• Title/Summary/Keyword: skim milk-based medium

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High Cell Density Cultivation of Bifidobacterium longum Using a Calcium Carbonate-Alginate Beads System

  • Yu, Won-Kyu;Kim, Ji-Youn;Lee, Ki-Yong;Heo, Tae-Ryeon
    • Journal of Microbiology and Biotechnology
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    • v.12 no.3
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    • pp.444-448
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    • 2002
  • A $CaCO_3$-alginate beads system was developed for high cell density cultivation of Bifidobacterium longum and the cost-effective media were also screened. In batch process with $CaCO_3$, beads, two strains of B. longum showed both the highest viable cells and optical density in TPY medium, resulting in maximum optical density and viable cell counts of 12.40, $2.22{\times}10^10$ cfu/ml for B. longum ATCC 15707 and 13.71, $3.93{\times}10^10$ cfu/ml for B. longum HLC 3742. Released size distribution, according to $CaCO_3$-alginate bead size preparation, was smaller than others. These results were also examined by observing their morphology. The skim milk-based medium was most adequate to cultivate B. longum as the cheapest medium, and $10\%$ skim milk supplemented with $2\%$ glucose and $1\%$ yeast extract was a suitable medium, supporting the growth to $5.57{\times}10^10$ cfu/ml for ATCC 15707 and $6.82{\times}10^9$ cfu/ml for HLC 3742. During the long-term storage at $4^{\circ}C\;and\;-20{\circ}C$, B. longum cultivated with $CaCO_3$ beads had the highest stability. Consequently, $CaCO_3$-alginate beads buffer was found to be useful not only to cultivate B. longum but also to preserve cultures.

Isolation from Chungkookjang and Characterization of a Bacterium Producing an Extracellular Protease of High Specific Activity (청국장으로부터 고 비활성 세포외 Protease 생산 세균의 분리 및 동정)

  • Park, Hee-Jin;Park, Heui-Dong
    • Food Science and Preservation
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    • v.17 no.3
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    • pp.410-417
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    • 2010
  • Several extracellular protease-producing bacteria were isolated from Chungkookjang, a traditional Korean food of fermented soybeans, on skim milk agar plates. Among these bacteria, strain D14 exhibited the highest production (15.2 U/mL) and specific activity (40.0 U/mg protein) of extracellular protease activity as assessed on growth in a protease induction medium composed of 1% (w/v) soluble starch, 1.5% (w/v) skim milk, 0.5% (w/v) yeast extract, and 2% (w/v) NaCl. The bacterium was identified as Bacillus subtilis based on morphological and physiological characteristics and 16S rDNA sequence. A BLAST search of 16S rDNA sequences revealed that the isolate was most closely related to Bacillus subtilis subsp. subtilis strain NCIB 3610. The 16S rDNA sequence homology was 99.9%. Our isolate produced the highest level of protease when grown in a protease induction medium containing 1% (w/v) sorbitol and 0.5% (w/v) yeast extract. Fructose and glucose reduced enzyme production to 12.7% and 35.9%, respectively, of the level seen when the strain was grown in medium containing soluble starch. Soytone also reduced enzyme production to 61.4% of the level noted when the strain was grown in medium containing yeast extract.

Growth Characteristics of Polyporales Mushrooms for the Mycelial Mat Formation

  • Bae, Bin;Kim, Minseek;Kim, Sinil;Ro, Hyeon-Su
    • Mycobiology
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    • v.49 no.3
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    • pp.280-284
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    • 2021
  • Mushroom strains of Polyporales from the genera Coriolus, Trametes, Pycnoporus, Ganoderma, and Formitella were explored in terms of mycelial growth characteristics for the application of mushroom mycelia as alternative sources of materials replacing fossil fuel-based materials. Among the 64 strains of Polyporales, G. lucidum LBS5496GL was selected as the best candidate because it showed fast mycelial growth with high mycelial strength in both the sawdust-based solid medium and the potato dextrose liquid plate medium. Some of the Polyporales in this study have shown good mycelial growth, however, they mostly formed mycelial mat of weak physical strength. The higher physical strength of mycelial mat by G. lucidum LBS5496GL was attributed to its thick hyphae with the diameter of 13 mm as revealed by scanning electron microscopic analysis whereas the hyphae of others exhibited less than 2 mm. Glycerol and skim milk supported the best mycelial growth of LBS5496GL as a carbon and a nitrogen source, respectively.

Production of the Extracellular Alkaline Proteinase by Yarrowia Lipolytica 504D (Yarrowia lipolytica 504D의 Extracellular Alkaline Proteinase 생산성)

  • 유춘발;김창화;김태곤
    • Journal of Life Science
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    • v.8 no.3
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    • pp.333-338
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    • 1998
  • Productivity of alkaline proteinase from Yarrowia lipolytica 504D was investigated. For the production fo the enzyme, hemoglobin was the best nitogen source, however, casein and skim milk were also good. All carbon sources inhibited strongly the producitivity of the enzyme. Yeast extract increased the productivity of the enzyme to 220%, but almost mineral salts except monovalant ions decreased it. Based on these results, optimal medium was composed of 1.2% casein, 0.2% glucose, 0.16% yeast extract, and 0.1% ammonium sulfate. the best condition for the production of the enzyme was observed at pH 9 and $20^{\circ}C$ for 42 hours.

