• Title/Summary/Keyword: Gly45Asp

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Site-Directed Mutagenesis Studies with Restriction Endonuclease EcoRV to Identify the Role of Ile91 in Recognition and Catalysis

  • Moon, Byung-Jo;Vipond, I. Barry;Halford, Stephen E.
    • BMB Reports
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    • v.29 no.2
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    • pp.99-104
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    • 1996
  • Site-directed substitutions were made to change the Ile91 of restriction endonuclease EcoRV to either Val, Ala or Gly to identify the role of Ile91 in recognition and catalysis, since substitution of Ile91 with Leu afforded dramatic effects on the activity and properties of restriction endonuclease EcoRV. These changes alter the size of the hydrophobic side chain at position 91 and thus might have revealed the reason for the altered phenotype of Ile91Leu. However, the properties of Ile91Val and Ile91Ala mutants were much like wild type EcoRV, in both activity and metal ion preference. Ile91Gly had very little activity with either $Mg^{2+}$ or $Mn^{2+}$ as cofactors. To try to understand the unusual $Mn^{2+}$ profile of the Ile91Leu mutant, two double mutants, Ile91Leu;Asp90Asn and Ile91Leu;Glu45Met were created. Both double mutants were seriously disabled by the second amino acid change. Ile91Leu;Glu45Met had some residual activity in the $Mn^{2+}$ reaction buffer, whereas the Ile91Leu;Asp90Asn displayed no detectable activity.

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Characteristics of bitter peptides from Doenjang (된장의 쓴맛 펩타이드 특성)

  • 홍혜정;이혜수
    • Korean journal of food and cookery science
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    • v.10 no.1
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    • pp.45-50
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    • 1994
  • Conventional Doenjang, three kinds of Improved Doenjang(A; koji: soybean: salt=53: 100: 33, B; koji: soybean: salt=100: 100: 40, C; koji: soybean: salt= 200: 100: 40) prepared with different ratio of koji and salt were made to study the changes in the general contents, characteristics bitter peptides, correlations between bitterness and overall eating quality. 1. Total nitrogen contents increased a little, and amino nitrogen contents in all samples increase markedly. Especially, Amino nitrogen contents of conventrional Doenjang increased more than others. Reducing sugars of doenjang prepared with Asp. oryzae were higher than conventional Doenjang and increased throughout the aging period and Doenjang prepared with Asp. oryzae were more acidic. 2. To characterize bitter peptides in fermented Doenjang, peptides were extracted with 2: 1(v/v) chloroform-methanol and separated by Gel chromato-graphy with Sephadex and TLC. After Gel chromatography and TLC, each fraction examined presence of bitterness and evaluated intensity of bitterness. Amino acid composition of the fractions showing bitter tastes were as follows. Conv. peak 1-1 Trp-(Asp, Arg, Thr, Ser, Glu, Pro)-Phe Imp. A peak 1 Trp-(Glu, Val, Arg, lie)-Phe Imp. B peak 1 Trp-(lie, Pro, Asp, Lys, Val, Glu)-Trp. Imp. C peak 1-2 Trp-(Try, Thr, Glu, Pro, Gly)-Phe 3. Sensory evaluation revealed that correlation coefficient between bitterness and overall eating guality was not high.

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Evaluation of the Apparent Ileal Digestibility (AID) of Protein and Amino Acids in Nursery Diets by In vitro and In vivo Methods

