• Title/Summary/Keyword: fermentation media

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Bacterial Stringent Signal Directs Virulence and Survival in Vibrio cholerae.

  • Oh, Young Taek;Kim, Hwa Young;Yoon, Sang Sun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.8-8
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    • 2019
  • The stringent response (SR) is characterized as a bacterial defense mechanism in response to various growth-inhibiting stresses. It is activated by accumulation of a small nucleotide regulator, (p)ppGpp, and induces global changes in bacterial transcription and translation. Recent work from our group has shown that (p)ppGpp plays a critical role in virulence and survival in Vibrio cholerae. The genes, relA and relV, are involved in the production of (p)ppGpp, while the spoT gene encodes an enzyme that hydrolyzes it in V. cholerae. A mutant strain defective in (p)ppGpp production (i.e. ${\Delta}relA{\Delta}relV{\Delta}spoT$ mutant) lost the ability to produce cholera toxin (CT) and lost their viability due to uncontrolled production of organic acids, when grown with extra glucose. In contrast, the ${\Delta}relA{\Delta}spoT$ mutant, a (p)ppGpp overproducer strain, produced enhanced level of CT and exhibited better growth in glucose supplemented media via glucose metabolic switch from organic fermentation to acetoin, a neutral fermentation end product, fermentation. These findings indicates that (p)ppGpp, in addition to its well-known role as a SR mediator, positively regulates CT production and maintenance of growth fitness in V. cholerae. This implicates SR as a promising drug target, inhibition of which may possibly downregulate V. cholerae virulence and survival fitness. Therefore, we screened a chemical library and identified a compound that induces medium acidification (termed iMAC) and thereby loss of wild type V. cholerae viability under glucose-rich conditions. Further, we present a potential mechanism by which the compound inhibits (p)ppGpp accumulation. Together, these results indicate that iMAC treatment causes V. cholerae cells to produce significantly less (p)ppGpp, an important regulator of the bacterial virulence and survival response, and further suggesting that it has a therapeutic potential to be developed as a novel antibacterial agent against cholera.

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Effect of glucose Feeding Strategy on Biomass of Serratia marcescens in High Density Fed-Batch Fermentation (고밀도 유기식 배양에서 글루코스 공급 방법이 Serratia marcescens의 균체량에 미치는 영향)

  • Kim, Kwang;Lee, Sang-Rok;Shon, Jeong-Woo;Ji, Hong-Seok
    • KSBB Journal
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    • v.13 no.6
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    • pp.681-686
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    • 1998
  • Effect of glucose feeding strategy and initial concentration of glucose on Serratia marcescens ATCC 27117 in high cell density fed-batch fermentation was investigated. The final biomasses in batch, constant feeding, constant and exponentially feeding strategy at glucose starvation condition in fed-batch were 1.40, 5,07, 6,93 and 7.60 g/L at 40, 41, 24 and 40 hrs, respectively. Productivities of biomass were 0.035, 0.124, 0.289 and 0.190 g/L$.$h, respectively. As a result, constant feeding strategy at starvation condition was 1.5∼8.6 times higher than other strategies. The relationship between dissolved oxygen and glucose feeding times was good identified in exponential feeding strategy and constant feeding strategy at starvation condition. And high cell density cultivation was obtained when minimal media was used.

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Overproduction of Sodium Gluconate Using the Recombinant Aspergillus niger (재조합 Aspergillus niger에 의한 글루콘산나트륨의 산업적 생산)

