• Title/Summary/Keyword: Biochemical study

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Sustainable Production of Dihydroxybenzene Glucosides Using Immobilized Amylosucrase from Deinococcus geothermalis

  • Lee, Hun Sang;Kim, Tae-Su;Parajuli, Prakash;Pandey, Ramesh Prasad;Sohng, Jae Kyung
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1447-1456
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    • 2018
  • The amylosucrase encoding gene from Deinococcus geothermalis DSM 11300 (DgAS) was codon-optimized and expressed in Escherichia coli. The enzyme was employed for biosynthesis of three different dihydroxybenzene glucosides using sucrose as the source of glucose moiety. The reaction parameters, including temperature, pH, and donor (sucrose) and acceptor substrate concentrations, were optimized to increase the production yield. This study demonstrates the highest ever reported molar yield of hydroquinone glucosides 325.6 mM (88.6 g/l), resorcinol glucosides 130.2 mM (35.4 g/l) and catechol glucosides 284.4 mM (77.4 g/l) when 400 mM hydroquinone, 200 mM resorcinol and 300 mM catechol, respectively, were used as an acceptor substrate. Furthermore, the use of commercially available amyloglucosidase at the end of the transglycosylation reaction minimized the gluco-oligosaccharides, thereby enhancing the target productivity of mono-glucosides. Moreover, the immobilized DgAS on Amicogen LKZ118 beads led to a 278.4 mM (75.8 g/l), 108.8 mM (29.6 g/l) and 211.2 mM (57.5 g/l) final concentration of mono-glycosylated product of hydroquinone, catechol and resorcinol at 35 cycles, respectively, when the same substrate concentration was used as mentioned above. The percent yield of the total glycosides of hydroquinone and catechol varied from 85% to 90% during 35 cycles of reactions in an immobilized system, however, in case of resorcinol the yield was in between 65% to 70%. The immobilized DgAS enhanced the efficiency of the glycosylation reaction and is therefore considered effective for industrial application.

Biocidal Effects of Chlorine Dioxide on Isolated and Identified Pathogens from Nosocomial Environment - Biochemical and Technical Covergence (병원내 환경으로부터 분리 및 확인된 병원균에 대한 이산화염소의 살균 효과 - 생화학 및 기술 융합)

  • Song, Kyoung-Ju;Jung, Suk-Yul
    • Journal of Digital Convergence
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    • v.15 no.6
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    • pp.339-344
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    • 2017
  • In this study, microorganisms were isolated from nosocomial environment and are identified by biochemical analysis as the part of biochemical and technical convergence. Microorganisms were collected at intense care unit of general hospital located in Pyeongtak (2014.11.28. - 2014. 11. 30). Using a VITEK2 equipment of biochemical approaches, eleven microorganisms e.g., Micrococcus luteus (or M. lylae), Granulicatella adiacens (M. luteus or M. lylae), Staphylococcus caprae, Sphingomonas paucimobilis, Kocuria kristinae, G elegans, Aerococcus viridans (or Staphylococcus arlettae), Methylobacterium spp., Dermacoccus nishinomiyaensis (or Kytococcus sedentarius), Kocuria kristinae (or M. luteus, M. lylae), Pseudomonas oryzihabitans were identified. And then identified bacteria plates were applied with a plastic stick, so called with "FarmeTok (medistick/Puristic) to produce ClO2. ClO2-releasing plastic stick showed the very strong inhibition of bacterial growth with about 99.9%. There were no bacterial colonies on the ClO2-incubated plate. Taken together, it is suggested that chlorine dioxide should be very strong inhibitor to microorganisms of nosocomial infections.

Evidence of Tandem Repeat and Extra Thiol-groups Resulted in the Polymeric Formation of Bovine Haptoglobin: A Unique Structure of Hp 2-2 Phenotype