  • Cho, J.H.;Kim, I.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.7
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    • pp.1007-1010
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    • 2011
  • The objective was to evaluate in vitro prediction of ileal digestibility of protein and amino acids (AA) for current nursery pig diets (n = 10) by using pepsin and pancreatin incubations. To compare in vivo ileal digestibility, forty nursery pigs (4 pigs per diet) with an initial BW of $12.2{\pm}2.7$ kg were surgically equipped with T-cannula in the distal ileum. In all cases, the values of in vitro digestibility were higher than those of in vivo digestibility (p<0.05). With regard to the relationships of essential and non essential AA (CP), the $r^2$ value was 0.76. With regard to AA, high relationships were observed in Ile, Thr, and Gly (0.85, 0.83, and 0.89, respectively). Also, there was a lower relationship for Arg, Met, Ala, Asp, Glu, Pro, Ser, and Tyr with $R^2$ values of 0.56, 0.54, 0.40, 0.54, 0.45, 0.24, 0.49, and 0.35, respectively between in vitro and in vivo digestibility. The EAA relationship ($R^2$ = 0.71) was generally higher than that of NEAA ($R^2$ = 0.50) numerically. In conclusion, there were strong linear relationships between in vivo and in vitro ileal digestibility (CP, Ile, Thr, and Gly). In vitro prediction of ileal digestibility (CP, Ile, Thr, and Gly) seems to have significant potential for practical application.

Fermentation Process for Odor Removal of Oyster (Crassostrea gigas) Hydrolysate and Its Properties (이취 제거를 위한 굴 가수분해물의 발효공정과 제품의 특성)

  • Lee, Su-Seon;Park, Si-Hyang;Kim, Hyeun-A;Choi, Yeung-Joon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.4
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    • pp.542-550
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    • 2016
  • This study was carried out to investigate the optimal processing conditions for odor removal and maximal antioxidant effects of oyster (Crassostrea gigas) hydrolysate. The optimal hydrolysis conditions were 3.3% neutrase as the protease, $50^{\circ}C$ as the hydrolysis temperature, and 8.3 h as the hydrolysis time. Fish odor of enzymatic oyster hydrolysate was greatly reduced during Saccharomyces cerevisiae fermentation at $24^{\circ}C$ with 0.5% glucose. The protein content of the fermentation product from oyster hydrolysate powder was 25.7%, which contained the major amino acids Glu, Asp, Lys, Arg, Gly, and Ala, whereas Leu, Ala, Phe, Val, and Tau were abundant free amino acids. The important minor minerals were Zn and Fe. Toxicity against Chang cells was not observed in the fermentation product from the oyster hydrolysate up to $200{\mu}g/mL$. The results suggest that fermentation with S. cerevisiae could reduce the fish odor of enzymatic oyster hydrolysate. The hydrolysate has potential application as a food ingredients and nutraceutical.

Association of the G134A and G184C Polymorphisms in the CYP1A1 Gene with Lung Cancer Incidence

  • Ryu, Doug-Young;Huang, Ming-Ai;Park, Chang-Bo;Chang, Soo-Im;Im, Ruth;Choi, Seong-Jin;Kim, Na-Young;Park, In-Won;Choi, Byoung-Whui;Kim, Jae-Yeol;Shin, Jong-Wook;Choi, Jae-Chul;Choi, Byung-Sun;Park, Jung-Dock
    • Toxicological Research
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    • v.24 no.2
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    • pp.109-112
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    • 2008
  • The G184C and G134A single nucleotide polymorphisms(SNPs) of the CYP1A1 gene result in Ala62Pro and Gly45Asp substitutions, respectively. Here, we tested whether these SNPs are associated with an alteration in lung cancer incidence. We examined 80 Korean subjects with lung cancer and 240 age- and sex-matched controls. For each subject, the CYP1A1 gene was PCR amplified and sequenced. We observed that the odds ratio(OR) for lung cancer was 3.37 higher in subjects with the G184C polymorphism than in controls(95% confidence interval(CI), $0.89{\sim}12.73$, P=0.07). In contrast, the OR for lung cancer was 1.23 in subjects with the G134A polymorphism compared to controls(95% CI, $0.68{\sim}2.20$, P=0.49). The G184C polymorphism exacerbated the effects of smoking on lung cancer development. Gene-smoking interaction analyses revealed that past or present smokers with the G184C polymorphism had a higher incidence of lung cancer(OR, 24.72; 95% CI, $4.48{\sim}136.31$; P<0.01) than control smokers(OR, 6.65; 95% CI, $2.72{\sim}16.28$; P<0.01). However, there was only a slight difference in the ORs for lung cancer between control smokers and smokers with the G134A polymorphism. These findings suggest that the G184C polymorphism, but not the G134A polymorphism, is associated with an increased risk of lung cancer.

