• Title/Summary/Keyword: yeast selection

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Strain Improvement for Enhanced Production of Streptokinase and Streptodornase in Streptococcus sp.

  • HYUN, HYUNG-HWAN;YOON-BUM LEE;KYUNG-HWA SONG;JI-YOUNG JEON;HYUNE-HWAN LEE
    • Journal of Microbiology and Biotechnology
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    • v.7 no.2
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    • pp.101-106
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    • 1997
  • Strain improvement for the enhanced production of streptokinase and streptodornase in Streptococcus sp. ATCC 12449 was performed. Strain UB111, a hyperproductive mutant which was isolated by use of nitrosoguanidine and selection of colonies with large clear zones on DNase test agar plates supplemented with $1{\%}$ glucose and $0.5{\%}$ ammonium chloride, produced about 3 fold more streptokinase and streptodornase than the wild type when tested in shake flask fermentations. The enhanced production of both streptokinase and streptodornase was achieved by cultivating the mutant in a pH-controlled fermentor containing fermentation medium enriched with yeast extract ($2.1{\%}$). Under these conditions, the mutant produced 7300 units/ml of streptokinase and 800 units/ml of streptodornase.

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Characteristics of Growth, Pigment and Monacolin K Production by Monascus strains in Liquid Culture (액체배양에서의 홍국균의 생장, 색소 및 monacolin K 생산 특성)

  • Seo, Jin-Won;Kim, Chang Sup;Seo, Eun Jeong;Jeon, Che Ok;Choi, Hyung-Kyoon;Park, Youn-Je
    • KSBB Journal
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    • v.27 no.5
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    • pp.301-307
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    • 2012
  • The hyphal growth, production of color pigments and monacolin K by Monascus strains were investigated in liquid medium. Thirty five different strains were collected and cultured in potato dextrose yeast extract broth (PDYB), potato dextrose broth (PDB) and malt extract broth (MEB) medea at $25^{\circ}C$ for 7 days. The growth rates of most of strains were highest in PDYB medium. Growth rate as well as pigment production were influenced by suspension conditions of mycelia during liquid cultivation. Most of strains producing monacolin K corresponded to strains producing red pigment highly and showing more pH changes of liquid media. Monacolin K produced from strains was detected in culture broth as well as mycelia. Any citrinin was not detected in monacolin K producing strains. These results imply that the selection of the strains producing red pigment highly and showing more pH changes in liquid cultivation could be applied for primary screening of Monascus strains for preparation of red mold rice.

Development of FK506-hyperproducing strain and optimization of culture conditions in solid-state fermentation for the hyper-production of FK506

  • Mo, SangJoon;Yang, Hyeong Seok
    • Journal of Applied Biological Chemistry
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    • v.59 no.4
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    • pp.289-298
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    • 2016
  • FK506 hyper-yielding mutant, called the TCM8594 strain, was made from Streptomyces tsukubaensis NRRL 18488 by mutagenesis using N-methyl-N'-nitro-N-nitrosoguanidine, ultraviolet irradiation, and FK506 sequential resistance selection. FK506 production by the TCM8594 strain improved 45.1-fold ($505.4{\mu}g/mL$) compared to that of S. tsukubaensis NRRL 18488 ($11.2{\mu}g/mL$). Among the five substrates, wheat bran was selected as the best solid substrate to produce optimum quantities of FK506 ($382.7{\mu}g/g$ substrate) under solid-state fermentation, and the process parameters affecting FK506 production were optimized. Maximum FK506 yield ($897.4{\mu}g/g$ substrate) was achieved by optimizing process parameters, such as wheat bran with 5 % (w/w) dextrin and yeast extract as additional nutrients, 70 % (v/w) initial solid substrate moisture content, initial medium pH of 7.2, $30^{\circ}C$ incubation temperature, inoculum level that was 10 % (v/w) of the cell mass equivalent, and a 10 day incubation. The results showed an overall 234 % increase in FK506 production after optimizing the process parameters.

Transformation of a Filamentous Fungus Cryphonectria parasitica Using Agrobacterium tumefaciens

  • Park, Seung-Moon;Kim, Dae-Hyuk
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.3
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    • pp.217-222
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    • 2004
  • As Agrobacterium tumefaciens, which has long been used to transform plants, is known to transfer T-DNA to budding yeast, Saccharomyces cerevisiae, a variety of fungi were subjected to the A. tumefaciens-mediated transformation to improve their transformation frequency and feasibility. The A. tumefaciens-mediated transformation of chestnut blight fungus, Cryphonectria parasitica, is performed in this study as the first example of transformation of a hardwood fungal pathogen. The transfer of the binary vector pBIN9-Hg, containing the bacterial hygromycin B phosphotransferase gene under the control of the Aspergillus nidulans trpC promoter and terminator, as a selectable marker, led to the selection of more than 1,000 stable, hygromycin B-resistant transformants per 1${\times}$10$\^$6/ conidia of C. parasitica. The putative transformants appeared to be mitotically stable. The transformation efficiency appears to depend on the bacterial strain, age of the bacteria cell culture and ratio of fungal spores to bacterial cells. PCR and Southern blot analysis indicated that the marker gene was inserted at different chromosomal sites. Moreover, three transformants out of ten showed more than two hybridizing bands, suggesting more than two copies of the inserted marker gene are not uncommon.

