• Food Science of Animal Resources
• /
• v.44 no.3
• /
• pp.662-683
• /
• 2024

Thai-Native×Anglo-Nubian goat meat cooked by grilling (GR), sous vide (SV), and microwave (MW), was compared to fresh meat (Raw) in terms of flavor development. Non-volatile [i.e., free amino acids, nucleotide-related compounds, taste active values (TAVs) and umami equivalency, sugars, lipid oxidation, Maillard reaction products] and volatile compounds, were investigated. Notably, inosine monophosphate and Glu/Gln were the major compounds contributing to umami taste, as indicated by the highest TAVs in all samples. Raw had higher TAVs than cooked ones, indicating that heat-cooking removes these desirable flavor and taste compounds. This could be proportionally associated with the increase in aldehyde, ketone, and nitrogen-containing volatiles in all cooked samples. GR showed the highest thiobarbituric acid reactive substances (1.46 mg malonaldehyde/kg sample) and browning intensity (0.73), indicating the greatest lipid oxidation and Maillard reaction due to the higher temperature among all cooked samples (p<0.05). In contrast, SV and Raw exhibited similar profiles, indicating that low cooking temperatures preserved natural goat meat flavor, particularly the goaty odor. The principal component analysis biplot linked volatiles and non-volatiles dominant for each cooked sample to their unique flavor and taste. Therefore, these findings shed light on cooking method selection based on desirable flavor and preferences.

• Journal of Life Science
• /
• v.22 no.4
• /
• pp.499-507
• /
• 2012

Previous work has characterized myomodulin A (MMA, PMSMLRLamide) and myomodulin E (MME, GLQMLRLamide) purified from the central nervous systems of the sea hare, $Aplysia$ $Kurodai$, using the anterior byssus retractor muscle (ABRM) of the mussel, $Mytilus$ $edulis$. The amino acid sequences of MMA and MME were the same as those of the myomodulin family peptide found in other mollusks. In this study, we synthesized MME, its derivatives, and other neuropeptides to investigate the relationship between the structure and biological activity of MME. The primary structures of MME's derivatives, Des[$Gly^1$]-MME, Des[$Gly^1,Leu^2$]-MME, and Des[$Gly^1,Leu^2,Gln^3$]-MME, were LQMLRLamide, QMLRLamide, and MLRLamide, respectively. MMA and synthetic peptides were tested on ABRM in $M.$ $edulis$ as well as muscle preparations in $Achatina$ $fulica$. MME displayed an inhibitory effect on phasic contraction of the ABRM at $1{\times}10^{-9}$ M or higher. MME also had a relaxing effect on the catch-tension of AMRM at $1{\times}10^{-8}$ M. Both MMA and its analogs stimulated a contractile response on the crop and relaxed the catch-relaxing response on the penial retractor muscle of $A.$ $fulica$. These results suggest that MME and its analogs have modulatory effects on various muscles of mollusks. This study has also laid the groundwork for future neural and circuit modulation studies during animal behavioral changes.

• Korean Journal of Environmental Agriculture
• /
• v.12 no.2
• /
• pp.144-152
• /
• 1993

A bacterium which degrades efficiently synthetic detergents was isolated from the polluted waters, activated sludge of wastewater treatment plants or polluted soil. This bacterium showed considerably higher growth rate in the agar plate containing $2,000{\mu}g/ml$ of synthetic detergents than any other isolated strains, was identified as a Pseudomonas fluorescens or strains similar to it. The strain was named as a Pseudomonas fluorescens S1. Optimum pH and temperature for the growth of the Pseudomonas fluorescens S1 were pH 7.0 and $30^{\circ}C$, respectively. The strain was resistant to streptomycin and gentamycin, but sensitive to kanamycin. The strain was greatly resistant to zinc chloride, lead nitrate and copper sulfate, but unable to grow in the presence of relatively low concentrations of mercury chloride and silver nitrate. This strain utilized benzene, catechol, cyclohexane and xylene as a sole carbon source. The strain was well grown in the medium containing ABS 10,000${\mu}g$/ml. Degradation of ABS was 55% and 60% at 20${\mu}g$/ml and 100${\mu}g$/ml of ABS, respectively. Benzene ring was degraded 45% in 100${\mu}g$/ml of ABS. During the incubation of the strain in the medium containing ABS 100${\mu}g$/ml and COD 10,000${\mu}g$/ml for 4 days, degradation of ABS and COD were reduced to 40${\mu}g$/ml and 3,200${\mu}g$/ml, respectively. Total amino acid content of the Pseudomonas fluorescens S1 grown with 1,000${\mu}g$/ml of ABS was 115mg/g cell, whereas its content was decreased in the bacterium grown without synthetic detergent by 9.4%.

