• Title/Summary/Keyword: Microbial isolation

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Isolation of Protease Producing Microorganisms (단백질 분해효소 생산 균주 분리)

  • Kim, Gi Eun
    • Journal of Korean Society of Environmental Engineers
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    • v.36 no.4
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    • pp.265-270
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    • 2014
  • Protease producing microorganisms were isolated from many kinds of food waste and fermented foods, which contains high amount and variable kinds of degraded substances. Several microorganisms were identified by 16S rRNA full sequencing analysis methods. The activity of protease was analyzed and identified in variable conditions for the application. For industrial use for biowaste treatment some proteases were isolated, identified and selected from microbial cells. And the tests were carried for the further use. The protein degrading activity at low temperature is useful for the treatment of organic waste, which contains much proteins. By the protein degradation process the organic waste can be utilized in variable fields, for example from feedstuff supplement to fertilizer for agriculture. Bacterial cells with protease activity at low temperature were isolated and identified. The optimal conditions for microbial cultivation and protease production were studied.

Isolation, Physiological Characterization of Bacteriophages from Enhanced Biological Phosphorus Removal Activated Sludge and Their Putative Role

  • Lee, Sang-Hyon;Satoh, Hiroyasu;Katayama, Hiroyuki;Mino, Takashi
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.730-736
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    • 2004
  • This study aims at characterizing the bacteriophages isolated from activated sludge performing enhanced biological phosphorous removal (EBPR) to understand the interactions between the phage-host system and bacterial community. Sixteen bacterial isolates (E1-E16) were isolated as host bacterial strains from EBPR activated sludge for phage isolation. Forty bacteriophages based on their plaque sizes (2 plaques on E4, 4 on E8, 11 on E10, 5 on E14, 18 on E16) were obtained from filtered supernatant of the EBPR activated sludge. Each bacteriophage did not make any plaque on bacterial strains tested in this study except on its own host bacterial strain, respectively, indicating that the bacteriophages are with narrow host specificity. However, fourteen of the forty bacteriophages obtained in this study lost their virulent ability even on their own host bacteria. All of the lytic phages showed similar one-step growth patterns and had long latent period (about 9 hours) to reproduce their phage particles in their host bacterial cells. On the other hand, their probable burst sizes (6 to 48 per host cell) were large enough to actively lyse their host bacterial cells. Therefore, it could be implied that bacteriophages are also important members of the microbial community in EBPR activated sludge, and lytic phages directly decrease the population size of their host bacterial groups in EBPR activated sludge by lysis.

Laccase- and Peroxidase-Free Tyrosinase Production by Isolated Microbial Strain

  • Sambasiva Rao, K.R.S.;Tripathy, N.K.;Mahalaxmi, Y.;Prakasham, R.S.
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.207-214
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    • 2012
  • Laccase- and peroxidase-free tyrosinase has commercial importance in the production of L-3, 4-dihydroxyphenylalanine (L-DOPA), which is mainly used in the treatment of Parkinson's disease. In the present study, isolation of an actinomycetes microbial strain capable of producing only tyrosinase is reported. Among all soil isolates, three individual colonies revealed black color around the colony in the presence of tyrosine. Further screening for laccase and peroxidase activities using syringaldazine denoted that one of the isolates, designated as RSP-T1, is laccase and peroxidase negative and produces only tyrosinase. The microbe was authenticated as Streptomyces antibioticus based on 16S ribotyping. Effective growth of this isolate was noticed with the use of medium (pH 5.5) containing casein acid hydrolysate (10.0 g/l), $K_2HPO_4$ (5.0 g/l), $MgSO_4$ (0.25 g/l), L-tyrosine (1.0 g/l), and agar (15 g/l). The scanning electron micrograph depicted that the microbe is highly branched and filamentous in nature. The enzyme production was positively regulated in the presence of copper sulfate. The impact of different fermentation parameters on tyrosinase production depicted that the maximized enzyme titer values were observed when this isolate was grown at 6.5 pH and at $30^{\circ}C$ temperature under agitated conditions (220 rpm). Among all the studied physiological parameters, agitation played a significant role on tyrosinase production. Upon optimization of the parameters, the yield of tyrosinase was improved more than 100% compared with the initial yield.