  • 이선희;이현철;김대혁;양문식;정봉우
    • KSBB Journal
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    • v.13 no.2
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    • pp.214-219
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    • 1998
  • Polymerase chain reaction(PCR) was conducted to obtain the gene encoding glucose oxidase(GOD) from Aspergillus niger(ATCC 2110) and the DNA sequence determined was coincided with published GOD sequence from A. niger. Recombinant transforming vector containing GOD and hygromycin B(hyg.B) resistant gene(hph) was constructed and used for further transformation of A. niger ATCC 2110. Selectivity of hyg.B against A. niger differed depending on which media were used i.e., nutrient-rich media such as potato dextrose agar(PDA) and complete medium(CM) showed only 50% growth inhibition at 400 $\mu$m ml$^-1$ of hyg.B while the minimal media inhibited mycelial growth completely at 200 $\mu$m ml$^-1$ of hyg.B. Twenty to sixty putative transformants were isolated from the hyg.B-containing minimal top agar, transferred successively onto alternating selective and nonselective media for a mitotic stability of hyg.B resistance and, then, single-spored. Among the stable transformants, the transformant(GOD1-6) grown by flask culture showed the considerable increase of extracellular GOD activity, which was estimated to the degree of 50% - 100% comparing to that of wild type. Transformation of tGOD1-6 was resulted from integration of the vectors into heterologous as well as homologous regions of the A. niger genome. Southern blot analysis revealed that there were two independent integrations of vector into fungal genome and one into the GOD gene due to homologous recombination. In addition, GOD activity and sodium gluconate production when tGOD1-6 was fed-batch fermented were enhanced 11 fold and 2.25 fold, respectively, compared to that of the wild type.

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Cultural Condition for the Mycelial Growth of Phellinus igniarius on Chemically Defined Medium and Grains (화학합성배지 및 곡물을 이용한 Phellinus igniarius의 균사체 배양조건)

  • Jung, In-Chang;Kim, Seon-Hee;Kwon, Yong-Il;Kim, So-Yeun;Lee, Jong-Suk;Park, Shin;Park, Kyung-Sook;Lee, Jae-Sung
    • The Korean Journal of Mycology
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    • v.25 no.2 s.81
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    • pp.133-142
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    • 1997
  • The chemical media composition and culture conditions were optimized for mycelial growth of Phellinus igniarius 26005. The method of solid-state fermentation, cultivation of basidiomycetal strains in various grains, was developed. Media composition for optimal growth of Phellinus igniarius 26005 was made of 7.0% malt extract, 0.3% bacto soytone, and 0.2% yeast extract. The optimum condition for mycelial growth was $28^{\circ}C$ and pH 7.0, respectively. For the mass cultivation of mycelia, the hydrated grains with cold water, were put into the plastic bottle. The mycelial growth rate in the bottled grains was high in the early stage with inoculation of homogenized mycelium. The activity of mycelium was maintained by adding sterilized water in the middle of cultivation. The glucosamine content which determins the mycelial growth rate in solid material was in the order of job's tears>barley>black soybean>wheat>malt soybean>brown rice>sorghum>glutinous rice.

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Development of a Novel Medium with Chinese Cabbage Extract and Optimized Fermentation Conditions for the Cultivation of Leuconostoc citreum GR1 (폐배추 추출물을 이용한 Leuconostoc citreum GR1 종균 배양용 최적 배지 및 배양 조건 개발)

  • Moon, Shin-Hye;Chang, Hae-Choon;Kim, In-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.7
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    • pp.1125-1132
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    • 2013
  • In the kimchi manufacturing process, the starter is cultured on a large-scale and needs to be supplied at a low price to kimchi factories. However, current high costs associated with the culture of lactic acid bacteria for the starter, have led to rising kimchi prices. To solve this problem, the development of a new medium for culturing lactic acid bacteria was studied. The base materials of a this novel medium consisted of Chinese cabbage extract, a carbon source, a nitrogen source, and inorganic salts. The optimal composition of this medium was determined to be 30% Chinese cabbage extract, 2% maltose, 0.25% yeast extract, and $2{\times}$ salt stock (2% sodium acetate trihydrate, 0.8% disodium hydrogen phosphate, 0.8% sodium citrate, 0.8% ammonium sulfate, 0.04% magnesium sulfate, 0.02% manganese sulfate). The newly developed medium was named MFL (medium for lactic acid bacteria). After culture for 24 hr at $30^{\circ}C$, the CFU/mL of Leuconostoc (Leuc.) citreum GR1 in MRS and MFL was $3.41{\times}10^9$ and $7.49{\times}10^9$, respectively. The number of cells in the MFL medium was 2.2 times higher than their number in the MRS media. In a scale-up process using this optimized medium, the fermentation conditions for Leuc. citreum GR1 were tested in a 2 L working volume using a 5 L jar fermentor at $30^{\circ}C$. At an impeller speed of 50 rpm (without pH control), the viable cell count was $8.60{\times}10^9$ CFU/mL. From studies on pH-stat control fermentation, the optimal pH and regulating agent was determined to be 6.8 and NaOH, respectively. At an impeller speed of 50 rpm with pH control, the viable cell count was $11.42{\times}10^9(1.14{\times}10^{10})$ CFU/mL after cultivation for 20 hr - a value was 3.34 times higher than that obtained using the MRS media in biomass production. This MFL media is expected to have economic advantages for the cultivation of Leuc. citreum GR1 as a starter for kimchi production.