• Tuberculosis and Respiratory Diseases
• /
• v.59 no.3
• /
• pp.257-265
• /
• 2005

Background : Rifabutin (ansamycin) is a spiro-piperidyl rifamycin, which is highly active against Mycobacterium tuberculosis. It has been found that some clinical isolates of tubercle bacilli that are resistant to rifampicin are susceptible to rifabutin, with some patients with multi-drug resistant pulmonary tuberculosis having shown favorable clinical and bacteriological responses to the rifabutin. This study was conducted to find the proportion of rifabutin-susceptible strains among rifampicin-resistant isolates from Korean MDR-TB patients, and investigate the presence of specific rpoB mutations, which may confer resistance to rifampicin, but not to rifabutin. Methods : 201 rifampicin-resistant and 50 pan-susceptible M. tuberculosis isolates were randomly selected for this study. The isolates were retested at rifampicin and rifabutin concentrations of 0, 20, 40 and $80{\mu}g/ml$, respectively. The isolates that grew at and/or over a rifabutin concentration of $20{\mu}g/ml$ were judged rifabutin-resistant. The rpoB gene was extracted from the isolates, and then amplified for direct sequencing to investigate specific rpoB mutations that conferred rifabutin- susceptibility but rifampicin-resistance. Results : Out of the 201 rifampicin-resistant M. tuberculosis, 41 strains (20.4%) were susceptible to rifabutin using the absolute concentration method on Lowenstein-Jensen media. The rpoB mutation types that showed susceptibility to rifabutin were Leu511Pro, Ser512Arg, Gln513Glu, Asp516Ala, Asp516Gly, Asp516Val, Asp516Tyr, Ser522Leu, His526Asn, His526Leu, His526Cys, Arg529Pro and Leu533Pro. A reverse hybridization technique was able to detect 92.5% of the rifabutin-susceptible isolates, with a specificity of 96.1% among 195 M. tuberculosis isolates with the rpoB mutation. Conclusions : Around 20% of the rifampicin-resistant isolates in Korea showed susceptibility to rifabutin, which was associated with some specific mutations of rpoB. Rifabutin could be used for the treatment of MDR-TB patients, especially when drug susceptibility testing reveals susceptibility to rifabutin.

• Proceedings of the Korean Society of Crop Science Conference
• /
• 2017.06a
• /
• pp.36-36
• /
• 2017

Out of twenty common protein amino acids, there are many kinds of non protein amino acids (NPAAs) that exist as secondary metabolites and exert ecological functions in plants. Mimosine (Mim), one of those NPAAs derived from L. leucocephala acts as an iron chelator and reversely block mammalian cell cycle at G1/S phases. Cysteine (Cys) is decisive for protein and glutathione that acts as an indispensable sulfur grantor for methionine and many other sulfur-containing secondary products. Cys biosynthesis includes consecutive two steps using two enzymes-serine acetyl transferase (SAT) and O-acetylserine (thiol)lyase (OASTL) and appeared in plant cytosol, chloroplast, and mitochondria. In the first step, the acetylation of the ${\beta}$-hydroxyl of L-serine by acetyl-CoA in the existence of SAT and finally, OASTL triggers ${\alpha}$, ${\beta}$-elimination of acetate from OAS and bind $H_2S$ to catalyze the synthesis of Cys. Mimosine synthase, one of the isozymes of the OASTLs, is able to synthesize Mim with 3-hydroxy-4-pyridone (3H4P) instead of $H_2S$ for Cys in the last step. Thus, the aim of this study was to clone and characterize the cytosolic (Cy) OASTL gene from L. leucocephala, express the recombinant OASTL in Escherichia coli, purify it, do enzyme kinetic analysis, perform docking dynamics simulation analysis between the receptor and the ligands and compare its performance between Cys and Mim synthesis. Cy-OASTL was obtained through both directional degenerate primers corresponding to conserved amino acid region among plant Cys synthase family and the purified protein was 34.3KDa. After cleaving the GST-tag, Cy-OASTL was observed to form mimosine with 3H4P and OAS. The optimum Cys and Mim reaction pH and temperature were 7.5 and $40^{\circ}C$, and 8.0 and $35^{\circ}C$ respectively. Michaelis constant (Km) values of OAS from Cys were higher than the OAS from Mim. Inter fragment interaction energy (IFIE) of substrate OAS-Cy-OASTL complex model showed that Lys, Thr81, Thr77 and Gln150 demonstrated higher attraction force for Cys but 3H4P-mimosine synthase-OAS intermediate complex showed that Gly230, Tyr227, Ala231, Gly228 and Gly232 might provide higher attraction energy for the Mim. It may be concluded that Cy-OASTL demonstrates a dual role in biosynthesis both Cys and Mim and extending the knowledge on the biochemical regulatory mechanism of mimosine and cysteine.

