• Title/Summary/Keyword: rhamnose

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Metabolic Engineering of Rational Screened Saccharopolyspora spinosa for the Enhancement of Spinosyns A and D Production

  • Jha, Amit Kumar;Pokhrel, Anaya Raj;Chaudhary, Amit Kumar;Park, Seong-Whan;Cho, Wan Je;Sohng, Jae Kyung
    • Molecules and Cells
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    • v.37 no.10
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    • pp.727-733
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    • 2014
  • Spinosyns A and D are potent ingredient for insect control with exceptional safety to non-target organisms. It consists of a 21-carbon tetracyclic lactone with forosamine and tri-Omethylated rhamnose which are derived from S-adenosyl-methionine. Although previous studies have revealed the involvement of metK1 (S-adenosylmethionine synthetase), rmbA (glucose-1-phosphate thymidylyltransferase), and rmbB (TDP-D-glucose-4, 6-dehydratase) in the biosynthesis of spinosad, expression of these genes into rational screened Saccharopolyspora spinosa (S. spinosa MUV) has not been elucidated till date. In the present study, S. spinosa MUV was developed to utilize for metabolic engineering. The yield of spinosyns A and D in S. spinosa MUV was $244mgL^{-1}$ and $129mgL^{-1}$, which was 4.88-fold and 4.77-fold higher than that in the wild-type ($50mgL^{-1}$ and $27mgL^{-1}$), respectively. To achieve the better production; positive regulator metK1-sp, rmbA and rmbB genes from Streptomyces peucetius, were expressed and co-expressed in S. spinosa MUV under the control of strong $ermE^*$ promoter, using an integration vector pSET152 and expression vector pIBR25, respectively. Here-with, the genetically engineered strain of S. spinosa MUV, produce spinosyns A and D up to $372/217mgL^{-1}$ that is 7.44/8.03-fold greater than that of wild type. This result demonstrates the use of metabolic engineering on rationally developed high producing natural variants for the production.

Physicochemical Properties of Rice Kernels Affected on Palatability (식미에 영향을 미치는 쌀 곡립의 이화학적 특성)

  • Yoon, Mi-Ra;Kim, Chae-Eun;Koh, Hee-Jong;Kang, Mi-Young
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.1
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    • pp.45-50
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    • 2007
  • This study was investigated the relationship between palatability and physicochemical properties of rice kernels from two rice cultivars, including Gopumbyeo and Palgongbyeo. Major fatty acids of rice were oleic(35% in Gopum, 33.2% in Palgong, respectively) and linoleic acids (42.9% in Gopumbyeo, 40.7% in Palgongbyeo, respectively). There were significant differences in composition of amino acids and content of sugar from non-starch polysaccharide between two rice cultivars. However, no difference was found in mineral contents between Gopumbyeo and Palgongbyeo. In Rapid Visco Analyzer examination, pasting temperature and breakdown of Gopumbyeo were lower than those of Plagongbyeo. In X-ray diffraction patterns of starches separated from two rice cultivars, traditional A type and there was no difference in crystalline of rice starch. We could also obtain the results that Goupumbyeo, known to have good eating quality, had higher palatability value than Palgongbyeo.

Immunostimulating Activites of Polysaccharide Fractions isolated from Aster scaber Thunb. (참취에서 분리한 다당의 면역자극 활성)

  • Sung, Su-Kyung;Rhee, Young Kyung;Cho, Chang-Won;Kim, Eun Young;Kang, Dong-Zhou;Hong, Hee-Do
    • The Korean Journal of Food And Nutrition
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    • v.28 no.5
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    • pp.821-828
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    • 2015
  • ASW0 is a polysaccharide derived from the perennial herb Aster scaber Thunberg. We isolated ASW0, a fraction of crude polysaccharide, by means of ethanol precipitation and dialysis after hot water extraction to investigate its physicochemical properties and immunostimulatory effects. ASW0 contains neutral sugar (45.7%), acidic sugar (51.6%), protein (2.3%), and 2-keto-3-deoxy-D-manno-octonate (KDO) (0.4%). The neutral sugar in ASW0 (in mole percentage) was mainly composed of arabinose (34.5 mol%), glucose (31.1 mol%), galactose (14.9 mol%), and rhamnose (8.1 mol%), which are characteristic of pectic polysaccharides. ASW0 also contained small amounts of xylose, mannose, and fucose. The anti-complementary activity of ASW-0 was similar to that of polysaccharide K (used as positive control). ASW0 exhibited no cytotoxicity in RAW 264.7 macrophages and dramatically increased nitric oxide (NO) production in a dose dependent manner ($0.3{\sim}30{\mu}g/mL$). Also, macrophages stimulated with ASW0 showed enhanced production of immunostimulatory cytokines such as interleukin-6 (IL-6) and tumor necrosis factor alpha ($TNF-{\alpha}$) in a dose dependent manner. These results suggest that the ASW0 have a potent immunostimulatory effect and can be used as a natural immune health ingredient.