Analysis of Total Bacteria, Enteric Members of γ-proteobacteria and Microbial Communities in Seawater as Indirect Indicators for Quantifying Biofouling

  • Lee, Jin-Wook;Kim, Sung-Min;Jung, Ji-Yeon;Oh, Byung-Soo;Kim, In S.;Hong, Soon-Kang
    • Environmental Engineering Research
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    • v.14 no.1
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    • pp.19-25
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    • 2009
  • In this study, total bacteria, enteric members of the $\gamma$-proteobacteria, and microbial communities in seawater were analyzed as indirect indicators for quantifying biofouling. Biomass in seawater can significantly affect feed water pretreatment and membrane biofouling of reverse osmosis desalination processes. The purpose of this paper is to investigate microbiological quantity and quality of seawater at the potential intake of a desalination plant. For this analysis, the total direct cell count (TDC) using 4'-6-diamidino-2-phenylindole (DAPI)-staining and DNA-based real-time PCR were used to quantify the total bacteria and relative content of enteric members of $\gamma$-proteobacteria in seawater, respectively. In addition, microbial communities were examined using 16S rRNA gene cloning and bacterial isolation to identify the most abundant bacteria for a further biofouling study. The experimental results of this study identified about $10^6$ cells/mL of (total) bacteria, $10^5$ 16S rRNA gene copies/mL of enteric $\gamma$-proteobacteria, and the presence of more than 20 groups of bacteria.

Studies on Antibiotic Producers of Korean Soil Microbes(II) -Isolation and Antibiotic Activity of Streptomyces Strain DMC-64- (한국(韓國) 토양균(土壤菌)중 항생물질(抗生物質) 생성균(生成菌)에 관한 연구(硏究)(제 2 보)(第 2 報) -스트렙토마이세스속(屬) 균주(菌株) DMC-64 호(號)의 분리(分離) 및 항균작용(抗菌作用)-)

  • Kim, Jung-Woo;Choi, Eung-Chil;Kim, Byong-Kak
    • The Korean Journal of Mycology
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    • v.12 no.3
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    • pp.85-92
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    • 1984
  • To isolate antibiotic-producing microorganisms from Korean soils, microbes were isolated from the soil samples and screened for antibacterial activity. A strain which was isolated from the soil sample collected in Choong Chung Book Do had a high antibacterial activity against gram-positive bacteria. The examination of morphological and physiological characteristics of that strain according to the International Streptomyces Project methods showed that it was one of Streptomyces species. After the antibacterial constituent of the strain was produced in submerged culture method, it was isolated and purified by XAD-2 and CM-Sephadex column chromatography. And it was found to be one of quinone type antibiotics.

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Isolation, Purification, and Partial Characterization of an AMP Deaminase from Saccharomyces cerevisiae D

  • Kim, Myung-Hee;Lee, Jung-Kee;Kim, Hyung-Kwoun;Oh, Tae-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.9 no.4
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    • pp.429-435
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    • 1999
  • An adenosine 5'-monophosphate deaminase (AMP aminohydrolase, EC 3.5.4.6) was purified to homogeneity from the cell-free extract of Saccharomyces cerevisiae DKCTC7248. The molecular mass of subunit was estimated to be 80 kDa on SDS-PAGE, and that of the holoenzyme was shown to be 240 kDa by gel filtration. The isoelectric point of the enzyme (AMP deaminase D) was determined to be 6.2. The AMP deaminase D was specific towards AMP with an apparent $K_m$ value of 4.1 mM and a Hill coefficient, $n_H$, of 2.2. Both ATP and ADP were positive allosteric effectors of the AMP deaminase D: The apparent $K_{m}$ was decreased to 1.6 mM and 3.3 mM in the presence of 0.1 mM ATP and ADP, respectively, lowering $n_{H}$ to 1.0. Univalent cations like $K^+, Na^+ and Li^ +$ activated the enzyme but some divalent cations such as $Cu^{ 2+} and Cd^{2+}$ showed strong inhibitory effects. This enzyme displayed optimum activity at $30^{\circ}C$ and pH 7.0. In addition, it was stable up to $45^{\circ}C$ and over a wide pH range(pH 5.5-9.0). Amino acid sequences of its N-terminal region were analyzed to be ADYKMQMFADDA.

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Evaluation of Selective Media for Isolation of Staphylococcus aureus from Agricultural Products (농산물 중 Staphylococcus aureus의 분리를 위한 선택배지 평가)

  • Kim, Se-Ri;Lee, Seo-Hyun;Seo, Min-Kyoung;Kim, Won-Il;Park, Kyeong-Hun;Yun, Hye-Jeong;Yoon, Yo-Han;Yoo, Soon-Young;Ryu, Kyoung-Yul;Yun, Jong-Chul;Kim, Byung-Seok
    • Journal of Food Hygiene and Safety
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    • v.27 no.2
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    • pp.169-175
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    • 2012
  • Five kinds of selective media, such as mannitol salt agar (MSA), Baird-Parker agar (BPA), Baird-Parker supplemented with rabbit plasma fibrinogen (BPA+RPF), CHROMagar Staphylococcus aureus (CSA), and Petrifilm Staph Express count system (Petrifilm), were compared to recommend the optimum selective media for isolation of Staphylococcus aureus from agricultural products. Seventy four target and non target bacteria were inoculated on five selective media to analyze sensitivity and specificity. In the recovery test of injured S. aureus cells, S. aureus was exposed to acid (1% lactic acid for 10 min), heat ($60^{\circ}C$ for 90s), and cold ($-20^{\circ}C$ for 1h) conditions. And artificially contaminated agricultural products (iceberg lettuce, green pepper, and cherry tomato) was enumerated on five selective media. The sensitivity of BPA+RPF, CSA, Petrifilm, MSA, and BPA were 100%, 100%, 100%, 90.5%, 90.5%, respectively. In addition, the specificity of BPA+RPF, CSA, MSA, BPA and Petrifilm were 100%, 100%, 84.6%, 75.0%, 67.3%, respectively. However, no difference among five selective media was observed in recovery on injured S. aureus cell and enumeration from agricultural products. This results suggest that BPA+RPF and CSA are the optimum media for detection of S. aureus from agricultural products.

