• Journal of Microbiology and Biotechnology
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• v.26 no.12
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• pp.2087-2097
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• 2016

Most cold-adapted enzymes possess higher $K_m$ and $k_{cat}$ values than those of their mesophilic counterparts to maximize the reaction rate. This characteristic is often ascribed to a high structural flexibility and improved dynamics in the active site. However, this may be less convincing to cold-adapted metabolic enzymes, which work at substrate concentrations near $K_m$. In this respect, cold adaptation of a shikimate kinase (SK) in the shikimate pathway from psychrophilic Colwellia psychrerythraea (CpSK) was characterized by comparing it with a mesophilic Escherichia coli homolog (EcSK). The optimum temperatures for CpSK and EcSK activity were approximately $30^{\circ}C$ and $40^{\circ}C$, respectively. The melting points were $33^{\circ}C$ and $45^{\circ}C$ for CpSK and EcSK, respectively. The ${\Delta}G_{H_2O}$ (denaturation in the absence of denaturing agent) values were 3.94 and 5.74 kcal/mol for CpSK and EcSK, respectively. These results indicated that CpSK was a cold-adapted enzyme. However, contrary to typical kinetic data, CpSK had a lower $K_m$ for its substrate shikimate than most mesophilic SKs, and the $k_{cat}$ was not increased. This observation suggested that CpSK may have evolved to exhibit increased substrate affinity at low intracellular concentrations of shikimate in the cold environment. Sequence analysis and homology modeling also showed that some important salt bridges were lost in CpSK, and higher Arg residues around critical Arg 140 seemed to increase flexibility for catalysis. Taken together, these data demonstrate that CpSK exhibits characteristics of cold adaptation with unusual kinetic parameters, which may provide important insights into the cold adaptation of metabolic enzymes.

• Microbiology and Biotechnology Letters
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• v.9 no.3
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• pp.145-152
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• 1981

Microorganisms from the waste water of starch industry, were isolated and a strain, Y-124, possessing a powerful enzymic activity was selected and identified as a member of the genus Bacillus. The ideal cultural condition for the formation of $\alpha$-amylase form Bacillus Y-124 and its preservation was investigated in connection with the biotechnological and industrial approach to the bulk enzyme production. High yield of $\alpha$-amylase was observed in medium containing casein as well as calcium pantothenate in this work. Calcium ions were found to have an effect in forming this particular enzyme. Ammonium phosphate dibasic was an important inorganic nitrogen source for the formation of $\alpha$-amylase. And preservation of this enzyme was greatly affected by calcium or sodium salts. The addition of calcium carbonate or sodium sulfate presented the most effective result for the prevention of its denaturation to various factors. The above data was obtained with crude enzyme preparation.

• Journal of Life Science
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• v.24 no.4
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• pp.428-436
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• 2014

Loop-mediated isothermal amplification (LAMP) technique relies on autocycling strand displacement DNA sysnthesis without template denaturation steps under isothermal conditions. LAMP has more advantages than modern PCR, as it requires only basic laboratory equipment like an isothermal water bath, oven, and heating cabinet. Hence, in this study, five random primers were designed with Streptococcus parauberis, shikimate kinase Arok gene sequences (Genbank accession number: CP0024711). Two primers were selected based on the ladder pattern. Other optimum reaction conditions like temperature, time, and sensitivity were established and confirmed with conventional SYBR-green PCR. Results confirmed that the designed random primers were species specific, without any non-target DNA amplification under isothermal conditions. Hence, this study suggests that LAMP techniques could be used in the diagnosis of fish pathogen, specifically S. parauberis.