• Reproductive and Developmental Biology
• /
• v.34 no.1
• /
• pp.33-40
• /
• 2010

Equine chorionic gonadotropin (eCG) is a heavily glycosylated glycoprotein composed of non-covalently linked $\alpha$- and $\beta$-subunits. To study the function and signal transduction of tethered recombinant-eCG (rec-eCG), a single chain eCG molecule was constructed, and the rec-eCG protein was prepared. In this study, we constructed 5 mutants (${\Delta}1$, ${\Delta}2$, ${\Delta}3$, ${\Delta}4$, and ${\Delta}5$) of rec-eCG using data about known glycoprotein hormones to analyze the role of specific follicle stimulating homone (FSH)-like activity. Three amino acids of certain specific sites were replaced with alanine. The expression vectors were transfected into CHO cells and subjected to G418 selection for 2~3 weeks. The media were collected and the quantity of secreted tethered rec-eCGs was quantified by ELISA. The LH- and FSH-like activities were assayed in terms of cAMP production by rat LH/CG and rat FSH receptors. Then, the metabolic clearance rate analyzed by the injection of rec-eCG (5 IU) into the tail vein was analyzed. The mutant eCGs (${\Delta}l$, ${\Delta}4$, and ${\Delta}5$) were transcripted, but not translated into proteins. Rec-eCG A2 was secreted in much lower amounts than the wild type. Only the rec-eCG ${\Delta}3$ ($\beta$-subunit: $Gln^{94}-Ile^{95}-Lys^{96}{\rightarrow}Ala^{94}-Ala^{95}-Ala^{96}$) was efficiently secreted. Although activity is low, its LH-like activity was similar to that of tethered $eCG{\beta\alpha}$. However, the FSH-like activity of rec-$eCG{\beta\alpha\Delta}3$ was completely flat. The result of the analysis of the metabolic clearance rate shoed the persistence of the mutant in the blood until 4 hours after the injection. After then, it almost disappeared at 8 hours. Taken together, these data suggest that 94~96 amino acid sequences in eCG $\beta$-subunit appear to be of utmost importance for signal transduction of the FSH receptor.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.2
• /
• pp.287-294
• /
• 2008

Three amino acid residues (His119, Glu164, and Glu338) in the active site of Thermus caldophilus GK24 ${\beta}$-glycosidase (Tca ${\beta}$-glycosidase), a family 1 glycosyl hydrolase, were mutated by site-directed mutagenesis. To verify the key catalytic residues, Glu164 and Glu338 were changed to Gly and Gln, respectively. The E164G mutation resulted in drastic reductions of both ${\beta}$-galactosidase and ${\beta}$-glucosidase activities, and the E338Q mutation caused complete loss of activity, confirming that the two residues are essential for the reaction process of glycosidic linkage hydrolysis. To investigate the role of His119 in substrate binding and enzyme activity, the residue was substituted with Gly. The H119G mutant showed 53-fold reduced activity on 5mM p-nitrophenyl ${\beta}$-D-galactopyranoside, when compared with the wild type; however, both the wild-type and mutant enzymes showed similar activity on 5mM p-nitrophenyl ${\beta}$-D-glucopyranoside at $75^{\circ}C$. Kinetic analysis with p-nitrophenyl ${\beta}$-D-galactopyranoside revealed that the $k_{cat}$ value of the H119G mutant was 76.3-fold lower than that of the wild type, but the $K_m$ of the mutant was 15.3-fold higher than that of the wild type owing to the much lower affinity of the mutant. Thus, the catalytic efficiency $(k_{cat}/K_m)$ of the mutant decreased to 0.08% to that of the wild type. The $k_{cat}$ value of the H119G mutant for p-nitrophenyl ${\beta}$-D-glucopyranoside was 5.l-fold higher than that of the wild type, but the catalytic efficiency of the mutant was 2.5% of that of the wild type. The H119G mutation gave rise to changes in optima pH (from 5.5-6.5 to 5.5) and temperature (from $90^{\circ}C\;to\;80-85^{\circ}C$). This difference of temperature optima originated in the decrease of H119G's thermostability. These results indicate that His119 is a crucial residue in ${\beta}$-galactosidase and ${\beta}$-glucosidase activities and also influences the enzyme's substrate binding affinity and thermostability.

