• Title/Summary/Keyword: yeast strains

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Cultivation of Lactic Acid Bacteria for the Development of Probiotic Products using Red Ginseng Starch (프로바이오틱스 개발을 위한 홍삼 전분을 활용한 유산균 배양)

  • Kim, Yeong-Su;Lee, Hwan;Kim, Do-Yeon;Kim, So-Young;Lee, Wan-Kyu;Lee, Sang-Myeong;Park, Jong-Dae;Shon, Mi-Yae
    • Journal of the East Asian Society of Dietary Life
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    • v.23 no.6
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    • pp.818-826
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    • 2013
  • To reduce the production cost of Lactobacillus, discarded red ginseng starch was collected from a factory of red ginseng extract in order to develop the Lactobacillus culture medium. According to the analysis of the gensenoside composition of red ginseng starch, the total gensenoside content of starch was 2.73 mg/g, and the gensenoside $Rb_1$, $Rb_2$ and $Rg_3$ contents were 0.1, 0.29 and 0.52 mg/g, respectively. For the preparation of the liquid media, red ginseng starch was added at rates of 0, 5, 10 and 20%. Further, Lactobacillus plantarum 15357 and Leuconostoc mesenteroides sub sp. strains were then inoculated to these prepared broths. With the red ginseng starch medium, the growth rates ($1.42{\times}10^7$ and $2.96{\times}10^{10}$ CFU/mL) and the final cell concentrations were higher than the MM medium ($1.0{\times}10^7$ CFU/mL). The optimal concentration of red ginseng starch and yeast extract as a medium were 20% and 10 g/L, respectively. Under these conditions, the cell mass of L. plantarum 15357 and L. mesenteroides sub sp. reached $5.11{\times}10^{10}$ and $8.17{\times}10^{10}$ CFU/mL. These results show a great possibility for the utilization of red ginseng starch as economic medium sources in the production of cell mass of lactic acid bacteria. This is the first trial of development of economic LAB growth medium using discarded red ginseng starch.

Changes in the quality characteristics of kimchi broths and growth of the isolated strains due to blue light emitting diode irradiation (Blue LED (Light emitting diode) 조사에 의한 배추김치와 백김치 발효액의 품질 특성 및 분리 균주의 생육 변화)

  • Oh, Yeong Ji;Hong, Jungil
    • Korean Journal of Food Science and Technology
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    • v.52 no.5
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    • pp.538-545
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    • 2020
  • Kimchi is a widely consumed traditional Korean food, and its probiotic properties have received great attention. In this study, changes in the quality characteristics of fermentation broths obtained from two types of Chinese cabbage kimchi (red with red pepper and white without red pepper) were assessed after the administration of blue light emitting diode (BLED) irradiation at 4℃; characteristics assessed included acidity, chromaticity, antioxidant activity, and growth modulation of isolated microorganisms. The pH of the white kimchi (WK) broth decreased with time; the decrease was delayed significantly under BLED irradiation (p<0.05). BLED irradiation decreased the L (lightness) and b (yellowness) values and increased a (redness) in WK, whereas the a and b values of the red kimchi (RK) broth increased with BLED irradiation. Growth stimulation of lactic acid bacteria by BLED irradiation was observed in both WK and RK. The numbers of yeast and mold were also increased in RK (p<0.05), but not in WK. There was no change observed in the scavenging activities against ABTS (2,2'-azido-bis(3-ethylbenzthiazoline-6-sulfonic acid) radicals in both kimchi broths after BLED treatment. The results of this study indicated that BLED irradiation could modulate the fermentation process and the quality characteristics of kimchi during storage.

