• Title/Summary/Keyword: solid culture medium

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The Pharmacological Activity of Coffee Fermented Using Monascus purpureus Mycelium Solid-state Culture Depends on the Cultivation Area and Green Coffees Variety (원산지 및 품종에 따라 조제된 홍국균 균사체-고체발효 원두커피의 생리활성)

  • Kim, Hoon;Yu, Kwang-Won;Lee, Jun-Soo;Baek, Gil-Hun;Shin, Ji-Young
    • Korean Journal of Food Science and Technology
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    • v.46 no.1
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    • pp.79-86
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    • 2014
  • In previous work, we fermented coffee beans using solid-state culture with various fungal mycelia to enhance the physiological activity of the coffee. The coffee fermented with Monascus sp. showed a higher physiological activity than non-fermented coffee or other coffees fermented with mushroom mycelium. The aim of this study was to characterize the various fermented coffees with respect to their area of cultivation and their variety using Monascus purpureus (MP) mycelium solid-state culture. Thirty types of green coffee beans, which varied in terms of their cultivation area or variety, were purchased from different suppliers and fermented with MP under optimal conditions. Each MP-fermented coffee was medium roasted and extracted further using hot water (HW) under the same conditions. Of the HW extracts, those derived from MP-Mandheling coffees had the highest yield (13.6-15.5%), and MP-Robusta coffee showed a significantly higher polyphenolic content (3.03 mg gallic acid equivalent/100 mg) and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) free radical scavenging activity (27.11 mg ascorbic acid equivalent antioxidant capacity/100 mg). Furthermore, in comparison to other MP-fermented coffees at $1,000{\mu}g/mL$, MP-Robusta coffee showed not only the most effective inhibition of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) production in LPS-stimulated RAW 264.7 cells (67.1% of that in LPS-stimulated control cells), but also an effective inhibition of lipogenesis in 3T3-L1 adipose cells (22.2% of that in differentiated control cells). In conclusion, these results suggest that Vietnam Robusta coffee beans solid-state fermented with MP mycelium are amenable to industrial applications as a functional coffee beverage or material.

Chinoketides A and B, Two New Antimicrobial Polyketides from the Endophytes of Distylium chinense with the "Black-Box" Co-culture Method

  • Lv, Meng-Meng;Tan, Ming-Hui;Lu, Li-Wen;Zhang, Rong-Hua;Guo, Zhi-Yong;Liu, Cheng-Xiong;Yang, Jin;Zou, Kun;Proksch, Peter
    • Natural Product Sciences
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    • v.24 no.3
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    • pp.159-163
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    • 2018
  • Two new polyketides, chinoketides A and B (1 - 2) with a known compound xylarphthalide A (3), were isolated from the solid medium of the endophytes from the leaves of the relic plant Distylium chinense with the "black-box" co-culture method, and the structures of two new compounds were elucidated by NMR, MS and CD spectra. And the absolute configurations of chinoketides A (1) and B (2) were determined as 2R,3R,8S and 5R,6S by calculating their ECD spectra to compare with the experimental CD spectra. Finally, the antimicrobial activities were evaluated to Erwinia carotovora sub sp. Carotovora (Jones) Bersey et al, and the results showed that compounds 1 - 3 displayed the antimicrobial activities with MIC value at 20.5, 30.4 and $10.2{\mu}g/mL$.

Plant Regeneration and Somatic Embryo Formation from Root-Derived Callus of Rice (벼 뿌리조직 유래의 캘러스로부터 체세포배 형성과 식물체 재분화)

  • 손재근;김경민;김종수
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.3
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    • pp.143-148
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    • 1995
  • The competence of callus formation and plant regeneration from root derived callus was higher in japonica cultivars than those of Tongil-type cultivars of rice. A japonica type cultivars Yeongdeogbyeo, showed the highest capacity (13%) for plant regeneration from root calli of 6 cultivars tested. The callus induced from seed and root tissues maintained higher capacity for plant regeneration during 7 passages of subculture on N$_{6}$ solid media at 2-week intervals. The maximum frequency (2 x 10$^{5}$ mL) of round cells and their cell colonies showed about 24 days after suspension culture of root-derived callus in N$_{6}$ medium with lmg/L 2,4-D, 300mg/L casein hydrolysate, 10mM L-proline, 20g/L sucrose and 30g/L sorbitol. The frequency of somatic embryo formation in suspension cultures of root-derived callus increased with prolonged advance of subculture time from 30 to 90 days, but their regenerative capacities decreased.