• Asian Pacific Journal of Cancer Prevention
• /
• v.16 no.12
• /
• pp.4915-4920
• /
• 2015

Background: : Familial adenomatous polyposis (FAP) is an autosomal dominant inherited disease mainly caused by mutations of the adenomatous polyposis coli (APC) gene with almost complete penetrance. These colorectal polyps are precancerous lesions that will inevitable develop into colorectal cancer at the median age of 40-year old if total proctocolectomy is not performed. So identification of APC germline mutations has great implications for genetic counseling and management of FAP patients. In this study, we screened APC germline mutations in Chinese FAP patients, in order to find novel mutations and the APC gene germline mutation characteristics of Chinese FAP patients. Materials and Methods: The FAP patients were diagnosed by clinical manifestations, family histories, endoscope and biopsy. Then patients peripheral blood samples were collected, afterwards, genomic DNA was extracted. The mutation analysis of the APC gene was conducted by direct polymerase chain reaction (PCR) sequencing for micromutations and multiplex ligation-dependent probe amplification (MLPA) for large duplications and/or deletions. Results: We found 6 micromutations out of 14 FAP pedigrees, while there were no large duplications and/or deletions found. These germline mutations are c.5432C>T(p. Ser1811Leu), two c.3926_3930delAAAAG (p.Glu1309AspfsX4), c.3921_3924delAAAA (p.Ile1307MetfsX13), c3184_3187delCAAA(p.Gln1061AspfsX59) and c4127_4126delAT (p.Tyr1376LysfsX9), respectively, and all deletion mutations resulted in a premature stop codon. At the same time, we found c.3921_3924delAAAA and two c.3926_3930delAAAAG are located in AAAAG short tandem repeats, c3184_3187delCAAA is located in the CAAA interrupted direct repeats, and c4127_4128 del AT is located in the 5'-CCTGAACA-3', 3'-ACAAGTCC-5 palindromes (inverted repeats) of the APC gene. Furthermore, deletion mutations are mostly located at condon 1309. Conclusions: Though there were no novel mutations found as the pathogenic gene of FAP in this study, we found nucleotide sequence containing short tandem repeats and palindromes (inverted repeats), especially the 5 bp base deletion at codon 1309, are mutations in high incidence area in APC gene,.

• Journal of Microbiology and Biotechnology
• /
• v.22 no.3
• /
• pp.292-300
• /
• 2012

We report the expression, purification, and characterization of L-asparaginase (AnsA) from Rhizobium etli. The enzyme was purified to homogeneity in a single-step procedure involving affinity chromatography, and the kinetic parameters $K_m$, $V_{max}$, and $k_{cat}$ for L-asparagine were determined. The enzymatic activity in the presence of a number of substrates and metal ions was investigated. The molecular mass of the enzyme was 47 kDa by SDS-PAGE. The enzyme showed a maximal activity at $50^{\circ}C$, but the optimal temperature of activity was $37^{\circ}C$. It also showed maximal and optimal activities at pH 9.0. The values of $K_m$, $V_{max}$, $k_{cat}$, and $k_{cat}/K_m$ were $8.9{\pm}0.967{\times}10^{-3}$ M, $128{\pm}2.8$ U/mg protein, $106{\pm}2s^{-1}$, and $1.2{\pm}0.105{\times}10^4M^{-1}s^{-1}$, respectively. The L-asparaginase activity was reduced in the presence of $Mn^{2+}$, $Zn^{2+}$, $Ca^{2+}$, and $Mg^{2+}$ metal ions for about 52% to 31%. In addition, we found that $NH_4{^+}$, L-Asp, D-Asn, and ${\beta}$-aspartyl-hydroxamate in the reaction buffer reduced the activity of the enzyme, whereas L-Gln did not modify its enzymatic activity. This is the first report on the expression and characterization of the L-asparaginase (AnsA) from R. etli. Phylogenetic analysis of asparaginases reveals an increasing group of known sequences of the Rhizobial-type asparaginase II family.

• Journal of Biosystems Engineering
• /
• v.29 no.1
• /
• pp.37-44
• /
• 2004

This study was conducted to develop a new drying method far reducing the drying cost and time and to investigate the drying characteristics of oak mushroom. A far infrared dryer of down draft air flow type used for this experiment can control the drying parameters, such as far infrared heater temperature and aeration velocity. The far infrared drying tests were performed at aeration velocities of 0.3 and 0.6m/s under the temperature of 90 and 100$^{\circ}C$ in for infrared heater, respectively. The results were compared and analyzed with those of an heated air drying method used as a control in terms of properties representing the drying characteristics. such as shrinkage rate, color, energy consumption amino acid components, drying rate and moisture ratio. The results obtained from this research can be summarized as follows. 1. The drying rate of far infrared drying was faster than that of heated air drying. With high temperature of far infrared heater and slow aeration velocity, the far infrared drying of down draft air flow type was superior to the heated air drying. 2. Most of far infrared drying conditions required less energy consumption than heated air drying. 3. The shrinkage rates of heated air drying and far infrared drying were decreased by 17.0% and 18.2∼19.8%, respectively. 4. The difference of color on oak mushroom surface before and after drying can be represented as $\Delta$E. $\Delta$E values of far infrared drying and heated air drying were 2.39∼4.55 and 6.77, respectively. 5. The amounts of free amino acids were higher in the far infrared than in the heated air drying. In addition the amounts of Gln and Glu generally were increased and those of Ala, Leu, and Val were decreased in order.