• Journal of Microbiology and Biotechnology
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• v.16 no.8
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• pp.1285-1291
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• 2006

The proline utilization (put) operon of Vibrio vulnificus consists of the putAP genes encoding a proline dehydrogenase and proline permease. The result of put-lux transcriptional fusion analysis suggests that the V vulnificus putAP operon is not autoregulated by the PutA protein. A putR null mutation decreased proline dehydrogenase activity and the level of the put transcripts, indicating that transcription of putAP is under the positive control of PutR. The deduced amino acid sequence of the putR was similar to those reported from other bacteria with high levels of identity. Chromatin IP and GST pull-down assays revealed that PutR specifically binds to the putAP promoter region in vivo, and interacts with CRP in vitro. Taken together, the results suggested that PutR exerts its effect on putAP expression by directly interacting with CRP bound to the upstream region of P$_{put}$.

• Journal of Environmental Science International
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• v.11 no.11
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• pp.1165-1172
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• 2002

The effects of salicylic acid(SA) and water deficit on growth and proline accumulation were investigated in cucumber(Cucurmis sativus L.) seedlings. Exogenous application of SA(100 $\mu$M-1 mM) led to a noticeable decrease in root and shoot growth, and dry weight of seedlings. Anatomical observation on leaf of cucumber revealed that the thickness of all leaf tissue components decreased in SA-treated plants. The effect was most pronounced on the width of the adaxial epidermis. In the separate effects of SA(0, 100, 500 and 1000 $\mu$M) and water deficit induced by PEG(0, 4.4, 7.0 and 9.6 %) on growth, the water deficit treatments had greater effects on growth traits than SA. Combinations of SA and PEG(SA+PEG) decreased shoot and root dry matter, and root length. Proline increased slightly in SA-treated seedlings, but exhibited a marked increase in water deficit application. Combinations of SA+PEG induced higher proline in both shoots and roots than SA stress alone. Shoots had higher proline than roots. Our data support a role of SA potentiating the osmotic stress response of germinating cucumber seedling.

• Journal of the Korean Chemical Society
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• v.34 no.1
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• pp.76-84
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• 1990

Optical isomers of dansyl amino acids were separated by a chiral mobile phase addition method. Two metha and para isomers of xylenyl-L-proline were prepared and used as the ligands of copper(Ⅱ) chelate to resolve the dansyl amino acids. Their elution behaviors were similar to those obtained from the addition of copper (Ⅱ) benzyl-L-proline chelate. The matrix effect of the mobile phase such as pH, concentration of buffer and compositions of organic solvent acetonitrile affected the optical resolution. The separation mechanism could be explained by a cis-trans effect of the ligand exchange reaction and hydrophobic interaction between the ternary complex and the stationary phase.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.14
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• pp.165-172
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• 1973

The sterile phenomenon is frequently found in the inter-species hybrids of ginseng as in other plants. It is known that among the hybrids between Panax Ginseng (PG) and Panax Quinquefolium (PQ), and between Panax Ginseng and Paxax Japonicus (PI), PG${\times}$PI is fertile only very rarely, while PG ${\times}$ PQ is always sterile. Therefore, in order to clarify the relationship between this sterility phenomenon and the metabolism of free amino acids, the changes of free amino acids through the formation of the flower organs and seeds of two hybrids, PG ${\times}$ PQ and PG ${\times}$ PI were investigated by thin layer chromatography. The results are summarized as follows: 1. Distinct differences in the quantity and number of free amino acids were recognized between PG ${\times}$ PQ, PG ${\times}$ PI and their parent plants. From the hybrid PG ${\times}$ PQ, 19 kinds of ninhyrin sensitive substances were detected in all. They were (1) 17 amino acids: alanine, valine, leucine, phenylalanine, proline, hydroxy-proline, serine, threonine, tyrosine, aspartic acid, glutamic acid, lysine, arginine, ${\gamma}$-amino butyric acid, ${\beta}$-alanine, cysteic acid and tryptophan, and (2) two amides: asparagine and glutamine. From the hybrid PG ${\times}$ PI, in addition to the above 19 substances, methionine and one unknown substance were detected. 2. Generally, alanine, as partie acid, glutamic acid, cysteic acid and asparagine were detected in large amounts in the two hybrids as in PG, PG and PJ but it was a noticeable fact concerning these two hybrids that the largest quantity of asparagine was found at microspore satge and pollen mature stage. 3. The decrease of cysteic acid in the two hybrids at the red ripened stage was the same as in PQ and PJ but opposite to the change in PG. The detection of methionine in PG ${\times}$ PJ was worthy of notice. 4. The change of proline was conspicuously different from that in their parent plants. It was detected as a trace of color at the micros pore stage while asparagine was detected in the greatest amount at that time. It is well known that the quantity of proline is closely related to the sterility of plant. This fact was also found true in the formation of ginseng seeds. It was reported as well that asparagine accumulated when proline decreased. 5. The deficiency of proline seemed to be closely related with the sterility of hybrids and with the degradation of pollen in anther. 6. The difference in the changes of free amino acids between the selfed lines of PG, PQ and PJ, and their hybrids seemed to be caused by the transformation of gene-action system by hybridization. On these phenomena along with proline metabolim and its physiological role in seed formation further studies are required.

