• Asian-Australasian Journal of Animal Sciences
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• v.20 no.10
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• pp.1517-1524
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• 2007

The [$^2H_5$]phenylalanine model was compared with the [1-$^{13}C$]leucine method to determine whole body protein synthesis (WBPS) and degradation (WBPD) in sheep fed at two levels. The animals were fed either 103 (M-diet) or 151 (H-diet) kcal $ME/kg^{0.75}/day$ once daily in a crossover design for 21 days each. The isotope dilutions were simultaneously conducted as a primed-continuous infusion of [$^2H_5$]phenylalanine, [$^2H_2$]tyrosine and [1-$^{13}C$]leucine on each dietary treatment. The WBPS and WBPD calculated from the [$^2H_5$]phenylalanine model were lower (p = 0.009 and p = 0.003, respectively) than those calculated from the [1-$^{13}C$]leucine method. The WBPS tended to be higher (p = 0.08) and WBPD was numerically higher (p = 0.33) for H-diet than M-diet in the [$^2H_5$]phenylalanine model, whereas the WBPS was numerically higher (p = 0.37) for H-diet and WBPS remained similar (p = 0.79) between diets in the [1-$^{13}C$]leucine method. However, the absolute values and the directions of WBPS as well as WBPD from M-diet to H-diet were comparable between the [$^2H_5$]phenylalanine model and [1-$^{13}C$]leucine method. Moreover, the values vary depending on the use of the respective amino acid contents in the carcass protein when calculating WBPS and WBPD. Therefore, it is concluded that the [$^2H_5$]phenylalanine model could be used as an alternative to the [1-$^{13}C$]leucine method for the determination of WBPS and WBPD in sheep.

• Journal of Microbiology and Biotechnology
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• v.25 no.6
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• pp.782-787
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• 2015

In this study, we developed an assay system for missense mutations in human phenylalanine hydroxylases (hPAHs). To demonstrate the reliability of the system, eight mutant proteins (F39L, K42I, L48S, I65T, R252Q, L255V, S349L, and R408W) were expressed in a mutant strain (pah^-) of Dictyostelium discoideum Ax2 disrupted in the indigenous gene encoding PAH. The transformed pah - cells grown in FM minimal medium were measured for growth rate and PAH activity to reveal a positive correlation between them. The protein level of hPAH was also determined by western blotting to show the impact of each mutation on protein stability and catalytic activity. The result was highly compatible with the previous ones obtained from other expression systems, suggesting that Dictyostelium is a dependable alternative to other expression systems. Furthermore, we found that both the protein level and activity of S349L and R408W, which were impaired severely in protein stability, were rescued in HL5 nutrient medium. Although the responsible component(s) remains unidentified, this unexpected finding showed an important advantage of our expression system for studying unstable proteins. As an economic and stable cell-based expression system, our development will contribute to mass-screening of pharmacological chaperones for missense PAH mutations as well as to the in-depth characterization of individual mutations.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.53 no.3
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• pp.423-431
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• 2020

This study was conducted to investigate in vivo and in vitro digestibility in juvenile Atlantic Bluefin tuna Thunnus thynnus. In vivo digestibility was compared between four experimental diets to determine the optimum dietary inclusion level of an enzyme-treated sardine fish meal (EFM) and sardine fish meal (FM). The experimental diets were as follows; EFM75 (75% EFM), EFM60 (60% EFM and 15% FM), FM75 (75% FM) and SL (frozen sand lance) as a raw fish feed. Feces of Bluefin tuna (90.3 g) were collected both by siphoning from a 700 L cage and by dissection in 69 ton concrete rearing tanks. For the siphoning method, protein digestibility was higher in the tuna fed SL diet than that of other groups. The lowest protein digestibility was observed in FM75. For the dissection method, protein digestibility was higher in tuna fed EFM75 diet than that of other groups. The lowest protein digestibility was observed in the EFM60 group. In vitro digestibility was compared in six protein sources to find an alternative source of EFM for the tuna feed. The highest in vitro digestibility was observed in EFM (92%) followed by low temperature FM (72%), meat meal (65%), feather meal (60%), sardine fish meal (57%) and poultry by-product meal (55%).

• Molecules and Cells
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• v.38 no.2
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• pp.180-186
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• 2015

Escherichia coli AcrAB-TolC is a multidrug efflux pump that expels a wide range of toxic substrates. The dynamic nature of the binding or low affinity between the components has impeded elucidation of how the three components assemble in the functional state. Here, we created fusion proteins composed of AcrB, a transmembrane linker, and two copies of AcrA. The fusion protein exhibited acridine pumping activity, suggesting that the protein reflects the functional structure in vivo. To discern the assembling mode with TolC, the AcrBA fusion protein was incubated with TolC or a chimeric protein containing the TolC aperture tip region. Three-dimensional structures of the complex proteins were determined through transmission electron microscopy. The overall structure exemplifies the adaptor bridging model, wherein the funnel-like AcrA hexamer forms an intermeshing cogwheel interaction with the ${\alpha}$-barrel tip region of TolC, and a direct interaction between AcrB and TolC is not allowed. These observations provide a structural blueprint for understanding multidrug resistance in pathogenic Gram-negative bacteria.

