• Journal of Life Science
• /
• v.19 no.9
• /
• pp.1226-1231
• /
• 2009

Enzyme substitution with recombinant phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) is currently being explored for treatment of phenylketonuria (PKU), an autosomal recessive genetic disorder with mutations of the gene encoding phenylalanine-4-hydroxylase (EC 1.14.16.1). However, oral administration of PAL is limited because of proteolytic digestion in the gastrointestinal tract. The aim of this study was to determine the biochemical properties of PAL and delinate the susceptibility of wild-type PAL to pancreatic proteolysis by exploring several mutants, and to develop therapeutic drugs with PAL for PKU. The specific activity of PAL was assayed and its optimal pH, temperature stability, and intestinal protease susceptibility were investigated. Its $V_{max}$ values for phenylalanine and tyrosine were 1.77 and $0.47{\mu}mol$/ min/mg protein, respectively, and its $K_m$ values were $4.77{\times}10^{-4}$ and $4.37{\times}10^{-4}\;M$, respectively. PAL showed an optimal pH at 8.5, corresponding to the average pH range of the small intestine. It showed no loss of activity at $-80^{\circ}C$ for 5 months and possessed 93.4% of its activity under $4^{\circ}C$ for 4 wks. PAL was susceptible to chymotrypsin digestion and, to a lesser extent, to trypsin, elastase, carboxypeptidase A, and B. The trypsin and chymotrypsin cleaving sites were mutated to investigate protection from pancreatic digestion and the specific activities of these mutants were evaluated. The six mutants displayed low specific activities compared to the wild-type, suggesting that the primary trypsin and chymotrypsin cleaving sites may be essential for catalytic reaction. The PAL mutants could therefore be applied as a pretreatment modality without susceptibility to proteolytic attack, however, additional modification for enhancing enzymatic activity is needed to reduce the Phe levels effectively.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2003.04a
• /
• pp.61-62
• /
• 2003

Clonal propagation of high-value forest trees through somatic embryogenesis (SE) has the potential to rapidly capture the benefits of breeding or genetic engineering programs and to improve raw material uniformity and quality. A major barrier to the commercialization of this technology is the low quality of the resulting embryos. Several factors limit commercialization of SE for Corsican pine, including low initiation rates, low culture survival, culture decline causing low or no embryo production, and inability of somatic embryos to fully mature, resulting in low germination and reduced vigour of somatic seedlings. The objective was to develop a Corsican pine maturation medium that would produce cotyledonary embryos capable of germination. Treatments were arranged in a completely randomized design. Data were analyzed by analysis of variance, and significant differences between treatments determined by multiple range test at P=0.05. Corsican pine (Pinus nigra var. maritima) cultures were initiated on modified !P6 medium. Modifications of the same media were used for culture multiplication and maintenance. Embryogenic cultures were maintained on the same medium semi solidified with 2.5 g/l Gelrite. A maturation medium, capable of promoting the development of Corsican pine somatic embryos that can germinate, is a combination of iP6 modified salts, 2% maltose, 13% polyethylene glycol (PEG), 5 mg!l abscisic acid (ABA), and 2.5 g/l Gelrite. After initiation and once enough tissue developed they were grown in liquid medium. Embryogenic cell suspensions were established by adding 0.951.05 g of 10- to 14-day-old semisolid-grown embryogenic tissue to 9 ml of liquid maintenance media in a 250ml Erlenmeyer flask. Cultures were then incubated in the dark at 2022$^{\circ}$C and rotated at 120 rpm. After 2.53 months on maturation medium, somatic embryos were selected that exhibited normal embryo shape. Ten embryos were placed horizontally on 20 ml of either germination medium ($\frac{2}{1}$strength Murashige and Skoog (1962) salts with 2.5 g/l activated charcoal) or same medium with copper sulphate adjusted to 0.25 mg/1 to compensate for copper adsorption by activated carbon. 2% and 4% maltose was substituted by 7.5% and 13% PEG respectively to improve the yield of the embryos. Substitution of' maltose with PEG was clearly beneficial to embryo development. When 2% of the maltose was replaced with 7.5% PEG, many embryos developed to large bullet-shaped embryos. At latter stages of development most embryos callused and stopped development. A few short, barrel-shaped cotyledonary embryos formed that were covered by callus on the sides and base. When 4% of the maltose was removed and substituted with 13% PEG, the embryos developed further, emerging from the callus and increasing yield slightly. Microscopic examination of the cultures showed differing morphologies, varying from mostly single cells or clumps to well-formed somatic embryos that resembled early zygotic embryos only liquid cultures with organized early-stag. A procedure for converting and acclimating germinants to growth in soil and greenhouse conditions is also tested. Seedling conversion and growth were highly related to the quality of the germinant at the time of planting. Germinants with larger shoots, longer, straighter hypocotyls and longer roots performed best. When mature zygotic embryos germinate the root emerges, before or coincident with the shoot. In contrast, somatic embryos germinate in reverse sequence, with the cotyledons greening first, then shoot emergence and then, much later, if at all, the appearance of the root. Somatic seedlings, produced from the maturation medium, showed 100% survival when planted in a field setting. Somatic seedlings showed normal yearly growth relative to standard seedlings from natural seed.

