• Title/Summary/Keyword: streptomyces

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Cloning of Autoregulator Receptor Gene form Saccharopolyspora erythraea IFO 13426 (Saccharopolyspora erythraea IFO 13426으로부터 Autoregulator Receptor Protein Gene의 Cloning)

  • 김현수;이경화;조재만
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.117-123
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    • 2003
  • For screening of autoregulator receptor gene from Saccharopolyspora erythraea, PCR was performed with primers of receptor gene designed on the basis of amino acid sequences of autoregulator receptor proteins with known function. PCR products were subcloned into the BamHI site of pUC19 and transformed into the E. coli DH5$\alpha$. The isolated plasmid from transformant contained the fragment of 120 bp, which was detected on 2% gel after BamHI treatment. The insert, 120 bp PCR product, was confirmed as the expected internal segment of gene encoding autoregulator receptor protein by sequencing. Southern and colony hybridization using Saccha. erythraea chromosomal DNA were performed with the insert as probe. The plasmid (pEsg) having 3.2 kbp SacI DNA fragment from Saccha. erythraea is obtained. The 3.2 kbp SacI DNA fragment was sequenced by the dye terminator sequencing. The nucleotide sequence data was analyzed with GENETYX-WIN (ver 3.2) computer program and DNA database. frame analyses of the nucleotide sequence revealed a gene encoding autoregulator receptor protein which is a region including KpnI and SalI sites on 3.2 kbp SacI DNA fragment. The autoregulator receptor protein consisting of 205 amino acid was named EsgR by author. In comparison with known autoregulator receptor proteins, homology of EsgR showed above 30%.

Identification and Characterization of a New Alkaline SGNH Hydrolase from a Thermophilic Bacterium Bacillus sp. K91

  • Yu, Tingting;Ding, Junmei;Zheng, Qingxia;Han, Nanyu;Yu, Jialin;Yang, Yunjuan;Li, Junjun;Mu, Yuelin;Wu, Qian;Huang, Zunxi
    • Journal of Microbiology and Biotechnology
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    • v.26 no.4
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    • pp.730-738
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    • 2016
  • est19 is a gene from Bacillus sp. K91 that encodes a new esterase. A comparison of the amino acid sequence showed that Est19 has typical Ser-Gly-Asn-His (SGNH) family motifs and could be grouped into the SGNH hydrolase family. The Est19 protein was functionally cloned, and expressed and purified from Escherichia coli BL21(DE3). The enzyme activity was optimal at 60℃ and pH 9.0, and displayed esterase activity towards esters with short-chain acyl esters (C2-C6). A structural model of Est19 was constructed using phospholipase A1 from Streptomyces albidoflavus NA297 as a template. The structure showed an α/β-hydrolase fold and indicated the presence of the typical catalytic triad Ser49-Asp227-His230, which were further investigated by site-directed mutagenesis. To the best of our knowledge, Est19 is a new member of the SGNH hydrolase family identified from thermophiles, which may be applicable in the industrial production of semisynthetic β-lactam antibiotics after modification.

Effects of Fowl Dropping, Saw Dust and Rice Hull on Soil Microflora in vitro (실내배양에서 생계분, 톱밥 및 왕겨 첨가가 토양미생물상에 미치는 효과)

  • Yang, Chang-Sool;Kong, Hye-Suk
    • Korean Journal of Soil Science and Fertilizer
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    • v.29 no.1
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    • pp.53-59
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    • 1996
  • This study was conducted to evaluate the effects of fowl dzopping. saw dust and rice hall on the soil microflora in vitro. The experiment was designed in seven treatments with the various organic materials and they were only soil (control). soil + fowl dropping (S+F), soil+fowl dropping+rice hull (S+F+R) soil+fowl dropping*saw dust (S+F+S). soil+chemical fertilizer (S+C.F), fowl dropping+rice hull (F+R) and fowl dropping+saw dust (F+S). All the samples of treatment were incubated in $28{\pm}2^{\circ}C$ condition and tested the activity of soil microflora for 84 days The activity of fungi, total bacteria, gram-negative bacteria and actinomycetes showed the highest values at, twenty-first day and the spore-forming bacteria was at forty-second day after incubation. The number of fungi and gram-negative bacteria showed the highest values in the treatment of F+S, the spore-forming bacteria and the actinomycetes were in the S+F+S. and the number of total bacteria was in the F+C.F., but in the treatment of F+R. all the microorganism except fungi showed the lowest values in their numbers. The composition ratio of dead bacteria was higher in the treatments of S+F+R and F+R than in those of others as 70% and 40% respectively. Actinomycetes isolated from the treatments of S+F and S+F+S were identified as Streptomyces sp.. Nocardia sp., Micromonospora sp. Actinomadura sp. and Saccharomonospora sp.

