generate the homolog arms which is a time-consuming, costly and inefficient step. In this study, we first optimized CRISPR-Cas protocol in BL21 strain and successfully deleted 10 kb gene from the genome in one round of editing. To further simplify the protocol, asymmetric homolog arms as PCR fragments was used. It can be obtained by one-step PCR reaction with two primers and purified with desalting columns. Unlike conventional homolog arms that are prepared through overlapping PCR, cloning to plasmid or annealing synthetic DNA fragments, our method significantly shortened the time taken and reduced the cost to prepare the homolog arms. To test the robustness of the optimized workflow, we successfully deleted 26 / 27 genes across BL21 genome. Noteworthy, gRNA design is important for CRISPR-Cas system and a general heuristic gRNA design was proposed in the study. To apply our established protocol, we targeted 16 genes and iteratively deleted 7 genes from BL21 genome. The resulting strain increased lycopene production from ~15,000 ppm to > 40,000 ppm. Our work has optimized the homolog arms design for gene deletion in BL21 strains. The protocol efficiently edited BL21 strain to improve lycopene production. The same workflow is applicable to all E. coli strain which would be useful for genome rewiring to further increase metabolite production in microbial cell factories.

Background Noroviruses are a major cause of epidemic and sporadic acute non-bacterial gastroenteritis worldwide. Unfortunately, the development of an effective norovirus vaccine has proven difficult and no prophylactic vaccine is currently available. Further research on norovirus vaccine development should be considered an absolute priority and novel vaccine candidates are needed. One of the recent approaches in safe vaccine development is the use of virus-like particles (VLPs). VLP-based vaccines show great immunogenic potential as they mimic the morphology and structure of viral particles without the presence of the virus genome. Results This study is the first report showing successful production of norovirus VLPs in the protozoan Leishmania tarentolae (L. tarentolae) expression system. Protozoan derived vaccine candidate is highly immunogenic and able to not only induce a strong immune response (antibody titer reached 104) but also stimulate the production of neutralizing antibo...

The efficient and sustainable disposal of waste represents one of the major challenges for the human beings in the next future. The thermochemical conversion of livestock manure to biochar has received much interest thanks to its greenhouse gases extenuation and amendment power. In this study, horse manure was processed under pyrolytic condition at different temperatures, heating rate and residence times. In the first phase, a set of preliminary analysis aimed to assess the physical and chemical properties of the horse manure were carried out. Analogously, after the pyrolysis, the biochar was analysed both from the chemical and physical point of view. The results showed that the major impact on biochar yield (average -32%), pH (average + 40%), SSA (average 27.8 m 2 /g), PAH content, C (average + 37.4%) H (average -53.1%) N (average -15.5%) S (average -65.9%) O (average -37%) content was due to the HHT (highest heating temperature). Residence time (RT) and heating rate (HR) do not display a noticeable trend for most of the parameters. This analysis permitted to assess a range of pyrolysis parameters to treat this kind of biomass: 600 • C (HHT), 10 • C/min (HR) and 2h (RT), in order to comply with EBC (European Biochar Certification) standard.

The glucosyltransferase UGT76G1 from Stevia rebaudiana is a chameleon enzyme in the targeted biosynthesis of the next-generation premium stevia sweeteners, rebaudioside D (Reb D) and rebaudioside M (Reb M). These steviol glucosides carry five and six glucose units, respectively, and have low sweetness thresholds, high maximum sweet intensities and exhibit a greatly reduced lingering bitter taste compared to stevioside and rebaudioside A, the most abundant steviol glucosides in the leaves of Stevia rebaudiana. In the metabolic glycosylation grid leading to production of Reb D and Reb M, UGT76G1 was found to catalyze eight different reactions all involving 1,3-glucosylation of steviol C 13 -and C 19 -bound glucoses. Four of these reactions lead to Reb D and Reb M while the other four result in formation of side-products unwanted for production. In this work, side-product formation was reduced by targeted optimization of UGT76G1 towards 1,3 glucosylation of steviol glucosides that are already 1,2-diglucosylated. The optimization of UGT76G1 was based on homology modelling, which enabled identification of key target amino acids present in the substrate-binding pocket. These residues were then subjected to site-saturation mutagenesis and a mutant library containing a total of 1748 UGT76G1 variants was screened for increased accumulation of Reb D or M, as well as for decreased accumulation of side-products. This screen was performed in a Saccharomyces cerevisiae strain expressing all enzymes in the rebaudioside biosynthesis pathway except for UGT76G1. Conclusions: Screening of the mutant library identified mutations with positive impact on the accumulation of Reb D and Reb M. The effect of the introduced mutations on other reactions in the metabolic grid was characterized. This screen made it possible to identify variants, such as UGT76G1 Thr146Gly and UGT76G1 His155Leu , which diminished accumulation of unwanted side-products and gave increased specific accumulation of the desired Reb D or Reb M sweeteners. This improvement in a key enzyme of the Stevia sweetener biosynthesis pathway represents a significant step towards the commercial production of next-generation stevia sweeteners.

Background Natural products are an important source of drugs and other commercially interesting compounds, however their isolation and production is often difficult. Metabolic engineering, mainly in bacteria and yeast, has sought to circumvent some of the associated problems but also this approach is impeded by technical limitations. Here we describe a novel strategy for production of diverse natural products, comprising the expression of an unprecedented large number of biosynthetic genes in a heterologous host. Results As an example, genes from different sources, representing enzymes of a seven step flavonoid pathway, were individually cloned into yeast expression cassettes, which were then randomly combined on Yeast Artificial Chromosomes and used, in a single transformation of yeast, to create a variety of flavonoid producing pathways. Randomly picked clones were analysed, and approximately half of them showed production of the flavanone naringenin, and a third of them produced ...

