The quality of brown rice vinegar that was produced via static culture (A) was compared with the quality of three types of domestic commercial brown rice vinegar (B, C, and D) and of three types of Japanese brown rice vinegar (E, F, and G). The results showed titratable acidity levels of 6.39%, 4.52-6.32%, and 4.51-4.89% in the static-cultured brown rice vinegar, the domestic commercial brown rice vinegars, and the Japanese brown rice vinegars, respectively. The pH levels were 3.28, 2.58-2.97, and 3.03-3.27 in the static-culture brown rice vinegar, the domestic commercial brown rice vinegars, and the Japanese brown rice vinegars, respectively, which show similar values of the static-culture brown rice vinegar and the Japanese brown rice vinegars. The total nitrogen (TN) values of the static-culture brown rice vinegar, the domestic commercial brown rice vinegars, and the Japanese brown rice vinegars were 0.24, 0.03-0.16, and 0.12-0.17, respectively, with the highest value for the static-culture brown rice vinegar, substantial differences among the domestic commercial brown rice vinegars, and similar contents among the Japanese brown rice vinegars. For free sugar, glucose was either detected only in a small quantity or not detected at all in the static-culture brown rice vinegar and the Japanese brown rice vinegars, which showed perfect fermentation. The glucose and maltose contents were higher in the domestic commercial brown rice vinegars. The organic acid content of the static-culture brown rice vinegar was similar to that of the Japanese brown rice vinegars. Therefore, the total acidity content, TN value, sensory property, and quality of the static-culture brown rice vinegar (A) were superior to those of the domestic and Japanese brown rice vinegars.
With strong biotin binding affinity ($K_D=10^{-14}M$), the tetrameric feature of streptavidin could be used to increase the antigen binding activity of a camel heavy chain (VHH) antibody through their fusion, here stained with biotinylated horseradish peroxidase and subsequent immunoassays ELISA and Western blot analysis. For this application, we cloned the streptavidin gene amplified from the Streptomyces avidinii chromosome by PCR, and this was fused to the gene of the 8B9 VHH antibody which is specific to green fluorescent protein (GFP) antigens. To express a soluble fusion protein in Escherichia coli, we used the pUC119 plasmid-based expression system which uses the lacZ promoter for induction by IPTG, the pelB leader sequence at the N-terminus for secretion into the periplasmic space, and six polyhistidine tags at the C-terminus for purification of the expressed proteins using an $Ni^+$-NTA-agarose column. Although streptavidin is toxic to E. coli because of its strong biotin binding property, this soluble fusion protein was expressed successfully. In SDS-PAGE, the size of the purified fusion protein was 122.4 kDa in its native condition and 30.6 kDa once denatured by boiling, suggesting the tetramerization of the monomeric subunit by non-covalent association through the streptavidin moiety fusing to the 8B9 VHH antibody. In addition, this fusion protein showed biotin binding activity similar to streptavidin as well as GFP antigen binding activity through both ELISA and Western blot analysis. In conclusion, the protein resulting from the fusion of an 8B9 VHH antibody with streptavidin was successfully expressed and purified as a soluble tetramer in E. coli; it showed both biotin and GFP antigen binding activity suggesting the possible production of a tetrameric and bifunctional VHH antibody.
