• Title/Summary/Keyword: yeast selection

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Selection of Yeast Mutant Strain with High RNA Content and Its High Cell-Density Fed-Batch Culture. (고함량 RNA 효모 변이주의 선별 및 고농도세포 유가배양)

  • 김재범;권미정;남희섭;김재훈;남수완
    • Microbiology and Biotechnology Letters
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    • v.30 no.1
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    • pp.68-72
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    • 2002
  • To obtain a yeast mutant with high RNA content and high growth rate, Saccharomyces cerevisiae MTY62 was mutated with ethylmethane sulfonate. Among the selected mutants that were sensitive to the high concentration of KCl, M40-10 strain was finally selected due to its rapid cell growth and high RNA content in the tube and baffled-flask cultures. In the batch culture of M40-10 mutant, the maximum specific growth rate ($\mu_{max}$) of $0.38 h^{-1}$ , RNA concentration of 3210 mg-RNA/1, and RNA content of 183 mg-RNA/g-DCW were obtained, which were 23%, 15%, and 12% increased levels, respectively, compared to those of MTY62 parent strain. The intermittent fed-batch culture of M40-10 strain resulted in the maximum cell concentration of 35.6 g-DCW/1, RNA concentration of 5677 mg/1, and RNA content of 160 mg-RNA/g-DCW. Through the constant fed-batch culture, the maximum cell concentration of 46.4 g-DCW/1, RNA concentration of 6270 mg-RNA/1, and RNA content of 135 mg-RNA/g-DCW were obtained. At the 20 h culture time in the fed-batch cultures of M40-10 strain, the cell and RNA concentrations were increased by 30% and 10%, respectively, over the parent strain MTY62. In addition, it was also found that the accumulated RNA within the mutant cell was not degraded until the end of fed-batch cultivation, indicating that the M40-10 cell is a mutant with weak acidic RNase activity.y.

Selection of koji and yeast strain for improvement of Choungju quality (청주의 주질 개선을 위한 국 및 효모의 선정과 그 발효 특성)

  • Shin, Cheol-Seung;Park, Yoon-Joong;Lee, Suk-Kun
    • Applied Biological Chemistry
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    • v.39 no.1
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    • pp.9-15
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    • 1996
  • To improve the quality of Choungju. a kind of rice wine, two different types of koji were prepared and compared : one from wheat bran with Aspergillus usamii mut. shirousami Y-79 and the other from rice with A. oryzae, and yeast strains from cereal wine mashes were newly isolated and applied for the brewing method. Levels of the related enzymes such as glucoamylase, ${\alpha}-amylase$ and acid protease in the wheat bran koji were higher than those in the rice koji, whereas vice versa in the case of acid carboxypeptidase. An amount of $2{\sim}3%$ wheat bran koji to the weight of total rice was adequate for saccharification of the mash and resulted in improved duality of the fermented mash, accompanied by decrease in koji ordor and amino acidity. When the solution of wheat bran koji and the isolated yeast strains were employed, the better Choungju taste was obtained in comparison with those fermented with Japanese sake yeasts, the strain K-7 and 9, due to the lower content of organic acids especially succinic acid. The amino acidity of the fermented mash was able to be controlled to some extent, when the rico types of koji and the isolated strains were employed, by changing the ratio of the two koji types. However, the application of the rice koji with the isolated strains was not desirable for the brewing process because organic acids were produced in excess and ethanol fermentation was retarded.

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Selection of Lactic Starter for the Improvement of Jeungpyun Manufacturing Process (증편 제조공정 개설을 위한 스타터 선발)

  • Moon, Hye-Joon;Chang, Hak-Gil;Mok, Chul-Kyoon
    • Korean Journal of Food Science and Technology
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    • v.31 no.5
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    • pp.1241-1246
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    • 1999
  • Jeungpyun has a unique sponge-like texture and sour taste imparted by the lactic acid and alcohol that are produced by the addition of Takju(turbid rice wine) as a starter. Its consumption, however. has been decreased due to the long preparation time, the difficulties in quality control and the offensive odor derived from the Takju. The present study was carried out in order to shorten the preparation time and to improve the quality of Jeungpyun. To achieve the objectives an appropriate commercial lactic acid starter was selected and a cofermentation system with yeast was developed. A starter containing Lactococcus lactis, Lactococcus cremoris and Lactococcus diacetylactis was selected based on the acid production rate and the quality of the produced sour taste. It took 3 hr for the lactic acid fermentation of rice slurry. The optimum addition levels of the lactic acid starter and yeast were 0.45% and 0.60%, respectively. The lactic acid fermented rice slurry was mixed with the rice slurry separately fermented for 2 hr by yeast, and cofermented for another 1 hr before steaming. Jeungpyun Prepared by the developed method was superior in quality to that Prepared by conventional method using Takju. The developed method reduced the preparation time more than 50% compared with the conventional method.

