• Applied Biological Chemistry
• /
• v.39 no.3
• /
• pp.177-182
• /
• 1996

Effects of 14 carbohydrates supplied as carbon sources on cell growth and sporulation of, and the production of insecticidal crystal proteins by Bacillus thuringiensis strains were investigated in liquid cultures. Strains grew well in media containing any one of the 14 carbohydrates supplied, reaching maximum cell densities of $10^7{\sim}10^8\;cells/ml$ in 16.7 to 22 hours after inoculation depending on the strain. Spores first appeared in 16.7 to 24.7 hours after inoculation, and 80% sporulation was reached in 28 to 51.3 hours after inoculation depending on the strain. No change in pH of media was observed after cell multiplication. The production of total protein was highest when supplied with sucrose and was lowest with starch. More insecticidal crystal proteins were produced when supplied with glucose, lactose, maltose, or sucrose. The amount of insecticidal crystal proteins produced by the strains was proportional to that of the total protein. The relative amount of individual insecticidal crystal protein species produced by B.t. kurstaki and B.t. israelensis was not influenced by the carbohydrates supplied.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.8 no.1
• /
• pp.117-121
• /
• 2004

Soluble proteins of silkworm hemolymph were investigated by means of isoelectric focusing (IEF). The protein spectra during ontogenesis of races and inter-races hybrids kept in Bulgaria was studied. A total of 51 protein bands in the hemolymph from fourth larval instar to imago were ascertained. Stage specific expression was established. The specific expression of some protein bands in the individual spectra manifest phenotype of gene determinate polymorphism (HP F, HP J, HP K, HP L, HP Q - in the zone with pH gradient 3.5-6.2 and HP K, HP L, HP N, HP P, HP T - in the zone with pH gradient 9.5 - 6.2). Breed specific expression was observed. On the basis of the obtained results, it was established that the investigated breeds are heterogeneous and the isoelectric focusing method is successful when specifying the inner-race and inter-race polymorphism in silkworm.

• Genomics & Informatics
• /
• v.11 no.1
• /
• pp.52-54
• /
• 2013

Proteins in DNAJ/K families are ubiquitous, from prokaryotes to eukaryotes, and function as molecular chaperones. For systematic phylogenomics of the DNAJ/K families, we developed the Eukaryotic DNAJ/K Database (EDD). A total of 12,908 DNAJs and 4,886 DNAKs were identified from 339 eukaryotic genomes in the EDD. Kingdom-wide comparison of DNAJ/K families provides new insights on the evolutionary relationship within these families. Empowered by 'class', 'cluster', and 'taxonomy' browsers and the 'favorite' function, the EDD provides a versatile platform for comparative genomic analyses of DNAJ/K families.

• Korean Journal of Food Science and Technology
• /
• v.30 no.1
• /
• pp.69-76
• /
• 1998

This study was investigated to determine the effect of additives and ionic strength on the functionality of extracted proteins in preblends in order to use less additive in restructured meat products. Preblends contained the combinations of sodium chloride (NaCl; 0, 4.5, 9.0%), sodium tripolyphosphate (STPP; 0, 2.5, 5.0%), and tetrasodium pyrophosphate (PP; 0, 2.44, 4.88%). The pH values increased linearly with increasing STPP and PP concentrations (p<0.01). In the equivalent ionic strengths, PP was more effective than STPP in increasing pH. Phosphate ions were more effective on total extractable protein (used 1 M NaCl buffer) than chloride ion at equivalent ionic strengths. Solubility was decreased by adding NaCl and increasing total extractable proteins. Meat sulfhydryl contents were high with increasing total extractable proteins. When protein extracts were heated at $65^{\circ}C$, 7 min, meat sulfhydryl contents decreased and surface hydrophobicity increased (p<0.01). However, all protein extracts showed no differences in SDS-PAGE pattern. In conclusion, PP is more effective than STPP in order to use less additive but there was no linear relationship between functionnal improvement and ionic strength.

• Korean Journal of Environmental Agriculture
• /
• v.28 no.4
• /
• pp.435-441
• /
• 2009

In the present study, we analyzed the defensin protein deduced from Korean radish (Raphanus sativus L.) seeds.To express the genes in E. coli, we constructed a recombinant expression vector with a defensin gene, named rKRs-AFP gene isolated from Korean radish seeds. Over expressed rKRs-AFP proteins was separated by SDS-PAGE to determine the purity, and protein concentration was determined by the Bradford method. Antifungal activity was assessed by disk assay method against the tested fungi. As a result, when 500 mL of cell culture were disrupted by sonicator, 32.5 mg total proteins were obtained. The purified protein showed a single band on SDS-PAGE with estimated molecular weight about 6 KDa, consistent with the molecular mass calculated from the deduced amino acid sequence. The purified rKRs-AFP protein showed remarkable antifungal activities against several fungi including Aspergillus niger, Botrytis cinerea causing the gray mold disease, and Candida albicans. In field tests using the purified rKRs-AFP protein, the protein showed the reducing activity of disease spot and the mitigating effect of spreading of disease like agrichemicals. The immuno-assay of rKRs-AFP protein showed that the purified protein entirely accumulated at B. cinerea cytoplasm through the hyphal septa shown by fluorescence imaging. There was no fluorescence inside the cell, when the hypha was incubated without the protein. These all results indicate that the recombinant rKRs-AFP proteins can be utilized as a potential antifungal drug to control harmful plant fungal pathogens.