Rice Fermentation by Korean Amylolytic Bifidobacterium spp. (한국인유래의 Amylolytic Bifidobacterium 에 의한 쌀발효)

  • Park, Jong-Hyun;Song, Hey-Kyung;Ahn, Jun-Bae;Ji, Geun-Eok;Mok, Chul-Kyoon
    • Korean Journal of Food Science and Technology
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    • v.29 no.3
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    • pp.581-587
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    • 1997
  • For bifidus fermentation food, gelatinized rice solution was fermented without liquefaction/saccharification by amylolytic Bifidobacterium spp. isolated from Korean. Eighteen amylolytic Bifidobcterium on the starch agar were isolated from 38 Korean and four strains were finally selected as good amylase producers. The most enzyme-producing strain of Bif. sp. FBD-12 secreted extracellular amylase of 0.17 U/mg and intracelluar amylase of 1.8 U/mg. Three strains of Bif. sp. FBD-12, Bif. sp. FBD-16 and Bif. sp. FBD-17 also showed good growth on pH controlled media by HCI/acetic acid to pH 5.0 while Bif. sp. FBD-6 was not so tolerant that viable cell counts reduced to $10^2\;CFU/mL$ times on the media. Initial cell number of $10^6\;CFU/mL$ for those strains reached to $10^9\;CFU/mL$ on the rice medium supplemented with yeast extract (0.2%) and cysteine (0.05%). Ascorbic acid instead of cysteine was added to the medium for improving off-flavour and the best growth was shown at 0.1% addition. Isolated soybean proteins (ISP) of 3% accelerated the growth of the strains. Maximum count of $10^9\;CFU/mL$ reached within 12 hour fermentation on the rice medium with ascorbic acid and isolated soybean protein instead of 32 hours on the cysteine medium, and total acidity increased from 0.5% to 1% on each media. Reducing sugar in the ascorbic acid/ISP cultures generally increased especially 2 mg/mL to 15.5 mg/mL for Bif. sp. FBD-6. From sensory evaluation, the products showed good acceptability so that it suggested possibility of development of bifidus-fermented rice food.

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Investigation of the Microbiological and Biochemical Properties of Kimchi in the Submerged Model System Designed for Fermented Sausages

  • Lee, Joo-Yeon;Kunz, Benno
    • Food Science of Animal Resources
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    • v.29 no.4
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    • pp.423-429
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    • 2009
  • The objective of this study was to investigate the potential of the application of lactic acid bacteria (LAB) from kimchi as a starter culture in the production of fermented sausages. To achieve this, a submerged model medium that contained LAB as part of a complex system of kimchi (0.5, 1.0, 1.0, 3.0, and 5.0%) and lyophilized kimchi powder (0.2 and 0.5%) was fermented for 120 h. During the fermentation period, the growth of total viable organisms and LAB, and the changes in the pH and the titratable acidity, were investigated. The initial LAB counts ranged from 6.4 to 7.7 Log CFU/mL for the kimchi media, and from 6.9 to 6.9 Log CFU/mL for the kimchi powder media. In all the kimchi batches, the LAB increased logarithmically, and the highest LAB counts (around 9 Log CFU/mL) were reached in 24 h. An evident lag phase of the LAB was observed in the kimchi powder samples and reached 8.8 Log CFU/mL in 8 h. The decrease in the pH and the formation of lactic acid were rapid in the kimchi batches, and reached pH values of 3.4-3.5 in 12 h. With these results, the LAB that was integrated with the addition of kimchi or kimchi powder demonstrated its potential utility as a substitute for starter culture.