Induced freezing tolerance and free amino acids perturbation of spinach by exogenous proline

  • Shin, Hyunsuk;Oh, Sewon;Kim, Daeil;Hong, Jeum Kyu;Yun, Jae Gil;Lee, Sang Woo;Son, Ki-Ho
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.357-363
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    • 2018
  • The objective of this study was to investigate whether exogenous proline (Pro) could confer freezing tolerance of spinach and determine fluctuations of free amino acids in spinach leaf tissues under freeze-induced stress. Treatment with Pro (10 mM) resulted in more accumulation of Pro (~2.6-fold) in Pro-treated spinaches compared to untreated ones. These Pro-pretreated spinaches were more freezing-tolerant, showing more turgid leaves and petioles compared to untreated controls. However, when spinaches pre-treated with or without Pro were subjected to freezing, there was no significant difference in overall amino acid contents, emphasizing the role of Pro as an osmoprotectant. Freezing stress prompted intensification of total amino acid contents irrespective of pretreatment with Pro. Asp, Glu, Ala, and Val were the most abundant free amino acids due to increased protein degradation and nitrogen mobilization for plant survival under freezing stress. Arg, a precursor for the synthesis of polyamines in plants, was profoundly enhanced under freezing stress. This implies that Arg plays an important role in modulating freezing tolerance. Gly, Leu, and Ile were maintained at relatively low levels in all treatments. However, Ser, Tyr, and Lys as primary constituents of dehydrins were accumulated under freezing stress, suggesting that they might play a role in increasing cryoprotective activity under freezing stress.

Chemical Compositions of the Green and Ripened Pumpkin(Cucurbita moschata Duch.) (미숙호박과 완숙호박의 화학성분)

  • Cho, Gyu-Seong
    • Korean Journal of Food Science and Technology
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    • v.29 no.4
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    • pp.657-662
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    • 1997
  • An attempt was made to investigate the chemical compositions of green pumpkin, edible flesh and seed of ripened pumpkin (Cucurbita moschata Duch.). The proximate compositions were moisture 9.34%, 11.98%, protein 12.70%, 13.38%, lipid 11.31%, 0.85%, carbohydrate 64.32%, 62.18%, fiber 6.31%, 4.54% and ash 6.05%, 7.76% in green and ripened pumpkin, respectively. The compositions of free sugar were glucose, fructose, sucrose, lactose and maltose in green and ripened pumpkin, respectively. During flesh growth, glucose, maltose and lactose was increased but sucrose and fructose was decreased in pumpkin. Pattern of 15 amino acid compositions in green and ripened pumpkin was shown to be of similarity. Major amino acids were glutamic acid, aspartic acid and alanine in green pumpkin and edible flesh of ripened pumpkin. And major amino acid in seed of ripened pumpkin were glutamic acid, arginine, aspartic acid and leucine. The predominant fatty acids were palmitic acid, linolenic acid, linoleic acid and oleic acid in green and ripened pumpkin, respectively. And those in seed of ripened pumpkin were linoleic acid, palmitic acid and oleic acid. The richest mineral contained in the green and ripened pumpkin was shown to be K and followed by Ca, Mg, Na and Fe in order.

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Mutations in Streptomycin Resistance Genes and Their Relationship to Streptomycin Resistance and Lineage of Mycobacterium tuberculosis Thai Isolates