Park, Kwang-Lai;Suga, Yuko;Hong, Seung-Gil;Lee, Chorong;Ahn, Minsil;Kim, Seok-Cheol;Hashimoto, Tomoyoshi
Journal of the Korea Organic Resources Recycling Association
/
v.24
no.4
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pp.77-83
/
2016
The objectives of this study was to investigate the difference between organic-farming and conventional-farming soils relatives to soil chemical properties and microbial flora. Fifteen soil sampling sites were chosen from the certified organic upland farm, considered with its location, crop and application of organic compost types. Soil chemical properties were analyzed by standard methods established by National Institute of Agricultural Sciences, Rural Development Administration. For the soil chemical properties, the values of pH were ranged from 4.5 to 7.3. The values of electrical conductivity (EC) in the sampling sites were below 2 dS/m of convention cultivation soil. For analyzing the microbial flora, the bacillus(16S rDNA) and cladothricosis(18S rDNA) were analyzed by using PCR-DGGE (Denaturing Gradient Gel Electrophoresis) in the soil of 15 sampling sites. Cluster analysis of biodiversity index was performed by using pattern of DGGE. DGGE patterns and clustering analysis of bacterial DNA from soil extracts revealed that the bacterial community was differentiated between less than 5 years and more than 5 years depending on the cultivation history. But there was no consistent tendency between cultivation history and regional trend in the case of molds. Therefore, it would be very effective to analyze bacterial clusters of organically cultivated soils in long - term cultivated soil for more than 5 years.
The rheological properties of ${\beta}-glucans$ isolated from non-waxy and waxy barley were investigated. ${\beta}-Glucan$ solutions showed pseudoplastic properties and their behaviors were explained by applying Power law model in the range of concentrations$(1{\sim}4%)$ and temperatures$(20{\sim}65^{\circ}C)$. The effects of temperature and concentration on the apparent viscosity at $700\;s^{-1}$ shear rate were examined by applying Arrhenius equation and power law equation, and their effect was more pronounced in waxy ${\beta}-glucan$ solutions. The activation energy for flow of ${\beta}-glucan$ solutions decreased with the increase of concentration, and the concentration-dependent constant A increased with the increase of temperature. The intrinsic viscosity of waxy ${\beta}-glucan$ was higher than that of non-waxy ${\beta}-glucan$. The transition from dilute to concentrate region occurred at a critical coil overlap parameter $C^*[{\eta}]=0.02.$ The slopes of non-waxy and waxy ${\beta}-glucan$ at $C[{\eta}] were similar, but the slope of waxy ${\beta}-glucan$ at $C[{\eta}]>C^*[{\eta}]$ was higher than that of non-waxy ${\beta}-glucan$. Dynamic viscoelasticity measurement showed that cross-over happened, and storage modulus was higher than loss modulus at frequency range above cross-over. ${\beta}-Glucan$ solutions formed weak gels after stored for 24 hr.
Beauveria bassiana (Cordycipitaceae, Hypocreales, Ascomycota) is an anamorphic fungus having a potential to be used as a biological control agent because it parasitizes a wide range of arthropod hosts including termites, aphids, beetles and many other insects. A number of bioactive secondary metabolites (SMs) have been isolated from B. bassiana and functionally verified. Among them, beauvericin and bassianolide are cyclic depsipeptides with antibiotic and insecticidal effects belonging to the enniatin family. Non-ribosomal peptide synthetases (NRPSs) play a crucial role in the synthesis of these secondary metabolites. NRPSs are modularly organized multienzyme complexes in which each module is responsible for the elongation of proteinogenic and non-protein amino acids, as well as carboxyl and hydroxyacids. A minimum of three domains are necessary for one NRPS elongation module: an adenylation (A) domain for substrate recognition and activation; a tholation (T) domain that tethers the growing peptide chain and the incoming aminoacyl unit; and a condensation (C) domain to catalyze peptide bond formation. Some of the optional domains include epimerization (E), heterocyclization (Cy) and oxidation (Ox) domains, which may modify the enzyme-bound precursors or intermediates. In the present study, we analyzed genomes of B. bassiana and its allied species in Hypocreales to verify the distribution of NRPS-encoding genes involving biosynthesis of beauvericin and bassianolide, and to unveil the evolutionary processes of the gene clusters. Initially, we retrieved completely or partially assembled genomic sequences of fungal species belonging to Hypocreales from public databases. SM biosynthesizing genes were predicted from the selected genomes using antiSMASH program. Adenylation (A) domains were extracted from the predicted NRPS, NRPS-like and NRPS-PKS hybrid genes, and used them to construct a phylogenetic tree. Based on the preliminary results of SM biosynthetic