The Suitable Condition for Mycelial Growth of Volvariella volvacea strains and Selection of the Superior strain (풀버섯의 균사배양 적합조건 및 우량균주 선발)

  • Jang, Myoung-Jun;Lee, Han-Bum;Kim, Jeong-Han;Lee, Yun-Hae;Ju, Young-Cheol
    • The Korean Journal of Mycology
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    • v.37 no.2
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    • pp.173-180
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    • 2009
  • This study was carried out to obtain basic data on mycelial growth characteristics for an artificial cultivation of Volvariella volvacea. Twelve URP primers were used to assess genetic relationship of V. volvacea and its result was divided into two groups. But there was nothing different in morphological characteristics in V. volvacea. Among five kinds of mushroom media, MCM medium was selected as the favorable culture medium. The optimal range of temperature and pH for mycelial growth on V. volvacea were 35oC and pH 6~8, respectively. Carbon sources had not an effect on 10 strains of V. volvacea, and nitrogen source for the optimum mycelial growth was yeast extract. Also, we selected GMVV 79004 as a superior strain of 10 strains V. volvacea based on the mycelial growth and yield.

Isolation of Debaryomyces hansenii and selection of an optimal strain to improve the quality of low-grade beef rump (middle gluteal) during dry aging

  • Yoonjeong Yoo;Hyemin Oh;Yohan Yoon
    • Animal Bioscience
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    • v.36 no.9
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    • pp.1426-1434
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    • 2023
  • Objective: The objective of this study was to evaluate the effect of Debaryomyces hansenii isolated from dry-aged beef on the tenderness and flavor attributes of low-grade beef during dry aging. Methods: Five D. hansenii strains were isolated from dry-aged beef samples. The rump of low-grade beef was inoculated with individual D. hansenii isolates and subjected to dry aging for 4 weeks at 5℃ and 75% relative humidity. Microbial contamination levels, meat quality attributes, and flavor attributes in the dry-aged beef were measured. Results: Of the five isolates, the shear force of dry-aged beef inoculated with SMFM201812-3 and SMFM201905-5 was lower than that of control samples. Meanwhile, all five isolates increased the total free amino acid, glutamic acid, serine, glycine, alanine, and leucine contents in dry-aged beef. In particular, the total fatty acid, palmitic acid, and oleic acid contents in samples inoculated with D. hansenii SMFM201905-5 were higher than those in control samples. Conclusion: These results indicate that D. hansenii SMFM201905-5 might be used to improve the quality of beef during dry aging.

Microbiological Hazard Analysis for HACCP System Application to Vinegared Pickle Radishes (식초절임 무의 HACCP 시스템 적용을 위한 미생물학적 위해 분석)

  • Kwon, Sang-Chul
    • Journal of Food Hygiene and Safety
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    • v.28 no.1
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    • pp.69-74
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    • 2013
  • This study has been performed for 150 days from February 1 - June 31, 2012 aiming at analyzing biologically hazardous factors in order to develop HACCP system for the vinegared pickle radishes. A process chart was prepared as shown on Fig. 1 by referring to manufacturing process of manufacturer of general vinegared pickle radishes regarding process of raw agricultural products of vinegared pickle radishes, used water, warehousing of additives and packing material, storage, careful selection, washing, peeling off, cutting, sorting out, stuffing (filling), internal packing, metal detection, external packing, storage and consignment (delivery). As a result of measuring Coliform group, Staphylococcus aureus, Salmonella spp., Bacillus cereus, Listeria Monocytogenes, E. coli O157:H7, Clostridium perfringens, Yeast and Mold before and after washing raw radishes, Bacillus cereus was $5.00{\times}10$ CFU/g before washing but it was not detected after washing and Yeast and Mold was $3.80{\times}10^2$ CFU/g before washing but it was reduced to 10 CFU/g after washing and other pathogenic bacteria was not detected. As a result of testing microorganism variation depending on pH (2-5) of seasoning fluid (condiment), pH 3-4 was determined as pH of seasoning fluid as all the bacteria was not detected in pH3-4. As a result of testing air-borne bacteria (number of general bacteria, colon bacillus, fungus) depending on each workplace, number of microorganism of internal packing room, seasoning fluid processing room, washing room and storage room was detected to be 10 CFU/Plate, 2 CFU/Plate, 60 CFU/Plate and 20 CFU/Plate, respectively. As a result of testing palm condition of workers, as number of general bacteria and colon bacillus was represented to be high as 346 $CFU/Cm^2$ and 23 $CFU/Cm^2$, respectively, an education and training for individual sanitation control was considered to be required. As a result of inspecting surface pollution level of manufacturing facility and devices, colon bacillus was not detected in all the specimen but general bacteria was most dominantly detected in PP Packing machine and Siuping machine (PE Bulk) as $4.2{\times}10^3CFU/Cm^2$, $2.6{\times}10^3CFU/Cm^2$, respectively. As a result of analyzing above hazardous factors, processing process of seasoning fluid where pathogenic bacteria may be prevented, reduced or removed is required to be controlled by CCP-B (Biological) and threshold level (critical control point) was set at pH 3-4. Therefore, it is considered that thorough HACCP control plan including control criteria (point) of seasoning fluid processing process, countermeasures in case of its deviation, its verification method, education/training and record control would be required.