Immune Enhancement of Hericium erinaceum Mycelium Cultured in Submerged Medium Supplemented with Ginseng Extract (수삼 추출물 첨가 액체배지에서 배양된 노루궁뎅이버섯 균사체의 면역활성 증진)

  • Kim, Hoon;Ra, Kyung-Soo;Hwang, Jong-Hyun;Yu, Kwang-Won
    • The Korean Journal of Food And Nutrition
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    • v.25 no.4
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    • pp.737-746
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    • 2012
  • 본 연구는 mushroom complete medium(MCM) 액체배지에 수삼 추출물(GE, $65^{\circ}Bx$)을 첨가하여 면역활성이 증진된 노루궁뎅이버섯(Hericium erinaceum) 균사체를 배양하고, 균사체로부터 활성다당성분을 분획하고자 하였다. MCM에 대하여 GE를 5, 10과 15%(v/v) 첨가한 액체배지에서 균사체를 배양하고, 각각의 조다당획분(HE-GE-5-CP, HE-GE-10-CP와 HE-GE-15-CP)으로 분획하여 면역활성을 측정한 결과, HE-GE-10-CP는 HE-GE-5-CP와 HE-GE-15-CP보다 높은 활성을 나타내었으며, GE를 첨가하지 않은 MCM에서 배양된 균사체 조다당획분(HE-CP)보다 유의적으로 증진된 면역활성을 나타내었다. 또한, HE-GE-10-CP의 DEAE-Sepharose CL-6B 분획물 중 가장 높은 활성을 나타낸 HE-GE-10-CP-II획분은 대조군인 HE-CP의 어떠한 획분보다도 유의적으로 높은 면역활성과 암 전이 억제활성을 나타내었다. 한편, 활성획분인 HE-GE-10-CP-II는 arabinose, rhamnose, galactose, glucose와 uronic acid(molar ratio; 0.34:0.26:0.99:1.00:0.39)로 구성되어 있으나, 대조군인 HE-CP의 동일용매 용출획분으로서 HE-GE-10-CP-II보다는 활성이 낮은 HE-CP-II는 fucose, mannose, galactose와 glucose(molar ratio; 0.32:0.55:1.00:0.96)를 함유하여 다른 구성당 분포를 나타내었다. 따라서 노루궁뎅이버섯 균사체 액체배양에서 수삼 추출물 첨가는 균사체의 구성당 변화를 통한 면역활성 증진에 관여하는 것으로 사료되어 기능성 소재 개발에 유용할 것으로 사료된다.

Influence of Physiological and Environmental Factors on Growth and Sporulation of an Antagonistic Strain of Trichoderma viride RSR 7

  • Jayaswal, R.K.;Singh, Rajesh;Lee, Youn-Su
    • Mycobiology
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    • v.31 no.1
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    • pp.36-41
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    • 2003
  • Influence of physiological and environmental factors on an antagonistic strain of Trichoderma viride RSR7 were studied optimize its biocontrol potential. The growth and sporulation of T. viride was greatly influenced by various carbon and nitrogen sources, and the environmental factors such as pH and temperature. The best growth and sporulation of T. viride was observed when sucrose, peptone and trehalose were supplemented in the medium as sole carbon sources. Rhamnose, pyruvic acid and sorbitol also supported a good growth. However, with these carbon sources the sporulation was poor. Growth and sporulation was also affected by various nitrogen sources. Growth and sporulation both were favoured by ammonium forms of nitrogen compared to nitrite or nitrate forms. Urea did not support either growth or sporulation. Among amino acids, glutamic acid, asparagine, leucine, aspartic acid, glutamic acid and alanine supported good growth as well as sporulation. T. viride was able to utilize large number of amino acids as sole nitrogen source. Proline was good for growth, but not for sporulation. Maximum growth and sporulation of T. viride was between pH 4.5 to 5.5. Temperatures between $20^{\circ}C\;and\;37^{\circ}C$ were good for both growth and sporulation of T. viride. At lower temperatures(i.e. below $20^{\circ}C$) growth and sporulation were inhibited. Based on the present study it may be concluded that T. viride RSR7 is capable of growing and sporulating with varied nutritional and environmental conditions and, therefore, this strain of T. viride may be useful as a biocontrol agent under diverse physiological and environmental conditions.

