• Journal of Plant Biotechnology
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• v.45 no.4
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• pp.315-321
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• 2018

Molecular and functional characterization of proteins and their levels is of great interest in understanding the mechanism of diverse cellular processes. In this study, we report on the convenient Escherichia coli-based protein expression system that allows recombinant of soluble proteins expression and cytosolic domain of membrane-localised kinases, followed by the detection of autophosphorylation activity in protein kinases. This approach is applied to regulatory proteins of Arabidopsis thaliana, including 14-3-3, calmodulin, calcium-dependent protein kinase, TERMINAL FLOWER 1(TFL1), FLOWERING LOCUS T (FT), receptor-like cytoplasmic kinase and cytoplasmic domain of leucine-rich repeat-receptor like kinase proteins. Our Western blot analysis which uses phospho-specific antibodies showed that five putative LRR-RLKs and two putative RLCKs have autophosphorylation activity in vitro on threonine and/or tyrosine residue(s), suggesting their potential role in signal transduction pathways. Our findings were also discussed in the broader context of recombinant expression and biochemical analysis of soluble and membrane-localised receptor kinases in microbial systems.

• Korean Journal of Agricultural Science
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• v.16 no.2
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• pp.201-211
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• 1989

This experiment was carried out to study the practical utility of Mucor rennet in making Blue cheese and to investigate the changes in the level of various nitrogen compounds. 1. The Mucor rennet cheese, the calf rennet cheese and the mixed rennet cheese did not show any significant difference in their yields. 2. The dry matter contents of Blue cheese was increased during the ripening and the levels of Mucor rennet did not have any influence in these respect. 3. The water soluble nitrogen contents of Blue cheese increased during ripening. On 0 day of ripening the Mucor rennet cheese contained water soluble nitrogen than the calf rennet cheese. On 40 days of ripening the mixed cheese contained less water soluble nitrogen than the calf rennet cheese. 4. Non protein nitrogen, peptone amino nitrogen, water soluble protein nitrogen, proteose nitrogen and peptone nitrogen appeared to contain similar levels of water soluble nitrogen. 5. The results of electrophoresis indicated a greater degredation on as-casein and ${\beta}$-casein in the Blue cheese made with Mucor rennet than in those made with calf rennet. On 60 days of ripening the mixed cheese more casein than did the Mucor rennet cheese. 6. The free amino acid contents of the Mucor rennet cheese was higher than the calf rennet cheese at 60 days of ripening.

• Journal of Bio-Environment Control
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• v.10 no.4
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• pp.244-250
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• 2001

This study was conducted to determine the change in freezing tolerance of Chinese cabbage (Brassica campestris L. ssp. penkinensis). Four-week old seedlings grown in a greenhouse at $25^{\circ}C$$\pm$0.5 were transferred to 5, 10 or 15$^{\circ}C$ condition for acclimation for one day, and then transferred to a lethal temperature of -3$^{\circ}C$ condition. Optimal acclimation temperatures were 5$^{\circ}C$ and 1$0^{\circ}C$. Freezing tolerance of leaf tissues was investigated during a week-long cold stress. Development of freezing tolerance was shown by survival ratio, and amounts of total RNA, soluble protein and soluble sugar. LT$_{50}$ value increased from -3 to -4$^{\circ}C$ after 5 days acclimation at 5 or 1$0^{\circ}C$, and this was considered to be the first indication of enhanced freezing tolerance. Plants, cold acclimated for five days, reached to a survival ratio of 60%. This increase in freezing tolerance was found to be associated with the increased levels of total RNA, soluble sugar and soluble protein. These metabolic changes imply the association of adjustment of growth and cell metabolism with low temperatures at the beginning of cold acclimation in chinese cabbage.e.

• Bulletin of the Korean Chemical Society
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• v.31 no.12
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• pp.3830-3833
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• 2010

• Korean Journal of Food Science and Technology
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• v.20 no.3
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• pp.447-452
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• 1988

In order to study effect of freezing rates on the quality changes such as pH, TBA value, free fatty acids and protein extractability, cylindrical chopped beef logs with 10cm of diameter and 10cm of height were frozen at three freezing rates(0.97cm/hr, 2.05cm/hr, 3.71cm/hr)using air blast freezer. Physicochemical changes of frozen meat were investigated during forzen storage at $-20^{\circ}C$ for 16weeks. Results on pH change showed $0.1{\sim}0.2unit$ increase at the 16th week of the frozen storage and the change was smaller with the increasing freezing rates. Free fatty acids content and TBA value also were increased during forzen storage, but they were minimal at 3.71cm/hr freezing rate. Correlation coefficient between TBA value and free fatty acids content were highly significant(r=0.804). After 16weeks of storage, extractibilities of salt soluble protein were decreased by 17.7%, 6.1% and 1.6% at freezing rates of 0.97, 2.05 and 3.71cm/hr, respectively. On the other hand, extractabilities of water soluble protein were decreased by 26.0%, 21.2% and 18.5%, respectively. The effect of freezing rates on the protein extractability appeared to be greater in salt soluble protein than in water soluble protein, but freezing denaturation was more rapid in water soluble protein.