  • Lai, Yi An;Lai, I Hsiang;Tseng, Chi Feng;Lee, James;Mao, Simon J.T.
    • BMB Reports
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    • v.40 no.6
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    • pp.1028-1038
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    • 2007
  • Human plasma Hp is classified as 1-1, 2-1, and 2-2. They are inherited from two alleles Hp 1 and Hp 2, but there is only Hp 1 in almost all the animal species. Hp 2-2 molecule is extremely large and heterogeneous associated with the development of inflammatory-related diseases. In this study, we expressed entire bovine Hp in E. coli as a $\alpha\beta$ linear form. Interestingly, the antibodies prepared against this form could recognize the subunit of native Hp. In stead of a complicated column method, the antibody was able to isolate bovine Hp via immunoaffinity and gelfiltration columns. The isolated Hp is polymeric containing two major molecular forms (660 and 730 kDa). Their size and hemoglobin binding complex are significantly larger than that of human Hp 2-2. The amino-acid sequence deducted from the nucleotide sequence is similar to human Hp 2 containing a tandem repeat over the $\alpha$ chain. Thus, the Hp 2 allele is not unique in human. We also found that there is one additional -SH group (Cys-97) in bovine $\alpha$ chain with a total of 8 -SH groups, which may be responsible for the overall polymeric structure that is markedly different from human Hp 2-2. The significance of the finding and its relationship to structural evolution are also discussed.

Morphological and biochemical differences in three Undaria pinnatifida populations in Korea

  • Park, Kwang-Jae;Kim, Bo-Yeon;Park, Seo-Kyoung;Lee, Jong-Hwa;Kim, Young-Sik;Choi, Han-Gil;Nam, Ki-Wan
    • ALGAE
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    • v.27 no.3
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    • pp.189-196
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    • 2012
  • Twelve morphological characters and the biochemical composition of Undaria pinnatifida f. distans sporophytes growing on the rocky shores of Jindo and Wando and on cultivation ropes in Kijang were measured to determine whether each population could be characterized by morphological features and biochemical composition. The goal of this study was to compare phenotypic variations between populations as they relate to environmental conditions. The sporophytes of the Kijang population were two times longer and 19 times heavier than those at Jindo. Sporophylls of the Jindo U. pinnatifida population were significantly smaller in length, width, frill number, and weight than those at Wando and Kijang. Kijang Undaria plants showed the highest contents of total protein, crude fiber, total amino acids, the amount of essential amino acids, the proportion of total unsaturated fatty acids, and eicosapentaenoic acid. However, the Jindo population showed the greatest content of carbohydrates, lipids, and minerals (Zn and Ca) of the three U. pinnatifida populations. In particular, Zn content of Jindo plants was 30 times greater than that of Kijang plants. Thus, the proximate composition, mineral composition, amino acids, and fatty acids of Undaria pinnatifida plants were distinguishable among the three representative Undaria populations evaluated. These results suggest that morphological and biochemical differences of the three U. pinnatifida populations can be attributed to differences in environmental conditions of their habitats.

Effects of Replacement of Fish Meal by Soy Protein Isolate on the Growth, Digestive Enzyme Activity and Serum Biochemical Parameters for Juvenile Amur Sturgeon (Acipenser schrenckii)

  • Xu, Q.Y.;Wang, C.A.;Zhao, Z.G.;Luo, L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.11
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    • pp.1588-1594
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    • 2012
  • An 8-wk experiment was conducted to evaluate the effect of replacing fish meal (FM) with soy protein isolate (SPI) on the growth, digestive enzyme activity and serum biochemical parameters of juvenile Amur sturgeon (Acipenser schrenckii). SPI was used to replace 0, 25, 50, 62.5, 75, 87.5, 100% of dietary FM and 100% replacement supplemented crystalline amino acid. Healthy sturgeon with an average initial weight of $26.38{\pm}0.24$ g were randomly assigned to 24 aquaria (8 treatments with triplicates each) at an initial stocking density of 11 fish per aquarium and cultured for 8 wks. The results showed that 75.00% or more substitution resulted in a poor weight gain rate, feed conversion ratio and survival rate compared to that of fish fed the control diet (p<0.05), whereas no significant differences were observed between diets of 25.00% to 62.50% substitution. Protease, lipase and amylase activity in foregut, mid-gut and hindgut were significantly (p<0.05) decreased by diets where SPI replacement levels were 62.50% or more. Levels of serum total protein (TP) and globulin decreased significantly from 21.03, 10.34 to 14.05, 5.63 g/L with the increasing dietary SPI (p<0.05), but alkaline phosphatase activity significantly increased (p<0.05). In addition, supplemental crystalline amino acid in the FM absence diet did not improve growth performance, intestine digestive enzyme activities and serum biochemical parameters. In conclusion, the results from this study showed adverse effects of inclusion of SPI in diets on growth performance, feed utilization and serum biochemical parameters in juvenile Amur sturgeon. Based on WGR and replacement ratio presented in this report, a 57.64% replacement level was recommended.

