Streptomycetes are industrially and pharmaceutically important bacteria that produce a variety of secondary metabolites including antibiotics. Streptomycetes have a complex metabolic network responsible for the production of secondary metabolites and the utilization of organic residues present in soil. In this study, we reconstructed a high-quality metabolic model for Streptomyces coelicolor A3(2), designated iMK1208, in order to understand and engineer the metabolism of this model species. In comparison to iIB711, the previous metabolic model for S. coelicolor, the predictive power of iMK1208 was enhanced by the recent insights that enabled the incorporation of an updated biomass equation, stoichiometric matrix, and energetic parameters. iMK1208 was validated by comparing predictions with the experimental data for growth capability in various growth media. Furthermore, we applied a strain-design algorithm, flux scanning based on enforced objective flux (FSEOF), to iMK1208 for actinorhodin overproduction. FSEOF results identified not only previously known gene overexpression targets such as actII-ORF4 and acetyl-CoA carboxylase, but also novel targets such as branched-chain α-keto acid dehydrogenase (BCDH). We constructed and evaluated the BCDH overexpression mutant, which showed a 52-fold increase in actinorhodin production, validating the prediction power of iMK1208. Hence iMK1208 was shown to be a useful and valuable framework for studying the biotechnologically important Streptomyces species using the principles of systems biology and metabolic engineering.

Our data suggest that HoLEP is effective in improving micturition, but de novo postoperative UI occurred in some patients although usually transient. Surgeons should be careful to not injure the bladder mucosa during morcellation.

Objectives:To determine the association of vesico-urethral anastomosis location (VUAL) with early recovery of urinary continence (UC) after radical prostatectomy (RP). Methods: A retrospective analysis of 678 patients who underwent RP was carried out. Patients were divided into three groups based on the VUAL as determined by postoperative cystography: group I -VUAL above the upper margin of the symphysis pubis (SP), group II -between the upper margin and the middle of the SP, and group III -below the middle of the SP. Early recovery of UC was defined as using no pads or an occasional security pad within 3 months. Recovery rates were compared between the groups and factors predicting an early recovery of UC were investigated. Results: Among all patients, 62.2% achieved an early recovery of UC. Patients in group I were younger, with a longer membranous urethra, greater percent of nerve sparing and shorter time to continence than those in groups II or III. Early recovery rates were 89.5%, 69.8% and 40.7% in group I, II and III, respectively (P < 0.001). VUAL remained an independent predictor of early recovery of UC (OR 3.2 for group I vs II and 10.8 for group I vs III [P < 0.001]) when adjusted for age, operative time, membranous urethral length and operation by surgeon with high surgical volume. Conclusion: VUAL represents an independent predictor of recovery of UC after RP. A higher VUAL is associated with a higher rate of early recovery of UC.

In silico model-driven analysis using genome-scale model of metabolism (GEM) has been recognized as a promising method for microbial strain improvement. However, most of the current GEM-based strain design algorithms based on flux balance analysis (FBA) heavily rely on the steady-state and optimality assumptions without considering any regulatory information. Thus, their practical usage is quite limited, especially in its application to secondary metabolites overproduction. In this study, we developed a transcriptomics-based strain optimization tool (tSOT) in order to overcome such limitations by integrating transcriptomic data into GEM. Initially, we evaluated existing algorithms for integrating transcriptomic data into GEM using Streptomyces coelicolor dataset, and identified iMAT algorithm as the only and the best algorithm for characterizing the secondary metabolism of S. coelicolor. Subsequently, we developed tSOT platform where iMAT is adopted to predict the reaction states, and successfully demonstrated its applicability to secondary metabolites overproduction by designing actinorhodin (ACT), a polyketide antibiotic, overproducing strain of S. coelicolor. Mutants overexpressing tSOT targets such as ribulose 5-phosphate 3-epimerase and NADP-dependent malic enzyme showed 2 and 1.8-fold increase in ACT production, thereby validating the tSOT prediction. It is expected that tSOT can be used for solving other metabolic engineering problems which could not be addressed by current strain design algorithms, especially for the secondary metabolite overproductions.

