A rational approach for predicting the dissolved oxygen profile in receiving waters

A practical method for predicting the dissolved oxygen (DO) profile in a stream receiving biodegradable waste is presented. In this method the BOD (accumulated O₂ uptake) curve is obtained using an open jug technique. The accumulated O₂ uptake curve thus developed is employed in numerical integration with the physical reaeration data for the receiving stream to predict the DO profile in the stream. In the present study, the method was examined using 10-liter open jar reactors to obtain the O₂ uptake curves, and the receiving stream was a 670-liter simulated stream apparatus which has been employed in previous studies on stream reaeration. The method was found to provide a fairly good prediction of the actual dissolved oxygen profile observed in the receiving stream. The effect of the reaeration constant, K₂ )agitation effect(, on the kinetics of O₂ uptake was also investigated and it was found that increased agitation (higher K₂ value) caused some increase in the accumulated oxygen uptake (BOD) curve with most of the increase coming after the so-called “plateau” area in the O₂ uptake curve, i.e., in the general case, after the low point along the DO sag curve.     

Metabolomic profile of cacao cell suspensions growing in blue light/dark conditions with potential in food biotechnology

As one of the effective and powerful methods for gene function analysis in plants, transient overexpression and virus-induced gene silencing (VIGS) system were developed for oriental melon (Cucumis melo var. makuwa Makino). Here, the full-length of CmLOX10, the conserved domains of CmLOX10 and CmPDS were isolated from oriental melon. The vectors for overexpression of CmLOX10, and the VIGS vector of CmLOX10 and CmPDS were constructed and transformed into agrobacterium, respectively. Especially, the sprout absorption method was performed on oriental melon, and CmPDS was selected as a reporter gene in VIGS. The expression level of CmLOX10 significantly increased in the oriental melon with the CmLOX10 overexpression construct, and lipoxygenases (LOXs) contributed to the leaf cell death in the oriental melon. The expression levels of CmLOX10 and CmPDS were significantly decreased in plants with the VIGS construct, and there was albino phenotype in CmPDS silencing leaves. In this study, CmLOX10 gene was silenced using Tobacco rattle virus based VIGS system firstly. These results proved that the transient expression technology provides an excellent tool to explore gene function in oriental melon.

     

A mathematical approach for the estimation of biomass production rate in solid state fermentation

The oxygen uptake rate (OUR) was studied in a solid state fermentation process of dried citrus peel with the strain Aspergillus niger QH-2 in order to obtain the growth estimation of the microorganism in the system. The relationship between OUR, the maintenance coefficient (m) and the yield for oxygen consumption Y_O2 allows the estimation of the biomass rate if we consider that both parameters are not constants in some periods of the process. It was estimated that in the first 24th the strain has an specific growth rate of 0.174 h^?1 with values for Y_O2 and m in the order of 2.84 g-cell/g-oxygen and 0.006 g-oxygen/g-cell ·h respectively.     

Taxol production in bioreactors: Kinetics of biomass accumulation, nutrient uptake, and taxol production by cell suspensions of Taxus baccata

The kinetics of biomass accumulation, nutrient uptake and taxol production of Taxus baccata cell suspensions were examined in three bioreactor configurations, viz. 250-mL Erienmeyerflasks, 1-L working volume pneumatically mixed (PMB), and stirred tank (STB) bioreactors. Qualitatively similar kinetics were observed in all three bioreactor types. Biomass accumulation and specific nutrient uptake rates exhibited biphasic characteristics. Carbohydrate uptake and biomass accumulation substantially ceased when phosphate was depleted from the medium. Phosphate was identified as a possible growth-limiting nutrient. Taxol accumulated exclusively in the second phase of growth. A maximum taxol concentration of 1.5 mg/L was obtained in the PMB which was fivefold greater than that obtained in the Erienmeyer flasks and the STB, but the relative kinetics of taxol production was the same in all three reactor types. Biomass yields were calculated from the kinetic data and a stoichiometry for biomass formation was evaluated. The similarity of kinetics in the three bioreactor configurations suggests that taxol production by T. baccata cell suspensions is amenable to scateup. (c) 1995 John Wiley & Sons, Inc.     