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Studies on the Proteolytic Enzyme Produced by Rhizopus japonicus S-62 (Rhizopus japonicus S-62가 생성(生成)하는 단백질분해효소(蛋白質分解酵素)에 관(關)한 연구(硏究))

  • Chung, Man-Jae
    • Korean Journal of Food Science and Technology
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    • v.9 no.1
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    • pp.31-35
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    • 1977
  • As a study on the acid protease production by Rhizopus japonicus S-62, the culture conditions for the enzyme production and characteristics of the crude enzyme were investigated. The results are summarized as follows: 1. The optimum conditions for solid culture on wheat bran were 48 hrs of culture period and $100{\sim}120%$ addition of tap water. 2. Of the several media, wheat bran medium was the most excellent in the enzyme production. 3. The addition of sucrose, fructose, $NH_4Cl$ and $NaH_2PO_4$ on wheat bran, respectively, increased largely the enzyme production. 4. The optimum pH for the enzyme action was pH $2.4{\sim}2.6$, the optimum temperature was about $40^{\circ}C$, and the stable pH range was $pH\;2.5{\sim}5.0$, the enzyme was stable below $40^{\circ}C$ and was inactivated abruptly above $40^{\circ}C$.

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Studies on the Cellulase produced by Myriococcum of albomyces (Myriococcum albomyces가 생산하는 Cellulase에 관한 연구)

  • Chung, Dong-Hyo
    • Applied Biological Chemistry
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    • v.14 no.1
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    • pp.59-97
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    • 1971
  • As a study on the cellulase of Myriococcum albomyces the culture media for enzyme formation and properties of its crude preparation were investigated and the crude enzyme preparation was further fractionated. The results are summarized as follows: 1. Wheat bran solid culture produced stronger activities of cellulase than rice bran or defatted soy bean meal solid culture. 2. Shaking culture with wheat bran, rice bran or defatted soy bean meal produced higher cellulase activities than solid culture with the corresponding media. 3. The enzyme formation was higher at $45^{\circ}C$ than at $37^{\circ}C$ or $50^{\circ}C$ regardless of the kind of culture medium. 4. The formation of CMCase activity was more promoted by organic nitrogen source than inorganic nitrogen source. 5. The formation of cellulase activities were increased 1.5 to 3.0-fold by adding CMC, Avicel or cellulose powder as an inducer into 5% wheat bran basal medium. 6. Cellulase production using a tank culture procedure with addition of CMC or Avicel as an inducer was the highest at fifth day and thereafter decreased slightly. 7. The crude enzyme preparation showed pH optimum in 4.0 to 4.5, and pH stability in the range of 3.5 to 8.0. Optimum temperature for the activity was $65^{\circ}C$ which was higher than among other cellulases and it was stable at $60^{\circ}C$ for 120 minutes. 8. Dialyzed crude enzyme was activated by $Ca^{++}$ and $Mg^{++}$, but inhibited by $Hg^{++}$, $Cu^{++}$ and $Ag^{+}$. 9. Four different types of cellulase, i. e., fraction I, fraction II-a, fraction II-b, and fraction III were purified from the culture filtrate of Myriococcum albomyces through a sequence of ammonium sulfate fractionation, and elution chromatography on DEAE-Sephadex A-25, Amberlite CG-25 type 2 and hydroxyapatite columns. 10. These four cellulase fractions were showed to be homogenous by electrophoresis and ultracentrifugation and also gave a typical ultraviolet absorption spectrum of protein. 11. Four purified fraction showed different specificity toward substrates, fraction I has a stronger activity toward Avicel, cellulose powder, and gauze than that of other cellulase fractions. Fraction II-a had a powerful activity toward cellobiose but it was almost inactive agaisnt fibrous cellulose contrary to fraction I. On the contrary, the main component fraction II-b had a fairly higher activity on CMC and Avicel. Activity of fraction II-b toward cellobiose was about one-third of that of fraction II-a and activity on Avicel was lower than that of fraction I. Fraction III had a more powerful activity in decreasing viscosity of CMC. 12. Final hydrolysis products of fibrous cellulose by each fraction were cellobiose and glucose. Whereas oligosaccharides were predominant in the early stage of hydrolysis, prolonged reaction produced more glucose than cellobiose. Fraction I and fraction II-a acted synergically on Avicel. 13. Optimum pH for the activities of cellulase fraction I, fraction II-a, fraction II-b and fraction III were found to be 5.5, 5.0, 4.0 and $4.0{\sim}4.5$, respectively. These fractions were found to be stable in the range of pH $3.0{\sim}7.5$. 14. Optimum temperature for the activities of fraction I, fraction II-a, fraction II-b, and fraction III were $50^{\circ}C$, $55^{\circ}C$, $60^{\circ}C$ and $55^{\circ}C$, respectively. No less of activity was found by heating 120 minutes at $55^{\circ}C$ and fraction II-a was more stable than the others at $60^{\circ}C$. 15. Fraction I and fraction II-b were activated by $Ca^{++}$ and $Mg^{++}$ but inhibited by $Hg^{++}$ and $Ag^{+}$.

