• Title/Summary/Keyword: Microbiology Quality

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Quality Control Probes for Spot-Uniformity and Quantitative Analysis of Oligonucleotide Array

  • Jang, Hyun-Jung;Cho, Mong;Kim, Hyung-Hoi;Kim, Cheol-Min;Park, Hee-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.19 no.7
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    • pp.658-665
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    • 2009
  • Quality control QC for spot-uniformity is a critical point in fabricating an oligonucleotide array, and quantification of targets is very important in array analysis. We developed two new types of QC probes as a means of confirming the quality of the uniformity of attached probes and the quantification of targets. We compared the signal intensities and fluorescent images of the QC and target-specific probes of arrays containing only target-specific probes and those containing both QC and target-specific probes. In a comparison of quality control methods, it was found that the arrays containing QC probes could check spot-uniformity or spot defects during all processes of array fabrication, including after spotting, after washing, and after hybridization. In a comparison of quantification results, the array fabricated by the method using QC probes showed linear and regular results because it was possible to normalize variations in spot size and morphology and amount of attached probe. This method could avoid errors originating in probe concentration and spot morphology because it could be normalized by QC probes. There were significant differences in the signal intensities of all mixtures (P<0.05). This result indicates that the method using QC probes is more useful than the ordinary method for quantification of mixed target. In the quantification of mixed targets, this method could determine a range for mixed targets of various amounts. Our results suggest that methods using QC probes for array fabrication are very useful to the quality control of spots in the fabrication processes of quantitative oligonucleotide arrays.

Heterologous Expression and Characterization of a Novel Exo-Polygalacturonase from Aspergillus fumigatus Af293 and Its Application in Juice Extraction

  • Chengwei Yang;Ting Zhang;Jing Zhu;Yunyi Wei;Furong Zhu;Zhong Cheng
    • Journal of Microbiology and Biotechnology
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    • v.33 no.4
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    • pp.533-542
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    • 2023
  • Exo-polygalacturonase (exo-PG) hydrolyzes pectin acids and liberates mono-galacturonate, which plays an important role in juice extraction, and has rarely been reported. Exo-PG (AfumExoPG28A) from Aspergillus fumigatus belongs to the glycoside hydrolase 28 family. In this study, its gene was cloned and the protein was expressed and secreted in Pichia pastoris with a maximal activity of 4.44 U/ml. The optimal temperature and pH of AfumExoPG28A were 55℃ and 4.0, respectively. The enzyme exhibited activity over almost the entire acidic pH range (>20.0% activity at pH 2.5-6.5) and remained stable at pH 2.5-10.0 for 24 h. The Km and Vmax values of AfumExoPG28A were calculated by the substrate of polygalacturonic acid as 25.4 mg/ml and 23.6 U/mg, respectively. Addition of AfumExoPG28A (0.8 U/mg) increased the light transmittance and juice yield of plantain pulp by 11.7% and 9%, respectively. Combining AfumExoPG28A (0.8 U/mg) with an endo-PG (0.8 U/mg) from our laboratory, the enzymes increased the light transmittance and juice yield of plantain pulp by 45.7% and 10%, respectively. Thus, the enzyme's potential value in juice production was revealed by the remarkable acidic properties and catalytic activity in fruit pulp.

Relationships between fruit-body development of Pleurotus ostreatus and environmental conditions in mushroom house

  • Jhune, Chang-Sung;Kong, Won-Sik;You, Young-Bok;Chun, Se-Chul
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.19-23
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    • 2003
  • Temperature conditions in the mushroom cultivating room affected color, yield, pinheading rate, growth rate and other characteristics of fruitbody. These results seemed to tell the quality of mushroom. Carbon dioxide gas generated from respiration of mushroom also made stipe length long and pilei size small. High concentration of carbon dioxide could make fruitbody abnormal or dead. Mycelial shapes in fruitbody inner tissue were different according to the part and the size of fruitbody.

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Expression of Codon Optimized β2-Adrenergic Receptor in Sf9 Insect Cells for Multianalyte Detection of β-Agonist Residues in Pork

