• The Plant Pathology Journal
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• 제31권3호
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• pp.212-218
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• 2015

In this study, we developed a species-specific PCR assay for rapid and accurate detection of three Xanthomonas species, X. axonopodis pv. poinsettiicola (XAP), X. hyacinthi (XH) and X. campestris pv. zantedeschiae (XCZ), based on their draft genome sequences. XAP, XH and XCZ genomes consist of single chromosomes that contain 5,221, 4,395 and 7,986 protein coding genes, respectively. Species-specific primers were designed from variable regions of the draft genome sequence data and assessed by a PCR-based detection method. These primers were also tested for specificity against 17 allied Xanthomonas species as well as against the host DNA and the microbial community of the host surface. Three primer sets were found to be very specific and no amplification product was obtained with the host DNA and the microbial community of the host surface. In addition, a detection limit of $1pg/{\mu}l$ per PCR reaction was detected when these primer sets were used to amplify corresponding bacterial DNAs. Therefore, these primer sets and the developed species-specific PCR assay represent a valuable, sensitive, and rapid diagnostic tool that can be used to detect three specific pathogens at early stages of infection and may help control diseases.

• 한국약용작물학회지
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• 제25권2호
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• pp.108-114
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• 2017

Background: The study was conducted to investigate the distributions of faecal bacteria in commercial oriental medicine herb products. Methods and Results: A survey was conducted on the microbial contamination levels and antimicrobial specificity of Bacillus cereus and other microbes using 106 oriental medicine herb products on sale in Seoul. Pouring and isolation methods such as standard plate counts were used to identify the bacteria. The isolated bacterias included coliforms, Bacillus spp., Enterococcus spp., Staphylococcus spp., Listeria spp.were identified by using gram staining and an API (analytical profile index) kit. Antimicrobial drugs discs were determined by CLSI (clinical and laboratory standards institute). Conclusions: The bacterial isolates present in the herbal medicines included 98 coliforms, 45 Bacillus spp., 29 Enterococcus spp., and 2 Listeria spp. Among these, there were nine Bacillus cereus strains, one Enterococcus faecium strain, and one Enterococcus faecalis strain present. The 9 Bacillus cereus strains were tested for susceptibility to 36 types of antibiotics products by the disc diffusion method. The strains showed resistance to 13 of these antibiotic products and semi-resistance to 5 antibiotic products. On the basis of these results, any oriental medicine herb product can be assumed to be contain resistant or semi-resistant bacterial strains. Therefore, we suggest prescribing guidelines and special management for the use of antibiotics in farms producing oriental medicine herb products.

• Journal of Microbiology
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• 제42권3호
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• pp.216-222
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• 2004

In a previous paper, the ogdH gene that encodes 2-oxoglutarat dehydrogenase was isolated from Salmonella typhimurium. The catalytic N-terminal region in the enzyme was found to be very specific for the Salmonella species. Therefore, the aim of the present study was to detect S. typhimurium in food sources using primers designed for OGDH-l and OGDH-2 which were based on the salmonella-specific region of the ogdH gene. A simple polymerase chain reaction (PCR) detection method was developed to detect low numbers of S. typhimurium in a chicken meat microbial consortium. Using the ogdH-specific primers under stringent amplification conditions and for gene probe analysis, fewer than 100 colony-forming units (CFUs) were detectable when pure cultures were employed. When the PCR assay was run on S. typhimurium-contaminated meat contents, only the positive meat samples containing as few as 200 CFUs reacted to the assay. The method employed for sample processing is simple and it was determined to provide a sensitive means of detecting trace amounts of S. typhimurium-specific sequences in the presence of mixed meat microbial populations. When compared with six representative intestinal gram-negative bacterial strains in foods, including Vibrio parahaemolyticus, V. vulnificus, Enterobacter cloacae, E. coli O157:H7, Pseudomonas aeruginosa, and Proteus sp., S. typhimurium had a unique and distinct PCR product (796 bp). In conclusion, the two OGDH primers were found to be rapid and sensitive detectors of Salmonella spp for the PCR method.