  • Hlaing, Yin Moe;Tongtawe, Pongsri;Tapchaisri, Pramuan;Thanongsaksrikul, Jeeraphong;Thawornwan, Unchana;Archanachan, Buppa;Srimanote, Potjanee
    • Tuberculosis and Respiratory Diseases
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    • v.80 no.2
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    • pp.159-168
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    • 2017
  • Background: Streptomycin (SM) is recommended by the World Health Organization (WHO) as a part of standard regimens for retreating multidrug-resistant tuberculosis (MDR-TB) cases. The incidence of MDR-TB in retreatment cases was 19% in Thailand. To date, information on SM resistance (SMR) gene mutations correlated to the SMR of Mycobacterium tuberculosis Thai isolates is limited. In this study, the mutations in rpsL, rrs, gidB, and whiB7 were investigated and their association to SMR and the lineage of M. tuberculosis were explored. Methods: The lineages of 287 M. tuberculosis collected from 2007 to 2011 were identified by spoligotyping. Drug susceptibility profiles were evaluated by the absolute concentration method. Mutations in SMR genes of 46 SM-resistant and 55 SM-susceptible isolates were examined by DNA sequencing. Results: Three rpsL (Lys43Arg, Lys88Arg, and Lys88Thr) and two gidB (Trp45Ter and Gly69Asp) mutations were present exclusively in the SM resistant M. tuberculosis. Lys43Arg rpsL was the most predominant SMR mutations (69.6%) and prevailed among Beijing isolates (p<0.001). No SMR-related mutation in was found rrs. The combination of rpsL and gidB mutations provided 76.1% sensitivity for detecting SMR in M. tuberculosis Thai isolates. whiB7 was not responsible for SMR in SM resistant isolates lacking rpsL and rrs mutations. The significance of the three gidB mutations, 276A>C, 615A>G, and 330G>T, as lineage signatures for Beijing and EAI were underscored. This study identified 423G>A gidB as a novel sub-lineage marker for EAI6-BGD1. Conclusion: Our study suggested that the majority of SMR in M. tuberculosis Thai isolates were responsible by rpsL and gidB polymorphisms constantly providing the novel lineage specific makers.

Processing of Enzymatic Hydrolysates from Conger eel Scrap (붕장어 가공잔사를 이용한 효소분해소재의 가공)

  • Kang Su Tae;Kong Chung Sik;Cha Yong Jun;Kim Jong Tae;Oh Kwang Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.3
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    • pp.259-264
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    • 2002
  • In order to develope nutritional and flavoring intermediate products, the optimal processing conditions for two stage enzyme hydrolysate (TSEH) from low-utilized conger eel scrap such as head and intestine were investigated. The optimal processing conditions for TSEH were revealed in temperature at $55^{\circ}C$ 3$\~$4 hours digestion with alcalase at the 1st stage, and 4 hours at $45{\~}50^{\circ}C$ digestion with neutrase at the 2nd stage. Among water extract, steam extract and enzyme hydrolysates of conger eel scrap, the present TSEH was superior to other extracts in terms of yield ana organoleptic taste such as harmonic umami and inhibition of fishy and greasy taste formation. From the results of chemical experiments and sensory evaluation, we may conclude that TSEH of conger eel scrap could be utilized as the flavoring intermediate materials for the fisheries products such as flavoring sauces, drinkable beverage and instant food materials.

Brewing of Acid-hydrolyzed Soy Sauce with Defatted Soybeans and Wheat Flour Koji (탈지대두와 밀가루 코오지를 이용한 산분해간장의 양조)

  • Sun, Sung-Kyun;Han, Eun-Mi;Lee, Taik-Soo;Lee, Myung-Whan
    • Applied Biological Chemistry
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    • v.30 no.2
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    • pp.147-152
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    • 1987
  • Soy sauce which was prepared with a mixture of defatted soybeans ant wheat flour koji added into acid-hydrolyzed soy sauce was evaluated for the chemical changes in levulinic acid, ethanol and amino acids composition during fermentation, Results showed that the highest reducing sugar content of $12.13{\sim}15.76%$ was found after $20{\sim}40\;days$ fermentation. The contents of total nitrogen, amino nitrogen, and ethanol increased with increase in fermentation time. A high levulinic acid content was fount at the initial fermentation period which was gradually decreased with further fermentation. The content of lactic acid also increased during fermentation. Amino acids such as Glu, Leu, Ala, Phe, Asp, Thr, Ser, Gly, Met, Tyr, Lys, His, Arg and Pro were detected in all tested groups. Higher values were found particularly for Glu, Leu, Ala and Phe and lower values for His, Tyr and Met. The most of amino acids increased in their contents after 120 days of fermentation.

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