The effect of space flight on the production of actinomycin D by Streptomyces plicatus

The effect of space flight on production of the antibiotic actinomycin D by Streptomyces plicatus WC56452 was examined onboard the US Space Shuttle mission STS-80. Paired space flight and ground control samples were similarly prepared using identical hardware, media, and inoculum. The cultures were grown in defined and complex media under dark, anaerobic, thermally controlled (20°C) conditions with samples fixed after 7 and 12 days in orbit, and viable residuals maintained through landing at 17 days, 15 h. Postflight analyses indicated that space flight had reduced the colony-forming unit (CFU) per milliliter count of S. plicatus and increased the specific productivity (pg CFU⁻¹) of actinomycin D. The antibiotic compound itself was not affected, but its production time course was altered in space. Viable flight samples also maintained their sporulation ability when plated on agar medium postflight, while the residual ground controls did not sporulate. Received 21 August 2001/ Accepted in revised form 30 July 2002     

Design of mineral medium for growth of <Emphasis Type="Italic">Actinomadura</Emphasis> sp. ATCC 39727, producer of the glycopeptide A40926: effects of calcium ions and nitrogen sources

Actinomadura sp. ATCC 39727 produces the glycopeptide antibiotic A40926, structurally similar to teicoplanin, with significant activity against Neisseria gonorrhoeae and precursor of the semi-synthetic antibiotic dalbavancin. In this study the production of A40926 by Actinomadura under a variety of growth conditions was investigated. The use of chemically defined mineral media allowed us to analyze the influence of carbon and nitrogen sources, phosphate, ammonium and calcium on the growth and the antibiotic productivity of Actinomadura. We confirm recent data [Gunnarsson et al. (2003) J Ind Microbiol Biotechnol 30:150–156] that low initial concentrations of phosphate and ammonium are beneficial for growth and A40926 production, and we provide new evidence that the production of A40926 is depressed by calcium, but promoted when l-glutamine or l-asparagine are used as nitrogen sources instead of ammonium salts.     

A novel conico-cylindrical flask aids easy identification of critical process parameters for cultivation of marine bacteria

A polymethylmethacrylate (PMMA) conico-cylindrical flask (CCF) with an inner arrangement consisting of eight equidistantly spaced rectangular strips mounted radially on a circular disk to provide additional surface area for microbial attachment and ports to allow air supply was employed for melanin production by Shewanella colwelliana and antibiotic production by Pseudoalteromonas rubra. The design allowed comparison of production between (1) CCF with hydrophobic surface (PMMA-CCF), (2) CCF with hydrophilic glass surface (GS-CCF), and (3) standard unbaffled Erlenmeyer flask (EF). Melanin production in the PMMA-CCF was higher by at most 33.5% and growth of S. colwelliana by at most 309.2% compared to the other vessels. Melanin synthesis was positively correlated with reactor surface area and hydrophobicity, suspended cell growth, and biofilm formation. Antibiotic production in the EF was higher by at most 83.3%, but growth of P. rubra was higher in the PMMA-CCF by at most 54.5% compared to the other vessels. A hydrophilic vessel surface, abundant air supply, but low shear stress enhanced antibiotic production. The CCF together with the EF allowed identification of the crucial parameters (vessel surface characteristics, growth, biofilm formation, and aeration) influencing productivity, knowledge of which in the initial stages of process development will facilitate informed decisions at the later phases.     

Microbial fed-batch production of 1,3-propanediol by Klebsiella pneumoniae under micro-aerobic conditions

The microbial production of 1,3-propanediol (1,3-PD) by Klebsiella pneumoniae under micro-aerobic conditions was investigated in this study. The experimental results of batch fermentation showed that the final concentration and yield of 1,3-PD on glycerol under micro-aerobic conditions approached values achieved under anaerobic conditions. However, less ethanol was produced under microaerobic than anaerobic conditions at the end of fermentation. The batch micro-aerobic fermentation time was markedly shorter than that of anaerobic fermentation. This led to an increment of productivity of 1,3-PD. For instance, the concentration, molar yield, and productivity of 1,3-PD of batch micro-aerobic fermentation by K. pneumoniae DSM 2026 were 17.65 g/l, 56.13%, and 2.94 g l^–1 h^–1, respectively, with a fermentation time of 6 h and an initial glycerol concentration of 40 g/l. Compared with DSM 2026, the microbial growth of K. pneumoniae AS 1.1736 was slow and the concentration of 1,3-PD was low under the same conditions. Furthermore, the microbial growth in fed-batch fermentation by K. pneumoniae DSM 2026 was faster under micro-aerobic than anaerobic conditions. The concentration, molar yield, and productivity of 1,3-PD in fed-batch fermentation under micro-aerobic conditions were 59.50 g/l, 51.75%, and 1.57 g l^–1 h^–1, respectively. The volumetric productivity of 1,3-PD under microaerobic conditions was almost twice that of anaerobic fed-batch fermentation, at 1.57 and 0.80 g l^–1 h^–1, respectively.     

