Chloroplasts are believed to be descended from certain cyanobacteria, which were taken up by phagocytosis into a host cell and lived there in a symbiotic relationship. In contrast to the current static concept on the chloroplast genome, its dynamism has been recently demonstrated: the chloroplast genome is active in intramolecular homolgous recombination, producing subgenomic circles when it obtains homolgous sequences via genetic transformation. Chloroplast tranformation in higher plants provides many advantages over nuclear transformation that include higher expression levels of transgenes, polycistronic expression of transgenes, and maternal transmission of transgenes. Tobacco has been used as a model for chloroplast genetic transformation. However, it is recently possible to transform the chloroplasts of other major food and economic crops including rice, soybean, and cotton. Chloroplast-transformed crops will be able to replace bioreactors using microorganisms for production of value-added proteins in future.
The Bacillus cereus group, also known as B. cereus sensu lato (B. cereus s.l.), is composed of various Bacillus species, some of which can cause diarrheal or emetic food poisoning. Several emerging highly heat-resistant Bacillus species have been identified, these include B. thermoamylovorans, B. sporothermodurans, and B. cytotoxicus NVH 391-98. Herein, we performed whole genome analysis of two thermotolerant Bacillus sp. isolates, Bacillus sp. B48 and Bacillus sp. B140, from an omelet with acacia leaves and fried rice, respectively. Phylogenomic analysis suggested that Bacillus sp. B48 and Bacillus sp. B140 are closely related to B. cereus and B. thuringiensis, respectively. Whole genome alignment of Bacillus sp. B48, Bacillus sp. B140, mesophilic strain B. cereus ATCC14579, and thermophilic strain B. cytotoxicus NVH 391-98 using the Mauve program revealed the presence of numerous homologous regions including genes responsible for heat shock in the dnaK gene cluster. However, the presence of a DUF4253 domain-containing protein was observed only in the genome of B. cereus ATCC14579 while the intracellular protease PfpI family was present only in the chromosome of B. cytotoxicus NVH 391-98. In addition, prophage Clp protease-like proteins were found in the genomes of both Bacillus sp. B48 and Bacillus sp. B140 but not in the genome of B. cereus ATCC14579. The genomic profiles of Bacillus sp. isolates were identified by using whole genome analysis especially those relating to heat-responsive gene clusters. The findings presented in this study lay the foundations for subsequent studies to reveal further insights into the molecular mechanisms of Bacillus species in terms of heat resistance mechanisms.
Journal of the Korean Society of Food Science and Nutrition
/
v.38
no.1
/
pp.76-82
/
2009
Wheat is the most widely cultivated cereal and an important source of dietary protein worldwide. Wheat allergy, defined as an adverse immunologic reaction to wheat, encompasses a broad spectrum of disorders with different pathomechanisms and clinical manifestation. The Nuruk, a traditional Korean Koji for brewing, was made with wheat flour and fermenting microbes such as bacteria, yeast and mold. The strains grown on Nuruk secrete various enzymes as amylase and protease. By the activation of such enzymes, starch and proteins in Nuruk are hydrolyzed to sugar and amino acid. Therefore, it is supposed to reduce allergic proteins in wheat. To study quality properties and degradation degree of allergenicity in Nuruk by fermentation, we investigated the changes of general ingredients and allergenicity in Nuruk during fermentation. Moisture contents was decreased from 24.2% to 13.6% during fermentation. Crude lipid and protein contents were gradually increased during fermentation. After 15 days of fermentation, reducing sugar and total sugar contents were reached its maximum level, and they were 27.45% and 39.00%, respectively. Acid and neutral protease activity were significantly increased during fermentation, but alkaline protease activity was not detected. ${\alpha}$-amylase activity was gradually increased and showed maximum level about 2,833.00 U/g after 15 days of fermentation. Glucoamylase activity was the highest level about 497.9 U/g after 10 days of fermentation. The increase of these proteolytic and saccharogenic enzyme activities will provide efficient condition for production of rice wine. Also, protein fractions were isolated from Nuruk, and degradation of these proteins during fermentation were confirmed by SDS-PAGE. IgE immunoblotting using patient's sera with wheat allergy was performed to confirm allergenic protein in Nuruk. These results as fermentation of Nuruk will provide a useful tool for developing safer wheat products to prevent wheat allergy.