• BMB Reports
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• 제45권2호
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• pp.59-70
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• 2012

Carbon catabolite repression (CCR) is a key regulatory system found in most microorganisms that ensures preferential utilization of energy-efficient carbon sources. CCR helps microorganisms obtain a proper balance between their metabolic capacity and the maximum sugar uptake capability. It also constrains the deregulated utilization of a preferred cognate substrate, enabling microorganisms to survive and dominate in natural environments. On the other side of the same coin lies the tenacious bottleneck in microbial production of bioproducts that employs a combination of carbon sources in varied proportion, such as lignocellulose-derived sugar mixtures. Preferential sugar uptake combined with the transcriptional and/or enzymatic exclusion of less preferred sugars turns out one of the major barriers in increasing the yield and productivity of fermentation process. Accumulation of the unused substrate also complicates the downstream processes used to extract the desired product. To overcome this difficulty and to develop tailor-made strains for specific metabolic engineering goals, quantitative and systemic understanding of the molecular interaction map behind CCR is a prerequisite. Here we comparatively review the universal and strain-specific features of CCR circuitry and discuss the recent efforts in developing synthetic cell factories devoid of CCR particularly for lignocellulose-based biorefinery.

• 한국미생물·생명공학회지
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• 제20권6호
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• pp.687-692
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• 1992

혼합배양중의 각 미생물의 생균수 측정은 순수배양보다 훨씬 복잡하다. 특히 두 균주의 크기가 비슷한 경우에는 사용할 수 있는 방법이 더 제한된다. 본 연구에서는 두 균의 크기가 비슷한 경우에도 적용될 수 있는 간단한 생균수 측정방법을 개발하였다. 미생물 배양액의 산소흡수속도(OUR)는 세포수에 비례하며 이때의 비례상수인 최대 비산소흡수속도( maximum specific OUR)를 알고 있으면 배양액의 OUR을 측정함으로써 간접적으로 생균수를 구할 수 있게된다. 혼합배양의 경우 산소흡수속도는 각 미생물의 호흡속도의 합이 되며, 각 미생물의 호흡속도가 서로 다르고 또한 온도의존성이 다르다면 호흡속도의 측정을 이용하여 각 생균수를 간접적으로 측정할 수 있다.

• 한국식품과학회지
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• 제23권6호
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• pp.689-694
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• 1991

우리나라 신라시대를 전후하여 일반적인 청량음료로 사용되었던 것으로 알려진 장수(?水)의 제조방법을 제민요술에 근거하여 재현하고 그 발효학적 특징을 규명 하려 하였다. 새로지은 뜨거운 쌀밥을 항아리에 담고 찬물을 부어 3일간 상온에서 발효한 후 윗물과 밥 절반을 떠낸 후 다시 새로지은 뜨거운밥과 찬물을 채워 넣는 일을 주기적으로 반복하면서 미생물균총의 조성을 관찰하였다. 밥과 물을 혼합하여 실온에 방치하면 3일 동안 총균수, 단백 질 분해균, 산생성균수가 지수적으로 증가하나 그 이후부터 다소 감소하며 6일 후에는 단백질 분해균수가 증가하여 부패현상을 나타낸다. 그러나 장수 제조법에 의하면 단백질 분해균이 크게 감소하고 산생성균이 지배균총이 된다. 장수에서 분리한 주요 산생성균은 Lactobacillus, Lactococcus, Pediococcus, Leuconostoc spp.였으며 이중 주요균으로 Lactococcus thermophilus, Lactobacillus coryniformis 및 Leuconostoc mesenterioides를 잠정 동정하였다. 분리한 유산균 들을 starter로 사용하여 장수를 제조한 결과 산생산력이 크고 풍미가 우수한 제품을 얻을 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제13권3호
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• pp.451-456
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• 2003

An expression vector system was developed for the secretory production of recombinant Bacillus stearothermophilus L1 lipase in Saccharomyces cerevisiae. The mature L1 lipase gene was fused to ${\alpha}-amylase$ signal sequence from Aspergillus oryzae for the effective secretion into the culture broth and the expression was controlled under GAL10 (the gene coding UDP-galactose epimerase of S. cerevisiae) promoter. S. cerevisiae harboring the resulting plasmid successfully secreted L1 lipase into the culture broth. To examine an optimum condition for L1 lipase expression in the fed-batch culture, L1 lipase expression was induced at three different growth phases (early, mid, and late-exponential growth phases). Maximum product on of L1 lipase (1,254,000 U/l, corresponding to 0.65/1) was found when the culture was induced at an early growth phase. Secreted recombinant L1 lipase was purified only through CM-Sepharose chromatography, and the purified enzyme showed 1,963 U/mg of specific activity and thermoalkalophilic properties similar to those reported for the enzyme expressed in Escherichia coli.