The impact of pH control on the volumetric productivity of mixed culture PHA production from fermented molasses

Polyhydroxyalkanoate (PHA) production via mixed microbial cultures (MMCs) can potentially decrease process operational costs as compared to conventional pure culture techniques. However, the volumetric productivity of PHA by MMCs must be augmented to increase its cost competitiveness. For this purpose, a three‐stage bioreactor system was operated in this study, with (i) anaerobic fermentation of molasses, (ii) culture selection, and (iii) PHA accumulation and harvesting stages. In stage 2, bioreactor operation with pH control at 8 led to twice the biomass concentration (up to 8 g VSS L^?1, where VSS is the volatile suspended solids) as compared to operation without pH control (maximum pH 9). No loss in the specific PHA storage efficiency was observed (PHA content up to 57.5% and PHA storage rate up to 0.27 Cmol PHA Cmol X^?1 h^?1, where X is the active biomass), thereby resulting in twice the volumetric PHA production rate. The limited biomass growth at the higher pH level was not due to nutrient limitation, but likely to a shift in the microbial community. It is hypothesized that the increased enrichment of Azoarcus at pH 8 led to higher PHA productivity. pH control in the culture selection stage can lead to enhanced PHA production from MMCs, improving the viability of the process.     

Competition-Mediated Antibiotic Induction in the Marine Bacterium Streptomyces tenjimariensis

Microbial competition for limiting natural resources within a community is thought to be the selective force that promotes biosynthesis of antimicrobial compounds The marine bacterium Streptomyces tenjimariensis produces the antibiotics istamycin A and B under select laboratory culture conditions; presumably these compounds serve an, ecological role under natural conditions. Here we report results of a novel marine microbial competion experiment that examined the impact of co-culture of marine bacteria on istamycin production by S. tenjimariensis. Twelve of the 53 bacterial species tested (i.e., 22.6%) induced Istamycin production; this antibiotic also inhibited growth of the competitor colonies. These results suggest that marine bacterial metabolites, serve an ecological role in countering competitive species.     

The effects of space flight on the production of monorden by Humicola fuscoatra WC5157 in solid-state fermentation

The effect of space flight on the production of the antibiotic monorden on two types of agar media, T8 and PG, by Humicola fuscoatra WC5157 was examined on board the US Space Shuttle mission STS-77 in May 1996. Paired space-flight and ground control samples were prepared using identical hardware, protocol, media, and inoculum. Inoculation occurred simultaneously for both groups 2.5 h after launch. The flight and ground samples were allowed to grow for the entire 10-day mission in a dark, thermally controlled (22 °C) environment. Post-flight HPLC analysis of the flight and ground sample extracts indicated that the production of monorden by H. fuscoatra WC5157 in the flight samples was higher than in the ground samples in both agar media. In the T8 medium, the production of monorden in the flight and ground samples was 11.6 ± 3.5 μg and 8.9 ± 1.1 μg respectively (30% increase). In the PG medium, the production of monorden in the flight and ground samples was 23.8 ± 3.3 μg and 8.2 ± 2.2 μg respectively (190% increase). The production of monorden in the flight and ground control samples was confirmed by HPLC-MS analysis. Received: 30 September 1997 / Received revision: 23 December 1997 / Accepted: 2 January 1998     

Study of docosahexaenoic acid production by the heterotrophic microalga <Emphasis Type="Italic">Crypthecodinium cohnii</Emphasis> CCMP 316 using carob pulp as a promising carbon source

