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

• Korean Journal of Food Science and Technology
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• v.53 no.6
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• pp.731-738
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• 2021

This study investigated the physicochemical properties of rice starch isolated from broken rice using alkaline steeping (AKL) and enzymatic digestion (ENZ) methods. Broken rice starch (BRS) by AKL and ENZ possessed crude protein contents (0.6-1.4%) acceptable to commercial products of native starch and belonged to an intermediate amylose rice starch. AKL-BRS and ENZ-BRS showed a typical A-type crystal packing arrangement with small variations in their relative crystallinity. ENZ-BRS exhibited higher gelatinization onset and peak temperatures, and a narrower gelatinization temperature range than AKL-BRS, indicating that annealing occurred in ENZ-BRS. Lower swelling power and solubility were generally observed in the ENZ-BRS. ENZ-BRS also showed slower viscosity development, higher peak and trough viscosities, and lower breakdown, final, and setback viscosities, compared to those in AKL-BRS. These results are ascribed to the annealing phenomenon in ENZ-BRS. Overall, BRS from cheap broken rice using AKL and ENZ could contribute to the expansion of rice starch utilization in food and non-food industries.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.37 no.1
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• pp.28-36
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• 1992

Fifty rice varieties were tested for alkali digestibility of milled rice grain at four different KOH levels, and twenty-four varieties selected were tested again for alkali digestibility at different degrading times and KOH levels. Gelatinization rate at several heating times and heating temperatures, and water absorption rate at 21$^{\circ}C$ and 77$^{\circ}C$ water temperatures were observed using rice samples of twenty-four varieties to clarify the relationship between alkali digestibility response, gelatinization rate and water absorption rate. Varietal difference of ADV in Japonica and Tongil tye rices was biggest at KOH 1.2%, but it was better to test at KOH 1.2% and 1.4% levels to know the exact alkali digestibility response of rice varieties. Rice varieties tested could be classified into three groups, low, intermediate and high, based on their alkali digestibility response at four KOH levels, and most of Korean cultivated rice varieties were belonged to intermediate or high ADV group. Varietal variation was also found in alkali degrading response at different soaking times in alkali solution. Low ADV varietal group showed higher gelatinization temperature and needed longer heating time for complete gelatinization compared with intermediate or high ADV group. Same trends was found between intermediate and high ADV groups, but varietal variation in the same ADV group was also found in gelatinization temperature and heating time needed for complete gelatinization of rice grain. Water absorption rate of low ADV group was lower than intermediate or high ADV group both at 21$^{\circ}C$ and 77$^{\circ}C$ water temperatures, and intermediate ADV group showed lower absorption rate than high ADV group only in initial water absorption stage at 21$^{\circ}C$.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.41 no.6
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• pp.427-434
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• 2008

Processing of collagen from yellowfin tuna (Thunnus albacares) abdominal skins was optimized by response surface methodology and central composite design. The values of independent variables at optimal conditions were NaOH concentration: 0.5 N, NaOH treatment time: 36.2 hr, pepsin concentration: 1:4.9 ratio (0.245%, w/v), and digestion time: 48.1 hr, respectively. The collagen content estimated under optimal conditions was 33.1%, and the actual experimental collagen content was 32.3%. Physicochemical properties of collagen from yellowfin tuna abdominal skin were investigated by amino acids analysis, SDS-PAGE, FT-IR, viscosity and denaturation temperature. Amino acids content of the collagen was 21.0%. SDS-PAGE pattern of the collagen showed two different $\alpha$-chain (${\alpha}_1$- and ${\alpha}_2$- chain), $\beta$-component and $\gamma$-component. The spectrum of FT-IR of the collagen showed wavenumber at 3,434, 1,650, 1,542 and $1,235\;cm^{-1}$ representing the regions of amide A, I, II and III, respectively. Relative viscosity of the collagen decreased continuously on heating up to $32^{\circ}C$, and the rate of decrease was retarded in the temperature range of $35-50^{\circ}C$. Denaturation temperature (Td) of the collagen solution (0.06%, w/v) was $31^{\circ}C$ and was lower than calf skin collagen ($35^{\circ}C$).

• Resources Recycling
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• v.24 no.2
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• pp.46-54
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• 2015

In this study, an applicability of thermal pretreatment for primary and secondary sludge, which are generated in a sewage treatment plant, was evaluated. The efficiency and charateristics was investigated with each sludge after pretreatment under the condition of $100{\sim}220^{\circ}C$ for 30 minutes. As the result, it was found that $SCOD_{Cr}$, $NH_4{^+}$, VFAs concentrations increased as the pre-treatment temperature increased. For COD solubilization, it was also highly dependent on an increase of temperature resulting in acceleration on hydrolysis and acid fermentation. In the BMP (Biochemical Methane Potential) experiment, for the primary sludge, it showed the higher biogas production rate at a temperature of $220^{\circ}C$, however, the effect was insignificant (5.6%). Whereas, for the secondary sludge, the increase on biogass production rate was 38.8% ($180^{\circ}C$) and this means that the secondary sludge is more suitable for an applicability of thermal pretreatment.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.45 no.3
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• pp.211-215
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• 2000