• Applied Biological Chemistry
• /
• v.16 no.1
• /
• pp.1-11
• /
• 1973

In order to obtain the basic informations on the production of single cell protein from ethanol, 145 yeast strains utilizing ethanol as a sole carbon source were isolated from 32 soil samples in Korea. A yeast strain showing the highest cell yield among the isolated strains was selected and identified. The optimum culture condition, utilization of other carbon sources and the cultural characteristics for the selected yeast, and the chemical analysis of the yeast cell composition, and utilization of ethanol by the selected yeast were investigated. All the culture was carried out in the shaking flasks. The results obtained were as follows: 1. The selected yeast strain was identified as Debaryomyces nicotianae-SNU 72. 2. The optimum composition of the medium for the selected yeast is : Ethanol 40 ml, Urea 0.5 g, Potassium phosphate (dibasic) 0.5 g, Ammoium phosphate (monobasic) 0.15 g, Magnesium sulfate 0.05 g, Calcium chloride 0.01g, Yeast extract 0.005 g, Tap water 1000 ml. 3. The optimum pH was 5.0-5.5, the optimum temperature $30-33^{\circ}C$ and the aerobic state was unimportant. 4. Utilization of methanol, n-propanol, iso-propanol, n-butanol, iso-butanol, tert-amyl alcohol and acetic acid by the selected yeast was very weak. So substitution of the subtrate was thought to be impossible. 5. Studies on the propagation of the yeast cells showed that the lag phase of the yeast cells lasted 16 hours, and the logarithmic growth phase extended 16 to 28 hours. The specific growth rate was about $0.19\;hr^{-1}$ and the doubling time was 3.6 hours during the logarithmic growth phase. 6. As the result of the chemical analysis of the dry yeast cells, the content rate of the crude protein was 55.19 %, the content of others was similar to the average content of the yeast component. 7. After 34 hours cultivation, under the optimum culture condition investigated, the dry cell yield against the amount of the added ethanol was 53.4 % (W/V%), the dry cell yield against the amount of the utilized ethanol was 73.6 % (W/V%), the evaporation rate of ethanol was about 19.1 %.

• Korean Journal of Soil Science and Fertilizer
• /
• v.33 no.2
• /
• pp.100-108
• /
• 2000

High sodium contents in food-waste compost(FWC) is the greatest limitation to recycle it to arable lands in Korea. The effects of the FWC application to paddy soil on the growth of rice plants, cationic balance in plants, and the sodicity of soil have been studied in pot trials. The effects of FWC application were compared with those of NaCl compound and swine manure compost(SMC) application. $Na_2O$ contents of FWC were high as 2.2%. Immediately after transplanting, rice plants in three treatments showed severe wilting in the order of 40Mg FWC $ha^{-1}$ > NPK+900kg $NaClha^{-1}$ > 20Mg FWC $ha^{-1}$. The high EC value and volatile acid contents of soil solution were regarded as the cause of severe wilting of young rice plants. Increase of NaCl application rate upto $900kgha^{-1}$ showed no significant reduction of dry matter yield at harvesting stage. Regardless of application rates FWC reduced the dry matter yield at harvesting stage, while SMC increased it with increase of application rates upto $40Mgha^{-1}$. In NPK+NaCl and FWC treatments, Na contents and equivalent ratio in plants increased linearly with increase of Na application rates. Between Na and K equivalent ratio negative correlation with high significance was shown. In contrast to much difference of Na, K, and Na/K equivalent ratio among treatments, little difference of Na+K indicated the physiological substitution of Na for K in rice plants. Na use efficiency in NPK+NaCl and FWC treatments showed 12-22%.

• Journal of the Mineralogical Society of Korea
• /
• v.19 no.1 s.47
• /
• pp.15-29
• /
• 2006

Synthetic Mn-tourmalines (tsilaisite) were obtained by hydrothermal synthesis under the condition of 2 Kbar, $375{\sim}700^{\circ}C$, and 50 day-run-time with complete substitution of Mg in dravite by Mn (Mn%=0, 25, 50, 75, and 100%). They are all 6 samples containing Mn-tourmaline with some amounts of albite, spessartine, rhodocrosite, phlogopite etc, showing different synthetic condition of temperature and Mn composition. Synthetic Mn-tourmalines are of site deficiency in X-site ($0.53{\sim}0.68$) more than that of natural ones (approx. $0.2{\sim}0.3$) and show Mn cations occupying Y-site less than expected with initial experiments, leading to failure in synthesis of end-member tsilaisite. Rietveld structural refinements reveal that $R_{wp}$ ($R_{p}/R_{exp}$) is in the range of 13.35 and 18.62%, $R_{B}$ and S (CofF) are $4.85{\sim}6.25%$ (S-18: 8.57%), $1.31{\sim}1.59$ (S-18: 1.81), respectively. Unit cell parameters (space group R3m, z=3) are ${\alpha}=15.8994\;{\AA}$ and $c=7.1846\;{\AA}$ in average (S-18: ${\alpha}=15.9491\;{\AA},\;c=7.1773\;{\AA}$). Average bond lengths of and are $2.67{\sim}2.69\;{\AA}$ (S-18: $2.65\;{\AA}$) and $2.00{\sim}2.02\;{\AA}$ (S-18: $1.96\;{\AA}$), respectively. Ditrigonality (${\delta}$) are in the range of 0.022 and 0.031 (S-18: 0.061), indicating degrading symmetry with increase of Mn content.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.43 no.4
• /
• pp.522-529
• /
• 2014