• Journal of Dairy Science and Biotechnology
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• v.35 no.4
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• pp.249-254
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• 2017

The purpose of this study was to investigate the acid tolerance, bile acid tolerance, and fermentation activity of lactic acid bacteria isolated from Kimchi in the presence of hydrolysates of whey protein concentrate. Kimchi isolates DK109, DK119, DK121, DK128, DK211, DK212, and DK215, which were identified as Lactobacillus sp., and L. casei DK128 showed the highest acid and bile acid tolerance. To produce whey hydrolysates, enzymes were added to a 10% (w/v) whey protein concentrate (WPC) solution at 1:50 (w/v, protein). The viabilities of the DK strains were determined in the presence of low pH and bile salts. Then, yogurt was produced via fermentation with L. casei DK128, an isolate from Kimchi, in the presence of the following additives: CPP, WPC, and WPC hydrolysates (WPCH) generated by alcalase (A) or neutrase (N). The produced yogurts were subjected to various analyses, including viable cell counts (CFU/mL), pH, titratable activity, and sensory testing. After 8 h of fermentation, the pH and titratable activity values of all test samples were 4.2 and 0.9, respectively. The viable counts of LAB were $3.49{\times}10^8$, $5.72{\times}10^8$, $7.01{\times}10^8$, and $6.97{\times}10^8$, for the Control, CPP, A, and N samples, respectively. These results suggest that whey proteins have potential as dietary supplements in functional foods and that WPCH could be used in yogurt as a low-cost alternative to CPP.

• Journal of Animal Reproduction and Biotechnology
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• v.38 no.4
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• pp.225-235
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• 2023

Background: Largemouth bass (Micropterus salmoides) is introduced species that has caused aquatic ecology activity both in vitro and in vivo were investigated for the possibility of application of the bass extract as an alternative feed ingredient. Methods: The bass oil was extracted using a 1-L supercritical extractor, while the protein was extracted from 250 g of bass dry matter, which was dissolved in 1 mL of H₂O at 50℃. Both oil and protein extracts were evaluated antioxidant activities and the level of DPPH radical scavenging assay and nitric oxide (NO) production assay with lipopolysaccharide response. Oral administration of 6.6 µL/g bass protein and 5.38 µL/g bass oil conducted for investigating serological and physiological effect. Results: DPPH radical scavenging showed similar radical scavenging ability of 50 µM of ascorbic acid at 200 ㎍ of protein and 10% of oil treatment. NO concentration was diminished by the treatment of bass oil. Oral administration of both bass oil and proteins to mice showed that the body weight increase rate of the bass oil treated group was significantly reduced by 1.55 g compared to the other groups. The number of white blood cells (WBC) was increased by 4.52 k/µL in the bass protein-treated group and 4.44 k/µL in the bass oil-treated group compared to the control group. However, the serum IgG level did not show a significant difference between the bass extract-treated groups and the control group. Conclusions: These studies demonstrate that both bass oil and proteins extracted from the bass not only provide excellent effects of antioxidant and immune activity but can also be used as functional food supplements.

• Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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• 2001.06a
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• pp.1273-1273
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• 2001