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Microbiological Characteristics of Low Salt Mul-kimchi (저염 물김치의 미생물균총 특성)

  • Oh, Ji-Young;Hahn, Young-Sook;Kim, Young-Jin
    • Korean Journal of Food Science and Technology
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    • v.31 no.2
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    • pp.502-508
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    • 1999
  • Microbiological characteristics of low salt Mul-kimchi was examined. Mul-kimchi was prepared by mixing of radish (25%), green onion (2.4%), red pepper (1.9%), garlic (1.9%) and salt (0, 0.2, 0.5, 1.0, 1.5, 2.0, 2.5, 2.5, 3.0%) in water and fermented at 4, 15 and $25^{\circ}C$ for 10 days, respectively. During fermentation period, total cell, Leuconostoc sp., Lactobacillus sp., Streptococcus sp., Pediococcus sp., coliform bacteria, gram (-) bacteria and yeast cell number were counted on their selection media. The microbes in Mul-kimchi were isolated and identified. Total cell number increased as salt concentration decreased and fermentation temperature increased. Lactic acid bacteria showed the highest number in 1.0% salt concentration. Yeast cell number increased with increase of salt concentration. Lactobacillus sp. were identified Lactobacillus plantarum and L. pentosus in Mul-kimchi containing $0.2{\sim}1.0%$ salt while those of Mul-kimchi containing 3.0% salt were Lactobacillus plantarum and L. brevis. The other lactic acid bacteria were identified Leuconostoc citrum, Leu.mes.ssp.mesenteroides/dextranicum and streptococcus facium in Mul-kimchi containing $0{\sim}3.0%$ salt while Pediococcus sp. was not detected. Gram-negative Aeromonas hydrophila, Pseudomonas fluorescens, Pseu. aureofaciens and yeast Candida pelliculosa, Cryptococcus laurentii were identified in the Mul-kimchi.

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Debaryomyces hansenii Strains from Valle De Los Pedroches Iberian Dry Meat Products: Isolation, Identification, Characterization, and Selection for Starter Cultures

  • Ramos, Jose;Melero, Yessica;Ramos-Moreno, Laura;Michan, Carmen;Cabezas, Lourdes
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1576-1585
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    • 2017
  • Yeasts, filamentous fungi, and bacteria colonize the surface of fermented sausages during the ripening process. The source of this microbiota is their surrounding environment, and is influenced by the maturing conditions and starter cultures. Debaryomyces hansenii was previously isolated from several dry-cured meat products and associated with the lipolytic and proteolytic changes that occur in these products, influencing their taste and flavor. Therefore, this study isolated the yeast microbiota present in the casing from different meat products ("lomo," "chorizo," and "$salchich{\acute{o}}n$") from the Valle de los Pedroches region in southern Spain. D. hansenii was by far the most abundant species in each product, as all 22 selected isolates were identified as D. hansenii by biochemical and/or molecular methods. In contrast, no yeasts were found in the meat batter. These data constitute the first study of the yeasts present in "lomo" sausages and particularly the highly appreciated Valle de los Pedroches "lomo" sausages. Furthermore, the resistance of these isolates to different pHs, temperatures, and saline stress was studied, together with their catabolic characteristics. Based on the results, certain isolates are proposed as valuable candidate starter cultures that could improve both the manufacture and the flavor of such dry-cured meat products, and provide an understanding of new mechanisms involved in stress tolerance. Applied medium-scale industrial tests are currently in progress.

Selection of Optimum Expression System for Production of Kringle Fragment of Human Apolipoprotein(a) in Saccharomyces cerevisiae

  • Cha Kwang Hyun;Kim Myoung Dong;Lee Tae Hee;Lim Hyung Kweon;Jung Kyung Hwan;Seo Jin Ho
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.6
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    • pp.523-527
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    • 2004
  • Recombinant Saccharomyces cerevisiae expression systems were developed to pro­duce a novel human anti-angiogenic protein called LK8, an 86 amino-acid kringle fragment pro­tein with three disulfide linkages. Galactose-inducible LK8 expression plasmid was constructed, and LK8 production levels by four S. cerevisiae strains were compared in order to select an op­timal host strain. S. cerevisiae 2805 was the most efficient among the strains tested. Elevating the LK8 gene copy number through multiple integration using 8-sequences as target sites re­sulted in more than a two-fold increase in the LK8 production level compared with the plasmid­based expression system. The maximum LK8 protein concentration of 25 mg/L was obtained from batch cultivation of the yeast transformant that harbors 16 copies of the LK8 gene. In con­clusion, the strain integrated with the multiple LK8 gene secreted the protein with relatively high yield, although, the increased LK8 gene dosage over 11 copies did not lead to further en­hancement in batch cultivations.