Formulations of Bacillus thuringiensis Insecticides by Liquid and Semi-Solid Fermentations. (액상 및 반고체배지 발효에 의한 Bacillus thuringiensis 살충제의 제조)

  • 이형환
    • Microbiology and Biotechnology Letters
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    • v.26 no.4
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    • pp.369-372
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    • 1998
  • Microbial insecticide formulations were prepared by liquid and semi-solid fermentations using Bacillus thuringiensis subsp. kurstaki, HL-106 (BTK-HL106), B. thuringiensis subsp. israelensis HL-63 (BTI-HL63) and B. sphaericus 1593 (BS-1593) strains. The liquid fermentation medium contained molasses 2%, dextrose 1.5%, peptone 2%, D-xylose 0.025%, CaCl$_2$ 0.1%, K$_2$HPO$_4$ 0.1%, KH$_2$PO$_4$ 0.1%, MgSO$_4$$.$7H$_2$O 0.03%, FeSO$_4$$.$7H$_2$O 0.002%, ZnSO$_4$$.$7H$_2$O 0.02%. The composition of the semi-solid fermentation medium was rice bran 45.2%, zeolite 31%, yeast powder 0.02%, corn powder 5%, dextrose 3%, lime 0.3%, NaCl 0.06%, CaCl$_2$ 0.02%, and H$_2$O 15.42%. Insecticide formulations produced in the liquid fermentation named BTK-HL106, BTI-HL63 and BS-1593 pesticides and those in the semi-solid fermentation were designated as BTK-HL106-1, BTI-HL63-1 and BS-1593-1 pesticides, respectively. The number of spore (endotoxin crystals) was 2.65${\times}$10$\^$9/ spores per $m\ell$ in the BTK-HL106 and 3.5${\times}$10$\^$10/ in the BTK-HL106-1 3.8${\times}$10$\^$9/ spores in the BTI-HL63 and 7.0${\times}$10$\^$10/ in the BTI-HL63-1, and 7.5${\times}$10$\^$9/ in the BS-1593 and 1.4${\times}$10$\^$10/ in the BS-1593-1. The spores in the BS-1593 formulation was produced two times more than the other formulations. The spores in the BTI-HL63-1 were contained twice than those in the BTK-HL106-1, and five times than those in the BS-1593-1. The results indicated that spore (endotoxin crystals) productions in the semi-solid fermentation increased about ten times than those in the liquid fermentations. $LC_{50}$s of the BTI-HL63 and BS-1593 were 4.5 $\mu\textrm{g}$, and those of the BTI-HL63-1 and BS-1593-1 were 1.5 $\mu\textrm{g}$. $LC_{50}$ of the BTK-HL106 was 1.5 mg and that of the BTK-HL106-1 was 0.9 mg. The $LC_{50}$s of the formulations in the semi-solid fermentations showed about two to three times higher than those in the liquid fermentations.

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Isolation, Characterization, and Molecular Cloning of the cDNA Encoding a Novel Phytase from Aspergillus niger 113 and High Expression in Pichia pastoris

  • Xiong, Ai Sheng;Yao, Quan-Hong;Peng, Ri-He;Li, Xian;Fan, Hui-Qin;Guo, Mei-Jin;Zhang, Si-Liang
    • BMB Reports
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    • v.37 no.3
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    • pp.282-291
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    • 2004
  • Phytases catalyze the release of phosphate from phytic acid. Phytase-producing microorganisms were selected by culturing the soil extracts on agar plates containing phytic acid. Two hundred colonies that exhibited potential phytase activity were selected for further study. The colony showing the highest phytase activity was identified as Aspergillus niger and designated strain 113. The phytase gene from A. niger 113 (phyI1) was isolated, cloned, and characterized. The nucleotide and deduced amino acid sequence identity between phyI1 and phyA from NRRL3135 were 90% and 98%, respectively. The identity between phyI1 and phyA from SK-57 was 89% and 96%. A synthetic phytase gene, phyI1s, was synthesized by successive PCR and transformed into the yeast expression vector carrying a signal peptide that was designed and synthesized using P. pastoris biased codon. For the phytase expression and secretion, the construct was integrated into the genome of P. pastoris by homologous recombination. Over-expressing strains were selected and fermented. It was discovered that ~4.2 g phytase could be purified from one liter of culture fluid. The activity of the resulting phytase was 9.5 U/mg. Due to the heavy glycosylation, the expressed phytase varied in size (120, 95, 85, and 64 kDa), but could be deglycosylated to a homogeneous 64 kDa species. An enzymatic kinetics analysis showed that the phytase had two pH optima (pH 2.0 and pH 5.0) and an optimum temperature of $60^{\circ}C$.