Biochemical Reactions on a Microfluidic Chip Based on a Precise Fluidic Handling Method at the Nanoliter Scale

  • Lee, Chang-Soo;Lee, Sang-Ho;Kim, Yun-Gon;Choi, Chang-Hyoung;Kim, Yong-Kweon;Kim, Byung-Gee
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.2
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    • pp.146-153
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    • 2006
  • A passive microfluidic delivery system using hydrophobic valving and pneumatic control was devised for microfluidic handling on a chip. The microfluidic metering, cutting, transport, and merging of two liquids on the chip were correctly performed. The error range of the accuracy of microfluid metering was below 4% on a 20 nL scale, which showed that microfluid was easily manipulated with the desired volume on a chip. For a study of the feasibility of biochemical reactions on the chip, a single enzymatic reaction, such as ${\beta}-galactosidase$ reaction, was performed. The detection limit of the substrate, i.e. fluorescein $di-{\beta}-galactopyranoside$ (FDG) of the ${\beta}-galactosidase$ (6.7 fM), was about 76 pM. Additionally, multiple biochemical reactions such as in vitro protein synthesis of enhanced green fluorescence protein (EGFP) were successfully demonstrated at the nanoliter scale, which suggests that our microfluidic chip can be applied not only to miniaturization of various biochemical reactions, but also to development of the microfluidic biochemical reaction system requiring a precise nano-scale control.

Improving the Productivity of Recombinant Protein in Escherichia coli Under Thermal Stress by Coexpressing GroELS Chaperone System

  • Kim, So-Yeon;Ayyadurai, Niraikulam;Heo, Mi-Ae;Park, Sung-Hoon;Jeong, Yong-Joo;Lee, Sun-Gu
    • Journal of Microbiology and Biotechnology
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    • v.19 no.1
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    • pp.72-77
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    • 2009
  • Here, we demonstrate that the overexpression of the GroELS chaperone system, which assists the folding of intracellular proteins and prevents aggregation of its biological targets, can enhance the thermotolerance of Escherichia coli strains and facilitate the production of recombinant protein under thermal stress. The overexpression of GroELS led to an about 2-fold higher growth rate of E. coli XL-1 blue than control at $45^{\circ}C$ and induced the growth of the strain even at $50^{\circ}C$, although the growth was not sustained in the second-round culture. The effect of GroELS overexpression was also effective on other E. coli strains such as JM109, $DH5{\alpha}$, and BL21. Finally, we have shown that coexpression of GroELS allows us to produce recombinant protein even at $50^{\circ}C$, a temperature at which the protein production based on E. coli is not efficient. This study indicates that the employment of the GroELS overexpression system can expand the range of environmental conditions for E. coli.

Examination of Correlations Between Several Biochemical Components and Powdery Mildew Resistance of Flax Cultivars

  • Aly, Aly A.;Mansour, Mahmoud T. M.;Mohamed, Heba I.;Abd-Elsalam, Kamel A.
    • The Plant Pathology Journal
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    • v.28 no.2
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    • pp.149-155
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    • 2012
  • A field trial was conducted in 2009/2010 and 2010/2011 growing seasons at Giza Agricultural Research Station to examine correlations between some biochemical componets and powdery mildews ($PM_s$) resistance in flax cultivars. Nine flax cultivars could be divided into five distinct groups, i.e., highly susceptible (Cortland and C.I. 2008), moderately susceptible (Giza 7, and Marshall), moderately resistant (Cass), resistant (Koto, Dakota and Wilden), and highly resistant (Ottowa 770B). The cultivars showed considerable variation in PM severity ranged from 8.05 on Ottowa 770B to 97.02% on Cortland. Total soluble proteins, total phenols, antioxidant enzymes (peroxidase and polyphenoloxidase), ascorbic acid, tocopherol, and malondialdehyde (MDA), were determined in uninfected leaves of the tested cultivars. Pearson's correlation coefficient was calculated to measure the degree of association between PM severity and each component. All components showed significant (P < 0.05) or highly significant (P < 0.01) negative correlation with PM severity except MDA, which showed positive correlation (P < 0.01). Linear regression analysis was used to evaluate the causal relationship between the biochemical components (independent variables) and PM severity (dependent variable). Coefficient of determination ($R^2$) values of the generated models ranged from 48.76 to 77.15%. Tocopherol, MDA, and proteins were the most important contributors to the total variation in PM severity as the $R^2$ values of their models were 71.78, 75.28, and 77.15%, respectively. The results of the present study suggest that tocopherol, MDA, and proteins in uninfected leaves can be used as biochemical markers to predict PM resistance in flax.