• Journal of Microbiology and Biotechnology
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• 제20권9호
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• pp.1351-1358
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• 2010

The demand for ${\beta}$-glucosidases insensitive to product inhibition is increasing in modern biotechnology, for these enzymes would improve the process of saccharification of lignocellulosic materials. In this study, a ${\beta}$-glucosidase gene that encodes a 442-amino-acid protein was isolated from a marine microbial metagenomic library by functional screening and named as bgl1A. The protein was identified to be a member of the glycoside hydrolases 1 family, and was recombinantly expressed, purified, and biochemically characterized. The recombinant ${\beta}$-glucosidase, Bgl1A, exhibited a high level of stability in the presence of various cations and high concentrations of NaCl. Interestingly, it was activated by glucose at concentrations lower than 400 mM. With glucose further increasing, the enzyme activity of Bgl1A was gradually inhibited, but remained 50% of the original value in even as high as 1,000 mM glucose. These findings indicate that Bgl1A might be a potent candidate for industrial applications.

• 한국식생활문화학회지
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• 제10권1호
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• pp.67-74
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• 1995

This study researched microbial change of quality according to the various phases of product flow of cooked pea and rice, cold cucumber and seaweed soup, soybean sprouts japchae feeding urban type of a commissary school and a satellite school in Daejeon area, also it suggested the possibility that the central commissary foodservice system can be established and utilized more developmental to identify its food of variation of temperature and state of safety unitl 3 hours after cooking for the case of delay of distribution and holding because of the satellite school of geographical location and traffic problem. The critical Control Points identified for each category of menu items were: Boiled pea and rice: inadequate distribution, holding and storing before assembly; Cold cucumber and seaweed soup: pre-preparation and post-preparation after cooking; Soybean sprouts japchae: Pre-preparation, post-preparation and storing. As the result of observation of the variation of temperature and microbial safety according to the delay of distribution and holding for each food, all of them were relatively safe until 3 hours after cooking, but cold cucumber and seaweed soup being stored for 3 hours, the value of E. coli is $10^3$ CFU/g. The variation of temperature was more extreme in soybean sprouts japchae than cooked pea and rice and cold cucumber and seaweed soup. It was proved that the stainless container was excellent and that adequate holding container should be used.

• Asian-Australasian Journal of Animal Sciences
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• 제20권3호
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• pp.399-404
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• 2007

To elucidate the efficacy of different levels of microbial-fermented soy proteins (FSP) on piglet performance, a total of 240 weaned piglets ($L{\times}Y{\times}D$, $22{\pm}3$ d of age, $5.16{\pm}0.07$ kg initial BW) were allotted to 4 treatment diets comprising control, FSP-3%, FSP-6% and FSP-9%. The fermented soybean product named $Pepsoygen^{(R)}$ was utilized for the study. There were 15 pigs per pen and 4 pens per treatment. The control diet contained 15% soybean meal (SBM), and SBM for the treatment diets was replaced at 3, 6 and 9% with FSP, respectively. Experimental diets were fed from 0 to 14 d (phase-I) after weaning and then a common commercial diet was fed from 15 to 35 d (phase-II). There was a linear (p<0.05) increase in ADG and ADFI at both phases of measurement. The feed to gain ratio was also improved, showing a linear (p<0.01) trend as the level of supplementation increased. Except for phosphorus, the digestibility of all other nutrients was improved linearly (p<0.05) in the FSP added diets. However, villous height and crypt depth were not affected by dietary treatments. No special effect on intestinal morphology was noticed between FSP-added and control diets. In conclusion, the growth, digestibility of nutrients and morphological changes in weaned pigs fed FSP showed improved performance at higher levels of supplementation.