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Bacterial core community in soybean rhizosphere (콩 근권의 핵심 세균 군집)

  • Lee, Youngmi;Ahn, Jae-Hyung;Choi, Yu-Mi;Weon, Hang-Yeon;Yoon, Jung-Hoon;Song, Jaekyeong
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.347-354
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    • 2015
  • Soybean is well known to be originated from Korea and far-east Asian countries, and studies of many root nodule bacteria associated with soybean have mainly-focused on nitrogen fixation, but much less study was carried out on bacterial community in the rhizosphere of soybean. In this study, we analyzed the bacterial community in rhizosphere of Korean soybean, Daepungkong using the pyrosequencing method based on the 16S rRNA gene to characterize the change of the rhizosphere community structure according to the growth stages of soybeans and to elucidate bacterial core community in rhizosphere of soybean. Our results revealed that bacterial community of rhizosphere soil differed from that of bulk soil and was composed of a total of 21 bacterial phyla. The predominant phylum in the rhizosphere of soybean was Proteobacteria (36.6-42.5%) and followed by Acidobacteria (8.6-9.4%), Bacteroidetes (6.1-10.9%), Actinobacteria (6.4-9.8%), and Firmicutes (5.7-6.3%). The bacterial core community in soybean rhizosphere was mainly composed of the operational taxonomic units (OTUs) belonging to the phylum Proteobacteria throughout all growth stages. The OTU00006 belonged to the genus Bradyrhizobium had the highest abundance and Steroidobacter, Streptomyces, Devosia were followed. These results show that bacterial core community in soybean rhizosphere was mainly composed of OTUs associated with plant growth promotion and nutrient cycles.

Effect of Dietary Fiber on the In Vitro Digestibility of Fish Protein (식이 섬유소가 어류단백 소화율에 미치는 영향)

  • Ryu, Hong-Soo;Park, Nam-Eun;Lee, Kang-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.3
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    • pp.255-262
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    • 1992
  • In vitro digestibility of filefish, protein was substantially decreased by fiber constituents in the follow-ing order : pectin (9.97%), gum karaya (7.03%), sodium alginate (6.12%),and cellulose (1.52%). The order of reduction by fibrous residues from vegetables ranked as follows : sea tangle (12.36%), Ro-maine lettuce (11.12%), perillar leaf (8.96%), and green pepper (5.15%). The inhibitory effect of the dietary fibers towards filefish protein digestion, expressed as soybean trypsin inhibitor equivalents, in-creased with added levels, but the inhibition differed with the sources of dietary fibers. Sea tangle and sodium alginate were most active in decreasing the concentration of essential amino acid from filefish protein hydrolysis. Sodium alginate exerted an inhibitory effect on the activity of trypsin, but the other fiber constituents did not have an inhibitory potency on trypsin and bacterial pretense (Streptomyces griceus). Results supported that dietary fiber components may reduce protein digestibility through the interaction of dietary fiber components with filefish protein.

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Diversity, distribution, and antagonistic activities of rhizobacteria of Panax notoginseng