gene prediction in B. bassiana, we analyzed the conserved gene orders of beauvericin and bassianolide biosynthetic gene clusters among the hypocrealean fungi. Reciprocal best blast hit (RBH) approach was performed to identify the regions orthologous to the biosynthetic gene cluster in the selected fungal genomes. A clear recombination pattern was recognized in the inferred A-domain tree in which A-domains in the 1st and 2nd modules of beauvericin and bassianolide synthetases were grouped in CYCLO and EAS clades, respectively, suggesting that two modules of each synthetase have evolved independently. In addition, inferred topologies were congruent with the species phylogeny of Cordycipitaceae, indicating that the gene fusion event have occurred before the species divergence. Beauvericin and bassianolide synthetases turned out to possess identical domain organization as C-A-T-C-A-NM-T-T-C. We also predicted precursors of beauvericin and bassianolide synthetases based on the extracted signature residues in A-domain core motifs. The result showed that the A-domains in the 1st module of both synthetases select D-2-hydroxyisovalerate (D-Hiv), while A-domains in the 2nd modules specifically activate L-phenylalanine (Phe) in beauvericin synthetase and leucine (Leu) in bassianolide synthetase. antiSMASH ver. 2.0 predicted 15 genes in the beauvericin biosynthetic gene cluster of the B. bassiana genome dispersed across a total length of approximately 50kb. The beauvericin biosynthetic gene cluster contains beauvericin synthetase as well as kivr gene encoding NADPH-dependent ketoisovalerate reductase which is necessary to convert 2-ketoisovalarate to D-Hiv and a gene encoding a putative Gal4-like transcriptional regulator. Our syntenic comparison showed that species in Cordycipitaceae have almost conserved beauvericin biosynthetic gene cluster although the gene order and direction were sometimes variable. It is intriguing that there is no region orthologous to beauvericin synthetase gene in Cordyceps militaris genome. It is likely that beauvericin synthetase was present in common ancestor of Cordycipitaceae but selective gene loss has occurred in several species including C. militaris. Putative bassianolide biosynthetic gene cluster consisted of 16 genes including bassianolide synthetase, cytochrome P450 monooxygenase, and putative Gal4-like transcriptional regulator genes. Our synteny analysis found that only B. bassiana possessed a bassianolide synthetase gene among the studied fungi. This result is consistent with the groupings in A-domain tree in which bassianolide synthetase gene found in B. bassiana was not grouped with NRPS genes predicted in other species. We hypothesized that bassianolide biosynthesizing cluster genes in B. bassiana are possibly acquired by horizontal gene transfer (HGT) from distantly related fungi. The present study showed that B. bassiana is the only species capable of producing both beauvericin and bassianolide. This property led to B. bassiana infect multiple hosts and to be a potential biological control agent against agricultural pests.
Physicochemical properties of commercial sweet potato starches manufactured by 7 different companies were investigated in comparison with corn and potato starches. Crude ash and protein content varied from 0.36 to 1.02%, and from 0.04 to 0.14% based on dry weight, respectively. The protein contents were relatively smaller than that of corn or potato starch. But whiteness of the sweet potato starches was less than that of corn or potato starch. Mean diameter of the sweet potato starch granules varied from 14.23 to $21.08\;{\mu}m$ depending on the company and all sweet potato starches showed bimodal size distributions. Pasting viscosity measured by Rapid Viscoanalyzer(RVA) also showed variations among the starches of different companies. The starch from D company in Korea had the lowest pasting temperature$(74.00^{\circ}C)$ whereas the starch from a phillippine company(P) did the highest one$(80.35^{\circ}C)$. The peak viscosity of sweet potato starches was higher than that of corn starch but lower than that of potato starch. The D company starch also showed the highest peak viscosity(2283 cp) among the starches tested. Paste breakdown by hot shearing ranged from 524 cp (S company) to 1279 cp (HL company). Textural properties of the starch gels appeared significantly different among the starches of different manufacturers. The greatest hardness of the gel was $137.90\;g_{f}$ at 1 day storage whereas the lowest value was $31.53\;g_{f}$. Except the starches from 2 companies (P and S), the sweet potato starches formed very soft and weak gels. P or S company starches formed the gels similar to potato starch. Syneresis by freeze-thawing treatments appeared less for sweet potato starch gels than that for corn starch gels, but greater than that for potato starch gel. The overall properties of the sweet potato starches varied by the manufacturing companies, and ranged between those of corn and potato starches.