Figure 1 : Illustration des produits de B. bruxellensis d'altération des vins. Parmi les altérations microbiennes des vins, la production des phénols volatils par la levure Brettanomyces bruxellensis (B. bruxellensis) est l'une des plus redoutées par le vinificateur et probablement l'une des plus décriées par le consommateur. En outre, les phénols volatils ne sont pas les seuls produits de B. bruxellensis qui peuvent nuire à la qualité du vin (Figure ) et qui font de cette levure l'ennemi numéro un des vinificateurs soucieux de produire un vin fin, fruité et de qualité. Initialement isolée de la bière (Clausen, 1905), la description de la levure B. bruxellensis dans le vin remonte au début de l'oenologie moderne (Agostino, 1950 ; Barret et al. 1950 ; Peynaud et Domercq, 1956). Mais ce n'est que relativement récemment que les microbiologistes du vin, sensibilisés par des consommateurs de plus en plus soucieux de la qualité du produit, se sont penchés sur cette problématique. Les premiers travaux menés sur la production du 4-éthylphénol et du 4-éthylgaïacol se sont focalisés sur B. bruxellensis. Rapidement B. bruxellensis fut décrite comme la seule espèce impliquée et son développement dans le vin comme la conséquence d'une contamination au chai et du non-respect des conditions élémentaires d'hygiène, si bien que les chais victimes de B. bruxellensis étaient aussitôt montrés du doigt. Depuis des études ont démontré que le problème des « Brett » était commun à de nombreux domaines malgré les efforts d'hygiène réalisés. La levure B. bruxellensis fait partie intégrante du consortium microbiologique oenologique (Medawar, 2003) au même titre que l'espèce Saccharomyces cerevisiae par exemple. Les études ont montré que B. bruxellensis était une levure particulièrement résistante aux contraintes oenologiques comme l'alcool, le SO2 et les bas pH. De plus, elle est peu exigeante d'un point de vue nutritionnel (Uscanga et al. 2000 ; Medawar, 2003) et s'adapte aussi bien à la présence d'oxygène qu'à son absence (Ciani et al. 2003 ; Du Toit et al. 2005). Les vinifications où les B. bruxellensis sont totalement absentes sont très rares, si bien que l'objectif n'est plus de n'avoir aucune B. bruxellensis dans son vin mais de limiter sa multiplication. La question de son origine reste toutefois posée. Pour cela plusieurs pistes ont été avancées, et, curieusement une origine au vignoble a longtemps était délaissée au profit du chai, probablement à cause de la difficulté d'étudier la microflore présente sur le raisin.

A comparative study of the calcium phosphate fouling process, with and without proteins, was carried out using both standard 316 2R stainless steel and 2R surfaces modified by TiN magnetron sputtering. Fouling behavior was assessed in a heat transfer flow cell operating in the turbulent flow regime. The fouling curves resulting from calcium phosphate deposition in the absence of proteins were substantially different from the ones obtained when protein was present. In this last case, two different fouling periods could be observed. The surface energy of the modified materials was found to affect the deposition parameters (rate of deposition and final amount of deposit) leading to higher amounts of deposit on higher energy surfaces in the absence of protein, while leading to less deposit in its presence. The standard 316 2R substrate proved to be less prone to fouling from protein-calcium phosphate solutions than the TiN modified surfaces. However, the same conclusion could not be dra...

The efficacy of lipase from Aspergillus niger MTCC 2594 as an additive in laundry detergent formulations was assessed using response surface methodology (RSM). A five-level four-factorial central composite design was chosen to explain the washing protocol with four critical factors, viz. detergent concentration, lipase concentration, buffer pH and washing temperature. The model suggested that all the factors chosen had a significant impact on oil removal and the optimal conditions for the removal of olive oil from cotton fabric were 1.0% detergent, 75 U of lipase, buffer pH of 9.5 and washing temperature of 25°C. Under optimal conditions, the removal of olive oil from cotton fabric was 33 and 17.1% at 25 and 49°C, respectively, in the presence of lipase over treatment with detergent alone. Hence, lipase from A. niger could be effectively used as an additive in detergent formulation for the removal of triglyceride soil both in cold and warm wash conditions.

Considering the actual consumer habits for precooked food, in the present work the combined drying (microwave and hot air convection) was carried out with pretreated osmotic dehydrated potatoes. The use of osmotic dehydration as a pretreatment in the food industry improves the product quality in terms of colour, flavour and texture. The advantage of the combined drying, microwave and convection with hot air aims at a better drying rate preserving the organoleptic properties of the product when is compared with the convective drying with hot air or microwave only. The osmotic dehydration during 1 or 2 hours and the salt concentration (5 or 10 % w/w) were analyzed, before the combined drying. Also, the results of the combined drying at different powers of 40, 50 and 60 %, after a pretreatment with osmotic dehydration during 1 hour was studied.

Recibido el 11 de Julio de 2011, aprobado el 1° de Agosto de 2011 El presente trabajo se enfocó en el estudio y análisis de las variables que afectan el secado de un producto hortícola como la papa, que se destinará a la elaboración de guisos. En primer lugar, el secado se llevó a cabo mediante la deshidratación osmótica y luego a través del secado combinado (microondas y convección con aire caliente). Se aplicaron diferentes modelos matemáticos a los datos experimentales a fin de representar los datos cinéticos obtenidos durante el proceso de secado.

Pineapple (Anana comosus) slices were dried by hot-air convective drying technique at fixed temperature (45, 60 and 75 • C) and constant air velocity of 1.5 m/s. The effect of drying conditions (drying time and air temperature) on the pineapple quality was evaluated. The quality of dehydrated pineapple was analyzed by color and texture changes, l-ascorbic acid loss and the ability of water uptake during rehydration procedure. Water uptake during rehydration was described by Page model. Statistical analysis of data revealed not significant difference (p > 0.05) among color and mechanical characteristics of pineapple samples dried at different drying temperatures to preset moisture content. Pineapple samples dried at 45 • C had better rehydration ability and more l-ascorbic acid retention than those obtained by air drying 75 • C. Hence, 45 • C drying temperature was best condition for pineapple quality preservation.