Lipid extraction preceding fatty acid methyl esters preparation for gas-liquid chromatography is time-consuming and cumbersome. We performed one-step extraction/methylation method with a mixture of methanol-heptane-benzene-DMP-H$_2$SO$_4$ without prior fat extraction. The simultaneous digestion and lipid transmethylation takes place at 8$0^{\circ}C$ in a single phase. After cooling till room temperature, two phases are formed. The upper one of the phases contains the fatty acid methyl esters ready for GLC. The fatty acid composition of major industrial crops obtained by the one step extraction/methylation method (method 1 and 2) was almost identical with the fatty acid composition of the pure fats extracted with hexane by the Soxtec instrument (method 3). Due to its simplicity, speed, and reduced organic solvent the one-step extraction/methylation method (method 1 and 2) should be useful to determine overall fatty acid composition, especially in situations where many samples have to be analyzed.

• Analytical Science and Technology
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• v.12 no.2
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• pp.130-135
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• 1999

A graphite furnace atomic absorption spectrometry (GFAAS) with matrix modification has been used to determine trace amounts of arsenic, lead and selenium in rice flour samples. A mixed solution of palladium and magnesium nitrate was used as a matrix modifier to convert the analyte elements into a phase of higher thermostability and to increase the volatility of concomitants in graphite furnace. Matrix modification effects by the mixed solution were investigated for several elements (As, Cd, Cu, Pb, Se, Zn). It has been found that the matrix modifier substantially increase the pyrolysis and atomization temperature, and absorbance for As, Pb and Se. The concentration of As, Pb and Se in rice flour samples were determined by standard addition method with Zeeman background correction after microwave acid digestion. In this method the characteristic concentrations of As, Pb and Se are 26 ng/g, 18 ng/g, 24 ng/g on the basis of dry sample respectively.

• Journal of the Korea Organic Resources Recycling Association
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• v.23 no.4
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• pp.11-20
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• 2015

In the present work, a basic study on the in-situ biogenic methane generation from low grade coal bed was conducted. Lignite from Indonesia was used as a sample feedstock. A series of BMP (Biochemical Methane Potential) tests were carried out under the different experimental conditions. Although nutrients and anaerobic digester sludge were added to the coal, the produced amount of methane was limited. Both temperature control and particle size reduction showed little effect on the increase of methane potential. When rice straw was added to lignite as an external carbon source, methane yield of 94.4~110.4 mL/g VS was obtained after 60 days of BMP test. The calorific value of lignite after BMP test decreased (4.5~12.1 %) as increasing the content of rice straw (12.5~50 wt % of lignite), implying that anaerobic digestion of rice straw led to partial degradation of lignite. Therefore, rice straw could be used as an external carbon source for the start-up of in-situ biogas generation from low grade coal bed.

• Microbiology and Biotechnology Letters
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• v.14 no.2
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• pp.181-186
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• 1986

A defective lambda transducing phase carrying acetyl CoA carboxylase gene (fabE) from Escherichia coli chromosome (72 min on the current linkage map) has been isolated. A restriction map of the chromosomal region from defective transducing phage was established by digestion with combination of the restriction enzymes. No cleavage site for the enzyme EcoRI was found in this region. Restriction fragments were cloned from defective transducing phage into high copy number plasmid vector pACYC184 to generate hybrid plasmids which were capable of complementation of fabE temperature sensitive mutation. We show here that the fabE gene is located on a 3.4 megadalton Bam HI-SalI fragment with a HindIII site, which lies within the 7.4 megadalton BglIIfragment, by complementation analysis.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.7
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• pp.951-957
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• 2015

Four rumen fistulated swamp buffaloes were randomly assigned according to a $4{\times}4$ Latin square design to investigate the effects of Eucalyptus (E. Camaldulensis) leaf meal (ELM) supplementation as a rumen enhancer on feed intake and rumen fermentation characteristics. The dietary treatments were as follows: T1 = 0 g ELM/hd/d; T2 = 40 g ELM/hd/d; T3 = 80 g ELM/hd/d; T4 = 120 g ELM/hd/d, respectively. Experimental animals were kept in individual pens and concentrate was offered at 0.3% BW while rice straw was fed ad libitum. The results revealed that voluntary feed intake and digestion coefficients of nutrients were similar among treatments. Ruminal pH, temperature and blood urea nitrogen concentrations were not affected by ELM supplementation; however, ELM supplementation resulted in lower concentration of ruminal ammonia nitrogen. Total volatile fatty acids, propionate concentration increased with the increasing level of EML (p<0.05) while the proportion of acetate was decreased (p<0.05). Methane production was linearly decreased (p<0.05) with the increasing level of ELM supplementation. Protozoa count and proteolytic bacteria population were reduced (p<0.05) while fungal zoospores and total viable bacteria, amylolytic, cellulolytic bacteria were unchanged. In addition, nitrogen utilization and microbial protein synthesis tended to increase by the dietary treatments. Based on the present findings, it is suggested that ELM could modify the rumen fermentation and is potentially used as a rumen enhancer in methane mitigation and rumen fermentation efficiency.