Genetic Diversity Studies and Identification of Molecular and Biochemical Markers Associated with Fusarium Wilt Resistance in Cultivated Faba Bean (Vicia faba)

  • Mahmoud, Amer F.;Abd El-Fatah, Bahaa E.S.
    • The Plant Pathology Journal
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    • v.36 no.1
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    • pp.11-28
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    • 2020
  • Faba bean (Vicia faba L.) is one of the most important legume crops in Egypt. However, production of faba bean is affected by several diseases including fungal diseases. Fusarium wilt incited by Fusarium oxysporum Schlecht. was shown to be the most common wilt disease of faba bean in Assiut Governorate. Evaluation of 16 faba bean genotypes for the resistance to Fusarium wilt was carried out under greenhouse conditions. Three molecular marker systems (inter-simple sequence repeat [ISSR], sequence related amplified polymorphism [SRAP], and simple sequence repeat [SSR]) and a biochemical marker (protein profiles) were used to study the genetic diversity and detect molecular and biochemical markers associated with Fusarium wilt resistance in the tested genotypes. The results showed that certain genotypes of faba bean were resistant to Fusarium wilt, while most of the genotypes were highly susceptible. The percentage of disease severity ranged from 32.83% in Assiut-215 to 64.17% in Misr-3. The genotypes Assiut-215, Roomy-3, Marut-2, and Giza2 were the most resistant, and the genotypes Misr-3, Misr-1, Assiut-143, Giza-40, and Roomy-80 performed as highly susceptible. The genotypes Assiut-215 and Roomy-3 were considered as promising sources of the resistance to Fusarium wilt. SRAP markers showed higher polymorphism (82.53%) compared with SSR (76.85%), ISSR markers (62.24%), and protein profile (31.82%). Specific molecular and biochemical markers associated with Fusarium wilt resistance were identified. The dendrogram based on combined data of molecular and biochemical markers grouped the 16 faba bean genotypes into three clusters. Cluster I included resistant genotypes, cluster II comprised all moderate genotypes and cluster III contained highly susceptible genotypes.

Relationship between Nutrient Intake and Biochemical Markers of Bone Metabolism in Korean Postmenopausal Women (폐경 후 여성의 영양소섭취수준에 따른 골대사 지표물질의 관련성 분석)

  • 이행신;이다홍;이다홍
    • Korean Journal of Community Nutrition
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    • v.6 no.5
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    • pp.765-772
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    • 2001
  • To delineate the relationship between the nutrient intake from diet and the serum biochemical markers of bone metabolism, 56 postmenopausal women of 50 to 77 years of age were recruited. The biochemical markers including osteocalcin, calcium, phosphorus, estradiol and free testosterone were measured in fasting blood. Bone mineral density(BMD) was measured also by dual energy X-ray absorptiometry, and the nutrient intake of earth individual subject was estimated by 24-hour recall of 3 days. The age of the subjects was 64.8 $\pm$ 7.7 years, and the BMDs of the subject were 0.86 $\pm$ 0.26g/$cm^2$(Lumbar spine), 0.60 $\pm$ 0.10g/$cm^2$ (Femoral neck), 0.49 $\pm$ 0.10g/$cm^2$(Trochanter), and 0.41 $\pm$ 0.14g/$cm^2$(Ward's triangle). There were no significant differences among age and nutrient intake level groups due to the small sample size. The biochemical markers showed certain degree of relationship with nutrient intake levels. The results were compared among 3 groups with different nutrient intake level classified by the percentage of Recommended Daily Allowances(RDA) for Koreans as follows low < 75% RDA, 75% RDA $\leq$ adequate < 125% RDA, high $\geq$ 125% RDA. The low energy and low riboflavin groups showed significantly higher serum osteocalcin levels than those of the high intake groups(p < 0.05). On the other hand, there was a trend for serum Ca level to be higher with high nutrient intake. In this case, protein and thiamin were the only nutrients that reached a statistical significance(p < 0.05). And the groups with low intake for protein and Ca showed significantly lower serum free testosterone levels than that of other intake groups(p < 0.05). This study suggests an important role of nutrient intake levels on blood biochemical markers of bone metabolism.

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