Background: Plant lignocellulosic biomass is an abundant, renewable feedstock for the production of biobased fuels and chemicals. Previously, we showed that iron can act as a co-catalyst to improve the deconstruction of lignocellulosic biomass. However, directly adding iron catalysts into biomass prior to pretreatment is diffusion limited, and increases the cost of biorefinery operations. Recently, we developed a new strategy for expressing iron-storage protein ferritin intracellularly to accumulate iron as a catalyst for the downstream deconstruction of lignocellulosic biomass. In this study, we extend this approach by fusing the heterologous ferritin gene with a signal peptide for secretion into Arabidopsis cell walls (referred to here as FerEX). The transgenic Arabidopsis plants. FerEX. accumulated iron under both normal and iron-fertilized growth conditions; under the latter (iron-fertilized) condition, FerEX transgenic plants showed an increase in plant height and dry weight by 12 and 18 %, respectively, compared with the empty vector control plants. The SDS-and native-PAGE separation of cell-wall protein extracts followed by Western blot analyses confirmed the extracellular expression of ferritin in FerEX plants. Meanwhile, Perls' Prussian blue staining and X-ray fluorescence microscopy (XFM) maps revealed iron depositions in both the secondary and compound middle lamellae cell-wall layers, as well as in some of the corner compound middle lamella in FerEX. Remarkably, their harvested biomasses showed enhanced pretreatability and digestibility, releasing, respectively, 21 % more glucose and 34 % more xylose than the empty vector control plants. These values are significantly higher than those of our recently obtained ferritin intracellularly expressed plants. This study demonstrated that extracellular expression of ferritin in Arabidopsis can produce plants with increased growth and iron accumulation, and reduced thermal and enzymatic recalcitrance. The results are attributed to the intimate colocation of the iron co-catalyst and the cellulose and hemicellulose within the plant cell-wall region, supporting the genetic modification strategy for incorporating conversion catalysts into energy crops prior to harvesting or processing at the biorefinery.

A new screening method for the detection and identification of GMO, based on the use of multiplex PCR followed by microarray, has been developed and is presented. The technology is based on the identification of quite ubiquitous GMO genetic target elements first amplified by PCR, followed by direct hybridisation of the amplicons on a predefined microarray (DualChip Ò GMO, Eppendorf, Germany). The validation was performed within the framework of a European project (Co-Extra, contract no 007158) and in collaboration with 12 laboratories specialised in GMO detection. The present study reports the strategy and the results of an ISO complying validation of the method carried out through an inter-laboratory study. Sets of blind samples were provided consisting of DNA reference materials covering all the elements detectable by specific probes present on the array. The GMO concentrations varied from 1% down to 0.045%. In addition, a mixture of two GMO events (0.1% RRS diluted in 100% TOPAS19/2) was incorporated in the S. Leimanis Á S. Hamels (&) Á J.

Fluidized bed dryers are used extensively for the drying of wet particulate and granular materials that can be fluidized with or without mechanical assistance. Furthermore, agitation contributes to achieving the fluid like behaviour of solids where rapid, easy transport and intimate fluid contacting are often the most important fluidization properties for industrial operations. Therefore, dryers that permit solid movement are frequently used. In this research work, we pretended to evaluate a new treatment procedure on image analysis methodology for determining residence time distribution on a continuous vibro-fluidized dryer. Knowing what is happening within the dryer, i.e., a complete velocity distribution map for the solid; makes it possible to predict the behaviour of this solid inside the dryer. In many cases, it is not necessary to have substantial knowledge about the flow, simply how long the individual particles stay in the dryer. The intention of this work was to determine the residence time of granular material using image analysis without any manual procedure. The material used was black beans, whereas tracers were black beans painted with white spray ink. The digital images were fitted in a specific computational program. The residence time values obtained by evaluated methodology were compared with ideal threshold image analysis and manual separation procedure values. The residence time distribution curves were determined for different vibration amplitudes. Residence time values obtained by adjusted method present lower mean relative deviation modulus than simple image analysis.

Production of frozen dough demands the usage of freezing/thawing resistant strains of baker&#39;s yeast that can adapt to low temperature and preserve fermentative performance, which is closely related to the lipid composition of their cells. In order to determine the effect of low temperature storage on the baker’s yeast, we have analysed lipids of compressed commercial baker’s yeast of Saccharomyces cerevisiae strain maintained at 4 °C (7 and 14 days) and -20 °C (7, 14 and 120 days). In the phospholipid composition of all groups the most abundant was phosphatidylcholine (42 – 56 % of total phospholipids), followed by phosphatidylinositol (21 – 28 %). Phosphatidylcholine to phosphatidylethanolamine ratio, one of the parameters of the adaptation capability to extracellular changes, was high (2.4 – 3.3). Sterol esters and ergosterol were the most abundant neutral lipids in all groups. Oleic and palmitoleic acids were by far the most abundant fatty acids in all groups (39 – 48 % and 3...

Opium poppy (Papaver somniferum L.) has two major products: alkaloids in the capsules and the seeds. It is known that opium poppy today contains alkaloid in great quantities. Opiate alkaloids and their synthetic derivatives are widely used in medicines which are produced in hundreds of tons for the medicine industry. The study aimed to screen the capsules of six Turkish opium poppy lines to evaluate their morphine percentage. The trial was carried out at the Experimental Fields of the Agronomy Department, Faculty of Agriculture of Ankara University, Turkey during three years. All seeds were sown during first week of October and harvested during second week of July during three years of experimentation to investigate the variation of morphine content. During first year hairless stem and strong capsule types were selected and sown followed by. Alkaloid analysis was performed at the laboratory of the alkaloid factory at Bolvadin province of Turkey in three years. Morphine percentage of six different lines were determined. The results showed that the average of morphine contents of lines ranged 0.731-0.851%, 0,813-1,147% and 0.815-1.142% during 2010, 2011 and 2012 respectively. In conclusion, these opium poppy especially L 1 and L 2 lines could serve as major source of morphine in future.