Nargenicin A1, an antibacterial produced by Nocardia sp. CS682 (KCTC 11297BP), demonstrates effective activity against various Gram-positive bacteria. Hence, we attempted to enhance nargenicin A1 production by utilizing the cumulative effect of synthetic biology, metabolic engineering and statistical media optimization strategies. To facilitate the modular assembly of multiple genes for genetic engineering in Nocardia sp. CS682, we constructed a set of multi-monocistronic vectors, pNV18L1 and pNV18L2 containing hybrid promoter (derived from ermE* and promoter region of neo ), ribosome binding sites (RBS), and restriction sites for cloning, so that each cloned gene was under its own promoter and RBS. The multi-monocistronic vector, pNV18L2 containing transcriptional terminator showed better efficiency in reporter gene assay. Thus, multiple genes involved in the biogenesis of pyrrole moiety (ngnN2, ngnN3, ngnN4, and ngnN5 from Nocardia sp. CS682), glucose utilization (glf and glk from Zymomonas mobilis), and malonyl-CoA synthesis (accA2 and accBE from Streptomyces coelicolor A3 (2)), were cloned in pNV18L2. Further statistical optimization of specific precursors (proline and glucose) and their feeding time led to ~84.9 mg/L nargenicin from Nocardia sp. GAP, which is ~24-fold higher than Nocardia sp. CS682 (without feeding). Furthermore, pikC from Streptomyces venezuelae was expressed to generate Nocardia sp. PikC. Nargenicin A1 acid was characterized as novel derivative of nargenicin A1 produced from Nocardia sp. PikC by mass spectrometry (MS) and nuclear magnetic resonance (NMR) analyses. We also performed comparative analysis of the anticancer and antibacterial activities of nargenicin A1 and nargenicin A1 acid, which showed a reduction in antibacterial potential for nargenicin A1 acid. Thus, the development of an efficient synthetic biological platform provided new avenues for enhancing or structurally diversifying nargenicin A1 by means of pathway designing and engineering.

Demand is increasing for mountain-cultivated Panax ginseng (MCG) because its quality is considered superior to that of fieldcultivated ginseng (FCG). However, MCG grows very slowly, and the factors that might affect this are unknown. In addition, little information is available about the physiological characteristics of its roots. Here, we investigated local soil environments and compared the histological and chemical properties of MCG and FCG roots. Average diameters, lengths, and fresh weights were much smaller in the former. Photosynthesis rates and root cambial activity also were reduced in the MCG tissues. Our analysis of soil from the mountain site revealed an extremely low phosphorus content, although those samples were ricINer in total nitrogen and organic matter than were the field soils. MCG roots also contained higher amounts of ginsenosides, and total accumulations increased with age. Moreover, ginsenoside Rh2, a red ginseng-specific compound, accumulated in the MCG roots but not in those from FCG plants. Interestingly, numerous calcium oxalate crystals were found in MCG roots, particularly in their rhizomes (i.e., short stems). Therefore, we can conclude from these results that low levels of the essential mineral phosphorus in mountain soils are a critical factor that retards the growth of mountain ginseng. Likewise, the high accumulation of calcium oxalate crystals in MCG roots might be an adaptation mechanism for survival in such a harsh local environment.

Eleutherococcus senticosus zygotic embryos were pretreated with 1.0 M mannitol or sucrose for 3-24 h. This pretreatment resulted in a high frequency of somatic-embryo formation on hormone-free medium. All the somatic embryos developed directly and independently from single epidermal cells on the surface of zygotic embryos after plasmolyzing pretreatment. Scanning electron microscopic observation revealed that the epidermal cells of hypocotyls rapidly became irregular and showed a random orientation before somatic-embryo development commenced. At the same time, the epidermal cells in the untreated control remained regular. Callose concentration determined by fluorometric analysis increased sharply in E. senticosus zygotic embryos after plasmolyzing pretreatment but remained low in the untreated control. Aniline blue fluorescent staining of callose showed that the plasmolyzing pretreatment of zygotic embryos resulted in heavy accumulation of callose between the plasma membrane and cell walls. On the basis of these results, we suggest that plasmolyzing pretreatment of zygotic embryos induces the accumulation of callose, and the interruption of cell-to-cell communication imposed by this might stimulate the reprogramming of epidermal cells into embryogenically competent cells and finally induce somatic-embryo development from single cells.

Acacia mangium is an evergreen fast-growing tropical tree, which can grow up to 30 m tall and 50 cm thick, under favorable conditions. It is a low-elevation species associated with rain forest margins and disturbed, well-drained acid soils. It is native to Papua, Western Irian Jaya and the Maluku islands in Indonesia, Papua New Guinea and north-eastern Queensland in Australia. Due to its rapid growth and tolerance of very poor soils, A. mangium was introduced into some Asian, African and western hemisphere countries where it is used as a plantation tree. A. mangium has good quality wood traits, such as a comparatively low proportion of parenchymatous cells and vessels, white and hard wood, and high calorific value. Therefore, it is useful for a variety of purposes, such as furniture, cabinets, turnery, floors, particleboard, plywood, veneer, fence posts, firewood, and charcoal. It is also being used in pulp and paper making because it has good pulp traits, with high yields of pulp, quality of kraft, and produces paper with good optical, physical and surface properties. Because there are significant provenance differences in growth rate, stem straightness, heartwood formation and frequency of multiple leaders, the productivity and quality also varies depending upon environmental conditions, so genetic improvement programmes have been undertaken in countries like Australia, India, Indonesia, Malaysia, the Philippines, Taiwan and Thailand. The programme includes provenance identifications and testing, plus tree selection and clonal multiplication, establishment of seed orchards and hybridization. The phenology, reproductive biology, fruit characteristics, silvicultural practices for cultivation, pest and diseases problems, production of improved planting stock, harvesting, wood properties and utilization have been discussed in this paper.