Two-stage heterotrophic and phototrophic culture strategy for algal biomass and lipid production

A two-stage heterotrophic and phototrophic culture strategy for algal biomass and lipid production was studied, wherein high density heterotrophic cultures of Chlorellasorokiniana serve as seed for subsequent phototrophic growth. The data showed growth rate, cell density and productivity of heterotrophic C.sorokiniana were 3.0, 3.3 and 7.4 times higher than phototrophic counterpart, respectively. Hetero- and phototrophic algal seeds had similar biomass/lipid production and fatty acid profile when inoculated into phototrophic culture system. To expand the application, food waste and wastewater were tested as feedstock for heterotrophic growth, and supported cell growth successfully. These results demonstrated the advantages of using heterotrophic algae cells as seeds for open algae culture system. Additionally, high inoculation rate of heterotrophic algal seed can be utilized as an effective method for contamination control. This two-stage heterotrophic phototrophic process is promising to provide a more efficient way for large scale production of algal biomass and biofuels.     

Salt-induced lipid changes in Catharanthus roseus cultured cell suspensions

Salt treatment strongly affected cell growth by decreasing dry weight. Exposure of Catharanthus roseus cell suspensions to increasing salinity significantly enhanced total lipid (TL) content. The observed increase is mainly due to high level of phospholipids (PL). Hundred mM NaCl treatment increased phospholipid species phosphatidylcholine (PC) and phosphatidylethanolamine (PE), whereas it reduced glycolipid ones monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) but not sulfoquinovosyldiacylglycerol (SQDG). Moreover, fatty acid composition was clearly modified when cells were cultured in the presence of 100 mM NaCl, whereas only few changes occurred at 50 mM. Salt treatment decreased palmitic acid (16:0) level and increased that of linolenic acid (18:2). Such effect was observed in phospholipid species PC and PE and in glycolipid DGDG. Double bond index (DBI) was enhanced more than 2-fold in fatty acids of either glycolipids or phospholipids from cells submitted to 100 mM NaCl. Free sterol content was also significantly enhanced, especially at 100 mM NaCl, whereas free sterols/phospholipids (St/PL) ratio was slightly decreased. All these salt-induced changes in membrane lipids suggest an increase in membrane fluidity of C. roseus cells.     

A rational approach to improving the biotechnological production of taxanes in plant cell cultures of Taxus spp.

Taxol is a complex diterpene alkaloid scarcely produced in nature and with a high anticancer activity. Biotechnological systems for taxol production based on cell cultures of Taxus spp. have been developed, but the growing commercial demand for taxol and its precursors requires the optimization of these procedures. In order to increase the biotechnological production of taxol and related taxanes in Taxus spp. cell cultures, it is necessary not only to take an empirical approach that strives to optimize in-put factors (cell line selection, culture conditions, elicitation, up-scaling, etc.) and out-put factors (growth, production, yields, etc.), but also to carry out molecular biological studies. The latter can provide valuable insight into how the enhancement of taxane biosynthesis and accumulation affects metabolic profiles and gene expression in Taxus spp. cell cultures.     

Environmental control of lipid and biomass production in two diatom species

Biomass and neutral lipid accumulation were examined in Chaetoceros muelleri var. subsalsum and Navicula saprophila using a multivariate, fractional factorial design. Variables included were conductivity, temperature, nitrogen concentration, silicon concentration, time (culture age), and alkalinity. Measured characteristics included nile red fluorescence (as a measure of neutral lipid content) and ash-free dry weight (AFDW). Nitrogen concentration was the variable with the greatest effect on neutral lipid and ash-free dry weight accumulation over the ranges tested. Increasing conductivity in the range examined had a significant, negative impact on neutral lipid accumulation in both of these strains, while increasing alkalinity had a positive effect on lipid and ash-free dry weight in both strains. Experimental designs such as those described here have great potential utility in biological systems with complex interactions.     