In this work, carob pulp syrup was used as carbon source in C. cohnii fermentations for docosahexaenoic acid production. In preliminary experiments different carob pulp dilutions supplemented with sea salt were tested. The highest biomass productivity (4 mg/lh) and specific growth rate (0.04/h) were observed at the highest carob pulp dilution (1:10.5 (v/v), corresponding to 8.8 g/l glucose). Ammonium chloride and yeast extract were tested as nitrogen sources using different carob pulp syrup dilutions, supplemented with sea salt as growth medium. The best results were observed for yeast extract as nitrogen source. A C. cohnii fed-batch fermentation was carried out using diluted carob pulp syrup (1:10.5 v/v) supplemented with yeast extract and sea salt. The biomass productivity was 420 mg/lh, and the specific growth rate 0.05/h. Under these conditions the DHA concentration and DHA production volumetric rate attained 1.9 g/l and 18.5 mg/lh respectively after 100.4 h. The easy, clean and safe handling of carob pulp syrup makes this feedstock a promising carbon source for large-scale DHA production from C. cohnii. In this way, this carob industry by-product could be usefully disposed of through microbial production of a high value fermentation product.     

Bottom-up regulation of bacterial growth in tropical phytotelm bromeliads

Evaluating the factors that regulate bacterial growth in natural ecosystems is a major goal of modern microbial ecology. Phytotelm bromeliads have been used as model ecosystems in aquatic ecology as they provide many independent replicates in a small area and often encompass a wide range of limnological conditions. However, as far as we know, there has been no attempt to evaluate the main regulatory factors of bacterial growth in these aquatic ecosystems. Here, we used field surveys to evaluate the main bottom-up factors that regulate bacterial growth in the accumulated water of tank bromeliads. Bacterial production, water temperature, water color, chlorophyll-a, and nutrient concentrations were determined for 147 different tank bromeliads in two different samplings. Bromeliad position and the season of sampling were also noted. Bacterial production was explained by ion ammonium concentration and water temperature, but the total variance explained was low (r ² = 0.104). Sampling period and bromeliad position were included in additional models that gave empirical support for predicting bacterial production. Bromeliad water tanks are extremely variable aquatic ecosystems in space (among bromeliads) and time (environmental conditions can change within hours), and it is well known that bacterial production responds rapidly to environmental change. Therefore, we concluded that several factors could independently regulate bacterial growth in phytotelm bromeliads depending on the characteristics of each bromeliad, such as location, amount of detritus, and ambient nutrient concentrations. A clear bottom-up limitation pattern of bacterial production in tropical phytotelm bromeliads was not found. Handling editor: Luigi Naselli-Flores     

The significance of viruses to mortality in aquatic microbial communities

A variety of approaches including enumeration of visibly infected microbes, removal of viral particles, decay of viral infectivity, and measurements of viral production rates have been used to infer the impact of viruses on microbial mortality. The results are surprisingly consistent and suggest that, on average, about 20% of marine heterotrophic bacteria are infected by viruses and 10–20% of the bacterial community is lysed daily by viruses. The effect of viruses on phytoplankton is less certain, but ca. 3% of Synechococcus biomass may be lysed daily. The fraction of primary productivity this represents depends upon the relative biomass and growth rate of Synechococcus. Virus enrichment experiments suggest that the productivity of eukaryotic phytoplankton would be ca. 2% higher in the absence of viruses. Overall, probably about 2–3% of primary productivity is lost to viral lysis. There is considerable variation about these estimates; however, they represent a starting point for incorporating viral-mediated processes into aquatic ecosystem models.     

Fermentative Production of Succinic Acid from n-Paraffin by Candida brumptii IFO 0731

An investigation of succinic acid production from n-paraffin under various culture conditions was carried out with Candida brumptii IFO 0731. Ammonium nitrogen was required for both cell growth and succinic acid production. Favorable culture conditions for succinic acid production were ascertained. The productivity was markedly increased by the additions of CaCO₃ and organic nutrients. Under the best condition, the largest quantity of succinic acid production, 23.6 mg/ml, was obtained in a 67% yield from super heavy n-paraffin after 8 days cultivation.     

Analysis of drought responsive proteins in wheat (Triticum durum) by 2D-PAGE and MALDI-TOF mass spectrometry

Drought is an abiotic stress that strongly influences plant growth, development and productivity. By proteomics study it is possible to identify the complex mechanism of water-stress response. The aim of this research was the analysis of wheat (Triticum durum) proteome changes under stress conditions by applying a severe water deficit treatment for 7 days. Stress-induced proteome changes were analyzed by two-dimensional gel electrophoresis in conjunction with matrix-assisted laser desorption ionization-time of flight mass spectrometry. Thirty-six protein spots showed a reproducible significant change between control and stressed samples. The reasonable implications in drought response of the identified proteins were discussed. Results provide new insights that can lead to a better understanding of the molecular basis of drought-sensitivity in plants. Therefore, the obtained data could suggest the development of drought resistant wheat varieties, in order to improve agricultural production in dry regions.     