The efficiency of the luminal fermentation process influences overall efficiency of luminal production, animal health and reproduction. Ruminant production systems have a significant impact on the global environment, as well. Animal wastes contribute to pollution of the environment as ammonia volatilized to the air and nitrate leached to ground water. Microbial protein synthesis in the rumen satisfies a large proportion of the protein requirements of animals. Quantifying the microbial synthesis is possible by using markers for lumen bacteria and protozoa such as nucleic acids, purine bases, some specific amino acids, or by isotopic $^{15}N,^{32}P,\;and\;^{35}S$ labelled feeds. All those methods require cannulated animals, they are time-consuming and some methods are very expensive as well. Many attempts have been made to find an alternative method for indirect measurement of microbial synthesis in intact animals. The present investigations aimed to assess possibilities of NIRS for prediction of purine nitrogen excretion and ruminal microbial nitrogen synthesis by NIR spectra of urine. Urine samples were collected from 12 growing sheep,6 of them male, and 6- female. The sheep were included in feeding experiment. The ration consisted of sorghum silage and protein supplements -70:30 on dry matter basis. The protein supplements were chosen to differ in protein degradability. The urine samples were collected daily in a vessel containing $60m{\ell}$ 10% sulphuric acid to reduce pH below 3 and diluted with tap water to 4 liters. Samples were stored in plastic bottles and frozen at $-20^{\circ}C$ until chemical and NIRS analysis. The urine samples were analyzed for purine derivates - allantoin, uric acid, xantine and hypoxantine content. Microbial nitrogen synthesis in the lumen was calculated according to Chen and Gomes, 1995. Transmittance urine spectra with sample thickness 1mm were obtained by NIR System 6500 spectrophotometer in the spectral range 1100-2500nm. The calibration was performed using ISI software and PLS regression, respectively. The following statistical results of NIRS calibration for prediction of purine derivatives and microbial protein synthesis were obtained.(Table Omitted). The result of estimation of purine nitrogen excretion and microbial protein synthesis by NIR spectra of urine showed accuracy, adequate for rapid evaluation of microbial protein synthesis for a large number of animals and different diets. The results indicate that the advantages of the NIRS technology can be extended into animal physiological studies. The fast and low cost NIRS analyses could be used with no significant loss of accuracy when microbial protein synthesis in the lumen and the microbial protein flow in the duodenum are to be assessed by NIRS.

• Korean Journal of Agricultural Science
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• v.43 no.5
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• pp.750-760
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• 2016

The use of antibiotics in Korean swine production has been changing to a restricted use of in-feed antibiotics. These antibiotics, which are also growth promoters, are powerful for disease control. Due to this issue, the swine industry is consistently looking for any kind of alternatives to antibiotics such as new feed ingredients, feed additives, feed formulation practices, or feeding methods to improve pig health and performance. In general, dietary factors provide bioavailable nutrients and/or affect physiological activity to modify the physiological condition, immune system, or microbial population of pigs to improve their performance and health. Thus, it is suggested that dietary factors may be important components in the growth and health management of pigs. Using an alternative grain feed such as rice, barley, and oats, low protein diets or low-high energy diets can be used as solutions to manage the effect of stress factors that cause growth and health problems at specific time points during the stages of pig production. Several studies support that these alternative feeds and dietary factors may improve pig growth and health by changes in intestinal conditions, immunity, or other physiological conditions compared with typical feed ingredients and diet management in pig production. Therefore, feed ingredients, low protein levels, and different energy contents in swine diets were reviewed to better understand how these dietary factors can contribute to improved pig performance and health under different physiological conditions.

• Journal of Plant Biotechnology
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• v.37 no.3
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• pp.262-268
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• 2010

Antibodies are powerful and versatile tools to play a critical role in the diagnosis and treatment of many diseases. Their application has been enhanced significantly with the advanced recombinant DNA and heterologonous expression technologies, allowing to produce immunotherapeutic proteins with improved biofunctional properties. However, with currently available technologies, mammalian cell-based therapeutic antibody production, as an alternative for production in humans and animals, is often not plentiful for passive immunotherapeutics in treatment of many diseases. Recently, plant expression systems for therapeutic antibodies have become well-established. Thus, plants have been considered to provide an attractive alternative production system for therapeutic antibodies, as plants have several advantages such as the lack of human pathogens, and low cost of upstream production and flexible scale-up of highly valuable recombinant glycoproteins. Recent advances in modification of posttranslational processing for human-like glycosylation in transgenic plants will make it possible that plant can become a suitable protein expression system over the animal cellbased current production system. This review will discuss recent advances in plant expression technology and issues for their application to therapeutic antibody production.

• Research in Plant Disease
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• v.17 no.2
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• pp.111-120
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• 2011

A bacterial artificial chromosome library of Pectobacterium carotovorum 35 was constructed to characterize the genome and to sequence its hrp region. The hrp cluster of P. carotovorum 35 consisted of 26 open reading frames in five operons. A promoter-based green fluorescent protein technology was used to identify the genes regulated by the alternative sigma factor, HrpL, in P. carotovorum 35. The majority of the selected clones contained the hrpJ operon promoter sequence, which harbors a hrp box, but no putative hrp boxes were detected within the promoter sequences of two other hrpL-regulated genes encoding for pectate lyase and large repetitive protein. Although the promoters of five other hrp operons also contained hrp boxes, their expression was not HrpL-dependent in the promoter-based selection in E. coli. However, transcriptional analysis showed that expression from all operons harboring hrp boxes, except for the hrpN operon, was reduced significantly in the hrpL mutant. The severity of soft-rot symptoms when the hrpL mutant was applied to the surface of tobacco leaves, mimicking natural infection, was greatly attenuated. These results indicate that the hrpL gene of P. carotovorum 35 may be involved in the development of soft-rot symptoms.