Capsicum spp in vitro liquid cell suspensions: A useful system for the production of capsaicinoids and polyphenols

Capsicum are among the most extensively cultivated and consumed plant species in the world, because of their unique pungency, aroma and colour. The typical burning sensation caused by chili peppers is due to the occurrence of a group of alkaloids named capsaicinoids. In the present study, the production of solid callus and cell suspensions from hypocotyl explants of three different chili pepper cultivars (Capsicum annuum L. cv. Mazzolino, Capsicum chinense Jacq. cv. Naga Morich and Pimenta de Neyde), was optimised. In addition, C. chinense cv. Naga Morich cell suspensions were supplemented with biotic elicitors (methyl-jasmonate and chitosan) and with precursors and intermediates of capsaicin biosynthesis (vanillin, phenylalanine and valine), and both cells and media were analysed for capsaicinoid, polyphenol, flavonoid contents and for antioxidant activity. This is the first report regarding capsaicinoid elicitation with pure chitosan and with a combination of precursors of both phenylpropanoid and valine pathways. Overall, the highest capsaicinoid levels were detected in cell extracts from cultures treated with 10 μM methyl-jasmonate and with a combination of phenylalanine and valine amino acids (100 μM each). The present results confirm the possibility of using hypocotyl chili pepper cell suspensions to produce high amounts of health beneficial metabolites.     

A survey of yeast from the Yarrowia clade for lipid production in dilute acid pretreated lignocellulosic biomass hydrolysate

Applied Microbiology and Biotechnology - Yarrowia lipolytica is an oleaginous yeast species that has attracted attention as a model organism for synthesis of single cell oil. Among over 50 isolates...     

The problems associated with high biomass levels in plant cell suspensions

The development of plant cell cultures as an alternative supply of phytochemicals has been difficult. Although there has been some very suitable targets, the yields of these compounds has remained low despite considerable efforts. One of the main constituents of a process is its productivity which is the sum of the process run time (growth rate), yield, and biomass levels. The effect of changes in all three of these components on productivity has been demonstrated.Of the three components making up productivity, biomass is perhaps the easiest to increase. However, high biomass levels will increase the viscosity, which will affect both mixing and oxygen supply. Therefore, this will require more vigorous mixing which may increase the shear within the bioreactor. All these parameters need further investigation at high biomass concentrations.     

Progress on lipid extraction from wet algal biomass for biodiesel production

Lipid recovery and purification from microalgal cells continues to be a significant bottleneck in biodiesel production due to high costs involved and a high energy demand. Therefore, there is a considerable necessity to develop an extraction method which meets the essential requirements of being safe, cost‐effective, robust, efficient, selective, environmentally friendly, feasible for large‐scale production and free of product contamination. The use of wet concentrated algal biomass as a feedstock for oil extraction is especially desirable as it would avoid the requirement for further concentration and/or drying. This would save considerable costs and circumvent at least two lengthy processes during algae‐based oil production. This article provides an overview on recent progress that has been made on the extraction of lipids from wet algal biomass. The biggest contributing factors appear to be the composition of algal cell walls, pre‐treatments of biomass and the use of solvents (e.g. a solvent mixture or solvent‐free lipid extraction). We compare recently developed wet extraction processes for oleaginous microalgae and make recommendations towards future research to improve lipid extraction from wet algal biomass.     