Treatment test for bovine mastitis by the determination of ATP based on firefly bioluminescence (Bioluminescence 반응에 의한 ATP측정을 이용한 젖소 유방염 치료에 관한 연구)

  • Kim, Tae-jong;Kim, Jong-bae;Lee, Seong-bae;Jeon, Young-soo
    • Korean Journal of Veterinary Research
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    • v.29 no.3
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    • pp.393-405
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    • 1989
  • This study was carried out to treatment test for bovine mastitis by the determination of adenosine triphosphate (ATP) based on firefly bioluminescence. The results obtained are followed; 1. In the susceptibility test, cephalothin which looks the most effective were sensitive to Staphylococcus sp. (72.3%), Micrococcus sp. (84.2%), Streptococcus sp. (72.7%) and Gram positive bacilli (72.7%), Gram negative bacilli were sensitive to gentamicin (92.3%) and Yeast-like-fungi was the most sensitive to clotrimazole, and nystatin in order. 2. When the number of bacteria, such as Staphylococcus aureus, Candida tropicalis isolated from the mastitis milk were counted by conventional agar plating technique, and compared with the concentration of bacterial ATP, it gave a good linear relationship. The content of ATP per Staphylococcus aureus, cell was 3.1fM and Candida tropicalis showed the high level of A TP (90fM). 3. The ATP assay was applied to the determination of minimal inhibitory concentration (MIC) of various antibiotics. When Staphylococcus aureus was incubated in the presence of different concentration of tetracycline, erythromycin, kanamycin and streptomycin sulfate and the growth was monitored by the conventional agar plating technique and ATP assay, both methods shown the same results that they were 1mcg/ml, 2mcg/ml, 6.25mcg/ml and 8mcg/ml, respectively. 4. For the determination of susceptibility of sensitive and resistant Staphylococcus au reus isolated for the milk with mastitis to tetracycline, erythromycin kanamycin and streptomycin sulfate, the minimum time required for the test was determined by the assay of ATP every 30 minutes during incubation of 3 hours at $37^{\circ}C$. ATP concentration time curve calculated on both resistant and sensitive strains incubated 3 hours as the optimum time for the determination of susceptibilities of various antibiotics exemed. The ATP concentration of each test broth (antibiotic containing), expressed as a percentage of its own control brith (antibioticfree) indicated values of 30% to be indicative of each antibiotic sensitivity. Single time point ATP assay carried out on the various sensitive and resistant of Staphylococcus aureus to antibiotics examined after 3 hours at $37^{\circ}C$ correlated exactly with disc diffusion and MIC. 5. In the cure of intramammary treatment of bovine mastitis in lactating quarters, the cure rate of Staphylococcal mastitis showed to cephalexin (80%), cloxacillin and gentamicin (70%), ampicillin and oxytetracycline (60%), and Streptococcal mastitis showed to cephalexin (85%), penicillin (80%), cloxacillin and oxytetracycline (75%), and ampicillin (70%), but intramammary antimycotic drug (clotrimazol) were only a little effect about fungal mastitis.

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Characterization of ${\alpha}$-amylase Producing Hybrid Constructed between Saccharomycopsis and Saccharomyces (Saccharomycopsis속과 Saccharomyces속의 잡종형성 균주에서 생산하는 ${\alpha}$-amylase의 특성)