• Preventive Nutrition and Food Science
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• v.10 no.4
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• pp.378-381
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• 2005

Actin and myosin solutions and fresh ground pork were irradiated with the electron beam (e-beam) at a dose of 0, 1.5, 3.0, 5.0 and 10 kGy. The changes in SDS-PAGE pattern of 2 proteins and the salt-soluble proteins extracted from ground pork after e-beam irradiation were monitored. When the myosin solution was irradiated with e-beam, myosin was degraded completely. Complete myosin degradations were observed even with the lowest dose (1.5 kGy) of e-beam treatment. Actin was degraded with the irradiation, but to a less extent than myosin was. The degradation of actin increased as the e-beam treatment increased from 1.5 to 10.0 kGy. Among the salt-soluble proteins extracted from ground pork, myosin was degraded gradually when the e-beam dose increased from 1.5 up to 10.0 kGy. Similar gradual increase in the degradation of actin also occurred with the increase of irradiation. Increases of 2 low molecular weight compounds (<29 kDa) were observed when the irradiation dose increased from 1.5 to 10.0 kGy. These 2 molecules are thought to be the breakdown products produced from the degradation of major salt-soluble proteins, myosin and actin. The salt-soluble protein content of ground pork did not change with the e-beam irradiation.

• Journal of Applied Biological Chemistry
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• v.51 no.3
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• pp.88-94
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• 2008

Three different protein extraction methods-phenol extraction, trichloroacetic acid (TCA) precipitation, and desalting/TCA precipitation-were compared to determine the optimal reproducible high resolution 2-dimensional (2-D) electrophoresis for each chungkugjang, doenjang, and kochujang samples. The soluble proteins from Chungkugjang extracted by phenol were separated with high reproducibility and resolution, and gained 1.75- to 3-fold more protein spots on 2-D gel than those from the other methods. On the contrary, the extracted proteins from doenjang and kochujang treated by desalting/TCA precipitation method showed about 1.5- to 3.3-fold more protein spots on 2-D gel. Using the established methods, the changes in the protein profiles of the fermented soy foods were monitored during the fermentation period by 2-DE. One of the major proteins in soy, $\beta$-conglycinin $\alpha$-subuint, and some proteins with unknown functions were localized on 2-D gel as the protease-resistant proteins throughout the fermentation period of doenjang. Changes in the protein profile monitored by the established methods can provide basic information on unfolding the mechanisms of the generation of biofunctional activity in the fermented soy foods.

• Journal of the Korean Society of Tobacco Science
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• v.17 no.1
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• pp.20-26
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• 1995

This study was carried out to obtain the basic data which include the change of the photosynthetic rate and protein content according to growth stage in the process of senescence of tobacco plant The photosynthetic rate was the maximum with 26.31$\mu$mol.CO2/m2.sec and stomatal resistance was the minimum with 0.2552cm/sec at 15th days after leaf emergence. However, after 50 days the photosynthesis was very little occurred. During leaf developments the number of chloroplast was increased and reached at the maximum at 25th days after emergence of leaf, thereafter, it was decreased gradually. The content of protein increased continuously and showed the highest value at 15th days after leaf emergence. The degradation rate of soluble protein was more rapid than that of insoluble protein at early stage of senescence. The range of decrement in the insoluble protein was low at late stage of senescence. The content of Rubisco, the key enzyme of photoamthesis, corresponded to about 50% of soluble protein and reached to the maximum at 150 days after leaf emergence. As the senescence progressed, the content of large subunit(UV) of Rubisco showed a tendency to decrease more rapidly than that of small subunit(SSU). The total amount of amino acids was the highest at 15th days after leaf emergence.

• International Journal of Industrial Entomology and Biomaterials
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• v.48 no.2
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• pp.86-97
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• 2024

Alkaline- or salt-assisted extractions have been widely used to extract edible insect proteins, however, there is a need for extraction techniques that balance cost-efficient production as well as preserving the protein properties. Mealworm proteins (Tenebrio molitor larvae) were extracted using three different extraction methods-alkali (AMP), salt (SMP), and water (WMP)-and then physicochemical and techno-functional properties were examined. AMP had high yield, protein, and amino acid contents, whereas WMP had high moisture, ash, and fat contents. SDS-PAGE showed a wide range of molecular weights in WMP whereas mostly low molecular weights were observed in AMP and SMP. AMP had poor protein solubilities compared to SMP and WMP across all pHs. AMP had enhanced water-holding capacity and emulsion stability, whereas WMP had improved oil-holding capacity and foaming properties. WMP formed a gel with and without the transglutaminase. The physicochemical and techno-functional properties demonstrated that water-soluble mealworm protein was superior to alkali-and salt-soluble mealworm proteins. Considering the cost efficiency and minimal impact on the environment as well, a cold press juicer could be utilized for mass production of mealworm protein compared to the conventional methods of protein extraction using alkali and salt.

• Korean Chemical Engineering Research
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• v.47 no.5
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• pp.624-629
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• 2009

It is the most important step to screen soluble and insoluble proteins when we attempt to improve the solubility of recombinant proteins through directed evolution approach. Here we show that the solubility of a recombinant protein in vivo can be examined by expressing the recombinant protein with beta-lactamase as a fusion partner. First we constructed an expression system which can produc a fusion protein with the C-terminal of beta-lactamase. Two soluble proteins, i.e. adenine deaminase and aspartate aminotransferase, and insoluble GlcNAc-2-epimerase were cloned into the developed expression vector, respectively. We investigated the effect of the expression of the three recombinant fusion proteins on the growth of E. coli, and confirmed that the solubilities of the recombinant proteins correlated with cell growth rates.