• Analytical Science and Technology
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• v.9 no.2
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• pp.134-138
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• 1996

Cadmium was transported through the special carrier system into the cells of cotyledons, hypocotyls, and roots spring radish young plants. The transport of cadmium was inhibited by metabolic inhibitor, like DNP. The $K_m$ values for cadmium were 0.7 ppm for cotyledons, 1.72 ppm for hypocotyls, and 0.3 ppm for roots, and $V_{max}$ for cadmium was $40ppm{\cdot}h^{-1}{\cdot}g{\cdot}fresh\;weight^{-1}$ for cotyledons, $313ppm{\cdot}h^{-1}{\cdot}g{\cdot}fresh\;weight^{-1}$ for hypocotyls, and $606ppm{\cdot}h^{-1}{\cdot}g{\cdot}fresh\;weight^{-1}$ for roots. Cadmium cannot prove to be inducer for proline accumulation. Therefore, proline accumulation cannot be used as a marker to test the level of heavy metal pollution.

• Plant Biotechnology Reports
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• v.4 no.1
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• pp.37-48
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• 2010

[ ${\Delta}^1$ ]pyrroline-5-carboxylate synthetase (P5CS) is a proline biosynthetic pathway enzyme and is known for conferring enhanced salt and drought stress in transgenics carrying this gene in a variety of plant species; however, the wild-type P5CS is subjected to feedback control. Therefore, in the present study, we used a mutagenized version of this osmoregulatory gene-P5CSF129A, which is not subjected to feedback control, for producing transgenic indica rice plants of cultivar Karjat-3 via Agrobacterium tumefaciens. We have used two types of explants for this purpose, namely mature embryo-derived callus and shoot apices. Various parameters for transformation were optimized including antibiotic concentration for selection, duration of cocultivation, addition of phenolic compound, and bacterial culture density. The resultant primary transgenic plants showed more enhanced proline accumulation than their non-transformed counterparts. This proline level was particularly enhanced in the transgenic plants of next generation ($T_1$) under 150 mM NaCl stress. The higher proline level shown by transgenic plants was associated with better biomass production and growth performance under salt stress and lower extent of lipid peroxidation, indicating that overproduction of proline may have a role in counteracting the negative effect of salt stress and higher maintenance of cellular integrity and basic physiological processes under stress.

• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.165-171
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• 2002

Follicular fluid (FF) contains an oocyte maturation inhibitor with unknown chemical properties. This study was carried out to chemically define the factor(s) inhibiting cumulus cell denudation (CD) and first polar body formation (PBF). Porcine FF (PFF) was extracted with methanol and the extract was serially separated using gel filtration on Superose 12 and Superdex columns. A Superdex fraction was derived with PITC and analyzed with an amino acid analysis column. The results obtained are as follows; PFF had an activity inhibiting both CD and PBF of porcine primary oocytes. Superdex fractions RV2.11 prepared from PFF exhibited an activity inhibiting CD and PBF. By amino acid analysis, the fraction RV2.11 appeared to be proline having the same activity inhibiting CD and PBF. In conclusion, PFF had oocyte maturation inhibitors, of which proline should inhibit CD and PBF.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.27 no.1
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• pp.60-65
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• 1982