Shin, Seung Yong;Kang, Moohee;Shinn, Young Jae;Cheong, Snons
Economic and Environmental Geology
/
v.53
no.1
/
pp.33-43
/
2020
CO2 geological storage is currently considered as the most stable and effective technology for greenhouse gas reduction. The saline formations for CO2 geological storage are generally located at a depth of more than 800 m where CO2 can be stored in a supercritical state, and an extensive impermeable cap rock that prevents CO2 leakage to the surface should be distributed above the saline formations. Trough analysis of seismic and well data, we identified the basalt flow structure for potential CO2 storage where saline formation is overlain by basalt cap rock around PZ-1 exploration well in the Southern Continental Shelf of Korea. To evaluate CO2 storage capacity of the saline formation, total porosity and CO2 density are calculated based on well logging data of PZ-1 well. We constructed a 3D geological grid model with a certain size in the x, y and z axis directions for volume estimates of the saline formation, and performed a property modeling to assign total porosity to the geological grid. The estimated average CO2 geological storage capacity evaluated by the U.S. DOE method for the saline formation covered by the basalt cap rock is 84.17 Mt of CO2(ranges from 42.07 to 143.79 Mt of CO2).
Generally there is no law and liability system which applies particulary to commercial space ventures. There are several international treaties and national statutes which deal with space ventures, but their impact on the liability of commercial space ventures has not been significant. Every state law in the United States will impose both tort and contract liability on those responsible for injuries or losses caused by defective products or by services performed negligently. As with the providers of other products and services, those who participate in commercial space ventures have exposure to liability in both tort and contract which is limited to the extent of the resulting damage The manufacturer of a small and cheap component which caused a satellite to fail to reach orbit or to operate nominally has the same exposure to liability as the provider of launch vehicle or the manufacturer of satellite into which the component was incorporaded. Considering the enormity of losses which may result from launch failure or satellite failure, those participated in commercial space ventures will do their best to limit their exposure to liability by contract to the extent permitted by law. In most states of the United States, contracts which limit or disclaim the liability are enforceable with respect to claims for losses or damage to property if they are drafted in compliance with the requirements of the applicable law. In California an attempt to disclaim the liability for one's own negligence will be enforceable only if the contract states explicitly that the parties intend to have the disclaimer apply to negligence claims. Most state laws of the United States will refuse to enforce contracts which attempt to disclaim the liability for gross negligence on public policy grounds. However, the public policy which favoured disclaiming the liability as to gross negligence for providers of launch services was pronounced by the United States Congress in the 1988 Amendments to the 1984 Commercial Space Launch Act. To extend the disclaimer of liability to remote purchasers, the contract of resale should state expressly that the disclaimer applies for the benefit of all contractors and subcontractors who participated in producing the product. This situation may occur when the purchaser of a satellite which has failed to reach orbit has not contracted directly with the provider of launch services. Contracts for launch services usually contain cross-waiver of liability clauses by which each participant in the launch agrees to be responsible for it's own loss and to waive any claims which it may have against other participants. The crosswaiver of liability clause may apply to the participants in the launch who are parties to the launch services agreement, but not apply to their subcontractors. The role of insurance in responding to many risks has been critical in assisting commercial space ventures grow. Today traditional property and liability insurance, such as pre-launch, launch and in-orbit insurance and third party liability insurance, have become mandatory parts of most space projects. The manufacture and pre-launch insurance covers direct physical loss or damage to the satellite, its apogee kick moter and including its related launch equipment from commencement of loading operations at the manufacture's plant until lift off. The launch and early orbit