Three beef steers fitted with permanent cannulae in the rumen and duodenum were used to determine the effects of protein supply from soyhulls (SH) and wheat bran (WB) on ruminal metabolism, blood metabolites, nitrogen metabolism, nutrient digestion and concentrations of soluble non-ammonia nitrogen (SNAN) in ruminal (RD) and omasal digesta (OD). In a 3${\times}$3 Latin square design, steers were offered rice straw and concentrates formulated either without (control) or with two brans to increase crude protein (CP) level (9 vs. 11% dietary DM for control and bran-based diets, respectively). The brans used were SH and WB that had similar CP contents but different ruminal CP degradability (52 vs. 80% CP for SH and WB, respectively) for evaluating the effects of protein degradability. Ruminal ammonia concentrations were higher for bran diets (p<0.01) than for the control, and for WB (p<0.001) compared to the SH diet. Similarly, microbial nitrogen and blood urea nitrogen were significantly increased (p<0.05) by bran and WB diets, respectively. Retained nitrogen tended (p<0.082) to be increased by SH compared with the WB diet. Intestinal and total tract CP digestion was enhanced by bran diets. In addition, bran diets tended (p<0.085) to increase intestinal starch digestion. Concentrations of SNAN fractions in RD and OD were higher (p<0.05) for bran diets than for the control, and for WB than for the SH diet. More rumendegraded protein supply resulting from a higher level and degradability of CP released from SH and WB enhanced ruminal microbial nitrogen synthesis and ruminal protein degradation. Thus, free amino acids, peptides and soluble proteins from microbial cells as well as degraded dietary protein may have contributed to increased SNAN concentrations in the rumen and, consequently, the omasum. These results indicate that protein supply from SH and WB, having a low level of protein (13 and 16%, respectively), could affect ruminal metabolism and nutrient digestion if inclusion level is relatively high (>20%).
Journal of the Korean Society of Food Science and Nutrition
/
v.42
no.11
/
pp.1727-1735
/
2013
This study is carried out to investigate the analysis of nutrition and antioxidants of soybean Chungkukjang powder (SCP), Yak-Kong Chungkukjang powder (YCP) and Yak-Kong Chungkukjang powder added black foods (YCBP) for the management of Alzheimer's disease. The water content of YCBP was higher than that of SCP and YCP. The soluble and crude protein contents of YCP were the highest among three Chungkukjang powders. In SCP, YCP and YCBP, the ratios of unsaturated and saturated fatty acids were 5.17, 5.76, and 5.78, respectively. The mineral content of SCP was higher than that of YCP and YCBP. Antioxidants analysis showed that the content of diadzein was the highest in YCP and genistein, and anthocyanin contents were higher than the others in YCBP. In conclusion, YCP and YCBP had higher contents than SCP in soluble and crude proteins, and unsaturated fatty acids, which are needed for composition and function of the brain tissue. Also, it was found that the contents of diadzein, genistein and anthocyanin with outstanding antioxidative ability were high. Thus, this study suggests that Chungkukjang powder, which is made with Yak-Kong, black sesame, black rice and sea tangle, can be utilized for in vivo experiment for the control of oxidative stress, reported as cause and therapy for Alzheimer's Disease.
Paik Soon-Young;Ra Kyung Soo;Cho Hoon Sik;Koo Kwang Bon;Baik Hyung Suk;Lee Myung Chul;Yun Jong Won;Choi Jang Won
Journal of Microbiology
/
v.44
no.1
/
pp.64-71
/
2006
To investigate the effects of the nucleotide sequences in Shine-Dalgarno (SD) and the spacer region (SD-ATG) on bovine growth hormone (bGH) gene expression, the expression vectors under the control of the T7 promoter (pT7-7 vector) were constructed using bGH derivatives (bGH1 & bGH14) which have different 5'-coding regions and were induced in E. coli BL21 (DE3). Oligonucleotides containing random SD sequences and a spacer region were chemically synthesized and the distance between the SD region and the initiation codon were fixed to nine bases in length. The oligonucleotides were annealed and fused to the bGH1 and bGH14 cDNA, respectively. When the bGH gene was induced with IPTG in E. coli BL21(DE3), some clones containing only bGH14 cDNA produced considerable levels of bGH in the range of $6.9\%\;to\;8.5\%$ of total cell proteins by SDS-PAGE and Western blot. Otherwise, the bGH was not detected in any clones with bGH1 cDNA. Accordingly, the nucleotide sequences of SD and the spacer region affect on bGH expression indicates that the sequences sufficiently destabilize the mRNA secondary structure of the bGH14 gene. When the free energy was calculated from the transcription initiation site to the +51 nucleotide of bGH cDNA using a program of nucleic acid folding and hybridization prediction, the constructs with values below -26.3 kcal/mole (toward minus direction) were not expressed. The constructs with the original sequence of bGH cDNA also did not show any expression, regardless of the free energy values. Thus, the disruption of the mRNA secondary structure may be a major factor regulating bGH expression in the translation initiation process. Accordingly, the first stem-loop among two secondary structures present in the 5'-end region of the bGH gene should be disrupted for the effective expression of bGH.