Fermented dairy products have long been associated with positive health benefits. The present study was undertaken to evaluate the physicochemical and sensory properties, viable probiotic counts, antioxidant activity and total phenolic content of probiotic yoghurt made by reconstituting of whole milk powder in aqueous fennel seed extract. Different concentrations of fennel aqueous seeds extract (2%, 4% and 6% w/v) were used as a substitute for water to reconstitute whole milk powder in formulations of yoghurt as functional additives. Interestingly, the use of aqueous extract of fennel seeds in the reconstituted yoghurt milk did not affect yoghurt composition (moisture, protein, fat and ash contents) compared to plain yoghurt. The titratable acidity significantly decreased after using aqueous fennel seed extract in the yogurt manufacture. In this regard, the titratable acidity value was 0.85 in the control yogurt at the fresh period and 1.14 after 21 days of storage, while this value...

The advent of 'omics' techniques bears significant potential for the assessment of the microbiological stability of foods. This requires the integration of molecular data with their implication for cellular physiology. Here we performed a comparative physiological and transcriptional analysis of Bacillus subtilis stressed with three different weak organic acids: the commonly used food preservatives sorbicand acetic-acid, plus the well-known uncoupler carbonyl cyanide-m-chlorophenyl hydrazone (CCCP). The concentration of each compound needed to cause a similar reduction of the growth rate negatively correlated with their membrane solubility, and positively with the concentration of undissociated acid. Intracellular acidification was demonstrated by expressing a pH-sensitive GFP derivative. The largest drop in intracellular pH was observed in CCCP-stressed cells and was accompanied by the transcriptional induction of the general stress response (GSR) and SigM regulon, responses known to be induced by acidification. The GSR was induced by acetate, but not by sorbate in mildly-stressed cells. Microarray analysis further revealed that all three acids activate transcriptional programs normally seen upon nutrient limitation and cause diverse responses indicative of an adaptation of the cell envelope. Based on the responses observed and the utilized pH measurements, the inhibitory effect of sorbic acid seems to be more focused on the cell membrane than that of acetic acid or CCCP.

Probiotics were defined as microbial strains that confer health benefits to their consumers. The concept has evolved during the last 20 years, and today metabolites produced by the strains, known as postbiotics, and even dead cells, known as paraprobiotics, are closely associated to them. The isolation of commensal strains from human microbiome has led to the development of next generation probiotics. This review aims to present an overview of the developments in the area of cancer prevention and treatment, intimately related to advances in the knowledge of the microbiome role in its genesis and therapy. Strain identification and characterization, production processes, delivery strategies, and clinical evaluation are crucial to translate results into the market with solid scientific support. Examples of recent tools in isolation, strain typification, quality control, and development of new probiotic strains are described. Probiotics market and regulation were originally developed in...

Во овој учебник по предметот Основи на органска хемија (за фармацевти) се објаснети шеснаесет поглавја во кои подробно се елаборирани најважните класи на органски соединенија како и начинот на нивно добивање, физичките и хемиските својства на секоја од класите на органските соединенија, како и употребата на органските соединенија во фармацијата. Овој учебник е наменет за студентите на Факултетот за медицински науки, отсек Фармација, со фонд на часови 3+2+2. За секоја класа од органските соединенија објаснети се најважните механизми, како и начинот на кој некои од нив може да се синтетизираат во лабораториски услови

Ракописот содржи девет поглавја во кои подробно се елаборирани јаглехидратите, мастите и маслата, протеините, витамините, аромите и мирисите како и основните класи на феромони. Исто така, во овој ракопис е објаснета анатомијата на вегетативните растителни органи и структурата и градбата на анималното ткиво, како и на технолошки значајни микроорганизми. Овој учебник е наменет за студентите на Техничко-технолошкиот факултет, отсек Прехранбена технологија, со фонд на часови 2+1+1. Во последното поглавје на овој учебник објаснети се четири лабораториски вежби за добивање на органски соединенија кои се користат во прехранбената технологија

The reaction system for octyl-β-glucoside synthesis catalysed by the almond-β-glu cosidase has been characterised. The monophasic octanol saturated with diff erent amounts of water served as a reaction medium. Both the glucose and the activated substrate p-nitrophenyl-β-glucoside were examined as glycon donors. The reverse hydrolysis and the transglycosylation were both used as reaction models for this enzymatically catalysed alkyl-β-glucoside synthesis. The rate of synthesis of octyl-β-glucoside (v S ), the rate of hydrolysis, i.e. the glucose formation (v H ) and the predicted yield (Y) were determined. The eff ect of water activity on the synthetic and hydrolytic activity of the enzyme was investigated. Both the rate of synthesis and the rate of hydrolysis increased with the increase of the water activity in the reaction system, showing their maximum values at the water activity close to the saturation level. Thus, the maximum ratio of v S /v H =0.165 was achieved at the water activity of 0.94. The predicted yields were 0.5, 0.75 and 14.19 % and were lower than the actually achieved yields of 19.45, 38 and 36.40 % at the water activities of 0.75, 0.84 and 0.94, respectively. The yield of octyl-β-glucoside in the reverse hydrolysis was only 15.2 %, i.e. 3.25 times lower than the yield obtained in the transglycosylation reaction with the water activity of 0.94. The solubility of glucose in pure octanol was only 1.5 mmol/L at the saturation level of 12 mmol/L in the presence of 10 mmol/L of p-nitrophenyl-β-glu coside, and it increased to 15.5 mmol/L in the presence of octyl-β-glucoside.