Can Nitrogen Fertilization Aid Restoration of Mature Tree Productivity in Degraded Dryland Riverine Ecosystems?

Restoration of riparian forest productivity lost as a consequence of flow regulation is a common management goal in dryland riverine ecosystems. In the northern hemisphere, dryland river floodplain trees often include one or another species of Populus, which are fast‐growing, nutrient‐demanding trees. Because the trees are phreatophytic in drylands, and have water needs met in whole or in part by a shallow water table, their productivity may be limited by nitrogen (N) availability, which commonly limits primary productivity in mesic environments. We added 20 g N m^?2 in a 2‐m radius around the base of mature Populus fremontii along each of a regulated and free‐flowing river in semiarid northwest Colorado, USA (total n = 42) in order to test whether growth is constrained by low soil N. Twelve years after fertilization, we collected increment cores from these and matched unfertilized trees and compared radial growth ratios (growth in the 3‐year post‐fertilization period/growth in the 3‐year pre‐fertilization period) in paired t tests. We expected a higher mean ratio in the fertilized trees. No effect from fertilization was detected, nor was a trend evident on either river. An alternative test using analysis of covariance (ANCOVA) produced a similar result. Our results underscore the need for additional assessment of which and to what extent factors other than water control dryland riverine productivity. Positive confirmation of adequate soil nutrients at these and other dryland riparian sites would bolster the argument that flow management is necessary and sufficient to maximize productivity and enhance resilience in affected desert riverine forests.     

Ethylene Is Responsible for a Disturbed Development of Plant Reproductive System under Conditions of Space Flight

The development of reproductive organs was studied on three dwarf cultivars of wheat Triticum aestivum and the fast-cycling Brassica rapa plants, grown under earth control conditions, during the space flight in the Mir orbital station, and in a earth experiment that simulated growth conditions during the space flight, including an elevated content of ethylene in the air (1 mg/m³ on average). We found that the embryological characteristics of the plants were not affected by space flight conditions. The elevated ethylene content in air resulted in some changes in the morphometric characteristics of inflorescences and a greater frequency of sterility similar under conditions of space flight and control earth experiment. We conclude that the abnormalities and modifications in the development of reproductive organs, induced by space flight conditions, were caused by a secondary factor, an elevated ethylene content in the cabin air, rather than by microgravity.     

Microbial inoculants with higher capacity to colonize soils improved wheat drought tolerance

Microbial inoculants have gained increasing attention worldwide as an eco-friendly solution for improving agriculture productivity. Several studies have demonstrated their potential benefits, such as enhanced resistance to drought, salinity, and pathogens. However, the beneficial impacts of inoculants remain inconsistent. This variability is attributed to limited knowledge of the mechanisms by which microbial inoculants affect crop growth and a lack of ecological characteristics of these inoculants that limit our ability to predict their beneficial effects. The first important step is believed to be the evaluation of the inoculant's ability to colonize new habitats (soils and plant roots), which could provide crops with beneficial functions and improve the consistency and efficiency of the inoculants. In this study, we aimed to investigate the impact of three microbial inoculants (two bacterial: P1 and P2, and one fungal: P3) on the growth and stress responses of three wheat varieties in two different soil types under drought conditions. Furthermore, we investigated the impact of microbial inoculants on soil microbial communities. Plant biomass and traits were measured, and high-throughput sequencing was used to characterize bulk and rhizosphere soil microbiomes after exposure to drought stress. Under drought conditions, plant shoot weight significantly increased (11.37%) under P1 treatments compared to uninoculated controls. In addition, total nitrogen enzyme activity increased significantly under P1 in sandy soil but not in clay soil. Importantly, network analyses revealed that P1, consisting of Bacillus paralicheniformis and Bacillus subtilis, emerged as the keystone taxa in sandy soil. Conversely, P2 and P3 failed to establish as keystone taxa, which may explain their insignificant impact on wheat performance under drought conditions. In conclusion, our study emphasizes the importance of effective colonization by microbial inoculants in promoting crop growth under drought conditions. Our findings support the development of microbial inoculants that robustly colonize plant roots for improved agricultural productivity.     