insurance covers the satellite for physical loss or damage from attachment of risk through to commissioning and for some period of initial operation between 180 days and 12 months after launch. The in-orbit insurance covers physical loss of or damage to the satellite occuring during or caused by an event during the policy period. The third party liability insurance covers the satellite owner' s liability exposure at the launch site and liability arising out of the launch and operation in orbit. In conclusion, the liability in commercial space ventures extends to any organization which participates in providing products and services used in the venture. Accordingly, it is essential for any organization participating in commercial space ventures to contractually disclaim its liability to the extent permitted by law. To achieve the effective disclaimers, it is necessary to determine the applicable law and to understand the requirements of the law which will govern the terms of the contract. A great deal of funds have been used in R&D for commercial space ventures to increase reliability, safety and success. However, the historical reliability of launches and success for commercial space ventures have proved to be slightly lower than we would have wished for. Space insurance has played an important role in reducing the high risks present in commercial space ventures.
The building process of any geological map involves linking sparse lithological outcrop information with equally sparse geometrical measurements, all in a single entity which is the preferred interpretation of the field geologist. The actual veracity of this interpretative map is partially dependent upon the frequency and distribution of geological outcrops compounded by the complexity of the local geology. Geophysics is commonly used as a tool to augment the distribution of data points, however it normally does not have sufficient geometrical constraints due to: a) all geophysical inversion models being inherently non-unique; and b) the lack of knowledge of the physical property contrasts associated with specific lithologies. This contribution proposes the combined use of geophysical edge detection routines and 'three point' solutions from topographic data as a possible approach to obtaining geological contact geometry information (strike and dip), which can be used in the construction of a preliminary geological model. This derived geological information should first be assessed for its compatibility with the scale of the problem, and any directly observed geological data. Once verified it can be used to help constrain the preferred geological map interpretation being developed by the field geologist. The method models the contacts as planar surfaces. Therefore, it must be ensured that this assumption fits the scale and geometry of the problem. Two examples are shown from folded sequences at the Bathurst Mining Camp, New Brunswick, Canada.
1. Within experimental chromatin, the total protein: DNA ratio did not vary in the same organs of control and irradiated rats. However, the amount of RNA and total protein associated with the DNA varied considerably among the different types of chromatin. In particular, the content of chromatin was the highest in the irradiated tissue, and the lowest in the chromatin control tissue. RNA and total protein ratio of chromatins from brain, liver, testis and spleen declined with experimental organs. 2. There was the same quantitative relationship between the amount of RNA and the amount histone-protein associated with DNA in each chromatin. 3. RNA:DNA ratio of chromatin showed a $1.5{\sim}2$ times increase in the irradiated organs except brain. However, RNA:DNA ratio was decreased in chromatin by irradiation. 4. Histone-protein:Residual protein ratio was greatly varied among the organs. However, the effect was not found by irradiation. 5. Priming activity of chromatins showed a higher value in testis and the activity was greater in organs with higher metabolic activity. 6. Inhibition of Actinomycin D observable in chromatin for testis, liver, spleen and brain declined without relationship between irradiated and non-irradiated conditions. Ammonium sulfate in DNA of chromatin from histone showed increased priming activity with dissociation by Electrostatics. It may give different effect of ammonium sulfate on stimulation by property of chromatins. 7. It is suggested that the results support a proposal that the higher sensitivity of radioactive in testis, spleen by irradiated showed a increase and decrease lower-sensitivity of radioactive from brain, liver than did priming activity under the radioactive conditions.
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