To increase both the production of mushrooms and the usages of the residues (used media) for roughage, the pinchips(PC) and sawdusts(SD) of three Korean oak species were tested as cultivating media for Lentinus edodes with varing the composition of the media. The results are as follows. 1. Organic acids(tannic acid and citric acid), which were added in the PC medium to enrich the quality of mushrooms, did not increase the growths of the mycelia and the quality. 2. The mushrooms were able to be harvested from six months after beginning the cultivation. No significant differences were found among the qualities of the mushrooms produced by various treatments. 3. The contents of crude proteins(1.82-4.55%) in the used media were higher than those in woods (c.a. 0.7%), and the total digestible nutrients (44.0-46.0%) in the used medium were not much different from those in rice straws (c.a.48.0%). 4. The contents of some essential amino acids (methionine in the used medium of the tannic acid added PC or S.D. and isoleucine, phenylalanine and lysine in the used medium of the untreated PC or SD) were higher than those in the control (raw media). The contents of a few other essential amino acids (threonine, valine and arginine) were lower in the used media. Most nonessential amino acids in the used media, particularly in the used bed-log, appeared to decrease than those in the control.
Nguyen, Ngoc Tuan;Lee, Kyoung;Kang, Ju Beom;Huang, Shir-Ly
Korean Journal of Microbiology
/
v.50
no.3
/
pp.210-215
/
2014
Pseudomonas nitroreducens TX1 was isolated from a rice field drainage in Taiwan. The bacterium is of special interest because of its capability to use a group of nonionic surfactants such as alkylphenol polyethoxylates even at high concentrations as a sole carbon source. In this study, a small cryptic circular plasmid, pTX1, was characterized from P. nitroreducens TX1. It is 2,286 bp in length with a GC content of 63.3% and harbors three open reading frames, $Rep_{pTX1}$ and functionally unidentified ORF1 and ORF2. The predicted $rep_{pTX1}$ gene product is homologous to Rep proteins of plasmids belonging to the pC194/pUB110 family, which is predominantly found in Gram-positive bacteria and is known to replicate by the rolling-circle mechanism. The copy number of pTX1 was estimated to be about 150 in each cell. Based on the genetic fingerprints and comparison with other plasmids, it is concluded that pTX1 replicates by a rolling circle mechanism which is rarely found for Pseudomonas plasmids.
Park, Sei Joon;Park, Jun Young;Hwang, Su Min;Seo, Myung Chul;Kim, Tae Wan
KOREAN JOURNAL OF CROP SCIENCE
/
v.59
no.3
/
pp.245-251
/
2014
Sorghum seed is traditionally used as health supplements and the secondary food mixed with rice in Korea. While the research of reserve protein in sorghum seed have been carried out in many countries used as major food, much less is known about reserve proteins of Korean local sorghum seeds. To obtain protein characteristics in 20 Korean local sorghum seed, quantitative content of reserve protein was determined after fractionation by modified 'Osbone' method and ${\alpha}$-kafirin of prolamin was determined by SDS-PAGE. Mean albumin, globulin, prolamin and glutelin contents based on total seed protein content of 20 Korean local sorghum seed were 6.2%, 0.9%, 57.9% and 35.1%, respectively. Sorghum cultivar with high prolamin were 'Whin-susu', 'Whin-Chalsusu', 'Whanggeum-Chalsusu', and 'Daepungshushu'. Sorghum cultivar with high ${\alpha}$-kafirin were 'Whin-susu', 'Geumsan-Chalsusu', 'Whin-Chalsusu', and 'Jangmok-susu'. Among the 20 varieties, 'Whin-susu' and 'Whin-Chalsusu' were selected as high ${\alpha}$-kafirin and prolamin sorghum cultivar, which showed 64.5 and 71.9% of prolamin contents, respectively.
Objectives: The purpose of the study was to compare intake of energy nutrients, physical characteristics, and the prevalence of metabolic syndrome according to protein intake group. Methods: Subjects were 827 men aged 40-65 years. The results presented were based on data from the 2012-2013 National Health and Nutrition Examination Survey and analyzed using SPSS. The odds ratio (OR) of metabolic syndrome was assessed according to the protein intake group and intake pattern of protein-rich foods. Results: The mean of protein intake was $73.96{\pm}0.71g$. According to level of protein intake, four groups (deficient, normal, excess 1, excess 2) were created and their percentages were 8.3%, 39.6%, 37.1%, and 15.0% respectively. The mean of daily energy intake was $2,312.33{\pm}24.08kcal$. It was higher in excess group 2 than in the deficiency group (p < 0.001). Moreover, the intake of all energy nutrients increased significantly with protein intake group (p < 0.001). The main contribution to daily protein included mixed grains ($10.96{\pm}0.32g$), milled rice ($7.14{\pm}0.30g$), chicken ($3.50{\pm}0.21g$), and grilled pork belly ($3.04{\pm}0.16g$). With regard to physical characteristics, and blood pressure and blood test results, only body mass index increased significantly according to protein intake groups (p < 0.05). The prevalence of metabolic syndrome in subjects was 38.5%, and there was no significant correlation with protein intake group. The OR of metabolic syndrome increased with protein intake, and was higher 4.452 times in excess group 2 than in the normal group (p < 0.05). Conversely, the OR of metabolic syndrome according to the frequency of protein-rich food intake did not show a significant correlation. Conclusions: The results of this study can be used as significant supporting data to establish guidelines for protein intake in middle-aged men.
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