Growth Inhibition of Polysaccharide Fraction in Cell Wall Components from Enterococcus faecalis 2B4-1 against Tumor Cell Lines (Enterococcus faecalis 2B4-1 세포벽 성분 중 Polysaccharide Fraction의 종양세포 증식억제 효과)

  • Park, Sang-Jin;Kim, Jeong-Hwan;Lee, Kyung-Ho;Yang, Jong-Beom;Baek, Young-Jin;Kim, Chang-Han
    • Microbiology and Biotechnology Letters
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    • v.27 no.1
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    • pp.8-14
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    • 1999
  • This study was developed to evaluate the growth inhibition effects of cell wall components of Enterococcus faecalis 2B4-1 obtained from feces of neonates against tumor cell lines. Polysaccharide fraction (PS) shown sensitive growth inhibition effect in the cell wall components was isolated and characterized. In growth inhibition effects, residue fractin of whole cell was shown sensitive level of percent survival about 30% when administrated at ehe concentration of 100${\mu}$g/ml, and that was more effective than that of supernatant fraction against the tumor cell lines, SNU-1, 3LL, FARROW and HEC-1-B. Sensitive growth inhibition effects against SNU-1, FARROW and HEC-1-B were performed by whole cell (WC) fraction from Ent. faecalis 2B4-1. Cytoplasm fractin (CP) of WC was shown non-inhibition effect, however, the other part of WC, precipitate of disrupted cell (PD), was sensitive against the tumor cell line mentioned above. Followed by separation to peptidoglycan fraction (PG) and polysaccharide fraction (PS) were all sensitive which the latter was shown more sensitive percent survival than the former. Composed sugars of polysaccharide fraction were determined to D-glucose, L-rhamnose and D-glucosamine, and the rate fo composition was calculated to about 1:1:1 by the data of elemental analysis, IR, TLC and HPLC.

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Studies on the Naringinase Produced by Aspergillus nidulans (Part I) Purification and Characteristics of Naringinase (Aspergillus nidulans 가 생산하는 Naringinase 관한 연구 (제2보) Naringinase의 정제 및 물리화학적 성질)

  • 변유량;문순옥;유주현
    • Microbiology and Biotechnology Letters
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    • v.6 no.2
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    • pp.65-73
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    • 1978
  • Naringinase extracted from the culture media of Aspergillus nidulans was purified, and the activity was proved to be stronger by 781 fold in the part of precipitation with ammonium sulfate, and column chromatography using DEAE-Sephadex A-25 and Sephadex G-100. Two fractions which had the same enzyme activity were isolated by the purification. Both fractions showed the highest enzymes activity under the reaction conditions of pH 5.0 and 40$^{\circ}C$. Molecular weight of fraction I and fraction II were estimated as 78,000 and 26,000 respectively. This indicated that fraction I would be trimer of fraction II. The enzyme was inhibited by glucose and rhammose, and the Km value was calculated to be 2.3${\times}$ 10$\^$-6/ g/ml.

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Isolation of a Novel Gellan-Depolymerizing Bacillus sp. Strain YJ-1

  • Jung, Yu-Jin;Park, Cheon-Seok;Lee, Hyeon-Gyu;Cha, Jae-Ho
    • Journal of Microbiology and Biotechnology
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    • v.16 no.12
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    • pp.1868-1873
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    • 2006
  • A novel microorganism that could degrade high molecular weight gellan was screened and isolated from soil. On gellan plate, the microorganism grew well and completely liquefied the plate. The gellan-degrading microorganism was isolated by pure culture on glucose and nutrient agar medium afterwards. The 16S rDNA sequence analysis and biochemical tests using an API 50CHB/20E kit revealed that the strain belonged to Bacillus sp. The isolate, named as Bacillus sp. YJ-1, showed optimum gellan-degrading activity in 0.5% gellan medium at pH 7.5 and 37$^{\circ}C$. The activity was measured and evaluated by the thiobarbituric acid and thin-layer chromatography method. Mass spectrometry revealed that the major gellan.. depolymerized product was an unsaturated tetrasaccharide consisting of $\Delta$4,5-glucuronic acid-(1$\rightarrow$4 )-$\beta$-D-glucose-(1$\rightarrow$4)- $\alpha$-L-rhamnose-(1$\rightarrow$3)-$\beta$-D-glucose, which is a dehydrated repeating unit of gellan, thus the enzyme was identified as gellan lyase. When the gellan was present in the medium, the gellan-degrading activity was much higher than that in glucose-grown cells. These results indicate that in the presence of gellan, Bacillus sp. YJ-1 is able to metabolize the gellan by inducing gellan-degrading enzymes that can degrade gellan into small molecular weight oligosaccharides, and then the gellan. depolymerized products are taken up by the cells and utilized by intracellular enzymes.