  • Liu, Yuan;Wang, Jian;Liu, Yang;Yang, Liting;Zhu, Xuran;Wang, Wei;Zhang, Jiaxiao;Wei, Dong
    • Journal of Microbiology and Biotechnology
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    • v.29 no.9
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    • pp.1470-1477
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    • 2019
  • ${\beta}_2$-adrenergic receptor (${\beta}_2-AR$) was expressed efficiently using Bac-to-Bac Baculovirus Expression System in Sf9 cells as a bio-recognition element for multianalyte screening of ${\beta}$-agonist residues in pork. Sf9 cells were selected as the expression system, and codon optimization of wild-type nucleic acid sequence and time-dependent screening of expression conditions were then carried out for enhancing expression level and biological activity. Under optimum conditions of multiplicity of infection (MOI) = 5 and 48 h post transfection, the protein yield was up to 1.23 mg/ml. After purification by chromatographic techniques, the purified recombinant protein was applied to develop a direct competitive enzyme-linked receptor assay (ELRA) and the efficiency and reliability of the assay was determined. The IC50 values of clenbuterol, salbutamol, and ractopamine were 28.36, 50.70, and $59.57{\mu}g/l$, and clenbuterol showed 47.61% and 55.94% cross-reactivities with ractopamine and salbutamol, respectively. The limit of detection (LOD) was $3.2{\mu}g/l$ and the relevant recoveries in pork samples were in the range of 73.0-91.2%, 69.4-84.6%, and 63.7-80.2%, respectively. The results showed that it had better performance compared with other present nonradioactive receptorbased assays, indicating that the genetically modified ${\beta}_2-AR$ would have great application potential in detection of ${\beta}$-agonist residues.

Changes of Enzyme Activities and Compositions of Abnormal Fruiting Bodies Grown under Artificial Environmental Conditions in Pleurotus ostreatus

  • Jang, Kab-Yeul;Cho, Soo-Muk;June, Chang-Sung;Weon, Hang-Yeon;Park, Jeong-Sik;Choi, Sun-Gyu;Cheong, Jong-Chun;Sung, Jae-Mo
    • Mycobiology
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    • v.33 no.1
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    • pp.30-34
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    • 2005
  • This study investigated the biochemical changes of abnormal fruiting bodies grown under artificial environmental conditions in P. ostreatus. Abnormal mushroom growth during cultivation damages the production of good quality mushroom. This study showed that different environmental conditions produced morphological changes in the fruiting bodies of P. ostreatus. The fruiting bodies with morphological changes were collected and examined for differences in biochemical properties, enzyme activities, and carbohydrates composition. The enzyme activities assay showed that glucanase and chitinase activities decreased when the temperature was below or above the optimum cultivation temperature for P. ostreatus. The biochemical compositions of the abnormal mushroom were significantly different from the normal fruiting bodies. It was suggested that the changes in the biochemical composition of abnormal mushroom were caused by the unfavorable environmental conditions during mushroom cultivation.

Studies on the Microbial Utilization of Agricultural Wastes (Part 11) Properties of Cellulolytic Enzyme Produced by a Cellulolytic Fungus Trichodrma sp. KI 7-2 and its Application to the Fermented Feed Production (농산폐자원의 미생물학적 이용에 관한 연구(제11보) Trichoderma sp KI 7-2가 생산하는 섬유소분해효소의 성질 및 발효사요에의 응용)

  • Bae, Moo;Lee, Gye-Jun;Tak, Sun-Mi;Kim, Byung-Hong
    • Microbiology and Biotechnology Letters
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    • v.6 no.1
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    • pp.1-8
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    • 1978
  • In order to develop the processes for the production of fermented feed from cellulosic agricultural by-product, cereal straw, by th action of cellulolytic fungus, the properties of the cellulolytic enzyme produced by Trichoderma sp. KI 7-2 was studied. A higher enzyme activity was obtained in the culture added by 1% rice or barley straw powder than in the culture of pure cellulose. The crude enzyme was prepared by precipitating from 20∼60% saturated ammonium sulphate of the culture supernatant. The optimum conditions for the enzyme reaction were temperature of of 50$^{\circ}C$ and pH 4.2. The crude enzyme was static at 50$^{\circ}C$ for two hours and at pH between 4 and 6. These properties were adopted for the fermented feed production, and several production. Thus, several processes of semisolid culture were devicced to up grade tile fermented feed and to develop into the acceptable quality.

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Isolation and Characterization of Lactic Acid Bacteria from Fermented Goat Milk in Tajikistan

  • Cho, Gyu-Sung;Cappello, Claudia;Schrader, Katrin;Fagbemigun, Olakunle;Oguntoyinbo, Folarin A.;Csovcsics, Claudia;Rosch, Niels;Kabisch, Jan;Neve, Horst;Bockelmann, Wilhelm;Briviba, Karlis;Modesto, Monica;Cilli, Elisabetta;Mattarelli, Paola;Franz, Charles M.A.P
    • Journal of Microbiology and Biotechnology
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    • v.28 no.11
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    • pp.1834-1845
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    • 2018
  • The lactobacilli associated with a fermented goat milk product from Tajikistan were isolated to characterize their technological properties and antibiotic resistances in order to assess their suitability for development as starter cultures. In this study, twenty three strains were identified by 16S rRNA sequencing as typical dairy-associated lactic acid bacterial strains, i.e. L. plantarum, L. pentosus, L. delbrueckii, L. helveticus and L. paracasei. These strains were generally susceptible to most antibiotics tested in this study and this allowed a selection of strains as safe starters. The draft genomes of four representative strains were sequenced and the number of contigs of the four assembled genomes ranged from 51 to 245 and the genome sizes ranged from 1.75 to 3.24 Mbp. These representative strains showed differences in their growth behavior and pH-reducing abilities in in vitro studies. The co-inoculation of these Lactobacillus spp. strains together with a yeast Kluyveromyces marxianus MBT-5698, or together with the yeast and an additional Streptococcus thermophilus MBT-2, led to a pH reduction to 3.4 after 48 h. Only in the case of fermentation inoculated with the co-culture, the viscosity of the milk increased noticeably. In contrast, fermentations with single strains did not lead to gelation of the milk or to a decrease in the pH after 24h. The results of this study provide a comprehensive understanding of the predominant lactobacilli related to Tajikistani fermented milk products.