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Development of Transgenic Plant (Codonopsis lanceolata Trautv.) Harboring a Bialaphos Resistance Gene, bar (Bialaphos 저항성 유전자 bar를 이용한 형질전환 더덕개발)

  • 조광수;장정은;류종석;권무식
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.4
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    • pp.281-287
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    • 1999
  • Codonopsis lanceolata ("Deoduck" in Korea) is a perennial herb, and belongs to family, Campanulaceae. Its taproot is used a good source of a wild vegetable as well as an herbaceous medicine. In this study, to develop a bialaphos-resistant transgenic Codonopsis, seed germination mechanism and somatic embryogenesis of the plant were investigated, and Agrobacterium-mediated transformation with bar gene encoding phosphinothricin acetyltransferase (PAT) was performed. Attempt were made to regenerate plant from cells via somatic embryogenesis. When the cotyledons, nodes and leaf disks were cultured on MS medium containing 2,4-D and zeatin, embryogenic calli were induced. Upon transferring the somatic embryos to N6 solid medium without plant growth regulators, they developed into plantlets under continuous illumination. All plants were dead on MS basal medium containing 10 mg/L phosphinothricin (PPT) and Basta, respectively. The explants did not produce calli in the medium containing 200 mg/L kanamycin. The explants were cocultured with Agrobacterium tumefaciens for 2 days, and transformants were selected in MS basal medium containing 1.0 mg/L 2,4-D, 100 mg/L kanamycin and 500 mg/L carbenicillin. After the selection, embryogenic calli were induced and then somatic embryos were produced by subsequent subculturing. The somatic embryos were germiated on N6 basal medium containing 200 mg/L kanamycin and 500 mg/L carbenicillin. PCR analysis showed that nptII and bar genes were introduced in the Deoduck transformants. After the confirmation of bar gene expression in RNA and protein level, the transgenic Deoduck will be used to study the genetics of filial generation with the herbicide control gene, bar.gene, bar.

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Factors Influencing Protoplast Isolation and Culture in Three Capsicum Species (세 가지 고추속 식물의 원형질체 분리 및 배양에 미치는 요인)

  • 임학태;염옥희;전익조;조미애;양승균
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.3
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    • pp.141-146
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    • 2001
  • Protoplasts were isolated from cotyledons, hypocotyls, and mesophyll tissues of three species of Capsicum species (C. anuumm, C. bacatuum, and C. chacoense). Combination of Cellulysin (1%) and Macero-zyme (0.25%) in 0.65 M sorbitol was found to be the most effective for the digestion of cell wall, regardless of the Capsicum species. Antioxidant MES (2-[N-Morpholino]ethanesulfonic acid) in the enzyme solution helped protoplasts overcome browning. After 5 days of initial culture, Cell division occurred in modified K8p medium containing 1~5 mg/L zeatin, 0.5 mg/L IAA, 0.1~0.5 mg/L TDZ, and 1 mg/L 2,4-D under continuous dark condition at $25^{\circ}C$. Semi-solid agarose culture method was more effective than liquid culture, and it also protected the cells from browning caused by polyphenolic compound released during protoplast culture. A total of 4000 calli were obtained from protoplast culture of different capsicum species. All of these calli were transferred to the 100 combinations of regeneration media using various plant growth regulators; TDZ, IAA, 2ip, BAP, NAA, and zeatin. These calli derived from protoplast of three species of capsicum were, however, not differentiated into shoots.

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Establishment of optimal conditions for micropropagation by node culture and multiple shoots formation from sucker explants of thornless Blackberry (Rubus fruticosus L. cv. BB21) (가시없는 블랙베리(Rubus fruticosus L. cv. BB21)의 근맹아를 이용한 다경유도와 절간배양을 통한 식물체 증식조건의 확립)