  • Yang, Young-Ki;Moon, Myeng-Nim;Lim, Chae-Young;Rhee, Young-Ha;Kim, Jeong-Ho
    • Korean Journal of Microbiology
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    • v.35 no.4
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    • pp.315-321
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    • 1999
  • This study has been performed to deveope a yeast strain having high ${\alpha}$-amylase production ability using nuclear transfer method. Hybrids formed between the strains of Saccharomyces fiburigera KCTC 7393 and Saccharomyces cerevisiae KCTC 7049 (tyr-, ura-)were obtained by nuclear transfer technique. Nuclei isolated from the wild type S. fiburigera strain were transfered into auxotrophic mutants S. cerevisiae and selected the hybrids showing an increased starch degrading capability were selected (MN-16). This transformant grew best and produced maximal ${\alpha}$-amylase activity on the medium containing 2% (V/V) soluble starch. ${\alpha}$-Amylase from MN-16 was purified electrophoretically homogenety and its properties were investigated. The enzyme was purified about 10.6 fold with an overall yield 9.7% from the culture medium by ammonium sulfate fractionation. DEAE-Sephacel column chromatography, and Sephacryl S-200 column chromatography. The purified enzyme showed a single band on SDS-polyacrylamide gel electrophoresis. The molecular weight of the ${\alpha}$-amylase was estimated to be 53,000 daltons by SDS-PAGE and by gel permeation chromatography on Sephacryl S-200. The purified enzyme showed the maximum activity at pH 5.5 and 40${\circ}C$. The km value for soluble starch was 2.5㎎/㎖. The enzyme activity increased in the presence of $Ca^{2+}, Co^{2+}, EDTA, Mg^{2+}, Mn^{2+}, Zn^{2+}$, but inhibited by $Cu^{2+}, Fe^{2+}$, and $Ni^{2+}$

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Secretory Overexpression of Clostridium Endoglucanase A in Saccharomyces cerevisiae Using GAL10 Promoter and Exoinulinase Signal Sequeice. (Saccharomyces cerevisiae에서 GAL 10 promoter와 exoinulinase 분비 서열을 이용한 Clostridium endoglucanase A의 과발현·분비)

  • Lim, Myung-Ye;Lee, Jin-Woo;Lee, Jae-Hyung;Kim, Yeon-Hee;Seo, Jin-Ho;Nam, Soo-Wan
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1248-1254
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    • 2007
  • The secretory overexpression of Clostridium thermocellum endoglucanase A gene (celA) was examined in Saccharomyces cerevisiae using Kluyveromyces marxianus exoinulinase (INU1) signal sequence and GAL10 promoter. The two plasmids, pYEG-CT1 with its own signal sequence, and pYInu-CT1 with INU1 signal sequence were introduced to S. cerevisiae SEY2102 and S. cerevisiae 2805 host strains, respectively, and then each transformant was selected on the synthetic defined media lacking uracil. The expression level and secretion efficiency of endoglucanase A was increased by $18{\sim}22%$ and 11%, respectively, by INU1 signal sequence over celA signal sequence. By considering the high level of expression (361 unit/I), plasmid stability (89%), and secretion efficiency (70%), S. cerevisiae 2805 harboring plasmid pYInu-CT1 was selected as the opti-mal host vector system for the production of cellulose-degrading enzyme and recombinant yeast probiotic. The total expression and secretion efficiency of endoglucanase A was 418 unit/l and 73%, respectively, in the batch fermentation of S. cerevisiae 2805/pYlnu-CT1 on galactose medium. The mo-lecular weight of secreted endoglucanase A was found to be greater than 100 kDa, presumably due to the N-linked glycosylation.

Development of Microbial Inoculant Using By-product of Oriental Herbal Medicine (한약재박을 이용한 미생물제제의 개발)