Improvement of Decarboxylating Agar Medium for Screening Biogenic Amine-Producting Bacteria in Kimchi

  • Mah, Jae-Hyung;Shin, Soon-Young;Lee, Heung-Shick;Cho, Hong-Yon;Hwang, Han-Joon
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.491-496
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    • 2001
  • A modification of decarboxylating agar medium as described by niven was performed to improve the detection method of biogenic amine-producing bacteria and to eliminate the false-positive. A total of 120 bacterial strains isolated from kimchi were used to evaluate different dicarboxylating agar media and for screening biogenic amines. Potential false-positives ranged from approximately 66 to 79% of the strains tested in the already well-known media. In our improved medium, none of the 120 strains showed the potential false-positives. There was a good agreement (81.7%-87.5%) between the results obtained by the improved medium and by HPLC analysis. Consequently, this medium was greatly improved in screening biogenic amine-producing bacteria and discarding false-positives. Of the 120 kimchi isolates, 14.2, 18.3, 37.5, and 0.8% were found by HPLC to be the producers of histamine, tyramine, putrescine (as a form of spermine), and cadaverine, respectively. The proportion of biogenic amine producer during kimchi fermentation increased to a maximum at an immature period and decreased thereafter.

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Mannitol Production by Leuconostoc citreum KACC 91348P Isolated from Kimchi

  • Otgonbayar, Gan-Erdene;Eom, Hyun-Ju;Kim, Beom-Soo;Ko, Jae-Hyung;Han, Nam-Soo
    • Journal of Microbiology and Biotechnology
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    • v.21 no.9
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    • pp.968-971
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    • 2011
  • Leuconostoc genus, which comprise heterofermentative lactic acid bacteria, reduces fructose to mannitol by recycling intracellular NADH. To evaluate the mannitol productivities of different Leuconostoc species, 5 stock cultures and 4 newly isolated strains were cultivated in MRS and simplified media containing glucose and fructose (1:2 ratio). Among them, L. citreum KACC 91348P, which was isolated from kimchi, showed superior result in cell growth rate, mannitol production rate, and yield in both media. The optimal condition for mannitol production of this strain was pH 6.5 and $30^{\circ}C$. When L. citreum KACC was cultured in simplified medium in a 2 l batch fermenter under optimal conditions, the maximum volumetric productivity was 14.83 $g{\cdot}l^{-1}h^{-1}$ and overall yield was 86.6%. This strain is a novel and efficient mannitol producer originated from foods to be used for fermentation of fructose-containing foods.

Optimization of Organic Compounds and Hydrogen Production in Dark Fermentation using Chlamydomonas reinhardtii (Chlamydomonas reinhardtii를 이용한 암반응에서의 수소 및 유기물 생산 최적화)

  • 공경택;심상준;박대원;김미선;박태현
    • KSBB Journal
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    • v.18 no.1
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    • pp.51-54
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    • 2003
  • The objective of this study was to optimize culture conditions and to produce hydrogen and organic compounds using microalga Chlamydomonas reinhardtii. First of all, C. reinhardtii UTEX 90 was chosen from the three kinds of strains in terms of their hydrogen and organic compound productivity. The optimum $\textrm{CO}_2$ concentration range of C. reinhardtii UTEX 90 was 1to 3%. We tested two medium, which are popular in this microalga culture; Brostol's medium and TAP medium (8). The cell growth in TAP medium was found to be higher than a Brostol's medium. Optimum culture with 3% of $\textrm{CO}_2$ in TAP medium produced the most hydrogen ($0.5\mu$ mol/ mg DCW), though Bristol's medium produced twice as much total organics.