A promising approach on biomass accumulation and withanolides production in cell suspension culture of Withania somnifera (L.) Dunal

Withanolide is one of the most extensively exploited steroidal lactones, which are biosynthesized in Withania somnifera. Its production from cell suspension culture was analyzed to defeat limitations coupled with its regular supply from the plant organs. In order to optimize the different factors for sustainable production of withanolides and biomass accumulations, different concentrations of auxins or cytokinins and their combinations, carbon sources, agitation speed, organic additives and seaweed extracts was studied in cell suspension culture. Maximum biomass accumulation (16.72 g fresh weight [FW] and 4.18 g dry weight [DW]) and withanolides production (withanolide A 7.21 mg/g DW, withanolide B 4.23 mg/g DW, withaferin A 3.88 mg/g DW and withanone 6.72 mg/g DW) were achieved in the treatment of Gracilaria edulis extract at 40 % level. Organic additive l-glutamine at 200 mg/l in combination with picloram (1 mg/l) and KN (0.5 mg/l) promoted growth characteristics (11.87 g FW and 2.96 g DW) and withanolides synthesis (withanolide A 5.04 mg/g DW, withanolide B 2.59 mg/g DW, withaferin A 2.36 mg/g DW and withanone 4.32 mg/g DW). Sucrose at 5 % level revolved out to be a superior carbon source yielded highest withanolides production (withanolide A 2.88 mg/g DW, withanolide B 1.48 mg/g DW, withaferin A 1.35 mg/g DW and withanone 2.47 mg/g DW), whereas biomass (7.28 g FW and 1.82 g DW) was gratefully increased at 2 % level of sucrose in cell suspension culture. This optimized protocol can be utilized for large scale cultivation of W. somnifera cells in industrial bioreactors for mass synthesis of major withanolides.     

A combined fermentative-chemical approach for the scalable production of pure E. coli monophosphoryl lipid A

Lipid A is the lipophilic region of lipopolysaccharides and lipooligosaccharides, the major components of the outer leaflet of most part of Gram-negative bacteria. Some lipid As are very promising immunoadjuvants. They are obtained by extraction from bacterial cells or through total chemical synthesis. A novel, semisynthetic approach to lipid As is ongoing in our laboratories, relying upon the chemical modification of a natural lipid A scaffold for the fast obtainment of several other lipid As and derivatives thereof. The first requisite for this strategy is to have this scaffold available in large quantities through a scalable process. Here, we present an optimized fed-batch fermentation procedure for the gram-scale production of lipid A from Escherichia coli K4 and a suitable phenol-free protocol for its purification. A study for regioselective de-O-phosphorylation reaction was then performed to afford pure monophosphoryl lipid A with an attenuated endotoxic activity, as evaluated by cytokine production in human monocytic cell line THP-1 in vitro. The reported method for the large-scale obtainment of monophoshoryl lipid A from the fed-batch fermentation broth of a recombinant strain of E. coli may permit the access to novel semisynthetic lipid A immunoadjuvant candidates.     

Mass propagation and genetic improvement of forest trees for biomass production by tissue culture

Wood derived from forest trees can serve as a major alternative source of energy and fuel because of the current energy shortage and increase in price of oil and natural gas. Normally, trees take several years to grow and produce seeds. But, in recent years, ‘test tube trees’ have been produced in large numbers (as many as 3000 plants per year) from one seedling using tissue culture by treating a few cells of a tree with specific chemical substances. Tissue culture is a promising technique for mass production of large numbers of superior trees, derived through genetic improvement, and may prove widely applicable to trees which show promise as energy sources.Three selected tree genera, viz. Sapium sebiferum (Chinese Tallow), Leucaena leucocephala (giant ipil-ipil, a tropical legume) and Copaifera multijuga (Copaiba tree from Brazil) have been studied because of their potential usefulness for biomass production. Regeneration of vegetatively produced plantlets has been achieved from embryos and callus cells grown in specific culture medium for two of the above genera. High yields of protoplasts have been obtained isolated from cells of different plant parts and grown as calli. Conditions which will enable callus derived from protoplasts to undergo in vitro regeneration, plantlet formation and eventually growth into plants are being investigated.