  • Lee, Kang Seop;Kim, Hyo Jin;Park, Dae Hyun;Oh, Seung Cheol;Cho, Han Jig;Kim, Ee Youb
    • Journal of Plant Biotechnology
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    • v.45 no.2
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    • pp.110-116
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    • 2018
  • This study was conducted to develop a simple, rapid, and reliable method for in vitro propagation of disease-free and true-to-type clones from sucker explants of thornless blackberry (Rubus fruticosus L. ${\times}$ R. parvifolius L.). To induce multiple shoots, the sucker explants were sterilized in 1% NaOCl solution, and then were aseptically cultured on the full and 1/2 MS solid medium supplemented with BAP (0.1, 0.5, 1.0, 2.0 mg/L). After six weeks of culture, the highest frequency (85.4%) of shoot formation from sucker explants was obtained on the full-strength MS medium with 1.0 mg/L BAP. Node explants obtained from multiple shoots were cultured on the various media of full- or half-strength of AD, B5, MS, SH, QL, WPM media, respectively. After 30 days of culture, plant growth was good on the half-AD, half-QL medium. After 90 days of culture, plant growth was good on the full MS and full SH medium. The survival rate of the plantlets after transfer to plastic pots containing soil mixture (sand: soil: vermiculite was 1:1:1, vol.) in the greenhouse was 98%. The results indicate that a multiple-shoot procedure can be applied for an efficient mass propagation of Rubus fruticosus L. ${\times}$ R. parvifolius L.

Oligotrophic Media Compared with a Tryptic Soy Agar or Broth for the Recovery of Burkholderia cepacia Complex from Different Storage Temperatures and Culture Conditions

  • Ahn, Youngbeom;Lee, Un Jung;Lee, Yong-Jin;LiPuma, John J.;Hussong, David;Marasa, Bernard;Cerniglia, Carl E.
    • Journal of Microbiology and Biotechnology
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    • v.29 no.10
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    • pp.1495-1505
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    • 2019
  • The Burkholderia cepacia complex (BCC) is capable of remaining viable in low-nutrient environments and harsh conditions, posing a contamination risk in non-sterile pharmaceutical products as well as a challenge for detection. To develop optimal recovery methods to detect BCC, three oligotrophic media were evaluated and compared with nutrient media for the recovery of BCC from autoclaved distilled water or antiseptic solutions. Serial dilutions ($10^{-1}$ to $10^{-12}CFU/ml$) of 20 BCC strains were inoculated into autoclaved distilled water and stored at $6^{\circ}C$, $23^{\circ}C$ and $42^{\circ}C$ for 42 days. Six suspensions of Burkholderia cenocepacia were used to inoculate aqueous solutions containing $5{\mu}g/ml$ and $50{\mu}g/ml$ chlorhexidine gluconate (CHX) and $10{\mu}g/ml$ benzalkonium chloride (BZK), and stored at $23^{\circ}C$ for a further 199 days. Nutrient media such as Tryptic Soy Agar (TSA) or Tryptic Soy Broth (TSB), oligotrophic media (1/10 strength TSA or TSB, Reasoner's $2^{nd}$ Agar [R2A] or Reasoner's $2^{nd}$ Broth [R2AB], and 1/3 strength R2A or R2AB) were compared by inoculating these media with BCC from autoclaved distilled water and from antiseptic samples. The recovery of BCC in water or antiseptics was higher in culture broth than on solid media. Oligotrophic medium showed a higher recovery efficiency than TSA or TSB for the detection of 20 BCC samples. Results from multiple comparisons allowed us to directly identify significant differences between TSA or TSB and oligotrophic media. An oligotrophic medium pre-enrichment resuscitation step is offered for the United States Pharmacopeia (USP) proposed compendial test method for BCC detection.

Effect of Thidiazuron on Callus and Multiple Shoot Formation in Shoot-tip Culture of Hibiscus syriacus L. 'Honghwarang' (Thidiazuron이 무궁화 '홍화랑' 품종의 정단배양으로부터 Callus형성과 Multiple Shoot형성에 미치는 효과)

  • Kim, Eun Kyoung;Yoo, Yong Kweon;Kim, Ki Sun
    • Horticultural Science & Technology
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    • v.16 no.4
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    • pp.520-524
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    • 1998
  • This study was carried out to investigate the effect of thidiazuron(TDZ) on callus and shoot primordia formation, to determine the most optimum multiple shoot induction medium, and to obtain the plantlets on solid medium via shoot organogenesis. TDZ 0.01 mg/L in MS medium was most effective on callus formation, and BA 0.1 mg/L was most effective on shoot growth, while TDZ 0.01 mg/L was most effective on callus formation. TDZ 0.001 mg/L was most effective in shoot primordia formation. Shoot tips were cultured with TDZ 0.01 mg/L for 8 weeks and induced callus was transferred to regeneration medium containing TDZ 0.001 mg/L. After 4 weeks induced shoot primordia were resubcultured at growth regulator-free medium for 4 weeks. The induced multiple shoots rooted more efficiently at NAA 1.0, 5.0 mg/L, or IBA 5.0 mg/L.

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