Sea tangle (Laminaria japonica) is well known as having anti-diabetic and hypolipidemic effects in animals as well as in humans. In this study, we evaluated the effects of sea tangle-added patty on postprandial blood glucose and lipid profiles in borderline-hypercholesterolemic (cholesterol ${\geq}200$ mg/dL) adults. Eleven subjects voluntarily participated in the experiment, and each subject provided written consent. Experimental patty (E) was made by adding 2.25 g of sea tangle powder as a substitution to 1.125 g each of pork and chicken. In the first week, 200 g of Control patty (C) was provided to each subject, who had fasted more than 12 hours. In the second week, the same amount of E patty was supplied under the same conditions. Serum glucose levels increased significantly less at 30, 60, and 120 min after consumption of E patty compared to the levels at all time points after eating C patty. Thus, the change in the area under curve (${\Delta}AUC$) of serum glucose levels through 120 minutes was lower when consuming E patty compared to C patty. Although serum C-peptide concentrations were not significantly different at all time points between the two patties, the ${\Delta}AUC$ of serum C-peptide concentrations through 120 minutes was lower when consuming E patty compared to C patty. However, there were no differences in serum levels of triglyceride, total cholesterol, LDL-C, and HDL-C at 30, 60, 120, 180, and 240 min between the two patties. Further, each ${\Delta}AUC$ of these lipid levels through 240 minutes was not significantly different between the two patties. The results indicate that sea tangle-added patty may decrease postprandial plasma glucose concentrations and reduce insulin secretion, although it might not ameliorate serum lipid profiles in adults with borderline-hypercholesterolemia.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.33 no.4
• /
• pp.723-729
• /
• 2004

To develop foods using dried persimmon, hot-water extracts of dried persimmons were added into strong wheat flour in the ratio of 10, 20, 30, and 40% and the quality characteristics of baked products were investigated. The proximate compositions of dried persimmons hot-water extracts were 70.37% of moisture, 1.72% of crude protein, 0.18% of crude lipids, 1.99% of crude ash and 4.37% of crude fiber, respectively. With increasing the addition of dried persimmons hot-water extracts, moisture content of the added breads was increased to 47.20% from 41.12% of non-added bread and water activity to 0.576 from 0.495. Water binding capacity of the added dried persimmons hot-water extracts rasied weight of the bread but decreased specific volume. In the hunter's color values the more dried persimmons hot-water extracts was added, the lower L vaules we, and the higher a and b values we. In the texture property, the addition of dried persimmons hot-water extracts increased hardness, gumminess and chewiness, while decreased adhesiveness and fracturability. Judging from texture, taste and overall acceptability of the product, recommended substitution level for hot-water extracts of dried persimmons in bread was 30% or less.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.68 no.4
• /
• pp.422-430
• /
• 2023

The nutrient-rich and climate-resilient finger millet (Eleusine coracana (L.) Gaertn.) is a relatively new crop on the agricultural landscape. The present study explores the agronomic characteristics and antioxidant activities of grains and cookies produced from 'Finger1ho,' which was the first finger millet variety developed in South Korea. With heightened calcium content (314 mg/100 g) and polyphenol levels, 'Finger1ho' exhibited superior radical scavenging activities compared to other millets. The investigation assessed the impact of whole finger millet flour at varying concentrations (0, 10, 30, 50, and 100%) on cookie properties. Increasing the substitution of finger millet flour in the cookie formulation resulted in a notable rise in calcium content, ranging from 1.8 times at 10% to an impressive 10.8 times at 100%, surpassing the levels found in conventional wheat cookies. Conversely, the sodium (Na) levels in finger millet cookies demonstrated minimal variations, presenting a potentially favorable aspect in addressing the high Na intake prevalent in the South Korean diet. Notably, the antioxidant activity, assessed through ABTS and DPPH radical scavenging assays, exhibited a significant elevation compared to the control. This increase in antioxidant activity was directly proportional to the quantity of finger millet incorporated (p<0.001), indicating the potential health benefits associated with higher levels of finger millet in the cookie formulation. This study highlights finger millet's potential as a beneficial ingredient, enhancing both consumer acceptability and the functional attributes of cookies. Notably, cookies with 10% to 50% added finger millet exhibited significantly superior quality characteristics compared to controls, suggesting promising avenues for health-functional cookie development.