Background: Pseudomonas putida is a promising candidate for the industrial production of biofuels and biochemicals because of its high tolerance to toxic compounds and its ability to grow on a wide variety of substrates. Engineering this organism for improved performances and predicting metabolic responses upon genetic perturbations requires reliable descriptions of its metabolism in the form of stoichiometric and kinetic models. In this work, we developed kinetic models of P. putida to predict the metabolic phenotypes and design metabolic engineering interventions for the production of biochemicals. The developed kinetic models contain 775 reactions and 245 metabolites. Furthermore, we introduce here a novel set of constraints within thermodynamicsbased flux analysis that allow for considering concentrations of metabolites that exist in several compartments as separate entities. We started by a gap-filling and thermodynamic curation of iJN1411, the genome-scale model of P. putida KT2440. We then systematically reduced the curated iJN1411 model, and we created three core stoichiometric models of different complexity that describe the central carbon metabolism of P. putida. Using the medium complexity core model as a scaffold, we generated populations of large-scale kinetic models for two studies. In the first study, the developed kinetic models successfully captured the experimentally observed metabolic responses to several single-gene knockouts of a wild-type strain of P. putida KT2440 growing on glucose. In the second study, we used the developed models to propose metabolic engineering interventions for improved robustness of this organism to the stress condition of increased ATP demand. The study demonstrates the potential and predictive capabilities of the kinetic models that allow for rational design and optimization of recombinant P. putida strains for improved production of biofuels and biochemicals. The curated genome-scale model of P. putida together with the developed large-scale stoichiometric and kinetic models represents a significant resource for researchers in industry and academia.

Accurate determination of physiological states of cellular metabolism requires detailed information about metabolic fluxes, metabolite concentrations and distribution of enzyme states. Integration of fluxomics and metabolomics data, and thermodynamics--based metabolic flux analysis contribute to improved understanding of steady--state properties of metabolism. However, knowledge about kinetics and enzyme activities though essential for quantitative understanding of metabolic dynamics remains scarce and involves uncertainty. Here, we present a computational methodology that allow us to determine and quantify the kinetic parameters that correspond to a certain physiology as it is described by a given metabolic flux profile and a given metabolite concentration vector. Though we initially determine kinetic parameters that involve a high degree of uncertainty, through the use of kinetic modeling and machine learning principles we are able to obtain more accurate ranges of kinetic parameters, and hence we are able to reduce the uncertainty in the model analysis. We computed the distribution of kinetic parameters for glucose--fed E. coli producing 1,4--butanediol and we discovered that the observed physiological state corresponds to a narrow range of kinetic parameters of only a few enzymes, whereas the kinetic parameters of other enzymes can vary widely. Furthermore, this analysis suggests which are the enzymes that should be manipulated in order to engineer the reference state of the cell in a desired way. The proposed approach also sets up the foundations of a novel type of approaches for efficient, non--asymptotic, uniform sampling of solution spaces.

Pearl millet is an underutilized and drought-resistant crop that is mainly used for animal feed and fodder. Starch (70%) is the main constituent of the pearl millet grain; this starch may be a good substitute for major sources of starch such as corn, rice, potatoes, etc. Starch was isolated from pearl millet grains and modified with different physical treatments (heat-moisture (HMT), microwave (MT), and sonication treatment (ST)). The amylose content and swelling capacity of the starches decreased after HMT and MT, while the reverse was observed for ST. Transition temperatures (onset (To), peak of gelatinization (Tp), and conclusion (Tc)) of the starches ranged from 62.92-76.16 • C, 67.95-81.05 • C, and 73.78-84.50 • C, respectively. After modification (HMT, MT, and ST), an increase in the transition temperatures was observed. Peak-viscosity of the native starch was observed to be 995 mPa.s., which was higher than the starch modified with HMT and MT. Rheological characteristics (storage modulus (G ) and loss modulus (G )) of the native and modified starches differed from 1039 to 1730 Pa and 83 to 94 Pa; the largest value was found for starch treated with ST and HMT. SEM showed cracks and holes on granule surfaces after HMT as well as MT starch granules. Films were prepared using both native and modified starches. The modification of the starches with different treatments had a significant impact on the moisture, transmittance, and solubility of films. The findings of this study will provide a better understanding of the functional properties of pearl millet starch for its possible utilization in film formation.

The advancements in genome editing techniques over the past years have rekindled interest in rational metabolic engineering strategies. While Metabolic Control Analysis (MCA) is a well-established method for quantifying the effects of metabolic engineering interventions on flows in metabolic networks and metabolite concentrations, it does not consider the physiological limitations of the cellular environment and metabolic engineering design constraints. We report here a constraint-based framework, Network Response Analysis (NRA), for rational genetic strain design. NRA is cast as a Mixed-Integer Linear Programming problem that integrates MCA, Thermodynamically-based Flux Analysis (TFA), biologically relevant constraints, as well as genome editing restrictions into a comprehensive platform for identifying metabolic engineering targets. We show that the NRA formulation and its core constraints are equivalent to the ones of Flux Balance Analysis (FBA) and TFA, which allows it to be used for a wide range of optimization criteria and with various physiological constraints. We also show how the parametrization and introduction of biological constraints enhance the NRA formulation compared to the classical MCA approach, and we demonstrate its features and its ability to generate multiple alternative optimal strategies given several user-defined boundaries and objectives. In summary, NRA is a sophisticated alternative to classical MCA for rational metabolic engineering that accommodates the incorporation of physiological data at metabolic flux, metabolite concentration, and enzyme expression levels.

Asparagus by-products were submitted to hydrothermal treatments to improve their solubility and antioxidant capacity. Four severity conditions were applied (logR o 3. 05, 3.64, 4.23, 4.82) and two fractions were isolated from each treatment. The solid fibrous residues were enriched in cellulose, and depleted of hemicelluloses and phenolics. Their antioxidant activities were higher than those presented for asparagus bioactive fibers (81-154 vs 21-26 mmol Trolox/Kg) and did not correlate with phenol content. The soluble liquors were rich in phenolics and also contained changeable amounts of mono-, oligo-, and polysaccharides as a function of severity. The majority of the released oligosaccharides (xylooligosaccharides) are considered as prebiotic and their percentage could be optimized by modifying hydrothermal treatment conditions. This fraction presented very high antioxidant activity (131-232 mmol Trolox/Kg), and had a moderate correlation with phenols. Melanoidins are suggested to be present in both fractions. A new fiber fraction with potential prebiotic activity and high antioxidant activity has been obtained from asparagus by-products. Its high solubility in water and a pleasant flavor broaden its applications in food formulations

A comparative study of methods for isolating cell wall material from olives (variety Hojiblanca) was performed. Treatment with cold (T-l) or hot (T-2) ethyl alcohol was followed by extraction into cold or hot water (water-soluble fraction). The insoluble residues were delignified. Hemicelluloses A and B were isolated and the neutral sugar composition and protein content of the different fractions were determined. In the delignified fractions following treatments T-1 and T-2, hemicelluloses A were composed of 87.35% and 87.37% xylose, respectively, indicating the presence of a pure xylan. In the non-delignified fraction, the content of hemicelluloses B was higher than in the delignifled fraction and its predominant sugar was glucose, although significant amounts of xylose, galactose and arabinose were also found. The study suggests that treatment T-1 gives better results than T-2 and that a higher degree of purification of hemicelluloses A is obtained from the delignified cell wall material, as well as a decrease in the proteins bound to the fractions.

