Hydrogen Production from Barley Straw and Miscanthus by the Hyperthermophilic Bacterium, Cadicellulosirupter bescii.

This work aimed to evaluate the feasibility of biohydrogen production from Barley Straw and Miscanthus. The primary obstacle in plant biomass decomposition is the recalcitrance of the biomass itself. Plant cell walls consist of cellulose, hemicellulose, and lignin, which make the plant robust to decomposition. However, the hyperthermophilic bacterium, Caldicellulosiruptor bescii, can efficiently utilize lignocellulosic feedstocks (Barley Straw and Miscanthus) for energy production, and C. bescii can now be metabolically engineered or isolated to produce more hydrogen and other biochemicals. In the present study, two strains, C. bescii JWCB001 (wild-type) and JWCB018 (ΔpyrFA Δldh ΔcbeI), were tested for their ability to increase hydrogen production from Barley Straw and Miscanthus. The JWCB018 resulted in a redirection of carbon and electron (carried by NADH) flow from lactate production to acetate and hydrogen production. JWCB018 produced ~54% and 63% more acetate and hydrogen from Barley Straw, respectively than its wild-type counterpart, JWCB001. Also, 25% more hydrogen from Miscanthus was obtained by the JWCB018 strain with 33% more acetate relative to JWCB001. It was supported that the engineered C. bescii, such as the JWCB018, can be a parental strain to get more hydrogen and other biochemicals from various biomass.

Structural features of a minimal intact methyltransferase of a type I restriction-modification system.

Type I restriction-modification enzymes are oligomeric proteins composed of methylation (M), DNA sequence-recognition (S), and restriction (R) subunits. The different bipartite DNA sequences of 2-4 consecutive bases are recognized by two discerned target recognition domains (TRDs) located at the two-helix bundle of the two conserved regions (CRs). Two M-subunits and a single S-subunit form an oligomeric protein that functions as a methyltransferase (M2S1 MTase). Here, we present the crystal structure of the intact MTase from Vibrio vulnificus YJ016 in complex with the DNA-mimicking Ocr protein and the S-adenosyl-L-homocysteine (SAH). This MTase includes the M-domain with a helix tail (M-tail helix) and the S1/2-domain of a TRD and a CR α-helix. The Ocr binds to the cleft of the TRD surface and SAH is located in the pocket within the M-domain. The solution- and negative-staining electron microscopy-based reconstructed (M1S1/2)2 structure reveals a symmetric (S1/2)2 assembly using two CR-helices and two M-tail helices as a pivot, which is plausible for recognizing two DNA regions of same sequence. The conformational flexibility of the minimal M1S1/2 MTase dimer indicates a particular state resembling the structure of M2S1 MTases.

Structural insights into the regulation of SigB activity by RsbV and RsbW.

Bacillus subtilis SigB is an alternative sigma factor that initiates the transcription of stress-responsive genes. The anti-sigma factor RsbW tightly binds SigB to suppress its activity under normal growth conditions and releases it when nonphosphorylated RsbV binds to RsbW in response to stress signals. To understand the regulation of SigB activity by RsbV and RsbW based on structural features, crystal structures and a small-angle X-ray scattering (SAXS) envelope structure of the RsbV-RsbW complex were determined. The crystal structures showed that RsbV and RsbW form a heterotetramer in a similar manner to a SpoIIAA-SpoIIAB tetramer. Multi-angle light scattering and SAXS revealed that the RsbV-RsbW complex is an octamer in solution. Superimposition of the crystal structure on the SAXS envelope structure showed that the unique dimeric interface of RsbW mediates the formation of an RsbV-RsbW octamer and does not prevent RsbV and SigB from binding to RsbW. These results provide structural insights into the molecular assembly of the RsbV-RsbW complex and the regulation of SigB activity.

Effective University Facility Management Plan Proposal Reflecting the Needs of the Main Users.