  • Fan, Ze-Yan;Miao, Cui-Ping;Qiao, Xin-Guo;Zheng, You-Kun;Chen, Hua-Hong;Chen, You-Wei;Xu, Li-Hua;Zhao, Li-Xing;Guan, Hui-Lin
    • Journal of Ginseng Research
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    • v.40 no.2
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    • pp.97-104
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    • 2016
  • Background: Rhizobacteria play an important role in plant defense and could be promising sources of biocontrol agents. This study aimed to screen antagonistic bacteria and develop a biocontrol system for root rot complex of Panax notoginseng. Methods: Pure-culture methods were used to isolate bacteria from the rhizosphere soil of notoginseng plants. The identification of isolates was based on the analysis of 16S ribosomal RNA (rRNA) sequences. Results: A total of 279 bacteria were obtained from rhizosphere soils of healthy and root-rot notoginseng plants, and uncultivated soil. Among all the isolates, 88 showed antagonistic activity to at least one of three phytopathogenic fungi, Fusarium oxysporum, Fusarium solani, and Phoma herbarum mainly causing root rot disease of P. notoginseng. Based on the 16S rRNA sequencing, the antagonistic bacteria were characterized into four clusters, Firmicutes, Proteobacteria, Actinobacteria, and Bacteroidetesi. The genus Bacillus was the most frequently isolated, and Bacillus siamensis (Hs02), Bacillus atrophaeus (Hs09) showed strong antagonistic activity to the three pathogens. The distribution pattern differed in soil types, genera Achromobacter, Acidovorax, Brevibacterium, Brevundimonas, Flavimonas, and Streptomyces were only found in rhizosphere of healthy plants, while Delftia, Leclercia, Brevibacillus, Microbacterium, Pantoea, Rhizobium, and Stenotrophomonas only exist in soil of diseased plant, and Acinetobacter only exist in uncultivated soil. Conclusion: The results suggest that diverse bacteria exist in the P. notoginseng rhizosphere soil, with differences in community in the same field, and antagonistic isolates may be good potential biological control agent for the notoginseng root-rot diseases caused by F. oxysporum, Fusarium solani, and Panax herbarum.

Studies on the Improvements of Functional Properties of Sardine Protein by Plastein Reaction (Plastein반응을 이용한 정어리 단백질의 기능성 개선에 관한 연구 3. Plastein의 기능성 및 소화율)

  • Kim, Se-Kwon;Kwak, Dong-Chae;Cho, Duck-Jae;Lee, Eung-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.17 no.4
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    • pp.312-319
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    • 1988
  • The functional properties of plasteins have been compared with those of sardine protein concentrate and egg albumin. The solubility of plasteins was higher than that of FPG and the glu-plastein had 84% solubility in the range of pH 3-10. The dispersibility of plasteins was lower than that of egg albumin, however those of plasteins was higher than that of sardine protein concentrate. The water holding capacity of plasteins was higher than that of egg albumin. Lipid absorption of leu-papain plastein was the highest, holding 2.2m119, and that of the other plastein was higher than that of egg albumin. The emulsifying activity of leu-papain plastein was the highest, holding 66.4%, and that of glu-papain plastein was the lowest, holding 51.2%, The emulsifying stability of plasteins was similar to that of the emulsifying activity. The foaming capacitt of leu-papain plastein was the highest, holding 460%, and those of the other plasteins was higher than that of egg albumin. The foaming stability of plasteins was superior to that of egg albumin. The viscosity of plasteins was lower than that of see albumin. The in vitro digestibility of plasteins was 67.6-78.0% range. The digestibility by four pretense were somewhat lower in the glu-papain plastein than in the FPG. The digest of plasteins treated with the microbiol pretense such as molsin and pretense(from Streptomyces griceus), which had a storage broth taste.

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Functional Role of $^{60}RR^{61}$ in 23S rRNA Methylation, Which is in N-Terminal End Region of ErmSF (ErmSF의 N-Terminal End Region에 존재하는 $^{60}RR^{61}$의 23S rRNA Methylation에서의 역할)

  • Jin, Hyung-Jong
    • Korean Journal of Microbiology
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    • v.44 no.3
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    • pp.193-198
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    • 2008
  • ErmSF is one of the proteins which are produced by Streptomyces fradiae to avoid suicide by its autogenous macrolide antibiotic, tylosin and one of ERM proteins which are responsible for transferring the methyl group to $A_{2058}$ (Escherichia coli coordinate) in 23S rRNA, which reduces the affinity of MLS (macrolide-lincosamide-streptogramin B) antibiotics to 23S rRNA, thereby confers the antibiotic resistance on microorganisms ranging from antibiotic producers to pathogens. ErmSF contains an extra N-terminal end region (NTER), which is unique to ErmSF and 25% of amino acids of which is arginine known well to interact with RNA. Noticeably, arginine is concentrated in $^{58}RARR^{61}$ and functional role of each arginine in this motif was investigated through deletion and site-directed mutagenesis and the activity of mutant proteins in cell R60 and R61 was found to play an important role in enzyme activity through the study with deletion mutant up to R60 and R61. With the site-directed mutagenesis using deletion mutant of 1 to 59 (R60A, R61A, and RR60, 61AA), R60 was found more important than R61 but R61 was necessary for the proper activity of R60 and vice versa. And these amino acids were presumed to assume a secondary structure of $\alpha$-helix.