Protoplast Regeneration, Reversion and Isolation of Auxotrophic Mutants in Flammulina velutipes (팽이버섯 (Flammulina velutipes)원형질체(原形質體)의 재생(再生), 환원(還元) 및 영양요구성(營養要求性) 균주선발(菌株選拔))

  • Shin, Gwan Chull;Park, Jong Seung;Yoo, Young Bok;Yeo, Un Hyung
    • Korean Journal of Agricultural Science
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    • v.15 no.1
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    • pp.15-22
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    • 1988
  • Factors affecting the regeneration, reversion of protoplasts from mycelium of F. velutipes were investigated and the selection of auxotrophic mutants from protoplasts of F. velutipes was performed. PDP medium stabilized with 0.6M sucrose was suitable for the regeneration of protoplasts, and regeneration frequency was 0.47-1.32. The regeneration frequency of protoplasts was increased when nutrients were added to the regeneration medium. Especially, yeast extract was the most effective to regeneration of protoplasts. Regeneration pattern of protoplasts was formation of germ tubes from bud-like cells. 13-18% of monokaryotic strains was appeared from reverted protoplasts. Five of auxotrophic mutants were isolated from strains showed survival frequency of 1.9-16.

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Effect of a Common Medium on the Growth of Nitrogen Fixer Rhizobium and Phosphate Solubilizer Bacillus megaterium (질소고정균(Rhizobium)과 인산가용화균(Bacillus megaterium)의 동시배양을 위한 배양조건 탐색)

  • Poonguzhali, Selvaraj;Thangaraju, Muthu;Ryu, Jyung-Hyun;Madhaiyan, Munusamy;Chung, Keun-Yook;Sa, Tong-Min
    • Korean Journal of Soil Science and Fertilizer
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    • v.38 no.1
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    • pp.8-14
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    • 2005
  • Mass culturing of two beneficial organisms used as biofertilizers for crops would reduce the risks in production and minimize the capital involved and this demands appropriate media that supports both organism and also selection of organisms that are not antagonistic to each other. A study was initiated to culture a nitrogen fixer (Rhizobium) and phosphate solubilizer (Bacillus megaterium) in a single medium and to study their growth patterns and shelf life in carrier. The growth of Rhizobium and Bacillus megaterium was assessed in different media and a slight modification in the traditional yeast extract mannitol media promoted the growth of both the organisms. The growth of the individual organisms in the modified medium was assessed by estimating the population at regular intervals and compared to their original medium. Maximum population of Rhizobium and phosphobacteria was at 60 hr when the phosphiobacteria inoculation of later was after 48 hr of Rhizobium inoculation. The shelf life of the individual inoculants in the inoculant containing both the organism in a sterile carrier base revealed no significant differences compared to individual organisms inoculated in a sterilized carrier. The population of both organisms in carrier based mixed inoculant remained at $10^8$ cells till 90 days.

Manufacturing of Wine with Watermelon (수박을 이용한 발효주의 제조)

  • Hwang, Young;Lee, Ki-Kwon;Jung, Gi-Tai;Ko, Bok-Rae;Choi, Dong-Chil;Choi, Yeong-Geun;Eun, Jong-Bang
    • Korean Journal of Food Science and Technology
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    • v.36 no.1
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    • pp.50-57
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    • 2004
  • Selection of yeast strains, optimum conditions for alcohol fermentation, sterilization methods, and additives for improving wine quality were investigated to manufacture watermelon wine. Eight yeast strains exhibited significant alcohol fermentation, among which KWS 06 was selected for watermelon wine fermentation, because watermelon wine made by this strain showed the best overall acceptability in sensory evaluation. Sucrose was determined as the best saccharide for alcohol fermentation among sucrose, corn syrup, glucose, fructose, and lactose. Optimum concentration of soluble solid and $(NH_{4})_{2}HPO_{4}$ of nitrogen source were $24^{\circ}Brix and 0.2%, respectively. Addition of raspberries and omija increased wine flavor and alcohol production, respectively, with optimum alcohol production, taste, and color achieved with addition of 20 g/L raspberries and 10 g/L omija. Best sensory quality was obtained by addition of 0.04 % watermelon flavorant to the juice.