As the low birthrates in Korea intensify, the average school-age population is showing great reduction and the resources used for admission to the university are gradually decreasing as well, due to the continuing economic downturn. Therefore, in order for the university to remain competitive and keep up with the fast paced changes, it is very significant that they induce qualitative growth of university education and facilities through extensive and thorough analysis on the university facility management (UFM) for students who are the main users of university facilities. However, research done on university facilities in Korea has been focused mainly on space management, while the effective UFM reflecting in-depth opinions on the user class has been quite inadequate. Therefore, in order to improve student satisfaction and improve the efficiency of UFM, it is crucial to understand the importance and status of the UFM items. This way, an initial plan for improvement of UFM considering the priorities that actively reflect the opinions of the students is prepared in this research. For the UFM items, the eight classifications of UFM, which can be evaluated by users, and the details of the classifications are used for this research. For the UFM strategies, the first 176 performance indicators (PIs) are collected, consolidated, and deleted. Finally, eight UFM strategies are derived. In order to find out which UFM items need more focus on, importance-performance analysis (IPA) is conducted. The priority of management is determined by where each factor is located on the grid. Additionally, multiple linear regression analysis is conducted to examine the effect of the importance value on the UFM items on the importance value on the UFM strategies. Finally, the average values of importance for the strategies of UFM of two groups are compared and analyzed. As a result of the stages listed above, this research attempts to provide basic data on preparing the UFM plan. Therefore, it is possible to apply the method that reflects the needs of actual users in preparing future UFM plans throughout the research methods proposed in this research.

Anion Exchange Membranes Obtained from Poly(arylene ether sulfone) Block Copolymers Comprising Hydrophilic and Hydrophobic Segments.

The anion exchange membrane may have different physical and chemical properties, electrochemical performance and mechanical stability depending upon the monomer structure, hydrophilicity and hydrophobic repeating unit, surface form and degree of substitution of functional groups. In current work, poly(arylene ether sulfone) (PAES) block copolymer was created and used as the main chain. After controlling the amount of NBS, the degree of bromination (DB) was changed in Br-PAES. Following that, quaternized PAES (Q-PAES) was synthesized through quaternization. Q-PAES showed a tendency of enhancing water content, expansion rate, ion exchange capacity (IEC) as the degree of substitution of functional groups increased. However, it was confirmed that tensile strength and dimensional properties of membrane reduced while swelling degree was increased. In addition, phase separation of membrane was identified by atomic force microscope (AFM) image, while ionic conductivity is greatly affected by phase separation. The Q-PAES membrane demonstrated a reasonable power output of around 64 mW/cm2 while employed as electrolyte in fuel cell operation.

Chemically Denatured Structures of Porcine Pepsin using Small-Angle X-ray Scattering.

Porcine pepsin is a gastric aspartic proteinase that reportedly plays a pivotal role in the digestive process of many vertebrates. We have investigated the three-dimensional (3D) structure and conformational transition of porcine pepsin in solution over a wide range of denaturant urea concentrations (0-10 M) using Raman spectroscopy and small-angle X-ray scattering. Furthermore, 3D GASBOR ab initio structural models, which provide an adequate conformational description of pepsin under varying denatured conditions, were successfully constructed. It was shown that pepsin molecules retain native conformation at 0-5 M urea, undergo partial denaturation at 6 M urea, and display a strongly unfolded conformation at 7-10 M urea. According to the resulting GASBOR solution models, we identified an intermediate pepsin conformation that was dominant during the early stage of denaturation. We believe that the structural evidence presented here provides useful insights into the relationship between enzymatic activity and conformation of porcine pepsin at different states of denaturation.

Development of Reproducible EST-derived SSR Markers and Assessment of Genetic Diversity in Panax ginseng Cultivars and Related Species.

Little is known about the genetics or genomics of Panax ginseng. In this study, we developed 70 expressed sequence tag-derived polymorphic simple sequence repeat markers by trials of 140 primer pairs. All of the 70 markers showed reproducible polymorphism among four Panax speciesand 19 of them were polymorphic in six P. ginseng cultivars. These markers segregated 1:2:1 manner of Mendelian inheritance in an F2 population of a cross between two P. ginseng cultivars, 'Yunpoong' and 'Chunpoong', indicating that these are reproducible and inheritable mappable markers. A phylogenetic analysis using the genotype data showed three distinctive groups: a P. ginseng-P. japonicus clade, P. notoginseng and P. quinquefolius, with similarity coefficients of 0.70. P. japonicus was intermingled with P. ginseng cultivars, indicating that both species have similar genetic backgrounds. P. ginseng cultivars were subdivided into three minor groups: an independent cultivar 'Chunpoong', a subgroup with three accessions including two cultivars, 'Gumpoong' and 'Yunpoong' and one landrace 'Hwangsook' and another subgroup with two accessions including one cultivar, 'Gopoong' and one landrace 'Jakyung'. Each primer pair produced 1 to 4 bands, indicating that the ginseng genome has a highly replicated paleopolyploid genome structure.