• BMB Reports
• /
• v.45 no.11
• /
• pp.665-670
• /
• 2012

The insect midgut epithelium is generally lined with a unique chitin and protein structure, the peritrophic membrane (PM), which facilitates food digestion and protects the gut epithelium. We used gel electrophoresis and mass spectrometry to identify the extracted proteins from the silkworm PM to obtain an in-depth understanding of the biological function of the silkworm PM components. A total of 305 proteins, with molecular weights ranging from 8.02 kDa to 788.52 kDa and the isoelectric points ranging from 3.39 to 12.91, were successfully identified. We also found several major classes of PM proteins, i.e. PM chitin-binding protein, invertebrate intestinal mucin, and chitin deacetylase. The protein profile provides a basis for further study of the physiological events in the PM of Bombyx mori.

• Parasites, Hosts and Diseases
• /
• v.53 no.1
• /
• pp.85-93
• /
• 2015

Proteomic tools allow large-scale, high-throughput analyses for the detection, identification, and functional investigation of proteome. For detection of antigens from Haemaphysalis longicornis, 1-dimensional electrophoresis (1-DE) quantitative immunoblotting technique combined with 2-dimensional electrophoresis (2-DE) immunoblotting was used for whole body proteins from unfed and partially fed female ticks. Reactivity bands and 2-DE immunoblotting were performed following 2-DE electrophoresis to identify protein spots. The proteome of the partially fed female had a larger number of lower molecular weight proteins than that of the unfed female tick. The total number of detected spots was 818 for unfed and 670 for partially fed female ticks. The 2-DE immunoblotting identified 10 antigenic spots from unfed females and 8 antigenic spots from partially fed females. Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF) of relevant spots identified calreticulin, putative secreted WC salivary protein, and a conserved hypothetical protein from the National Center for Biotechnology Information and Swiss Prot protein sequence databases. These findings indicate that most of the whole body components of these ticks are non-immunogenic. The data reported here will provide guidance in the identification of antigenic proteins to prevent infestation and diseases transmitted by H. longicornis.

• Journal of Plant Biology
• /
• v.22 no.4
• /
• pp.95-100
• /
• 1979

As a part of the studies on the regulatory mechanism of gene expression by $GA_{3}$ , the effects of $GA_{3}$ on the protein biosynthesis and phosphorylation in maize seedlings were investigated. 1. The optimum concentration of $GA_{3}$ for the stimulation of the protein biosynthesis was 0.3mM. 2. The protein biosynthesis was remarkably increase by $GA_{3}$ during the germination. The reason for the decrease in the protein biosynthesis by 48hrs. after germination seems to be a staggered gene expression, and/or increases in protease and RNase activities. 3. The ratio of the amount of the newly synthesized protein in germinating seeds treated with $GA_{3}$ to the amount of proteins secreted into the endosperm was similar to that ratio in control. According to this result, it seems that $GA_{3}$ stimulates only the expression of certain definite genes. 4. By the treatment with $GA_{3}$, the rates of biosynthesis and phosphorylation of proteins were increased up to about 1.5 times during germination and 6 times by 72hrs. after germination, respectively. The ratio of the total soluble proteins to the phosphorpoteins considerably increased in the early germination stage (24hrs.) but decreased after 24hrs. According to the above mentioned results, the stimulation of the phosphorylation of proteins of $GA_{3}$ seems to be attributed to the increases in the activities of protein kinases.

• Environmental Engineering Research
• /
• v.21 no.1
• /
• pp.22-28
• /
• 2016

The aim of this study was to recycle the waste livestock blood as one of the waste biomass by turning proteins, the main constituent of blood, into effective biological resources like amino acid. Ultrasonic technology was applied to solubilize the proteins in the waste livestock blood. And of the multiple ultrasonic frequencies tested, 20 kHz was confirmed to yield the highest solubilization rate. The optimum pretreatment conditions were determined to be 30-min treatment at an ultrasonic irradiation density of 0.5 W/mL, which resulted in a solubilization rate of 96.01%. Also, a gel permeation chromatography (GPC) confirmed that a large amount of proteins were solubilized, and in an experiment where ultrasonic treatment was applied to kill bacteria, death rates of general bacteria and total coliforms were found to be reduced by 99.93% and 100%, respectively. Based on these results, ultrasonic technology was confirmed to be a crucial part of treating and recycling the proteins in waste livestock blood.

• YAKHAK HOEJI
• /
• v.54 no.5
• /
• pp.377-386
• /
• 2010

Single-dimensional (1-D) and two-dimensional (2-D) LC methods were utilized to separate peptides from various sources followed by MS/MS analysis. Two-dimensional ultra-high performance liquid chromatography is a useful tool for proteome analysis, providing a greater peak capacity than 1-D LC. The most popular 2-D LC approach used today for proteomic research combines strong cation exchange and reversed-phase LC. We have evaluated an alternative mode for 2-D LC of peptides using 2-D RP-RP nano UPLC Q-TOF Mass Spectrometry, employing reversed-phase columns in both separation dimensions. As control experiments, we identified 129 proteins in 1-D LC and 322 proteins in 2-D LC from E. coli extract peptides. Furthermore, we applied this method to rat primary hepatocyte and a total of 170 proteins were identified from 1-D LC, and 527 proteins were identified from all 2-D LC system. The in-depth protein profiling established by this 2-D LC MS/MS from rat primary hepatocyte could be a very useful reference for future applications in regards to drug induced liver toxicity.