• Journal of the Korean Wood Science and Technology
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• v.29 no.3
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• pp.112-122
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• 2001

Laccase enzymes from Cerrena unicolor and Trametes versicolor were immobilized on the activated glass beads (CPG), silica gel (SG) and soil (SL). The heterogeneous matrices were activated by ${\gamma}$-aminopropyltriethoxysilane (APTES) and glutaraldehyde (GA), and their surfaces were coated by keratin (KER) on activated or non-activated CPG, SG and SL. The laccase activities were tested in the aqueous solution for the native and immobilized preparations using different pH and temperature conditions. By keratin coating on supports, in the cases of CPG-KER and SL-KER, the immobilization yield was increased from about 80% to 90%. Moreover, much less protein was immobilized in keratin coated matrices than in inorganic ones alone (e.g. on CPG-KER 57.6%, whereas on CPG alone 80.6%). Laccase immobilization on keratin coated inorganic matrices was generally more effective than that of non-coated matrices. Concerned to pH dependency, the optima pH for immobilized laccases generally shifted towards to higher values, 5.5-5.8 and even 5.9 in the case of keratin for C. unicolor and from 5.3 to 5.7 for T. versicolor, respectively, and decreased less gradually both in acidic and alkaline regions. The immobilized laccase was more stable against thermal denaturation. This seems particularly true at $75^{\circ}C$ in the case of C. unicolor, where the activity of immobilized enzyme is > 50% higher than that of the free enzyme. For T. versicolor the respective values were $65^{\circ}C$, and 50%.

• Korean Journal of Food Science and Technology
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• v.21 no.4
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• pp.536-541
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• 1989

In present paper, we investigated the quality stability of frozen seasoned anchovy meat products during storage at $-25\;{\pm}\;2^{\circ}C$. The pH and VBN contents of the products revealed a tendency to increase slightly during frozen storage 150 days. Viable cell counts and histamine contents of products are $0.8-2.6\;{\times}\;10^5/g,\;70.6-76.7 mg/100g$, respectively. In changes of fatty acid composition, percentage of polyenes such as eicosapentaenoic docosahexaenoic acid slightly decreased, while that of saturates and monoenes increased during frozen storage, The results of changes in POV, TBA values, color values, drips and salt extractable nitrogen contents during frozen storage showed that lipid oxidation and freeze denaturation of products could be retarded, and flavor could be enhanced by adding 0.2% sodium erythorbate and 12.8% emulsion curd.

• Journal of Life Science
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• v.14 no.2
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• pp.309-314
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• 2004

Cyclic AMP receptor protein (CRP) is involved in the transcriptional regulation of more than 100 genes in E. coli. CRP dimer is converted into active form via the sequential conformation change of cAMP binding pocket, hinge region and HTH DNA binding motif by binding of cAMP. The temperature gradient gel electrophoresis (TGGE) was applied to CRP protein to know whether it was an efficient technique to study the conformational transitions and the thermal stability. TGGE showed the unfolding process of wild-type and S83G CRP proteins with the temperature gradient set from 29 to 71$^{\circ}C$ on nondenaturing polyacrylamide gel. Melting temperature (Tm) was 57$\pm$1 and 55$\pm$1$^{\circ}C$ for wild-type and S83G CRP, respectively in acidic buffer[89.8 mM Glycine and 24 mM Boric acid (pH 5.8)].

• Fisheries and Aquatic Sciences
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• v.12 no.1
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• pp.6-15
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• 2009

Biochemical and functional properties of acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC) from rockfish skin were characterized. Yield of PSC (90.0%) was higher than that of ASC (63.2%). Both ASC and the PSC consisted of ${\alpha}1$ and ${\alpha}2$ chains, and $\alpha$-cross-linked components. According to the results of hydroxylation of proline and lysine, and FT-IR, no difference between the helical structure of ASC and PSC was identified. Thermal denaturation temperature (TDT) of ASC from rockfish skin was $22.8^{\circ}C$, the same as exhibited in PSC. Both ASC and PSC were higher in water absorption capacity (WAC) and oil absorption capacity (OAC) than other vegetable proteins. According to the results of emulsifying activity (EA) and cooking stability (CS), both ASC and PSC from rockfish skin were inferior compared to the commercial emulsifier (Tween-80). The results of FT-IR suggested that the structure of PSC was slightly different when compared to that of ASC. No differences in solubility were established between ASC and PSC from rockfish skin at various pH and NaCl concentrations.