Aktivit/it der Endoglucanase-Entwicklung der Olive w~ihrend der Reifung und des Weichwerdens und die Beziehung mit cellulolytischen Mikroorganismen Zusammenfassung. Die cellulolytischenEnzyme sind ffir viele Ver~nderungen w~ihrend des Reifens der Olive verantwortlich. Es wurde daher die Aktivit/it der Enzyme w~hrend der Entwicklung der Olive bei der Reifung und dem Weichwerden studiert, und zwar am Baum, nach der Ernte und nach dem Lagern, was auch in Beziehung zu der Gegenwart von Mikroorganismen steht. Es wurde die cellulolytische Aktivit~tsentwicklung wfihrend der vier Reife-Zust/inde (grfin, farbenwechselnd, schwarz und zartes schwarz) studiert und auf die Gesamtzahl der cellulolytischen Bakterien bezogen. Der Aktivitfits-Zuwachs wfihrend des Reifens und Lagerns gibt einen Hinweis, wie die normale Flora auf das Reifen der Oliven einwirkt. Summary. Cellulolytic enzymes are responsible for many of the changes associated with the ripening process in olive fruits (Olea europaea). Evolution of endoglucanase activity (EC 3.2.1.4.) was studied in olives during their ripening and softening, both on the tree and after being picked and stored, and was related to the presence of microorganisms. The time course of cellulolytic activity was measured for four stages of ripeness (green, turning colour, black and ripe black) and was related to the total number of cellulolytic bacteria. The increases in activity during ripening and storage gave an indication of how normal flora influences during the olive ripening process.

Zusammenfassung. Die Kenntnisse fiber das Reifen und Erweichen der Olive sind ffir Spanien sehr wichtig, weil es das Hauptland ffir die industrielle Bearbeitung der Speisenoliven ist. Man hat Cellulase-Aktivit/it nachgewiesen, die sich w/ihrend der weiteren Entwicklungsstadien vergr613erte. Dieser Cellulasen-Zuwachs hat einen wichtigen Einflul3 auf das Reifen und Weichwerden. Von den drei Enzymformen der Endoglucanase, der Exoglucanase und der/3-Glucosidase besch/iftigt sich diese Arbeit nur mit der ersteren.

The engineering of Saccharomyces cerevisiae strains for lactose utilization has been attempted with the intent of developing high productivity processes for alcoholic fermentation of cheese whey. A recombinant S. cerevisiae flocculent strain that efficiently ferments lactose to ethanol was previously obtained by evolutionary engineering of an original recombinant that displayed poor lactose fermentation performance. We compared the transcriptomes of the original and the evolved recombinant strains growing in lactose, using cDNA microarrays. Microarray data revealed 173 genes whose expression levels differed more than 1.5‐fold. About half of these genes were related to RNA‐mediated transposition. We also found genes involved in DNA repair and recombination mechanisms, response to stress, chromatin remodeling, cell cycle control, mitosis regulation, glycolysis and alcoholic fermentation. These transcriptomic data are in agreement with some of the previously identified physiological an...

This repor t was prep ared as an acco unt of Gove rnme nt spon sore d work. Neith er the Unite d Stat es, nor the Com miss ion, nor any pers on actin g on beha lf of the Com miss ion: 1 A. Make s any wrra nty or repre sent ation , expr esse d or impl ied, with resp ect to the accu - lacy. compl etene ss, or usefu lness of the infor matio li conta ined in this repor t, or that the Uie I of any infor mati on, appa ratua , meth od, or proc esB disc lose d in this repo rt may not infrin ge priva tely owned right e; or B. Assu mes any liabi lities with resp ect to the uee 4, or for dam ages resu lting from the 1 use of any Infor mati on, appa ratus , meth od, or proc ess discl oaed in this repor t. As used in the abov e, "pers on actin g on beha lf of the Com miss ion" inclu des any em-, ploye e or cont racto r of the Com miss ion, or emp loy6 e of auch cont racto r, to the exte nt that • such emp loyee or cont racto r of the Com miss lon, or emp loye e of such cont racto r prepa res, dise emin ates , or prov idee acce ss to, any infor mati oh purs uant to his emp loym ent or cont ract t A with the Com miss ion, or his emp loym ent with such Jont racto r.

Ascorbic acid deficiency has been associated with several health conditions. The objective of this study was to evaluate the content of ascorbic acid and the sensorial qualities of Mangifera indica and Myrciaria dubia yogurts. Four yogurt treatments were elaborated with different concentrations of these fruits (T1: 15% and 5%; T2: 15% and 10%; T3: 20% and 5% and T4: 20% and 10%) respectively, compared with a control treatment (CT: yogurt with 15% of Fragaria vesca). The ascorbic acid contents of the different treatments were determined by spectrophotometry, with values in the following order (T1: 63.2 mg/100 g; T2:114.3 mg/100 g; T3: 57.3 mg/100 g; T4: 115.1 and the control treatment CT:11.5 mg/100 g). The sensorial evaluation consisted of the application of a hedonic scale of 5 points (1: I dislike it very much; 2: I dislike it; 3: I neither like it nor dislike it; 4: I like it; 5: I like it a lot), results show evidence that the acidity level had a significant influence during the...