In order to observe the degree and response of drought-resistance and its physiological mechanism in barley and wheat seedling stage, 5 species (16 cultivars) were tested for the changes of nitrate reductase and protease activity and the accumulation of free proline, by being subjected to water stress by withholding watering for 8 days at 10 days (at the 3rd leaf stage) after emergence and by imposing water stress to the excised first leaf by polyethyleneglycol solution (osmotic potential, -20 bars) for 48 hours. The average rate of decrease of all cultivars was 42% in nitrate reductase activity and 73% in protease activity. But proline content in water stress was increased 10 folds more than that of control. The decrease4 rate of nitrate reductase activity in 5 species was in the order of wheat < rye < covered barley < naked barley < two-row barley: wheat being the lowest. The decreased rate of protease activity in 5 species was in the order of wheat > rye > two-row barley > covered barley > naked barley: wheat being the" heighest. The accumulated amount of free proline in 5 species by water stress was in the order of wheat > covered barley > rye > naked barley > two-row barley. And the increased ratio (folds) of free proline of water stress to control was in the order of rye(13) > wheat. covered barley(11) > naked barley(99) > two-row barley(7): rye being the highest. In terms of the enzymatic activity and the physiotically adaptive metabolism during the processing leading to drought-resistance, the degree of drought-resistance of 5 species to water stress at seedling stage was shown to be in the order of wheat > rye > covered barley > naked barley > two-row barley.

• Journal of Microbiology and Biotechnology
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• v.12 no.2
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• pp.318-326
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• 2002

The pathogenic marine bacterium Vibrio vulnificus is the causative agent of food-borne diseases such as life-threatening septicemia. To better understand this organism's strategies to survive osmotic stress, a mutant that was more sensitive to high osmolarity was screened from a library of mutants constructed by a random transposon mutagenesis. By a transposon-tagging method, putAP genes encoding a proline dehydrogenase and a proline permease were identified and cloned from V. vulnificus. The amino acid sequences deduced from nucleotide sequences of putAP from V. vulnificus were 38 to $59\%$ similar to those of PutA and PutP reported from other Enterobacteriaceae. Functions of putAP genes were assessed by the construction of mutants, whose putAP genes were inactivated by allelic exchanges. When proline as the sole carbon or nitrogen source was used, the putA mutant was not able to grow to the substantial level, revealing the proline dehydrogenase is the only enzyme for metabolic conversion of proline into other amino acids. Although the growth rate of the putP mutant on proline as the sole carbon or nitrogen source was significantly reduced, the mutant still grew. This indicated that at least one more proline permease is produced by V. vulnificus. The putP mutant decreased approximately $2-log_10$ CFU/ml after a hyperosmotic challenge, while the parent strain decreased approximately $l-log_10$ CFU/ml. This result suggests that the gene product of putP contributes to the osmotic tolerance of V. vulnificus.

• Applied Biological Chemistry
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• v.38 no.3
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• pp.218-223
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• 1995

Osmolytes accumulated in the osmotically stressed Yersinia enterocolitica ATCC 9610 were investigated using natural abundance $^{13}C$ NMR spectroscopy. Glycine betaine, one of the more common and most effective osmolytes found in nature, was the dominant osmolyte in osmotically stressed Y. enterocolitica cells. Glycine betaine concentration was 41.8 times higher (801.9 nmol/mg protein) in stressed cells than in unstressed cells (19.2 nmol/mg protein). Proline was the minor osmolyte, and its concentration was 284.8 nmol/mg protein. The effects of glycine betaine and related osmolytes on growth rate of osmotically stressed Y. enterocolitica were investigated to identify their ability as osmolytes for Y. enterocolitica. When glycine betaine and proline were added in MMA medium containing 2.5% NaCl, the growth rate with glycine betaine (1 mM) was 3.6 times higher than in control (no addition of osmolyte), and that with proline was 1.3 times higher. Dimethylglycine (5 mM) also increased the growth rate 3.1 folds. On the other hand, monomethylElycine had no effect on growth of osmotically stressed and unstressed Y. enterocolitica. When carnitine was added in MMA medium containing 2.5% NaCl, carnitine (5 mM) increased the growth rate 2.4 folds, but choline had no effect on growth of osmotically stressed Y. enterocolitica. The above results indicate that glycine betaine is the dominant osmolyte in osmotically stressed Y. enterocolitica, and proline, dimethylglycine and carnitine also act as minor osmolytes.