• Journal of Plant Biology
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• v.39 no.1
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• pp.41-48
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• 1996

Two forms of homo serine dehydrogenase have been isolated from 8-day-old cotyledons of Canavalin lineata by a heat denaturation, ammonium sulfate fractionation, DEAE-8ephacel ion exchange and Sephacryl 8-300 gel filtration chromatographies, and Pro cion red dye, Cibacron blue dye and Resource Q column chromatographies. The molecular weights of T -form (threonine-sensitive) and K-form(threonine- insensitive) were estimated to 230 kD and 135 kD, respectively. In the presence of 10 mM threonine, the activity of T-form was inhibited with almost 70%, but that of K-form was not at all. The Km values tor homo serine of T- and Kform were 1.6 mM and 0.3 mM, respectively. The Km values for NAD of T- and K-form were 2.34 mM and 0.03 mM, respectively. And Km values for NADP of two isozymes were the same as 0.01 mM. The activities of T- and K-form were markedly stimulated up to 4.9and 2.8-fold, respectively, by 400 mM KCI. The partial purified(gel filtration) enzymes(Tform and K-form) can be reversibly converted.verted.

• Food Science of Animal Resources
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• v.35 no.5
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• pp.585-596
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• 2015

Chicken breast dipped with citric acid (CA) was treated by sous vide processing and stored in a refrigerated state for 0, 3, 6, 9, and 14 d. A non-dipped control group (CON) and three groups dipped in different concentrations of citric acid concentration were analyzed (0.5%, 0.5CIT; 2.0%, 2CIT and 5.0%, 5CIT; w/v). Cooking yield and moisture content increased due to the citric acid. While the redness of the juice and meat in all groups showed significant increase during storage, the redness of the citric acid groups was reduced compared to the control group (p<0.05). The percentage of myoglobin denaturation (PMD) of the CA groups was also increased according to the level of CA during storage. Total aerobic counts, Enterobacteriaceae counts, volatile basic nitrogen and thiobarbituric acid reactive substances (TBARS) were generally lower in the citric acid-treated samples than in untreated ones, indicating extended shelf life of the cooked chicken breast dipped in citric acid solution. The shear force of the 2CIT and 5CIT groups was significantly lower (p<0.05). The findings indicated positive effects in the physicochemical properties and storage ability of sous vide chicken breast at 2% and 5% citric acid concentrations.

• Food Science of Animal Resources
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• v.36 no.5
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• pp.583-593
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• 2016

The aim of this study was to investigate the effects of morphological characteristics of porcine muscle fibers on growth performance, muscle fiber characteristics, and pork quality taken from the longissimus dorsi muscle. A total of 239 crossbred pigs (164 castrated males and 75 females) were used in this study. Experimental pigs were categorized by the total number of muscle fiber (TNF: High and Low) and cross sectional area of muscle fiber (CSAF: Large, Middle, and Small). Their combinations were classified into six groups (High-Large, HL; High-Middle, HM; High-Small, HS; Low-Large, LL; Low-Middle, LM; Low-Small, LS). The TNF and CSAF were significantly (p<0.05) correlated with growth rate and carcass productivity, while the only of the type I number had no meaningful relationships excluding the correlation with loin area (p<0.001). The proportion of type I area was positively correlated with pH_{45 min} while the proportion of type IIB area was negatively correlated with pH_{45 min} and pH_{24 h} (p<0.05). Drip loss and protein denaturation had strong relationships with the proportion of type IIB number or area. The HL group exhibited the greatest growth performance. In addition, the HL group had significantly greater values in protein solubility than the other groups. In conclusion, this study suggest that high TNF combined to large CSAF improve the ultimate lean meat productivity and assure normal meat quality simultaneously with increased both proportion of number and area of type I, type IIA muscle fibers and lowered proportion of number and area of type IIB.