Quorum sensing-based metabolic engineering of the precursor supply in Streptomyces coelicolor to improve heterologous production of neoaureothin

Streptomyces are important industrial bacteria that produce pharmaceutically valuable polyketides. However, mass production on an industrial scale is limited by low productivity, which can be overcome through metabolic engineering and the synthetic biology of the host strain. Recently, the introduction of an auto-inducible expression system depending on microbial physiological state has been suggested as an important tool for the industrial-scale production of polyketides. In this study, titer improvement by enhancing the pool of CoA-derived precursors required for polyketide production was driven in a quorum sensing (QS)-dependent manner. A self-sustaining and inducer-independent regulatory system, named the QS-based metabolic engineering of precursor pool (QMP) system, was constructed, wherein the expression of genes involved in precursor biosynthesis was regulated by the QS-responsive promoter, scbAp. The QMP system was applied for neoaureothin production in a heterologous host, Streptomyces coelicolor M1152, and productivity increased by up to 4-fold. In particular, the engineered hyperproducers produced high levels of neoaureothin without adversely affecting cell growth. Overall, this study showed that self-regulated metabolic engineering mediated by QS has the potential to engineer strains for polyketide titer improvement.     

Effect of azithromycin on enhancement of methane production from waste activated sludge

In the methane production from waste activated sludge (WAS), complex bacterial interactions in WAS have been known as a major contribution to methane production. Therefore, the influence of bacterial community changes toward methane production from WAS was investigated by an application of antibiotics as a simple means for it. In this study, azithromycin (Azm) as an antibiotic was mainly used to observe the effect on microbial changes that influence methane production from WAS. The results showed that at the end of fermentation, Azm enhanced methane production about twofold compared to control. Azm fostered the growth of acid-producing bacterial communities, which synthesized more precursors for methane formation. DGGE result showed that the hydrolysis as well as acetogenesis stage was improved by the dominant of B1, B2 and B3 strains, which are Clostridium species. In the presence of Azm, the total population of archaeal group was increased, resulting in higher methane productivity achievement.     

Hollow-fibre bioreactors compared to batch and chemostat culture for the production of a recombinant toxoid by a marine Vibrio

Bioreactor selection is important for maximising the productivity of recombinant organisms. In this paper a comparison is made between growth and recombinant protein synthesis in three types of bioreactor containing a marine Vibrio capable of heterologous expression and secretion of the non-toxic B-subunit pentamer of Escherichia coli heat-labile enterotoxin, EtxB. The heterologous gene was located on the plasmid pMMB68. Resistance to carbenicillin was used to select for plasmid-containing cells. In batch and continuous culture, volumetric productivities were highest when cells were grown in the presence of carbenicillin. Without antibiotic selection, the highest volumetric productivity (9.4 mg EtxB⁻¹ h⁻¹) was observed in hollow-fibre bioreactors, and the production phase could be maintained for over 50 h. The highest specific productivity under these conditions was found in batch culture, but the maximal production phase was only of 5 h duration. In hollow-fibre reactors the type of fibre used significantly affected productivity, both with regards to the maintenance of reactor integrity and by allowing passage of the recombinant toxoid through the selectively permeable membrane. Where contamination of the product with carbenicillin is to be avoided, these bioreactors are superior to batch or continuous culture. Received: 29 January 1997 / Received revision: 9 April 1997 / Accepted: 13 April 1997     

RELATION BETWEEN MONTHLY GROWTH OF FEROCACTUS ACANTHODES AND AN ENVIRONMENTAL PRODUCTIVITY INDEX

Most productivity studies use destructive harvesting methods, prohibiting continuous plant monitoring under various environmental conditions. Here a nondestructive procedure involving the displacement of areoles from the apex was utilized to study the growth of Ferocactus acanthodes, a common barrel cactus of the Sonoran Desert. Net CO₂ uptake measured under controlled conditions of temperature, water, and photosynthetically active radiation in the laboratory was used to indicate how F. acanthodes would respond to field values of these parameters. For example, net CO₂ uptake over 24 hr was maximal at day/night air temperatures of 23 C/14 C, the mean annual values in the field, and was approximately halved at 11 C/5 C and 32 C/23 C, the monthly extreme values in the field. An environmental productivity index (EPI), constructed as the product of indices for the three environmental variables, indicated the fraction of maximal CO₂ uptake expected. The monthly production of areoles on 33 plants was highly correlated with EPI (r² = 0.81). Areole production for individual plants, however, tended to be in pulses represented by Fibonacci numbers. EPI predicted an annual stem growth of 8% compared with 9 ± 3% measured previously in the field. Thus, morphological and physiological studies can be usefully combined and applied to indicate field productivity of F. acanthodes and, by extension, of other plants.