Solvent-tolerant Lipases and Their Potential Uses (유기용매 내성 리파아제와 그 이용가능성)

  • Joo, Woo Hong
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1381-1392
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    • 2017
  • This review described solvent-tolerant lipases and their potential industrial, biotechnological and environmental impacts. Although organic solvent-tolerant lipase was first reported in organic solvent-tolerant bacterium, many organic solvent-tolerant lipases are in not only solvent-tolerant bacteria but also solvent-intolerant bacterial and fungal strains, such as the well-known Bacillus, Pseudomonas, Streptomyces and Aspergillus strains. As these lipases are not easily inactivated in organic solvents, there is no need to immobilize them in order to prevent an enzyme inactivation by solvents. Therefore, the solvent-tolerant lipases have the potential to be used in many biotechnological and biotransformation processes. With the solvent-tolerant lipases, a large number insoluble substrates become soluble, various chemical reactions that are initially impossible in water systems become practical, synthesis reactions (instead of hydrolysis) are possible, side reactions caused by water are suppressed, and the possibility of chemoselective, regioselective and enantioselective transformations in solvent and non-aqueous systems is increased. Furthermore, the recovery and reuse of enzymes is possible without immobilization, and the stabilities of the lipases improve in solvent and non-aqueous systems. Therefore, lipases with organic-solvent tolerances have attracted much attention in regards to applying them as biocatalysts to biotransformation processes using solvent and non-aqueous systems.

Synthesis and Functional Properties of Plastein from the Enzymatic Hydrolysates of Filefish Protein. 3. Functional Properties of Plasteins (말쥐치육 단백질의 효소적 가수분해물을 이용한 Plastein의 합성 및 그 물성 , 3. Plastein의 기능성)

  • KIM Se-Kwon;LEE Eung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.20 no.6
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    • pp.582-590
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    • 1987
  • Plasteins were synthesized from a peptic filefish protein hydrolysate by papain, $\alpha-chymotrypsin$ and protease(from Streptomyces griceus) under the optimum conditions of previous paper. L-glutamic acid diethylester and L-leucine ethylester were incorporated into plastein during the plastein reaction by papain. The structural changes of freeze-dried filefish meat, peptic hydrolysate, FPC and plasteins were observed by Scanning Electron Microscopy(SEM). The functional properties of plasteins also were measured. The solubility of plasteins was higher than that of FPC and the Glu-plastein had $95\%$ solubility in the range of pH 3-10. The dispersibility of Glu-plastein and protease plastein was similar to that of egg albumin, but those of the other plasteins were lower. The water holding capacity of plasteins was lower than that of egg albumin and C. Lipid absorption of Leu-plastein was tile highest, holding 1.80 ml/g, and that of the other plasteins was similar to that of egg albumin. The emulsifying activity of Leu-plastein was the highest, holding $61.2\%$, and that of Glu-plastein was the lowest, holding $50.7\%$. The emulsifying stability of plasteins was similar to that of the emulsifying activity. The emulsifying capacity of Leu-plastein was 384 ml/g(the highest), but that of Glu-plastein and $\alpha-chymotrypsin$ plastein was 248 ml/g(the lowest). The Leu-plastein shelved the highest foaming capacity, $373\%$. The foaming capacity of other plasteins was higher than that of egg albumin. The foaming stability of plasteins was superior to that of egg albumin. The viscosity of plasteins was lower than that of egg albumin. The microstructure of $\alpha-chymotrypsin$ plastein by SEM wassimilar to that of papain plastein, but other plasteins showed differences in their microstructure. The microstructure of Glu-plastein had a smooth shape.

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