Isolation and Characterization of Mucous Exopolysaccharide (EPS) Produced by Vibrio furnissii Strain VB0S3

  • Bramhachari P.V.;Kishor P.B. Kavi;Ramadevi R.;Kumar Ranadheer;Rao, B. Rama;Dubey Santosh Kumar
    • Journal of Microbiology and Biotechnology
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    • v.17 no.1
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    • pp.44-51
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    • 2007
  • Marine bacterial strains were isolated trom coastal regions of Goa and screened for the strains that produce the highest amount of mucous expolysaccharide (EPS). Our screening resulted in the identification of the strain Vibrio furnissii VB0S3 (hereafter called VB0S3), as it produced the highest EPS in batch cultures during the late logarithmic growth phase. The isolate was identified as VB0S3 based on morphological and biochemical properties. Growth and EPS production were studied in mineral salts medium supplemented with NaCl (1.5%) and glucose (0.2%). The exopolymer was recovered from the culture supernatant by using three volumes of cold ethanol precipitation and dialysis procedure. Chemical analyses of EPS revealed that it is primarily composed of neutral sugars, uronic acids, and proteins. Fourier-transform infrared (FT-IR) spectroscopy revealed the presence of carboxyl, hydroxyl, and amide groups, which correspond to a typical heteropolymeric polysaccharide, and the EPS also possessed good emulsification activity. The gas chromatographic analysis of an alditol-acetate derivatized sample of EPS revealed that it was mainly composed of galactose and glucose. Minor components found were mannose, rhamnose, fucose, ribose, arabinose, and xylose. EPS was readily isolated from culture supernatants, which suggests that the EPS was a slime-like exopolysaccharide. This is the first report of exopolysaccharide characterization that describes the isolation and characterization of an EPS expressed by Vibrio surnissii strain VB0S3. The results of the study contribute significantly and go a long way towards an understanding of the correlation between growth and EPS production, chemical composition, and industrial applications of the exopolysaccharide in environmental biotechnology and bioremediation.

Banana Peel: A Green Solution for Metal Removal from Contaminated Waters

  • Arunakumara, Kkiu;Walpola, Buddhi Charana;Yoon, Min-Ho
    • Korean Journal of Environmental Agriculture
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    • v.32 no.2
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    • pp.108-116
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    • 2013
  • BACKGROUND: Certain crop-based waste materials have been recognized as cost-effective and highly efficient adsorbents for removal and recovery of different kind of heavy metals from aqueous solutions. The ability is strongly attributed to the carboxyl functional group of some pectin substances such as galacturonic acid often found in fruit peels. The present manuscript was aimed at assessing the potential applicability of banana peel for metal removal from contaminated waters. METHODS AND RESULTS: As revealed by laboratory investigations, banana peel contains pectin (10-21%), lignin (6-12%), cellulose (7.6-9.6%), and hemicelluloses (6.4-9.4%). The pectin extraction is reported to have glucose, galactose, arabinose, rhamnose, xylose, and galactouroninc acid. Several studies conducted under different conditions proved that banana peel is capable of adsorbing 5.71, 2.55, 28.00, 6.88, 7.97, and 5.80 mg/g of $Cd^{2+}$, $Co^{2+}$, $Cu^{2+}$, $Ni^{2+}$, $Pb^{2+}$, and $Zn^{2+}$, respectively, from aqueous solutions. Adsorption capacity is, however, dependent upon several factors including solution pH, dose of adsorbent and metal concentration, contact time and shaking speed. CONCLUSION(S): Since the annual world production of banana exceeds 100 million tons, about 40 million tons of banana peel (40% of total weight of the fresh fruit) remains vastly unused. Exploring a sound technology with banana peel would therefore, not only address the much needed sustainable tool for cleaning contaminated waters, but of course bring an additional value to the banana industry worldwide.