Survey on the Status of Microbial Contamination of Chicken Meats Collected from Poultry Processing Plants in Nationwide (우리나라 도계장 수거계육의 미생물학적 위생실태 조사)

  • Woo, Yong-Ku
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.186-192
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    • 2007
  • This study was conducted to survey the hygienic status of chicken meats on the microbial levels, which were collected from poultry processing plants located in the local provinces in nationwide including the JeJu island (n=15) in 1997. In particular, Salmonella spp., Campylobacter jejuni, and Listeria monocytogenes, which were retarded as one of the most important entero-pathogens relating to food home illness from poultry, were investigated on their isolation frequency including the other pathogens related on the food-borne illness. A total of 115 processed chickens were submitted on the present study. In general, the bacterial contamination frequency showed more or less lower $(10{\sim}100 cells)$ than those of sold on the retail and super markets and department stores because of lacking of cross-contamination incidences, depending on the total cells, Coliforms and Staphylococcal cells count. While, Salmonella species, Campylobacter jejuni, Listeria monocytogenes, and coagulase positive Staphylococcus aureus isolation frequency of chicken meats from slaughter houses were 58.3%, 37.4%, 43.5%, and 30.4%, in order. But the present microbial isolation data were a little lower levels than those of sold on the retail and super markets and famous department stores in Seoul and GyeongGi province at the same period. It seemed that the cross-contamination problems (including the human, environmental and instrumental factors) during the marketing stage (after the last processing procedure; rinsing step) had the major roles on the increasing of the microbial contamination frequency on the chicken meats after the slaughter houses.

Isolation of lysozyme producing bacteria capable of solubilizing microbial cells (미생물 용해가 가능한 Iysozyme 분비 균주의 분리 및 특성)

  • Guo, Pengfei;Seo, Sun-Keun;Zhang, Lei;Kim, Hyo-Sang;Oh, Young-Khee;Jahng, Deok-Jin
    • KSBB Journal
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    • v.23 no.3
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    • pp.187-192
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    • 2008
  • Lysozyme-producing microorganisms were isolated to obtain bacteria which can efficiently solubilize microbial cells. Cells of normal and chloroform-treated Escherichia coli and Micrococcus Iysodeikticus were used as model substrates to isolate lysozyme-producing microorganisms and investigate the efficiency of cell lysis. The culture supernatant of the isolate New1 (98% similarity of 16S rDNA sequence with Thermomonas haemolytica) showed different lytic characteristics for different substrates. Thermal treatment (autoclave) of substrate cells showed a significant effect on cell solubilization by culture supernatant of the New1. For autoclaved substrate cells, E. coli, M. Iysodeikticus and chloroform-treated E. coli were solubilized by 58.7%, 49.4% and 79.1%, respectively, in the culture supernatant of New1. The lytic activity of New1 was mainly caused by lysozyme produced by the isolate. It was also showed that New1 exhibited high protease activity and a little cellulase activity.

Studies on the Microbial Glucose Isomerase Part 1. Isolation and Characterization of Streptomyces species Producing Glucose Isomerase (미생물의 포도당 이성화효소에 관한 연구 (제1보) 포도당 이성화효소 생산균주의 분리 및 성질에 관하여)

  • Chung, Tai-Wha;Kim, Hyun U.;Moon H. Han
    • Microbiology and Biotechnology Letters
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    • v.4 no.4
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    • pp.138-144
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    • 1976
  • five strains of Streptomyces spp. with high Productivity of glucose isomerase (15-30 units/ml) were obtained among 280 microbial strains isolated from 150 soil samples. These strains produced glucose isomerase with xylose as an inducer. These 5 strains were also identified to be different strains of Streptomyces spp.:streptomyces sp. K-14, K-53, K-71, K-77 and K-733. It was found that Streptomyces sp. K-14 produced the highest enzyme activity. The spore chains of these strains were rectiflexible and spore surface was smooth except Steptomyces sp. K-77 and K-733, with spiny surface.

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