Influence of Pulsed Electric Field on Accumulation of Calcium in Lactobacillus rhamnosus B 442

  • Goral, Malgorzata;Pankiewicz, Urszula;Sujka, Monika;Kowalski, Radoslaw;Giral, Dariusz;Kozlowicz, Katarzyna
    • Journal of Microbiology and Biotechnology
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    • v.30 no.1
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    • pp.44-53
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    • 2020
  • Calcium is an element that performs many important functions in the human body. A study was conducted on the use of a pulsed electric field (PEF) to enrich cells of Lactobacillus rhamnosus B 442 in calcium ions. The highest concentration of calcium ions in bacterial cells (7.30 mg/g d.m.) was obtained at ion concentration of 200 ㎍/ml of medium and with the use of the following PEF parameters: field strength 3.0 kV/cm, exposure time 10 min, pulse width 75 ms and 20 h of culturing after which bacteria were treated with the field. Cell biomass varied in the range from 0.09 g/g d.m. to 0.252 g/g d.m., and the total number of bacteria ranged from 1010 CFU/ml to 1012 CFU/ml. Microscope photographs prove that calcium ions were situated within the cells of the bacteria, and electroporation contributed to an increase in the effectiveness of the ion bioaccumulation process. Samples containing calcium and subjected to electroporation displayed intensive fluorescence. The significance of this research was the possibility of using probiotic bacteria enriched with calcium ions for the production of functional food in subsequent studies.

Starter Cultures for Kimchi Fermentation

  • Lee, Mo-Eun;Jang, Ja-Young;Lee, Jong-Hee;Park, Hae-Woong;Choi, Hak-Jong;Kim, Tae-Woon
    • Journal of Microbiology and Biotechnology
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    • v.25 no.5
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    • pp.559-568
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    • 2015
  • Kimchi is a traditional Korean vegetable product that is naturally fermented by various microorganisms present in the raw materials. Among these microorganisms, lactic acid bacteria dominate the fermentation process. Natural fermentation with unsterilized raw materials leads to the growth of various lactic acid bacteria, resulting in variations in the taste and quality of kimchi, which may make it difficult to produce industrial-scale kimchi with consistent quality. The use of starter cultures has been considered as an alternative for the industrial production of standardized kimchi, and recent trends suggest that the demand for starter cultures is on the rise. However, several factors should be carefully considered for the successful application of starter cultures for kimchi fermentation. In this review, we summarize recent studies on kimchi starter cultures, describe practical problems in the application of industrial-scale kimchi production, and discuss the directions for further studies.

Lactobacillus plantarum을 용균시키는 Bacteriophoge SC921의 분리 및 특성

  • Yoon, Sung-Sik;Shin, Young-Jae;Choi, Hak-Jong;Her, Song;Oh, Doo-Hwan
    • Microbiology and Biotechnology Letters
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    • v.25 no.1
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    • pp.96-101
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    • 1997
  • Among the lactic flora responsible for the development of acidity and characteristic flavor of Kimchi which is a traditional fermented Chiness cabbage. Homofermentative Lactobacillus plantarum is rod-shaped and to be known to ewert major role during later fermentation period. Once this strain establishes main flaora in the Kimchi fermentation process, it gives rise to excess acid production to reduce the taste and quality of Kimchi during storage. As a primary work to increase the keeping quality using virulent Lactobaillus plantarum bacteriophages, it was isolated sucessfully from collected Kimchi samples and their characteristics were studied. The new isolated phage, named SC 921, adsorbed to its host without Ca$^{2+}$, and nearly eliminated at 60$\circ $C of heat treatment for 5 min. This phages were atable at pH 4~ 10 but inactivated below pH 3.0 or pH 11.0 above. The latent period, rise period, and burst size of this phage was 100 min, 120 min, 31$\pm $2pfu/ml, respectively. Electron micrograph showed the phages particles were unusually oval feature of head (dia 80~ 120 nm) without contractile tail.

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