팽나무버섯(Flammulina velelutipes)균의 액체배양 생장조건 규명(Growth condition of Flammulina velutipes in Liquid culture)

  • Hong, Seong-Jun;Lee, Won-Ho;Park, Gi-Beom;Park, Gi-Byeong;Seong, Jae-Mo
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.355-358
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    • 2001
  • Selection of optima] nutrient sources and cultural methods for liquid spawn culture of Flammulina velutipes were carried out. The optimal temperature and pH for mycelial growth of F. velutipes were $20^{\circ}C$ and 6.0 to 7.5. respectively. In the 250ml ${Delta}$-flask culture. the amount of inoculum and culture period for the optimal mycelial growth of F. velutipes were 3 mycelial disks(diametcr 6mm) and 6 days, respectively. For the mass production of submerged culture. the optimal inoculum amount and aeration rate of F. velutipes were 5%(inoculum vol/medium vol.) and l.0vvm(vol of air/vol. of medium/min), respectively.

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Isolation and Characterization of Transposon \ulcorner¨ªKm-Mediated Nonpathogenic Mutants of Xanthomonas campestris pv. vesicatoria (고추 세균성 반점병균의 비병원성 돌연변이체 분리 및 생리적 특성)

  • 윤영채;김용식;조용섭
    • Korean Journal Plant Pathology
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    • v.11 no.3
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    • pp.265-270
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    • 1995
  • Transposon mutation of Xanthomonas campestris pv. vesicatoria (Xcv) was induced by using transposon omegon ($\Omega$)-Km (Tn $\Omega$Km), which was confirmed by resistance to kanamycin (KMr), and nonpathogenic mutants were selected through the inoculation test on pepper plants. The mutagenesis frequency was about 6$\times$10-8, and 53 out of 2,000 Kmr bacterial colonies tested were nonpathogenic to the pepper cultivar Cheung-Hong. Optimum conditions for the Tn $\Omega$Km mutagenesis of Xcv were Luria Bertani (LB) broth medium for culture of Xcv, yeast extract-dextrose-CaCO3 (YDC) agar medium for selection of Tn $\Omega$Km-mediated mutants, and over 1 to 2 in the ratio of the donor (Escherichia coli S17-1 with the plasmid pJFF350 $\Omega$Km) and the recipient (Xcv) in the culture for the mutagenesis. One of the 4 nonpathogenic mutants (WNP1, WNP3, WNP4 and WNP5), which had been reconfirmed through the inoculation on pepper cv. Dabokgun, showed no differences in the production of exoenzymes such as protease and polygalacturonase and extracellular polysaccharides in vitro and the bacterial growth rate from those of the wild type of Xcv.

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Development of a Multicopy Integration Vector in Yarrowia lipolytica (Yarrowia lipolytica의 Multicopy Integration Vector 개발)

  • Kim, Jeong-Yoon;Woo, Moon-Hee;Ryu, Dewey D.Y.
    • Microbiology and Biotechnology Letters
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    • v.23 no.5
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    • pp.536-543
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    • 1995
  • Multicopy integration vector is a very useful vector system in that they can be integrated into chromosomal DNA in several copies and stably maintained under non-selective conditions. To develop a multicopy integration vector system in the yeast Yarrowia lipolytica, P-type ribosomal DNA was cloned from Y lipolytica. A HindIII-BglII fragment of the cloned rDNA and a promoterless URA3 gene were inserted into pGEM1, generating multicopy integration vectors, pMIYL-1 and pMIYL-2. The rDNA fragment is for targeted homologous recombination between the vector and the chromosomal DNA of Y. lipolytica, and the promoterless URA3 gene is a defective selection marker for inducing multicopy integration. pMIYL-1 and pMIYL-2 have an unique restriction enzyme site, KpnI, and two unique restriction enzyme sites, KpnI and EcoRI, repectively, which can be used for targeting of the vectors into the rDNA of Y. lipolytica chromosomal DNA. After transformation of the vectors into Y. lipolytica, copy number and stability were analyzed by Southern hybridization. The vectors were found to be present in less than 5 copies per cell and were stably maintained during growth in non-selective media.

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Morphological Feature Extraction of Microorganisms Using Image Processing

  • Kim Hak-Kyeong;Jeong Nam-Su;Kim Sang-Bong;Lee Myung-Suk
    • Fisheries and Aquatic Sciences
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    • v.4 no.1
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    • pp.1-9
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    • 2001
  • This paper describes a procedure extracting feature vector of a target cell more precisely in the case of identifying specified cell. The classification of object type is based on feature vector such as area, complexity, centroid, rotation angle, effective diameter, perimeter, width and height of the object So, the feature vector plays very important role in classifying objects. Because the feature vectors is affected by noises and holes, it is necessary to remove noises contaminated in original image to get feature vector extraction exactly. In this paper, we propose the following method to do to get feature vector extraction exactly. First, by Otsu's optimal threshold selection method and morphological filters such as cleaning, filling and opening filters, we separate objects from background an get rid of isolated particles. After the labeling step by 4-adjacent neighborhood, the labeled image is filtered by the area filter. From this area-filtered image, feature vector such as area, complexity, centroid, rotation angle, effective diameter, the perimeter based on chain code and the width and height based on rotation matrix are extracted. To prove the effectiveness, the proposed method is applied for yeast Zygosaccharomyces rouxn. It is also shown that the experimental results from the proposed method is more efficient in measuring feature vectors than from only Otsu's optimal threshold detection method.

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