The present work investigates the effect of feed concentration and magnitudes of applied pressure of olive-extracted oleuropein on permeate flux and membrane transport properties through concentration polarization in a nanofiltration (NF) membrane. The filtration of oleuropein model solution by an NF membrane using different feed concentrations (0.3, 0.9, or 2.7 kg m -3 ) enables the correlation of a transport model, comprised of concentration polarization and osmotic pressure models, to membrane retention performance, evaluated using the Spiegler-Kedem model. Moreover, osmotic pressure was observed and interpreted using the van't Hoff equation at different levels of feed concentration and applied pressure. Transport parameters (reflection coefficient, water and oleuropein permeability, and mass transfer coefficient) were correlated with experiment values by developing a mathematical model that reflects the flux decline by increasing osmotic pressure.

Research background. Retort processing is one of the most widely used methods of thermal inactivation that provides convenient, ready-to-eat foods. Although this technology remains widespread, it can be revamped through processing of novel ingredients such as gums. This article aims to investigate the effect of the hydrocolloids collagen, soy protein isolate, carrageenan and modified starch with different salt mass fractions on the retorted meat products. Experimental approach. Firstly, solutions of the added hydrocolloids of different salt mass fractions in order to stimulate the salting-in effect were studied. Lipid oxidation, syneresis and water activity were analysed during shelf life to find the best overall treatments. Lastly, sensory and texture analyses were then performed to assess the impact of the added hydrocolloids. Results and conclusions. Yield, cooking loss and water-holding capacity had better results when higher salt mass fractions with hydrocolloids were used. The...

Random mutagenesis is a powerful technique to obtain mutant proteins with different properties from the wild-type molecule. Error-prone PCR is often employed for random mutagenesis in bacterial protein expression systems, but has rarely been used in the methylotrophic yeast Pichia pastoris system, despite its significant advantages, mainly because large (μg-level) amounts of plasmids are required for transformation. We developed a quick and easy technique for random mutagenesis in P. pastoris by sequential Phi29 DNA polymerase-based amplification methods, error-prone rolling circle amplification (RCA) and multiple displacement amplification (MDA). The methodology was validated by applying it for random mutation of the gene encoding cellulase from the basidiomycete Phanerochaete chrysosporium (PcCel6A), a key enzyme in degradation of cellulosic biomass. In the error-prone RCA step, the concentrations of manganese ion (Mn(2+)) and cellulase gene-containing plasmid were varied, and the...

Plant-associated microbial communities play a pivotal role in agricultural productivity by enhancing crop yields, while reducing fertilizer dependency. Genome sequencing revolutionized the understanding of individual microbes, while metagenomics changed the game by analyzing genetic material directly from the environment, bypassing limitations of culturing microbes. These microbiomes encode metabolic potentials offering solutions for climate resilience, such as improved drought and salinity tolerance in crops. Advanced sequencing technologies enable high-throughput analysis to identify key functional genes involved in nitrogen fixation, phosphate solubilization, and pathogen suppression. Despite challenges in data complexity and microbial interactions, integrating genomic andmetagenomic insights holds potential for sustainable agriculture through microbiome engineering, precision farming, and microbial inoculants. This review explores the harnessing of the plant microbiome through genomic and metagenomic approaches as a cornerstone for sustainable agriculture.