  • Joo, Gil-Jae;Kim, Young-Mog;Woo, Cheol-Joo;Lee, Oh-Seuk;Kim, Joung-Woong;So, Jae-Hyun;Kwak, Yun-Young;Lee, Jong-Jin;Kim, Jin-Ho;Rhee, In-Koo
    • Applied Biological Chemistry
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    • v.48 no.3
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    • pp.201-206
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    • 2005
  • The development of microbial inoculant was conducted using a by-product of oriental herbal medicine. The constituent of the by-product, which was high in organic matter, was 11.3% of crude protein, 5.1% of crude lipid, 49.7% of NDF (neutral detergent fiber), and 33.8% of ADF (acid detergent fiber). Microorganisms isolated from the by-product of oriental herbal medicine were 35 species. Among them, 6 bacterial species, 4 fungal species, 2 actnomycetes species, and 1 yeast species were effective in the utilization of the by-products. The 13 strains screened were tested for the plant growth-promoting effect in soybean seedling. BL-333 strain was found to increase the soybean yield by about 23% as compared with control. The strain BL-333 was identified as Paenibacillus marcerans. P. marcerans BL-333 showed high anti-fungal activities against virulent fungi, especially Fusarium sp. and Collectotrichum sp. Yields of plants which were inoculated with microbial inoculant prepared with P. marcerans BL-333 and by-product of oriental herbal medicine were found to be higher than control by $3{\sim}24%$. The yield was especially promoted in lettuce, radish, chinese cabbage and cucumber plants.

Antimicrobial Activity of the Aerial Part (Leaf and Stem) Extracts of Cnidium officinale Makino, a Korean Medicinal Herb (천궁(Cnidium officinale Makino) 지상부(잎과 줄기) 추출물의 항균활성)

  • Jung, Dong-Sun;Lee, Na-Hyun
    • Microbiology and Biotechnology Letters
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    • v.35 no.1
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    • pp.30-35
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    • 2007
  • This study was carried out to investigate the usefulness of the aerial part of Cnidium officinale Makino as a bioactive material source. The aerial part(leaf and stem) of Cnidium officinale Makino was extracted with three kinds of solvents and determined their antimicrobial activities against several bacteria and yeast strains using the paper disc method and the microtiter dilution method. The extracts of the Cnidium offocinale aerial part exhibited the broad spectrum of antibacterial activity against Gram (+) and Gram (-) bacteria, including food-borne pathogens such as Listeria monocytogenes, Salmonella typhimurium, and Staphylococcus aureus. The extracts of Cnidium officinale also showed antifungal activity against Saccharomyces cerevisiae. The ethyl acetate extracts completely inhibited the growth of Staphylococcus aureus and Pseudomonas aerogenes, and moderately inhibited the growth of Escherichia coli and Enterobacter cloacae at the concentration of 0.5 mg/mL. However, water extract of Cnidium officinale exhibited lower antimicrobial activity than ethyl acetate and methanol extracts. The inhibitory effect of the ethyl acetate extract of Cnidium officinale Makino was not destroyed by heating at $100^{\circ}C$ for 30 min or at $121^{\circ}C$... for 15 min. These results suggest that the aerial part of Cnidium officinale Makino could be a useful source for a natural antimicrobial material.

The Isolation and Culture Characterization of a Lipolytic Enzyme Producing Strain from Meju (메주로부터 지질분해 효소 생산 균주의 분리 및 배양학적 특성)

  • Yun, Hye-Ju;Lee, You-Jung;Yeo, Soo-Hwan;Choi, Hye-Sun;Park, Hye-Young;Park, Heui-Dong;Baek, Seong-Yeol
    • Microbiology and Biotechnology Letters
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    • v.40 no.2
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    • pp.98-103
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    • 2012
  • For screening of useful enzymes producing microorganisms from Meju, we isolated high lipase producing strains and their lipolytic enzyme activities were then tested. The lipolytic enzyme activities of isolated microorganisms were therefore tested on the Y124 strain. The gene sequence analysis of ITS from Y124 strain revealed Yarrowia lipolytica. Lipase production by the Y124 strain was studied in media containing various carbon sources. The Y124 strain drastically increased lipolytic enzyme activity in YPO media containing olive oil, as well as in YPDO media containing both olive oil and glucose. Maximal lipase production was achieved in YPD (yeast extract-peptone-D-glucose) media containing 0.7% olive oil when cultured at $30^{\circ}C$ for 8 hrs. The lipase produced from the Y124 strain showed the highest activity in p-NPO (p-nitrophenyl octanoate ($C_8$)), amongst the various p-nitrophenyl esters.