The aim of this study was to develop sustainable methods to modify the structure of wheat bran used in food applications. The methods of interest were wet milling and enzymatic treatment. The effect of these methods on a product quality was tested in a pilot scale baking process. Wheat bran is composed of many nutritionally valuable components, such as dietary fibre, antioxidants and proteins. Due to its relatively low price, wheat bran holds a great potential as an ingredient to be added into food products. However, most of the previous studies have shown that the addition of wheat bran usually decreases the quality of food products. The focus of this study was on modifying the properties of wheat bran, so that it could be used as a filler ingredient. Besides the particle size reduction and enzymatic treatment, the stability of wheat bran-water suspensions was studied by following the phase separation. The aim of the modifications was to release valuable components from the wheat bran matrix, to concentrate the dietary fibre and to alter the physico-chemical properties of wheat bran. Wet milling and subsequent centrifugation increased the fibre content of the wheat bran fractions and increased the proportion of insoluble fibre. The enzymatic treatment increased the amount of reducing sugars in the soluble fraction after centrifugation. The highest solubilisation was obtained by using enzymes Depol 740L and Ecopulp TX200A. In addition, the phase stability of the wheat bran water mixtures increased in proportion to the level of milling. The effect of modified wheat bran on the quality of the breads was not as good as wanted due to the decreased volume and hardness of the test breads. Similar results were obtained both with dry and wet milled wheat bran fractions. The negative effect of wheat bran on the bread's structure was most clearly seen with 20 % addition level. As a conclusion, wheat bran could be used as a bakery ingredient and a source of dietary fibre, when the addition level is lower than 20 % (of the dry mass in dough). However, the protocol for particle size reduction and the chosen enzymes are of utmost importance. As the desired properties of bread seemed to decrease in proportion to added wheat bran, the effect of particle size, enzymatic hydrolysis and addition level would need further studies. wheat bran, particle size, wet milling, enzymatic treatment, baking, bread Tiivistelmä Tekijä Nimi Sivumäärä Aika Olli-Pekka Lehtinen Vehnäleseen muokkaus elintarvikesovellutuksiin märkäjauhatuksella ja entsymaattisella hydrolyysillä 63 sivua + 9 liitettä 20.5.2012 Tutkinto Insinööri (AMK) Koulutusohjelma Bio-ja elintarviketekniikka Suuntautumisvaihtoehto Tuotekehitys ja laadunvalvonta Ohjaajat Tutkija, Projektipäällikkö, Juhani Sibakov (VTT) Lehtori, Mikko Halsas (Metropolia Ammattikorkeakoulu) Tämän insinöörityön tarkoituksena oli kehittää menetelmiä vehnäleseen rakenteen muokkaamiseksi elintarvikesovellutuksia varten. Tutkittavat muokkausmenetelmät olivat vehnäleseen märkäjauhatus sekä entsymaattinen hydrolyysi. Muokkauksen vaikutuksia arvioitiin pilot-mittakaavan koeleivonnassa. Vehnälese sisältää monia ravitsemuksellisesti arvokkaita komponentteja kuten ravintokuitua, antioksidatiivisia komponentteja sekä proteiinia. Vehnälesettä on hyödynnetty suhteellisen vähän elintarvikkeissa, mutta oikeanlainen muokkaus voisi mahdollistaa sen käytön ravitsemuksellista arvoa lisäävänä raaka-aineena. Aiemmat tutkimukset ovat osoittaneet, että muokkaamattoman vehnäleseen lisäys heikentää elintarvikkeiden ominaisuuksia. Insinöörityössä tutkittiin partikkelikoon pienentämisen ja entsymaattisen hydrolyysin lisäksi muokatun vehnälese-vesi -suspension stabiiliutta faasierottumisen funktiona. Menetelmien tavoitteena oli vapauttaa vehnäleseen matriisista arvokkaita komponentteja, kuten ravintokuitua ja proteiinia sekä tarkastella muuttuvia fysikaaliskemiallisia ominaisuuksia. Märkäjauhatus lisäsi sentrifugoinnin jälkeisen kiinteän lesefraktion kuitupitoisuutta sekä liukenemattoman kuidun osuutta. Entsymaattinen hydrolyysi lisäsi vehnälese-vesi -suspension pelkistävien sokereiden pitoisuutta supernatanttiliuoksessa sentrifugoinnin jälkeen. Parhaan tuloksen antoivat entsyymit: Depol 740L ja Ecopulp TX200A. Vehnälese-vesi -suspension stabiilius parani myös jauhatusasteen myötä. Muokkausten vaikutus leivontaominaisuuksiin ei kuitenkaan antanut toivottua lopputulosta. Muokatun vehnäleseen lisäys huononsi leipien tilavuutta ja lisäsi kovuutta. Kun vehnäleseen lisäystaso oli 20 %, leipien ominaisuudet olivat merkittävästi huonommat verrattuna vehnäjauholeipään. Johtopäätöksenä voidaan todeta, että vehnälesettä voidaan käyttää leipomotuotteissa nostamaan kuitupitoisuutta, kunhan lisäystaso ei ylitä 20 % taikinan kuivapainosta. Vehnäleseen partikkelikoon pienentäminen ja entsyymien valinta vaikuttaa oleellisesti tuotteen ominaisuuksiin. Koska ominaisuudet huononevat oleellisesti suhteessa lisättyyn vehnäleseeseen, lisätutkimukset ovat tarpeen partikkelikoon pienentämisen, valittavan entsyymin ja tuotteeseen lisättävän vehnäleseen määrän optimoinnin osalta.

For renewable and sustainable bioenergy utilization with cost-effectiveness, electron-shuttles (ESs) (or redox mediators (RMs)) act as electrochemical "catalysts" to enhance rates of redox reactions, catalytically accelerating electron transport efficiency for abiotic and biotic electrochemical reactions. ESs are popularly used in cellular respiratory systems, metabolisms in organisms, and widely applied to support global lives. Apparently, they are applicable to increase power-generating capabilities for energy utilization and/or fuel storage (i.e., dye-sensitized solar cell, batteries, and microbial fuel cells (MFCs)). This first-attempt review specifically deciphers the chemical structure association with characteristics of ESs, and discloses redox-mediating potentials of polyphenolics-abundant ESs via MFC modules. Moreover, to effectively convert electron-shuttling capabilities from non-sustainable antioxidant activities, environmental conditions to induce electrochemical mediation apparently play critical roles of great significance for bioenergy stimulation. For example, pH levels would significantly affect electrochemical potentials to be exhibited (e.g., alkaline pHs are electrochemically favorable for expression of such electron-shuttling characteristics). Regarding chemical structure effect, chemicals with ortho-and para-dihydroxyl substituents-bearing aromatics own convertible characteristics of non-renewable antioxidants and electrochemically catalytic ESs; however, ES capabilities of metadihydroxyl substituents can be evidently repressed due to lack of resonance effect in the structure for intermediate radical(s) during redox reaction. Moreover, this review provides conclusive remarks to elucidate the promising feasibility to identify whether such characteristics are non-renewable antioxidants or reversible ESs from natural polyphenols via cyclic voltammetry and MFC evaluation. Evidently, considering sustainable development, such electrochemically convertible polyphenolic species in plant extracts can be reversibly expressed for bioenergy-stimulating capabilities in MFCs under electrochemically favorable conditions.

Engine oil used in automobiles is a threat to soil and water due to the recalcitrant properties of its hydrocarbons. It pollutes surrounding environment which affects both flora and fauna. Microbes can degrade hydrocarbons containing engine oil and utilize it as a substrate for their growth. Our results demonstrated that cell-free broth of Bacillus velezensis KLP2016 (Gram + ve, endospore forming; Accession number KY214239) recorded an emulsification index (E24%) from 52.3% to 65.7% against different organic solvents, such as benzene, pentane, cyclohexane, xylene, n-hexane, toluene and engine oil. The surface tension of the cell-free broth of B. velezensis grown in Luria–Bertani broth at 35 °C decreased from 55 to 40 mN m−1at critical micelle concentration 17.2 µg/mL. The active biosurfactant molecule of cell-free broth of Bacillus velezensis KLP2016 was purified by Dietheylaminoethyl-cellulose and size exclusion chromatography, followed by HPLC (RT = 1.130), UV–vis spectrophotometr...

at middle stage of storage, and those contents were slightly higher in OG than in CG and LCG fruits. Shelf life of kiwi fruit considerably decreased in OG fruits by increasing fruit decay and softening during storage. Fluorescence measurements showed a difference between the investigated samples, especially in their antioxidant status. The used statistical evaluation confirmed the obtained results. The quenching properties of these samples are correlated with their antioxidant properties and the amount of polyphenols and the decrease in fluorescence intensity.