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Lactobacillus casei YIT 9018의 Shuttle Vector 개발을 위한 분자유전학적 연구

  • Yoo, Min;Nam, Jin-Sik;Kwon, Oh-Sik;Baek, Young-Jin
    • Microbiology and Biotechnology Letters
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    • v.25 no.5
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    • pp.464-467
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    • 1997
  • A shuttle vector, pSHvec, was constructed for Lactobacillus casei (L. casei) YIT 9018 and JM1O9 by recombinant DNA technology. This vector contained the $\beta$-lactamase II gene from Bacillus cereus as a selection marker and replication origins for both Gram(+) and Gram(-) strains. It could transform the wild type L. casei YIT 9018 as well as E. coli JM109 and transformed cells were selected based on antibiotics resistance. The ability of L. casei YIT 9018 for curd formation in 11% skim milk was maintained even after transformation with pSHvec. The vector was stable as long as antibiotics were added to the medium. These results could contribute to the study of lactic acid bacteria for the industrial purpose at a genetic level.

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Development of Cereal Product Containing γ-Aminobutyric Acid Producing Lactic Acid Bacteria Using Electrostatic Spray Technology (Electrostatic Spray 기술을 이용한 GABA 생성 유산균 함유 곡류 제품 개발)

  • Jeong, Ji-Hee;An, Do-Kyun;Kim, Dong-Kyun;Kim, Kwang-Yup
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.8
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    • pp.979-985
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    • 2017
  • This study was carried out to investigate the production of ${\gamma}$-aminobutyric acid (GABA) by lactic acid bacteria and to manufacture GABA using rice bran extract-based optimum medium. Electrostatic spraying technology was used to add GABA into the cereals. The isolated Lactobacillus brevis CFM11 produced the highest GABA production up to a concentration of $2,002.93{\mu}g/mL$ when cultivated in MRS broth containing 0.8% monosodium glutamate (MSG). The production level of GABA was $585.80{\mu}g/mL$ in rice bran extract containing 0.4% MSG, 2% sucrose, 1% skim milk, and 0.2% magnesium sulfate. After electrostatic spraying of the cultured suspension onto rice, GABA concentration reached $228.10{\mu}g/g$ while untreated rice reached $32.23{\mu}g/g$. These results demonstrate that rice bran extract can be an economic commercial medium for GABA production as a substitute for MRS broth. This study demonstrates the novel application of electrostatic spraying of GABA into cereal products for the first time.

Selection and Cultural Characteristics of Whole Chicken Feather-Degrading Bacterium, Bacillus sp. SMMJ-2 (Whole Chicken Feather-Degrading Keratinolytic Protease 생산균주의 분리 및 특성)

  • Park Sung-Min;Jung Hyuck-Jun;Yu Tae-Shick
    • Microbiology and Biotechnology Letters
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    • v.34 no.1
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    • pp.7-14
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    • 2006
  • Feather, generated in large quantities as a byproduct of commercial poultry processing, is almost pure keratin, which is not easily degradable by common professes. Four strains, SMMJ-2, FL-3, NO-4 and RM-12 were isolated from soil for production of extracellular keratinolytic protease. They were identified as Bacillus sp. based on their morphological and physiological characteristics. They shown high protease activity on 5.0% skim milk agar medium and produced a substrate like mucoid on keratin agar medium. Bacillus sp. SMMJ-2 had a faster production time for producing keratinolytic protease than other strains. This strain did not completely degrade whole chicken feather for five days in basal medium but completely degraded whole chicken feather when supplied with nitrogen source for 40hours in keratinolytic producing medium ($0.7%\;K_{2}HPO_{4},\;0.2%\;KH_{2}PO_{4},\;0.1%$ fructose, 1.2% whole chicken feather, $0.01%\;Na_{2}CO_3$, pH 7.0). When supplied with chicken feather as nitrogen source, keratinolytic protease activity was 89 units/ml/min. When soybean meal was used as nitrogen source, the keratinolytic protease production reached a maximum of 106 units/ml/min after 48 hours under $30^{\circ}C$, 180 agitation. To isolate the keratinolytic protease, the culture filtrate was precipitated with $(NH_4)_{2}SO_4$ and acetone. The recovery rate of keratinolytic protease was about 96% after treatment with 50% acetone. The enzyme was stable in the range of $30{\sim}50^{\circ}C$ and pH $6.0{\sim}12.0$.

Isolation and Characterization of Protease Producing B. amyloliquefaciens JH-35 from Food Waste (음식물 쓰레기로부터 Protease를 생산하는 B. amyloliquefaciens JH-35의 분리 및 특성)

  • Yoo, Jae Hong;Joo, Jin Ho;Kim, Sung Gug;Jang, In-Hwan
    • Korean Journal of Environmental Agriculture
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    • v.35 no.4
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    • pp.294-301
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    • 2016
  • BACKGROUND: Recent studies have described the importance of microbes and enzymes that can compost food waste. This study was carried out to improve production of protease of isolated microbes from food waste. METHODS AND RESULTS: Seven bacteria isolated from various sources were screened for protease production by adding skim milk into the agar medium. About 7 microbes producing protease were tested, and strain JH-35 showed the highest protease activity among them. The strain was identified as Bacillus amyloliquefaciens JH-35 based on morphological, cultural, physiological characteristics and 16S rRNA. In the fermentation experiment, the assay B. amyloliquefaciens JH-35 showed the highest protease activity in the condition of 1% glucose, 1.5% yeast extract and 0.2%$ K_2HPO_4$. The optimal condition of culture with temperature $35^{\circ}C$, initial pH of 7 and shaking speed of 200 rpm and 24 hr. CONCLUSION: The protease of the B. amyloliquefaciens JH-35 had its activity at pH 7 and the optimal culture time was 24 hr. Also, B. amyloliquefaciens JH-35 was high salt tolerance. Our results suggest that B. amyloliquefaciens JH-35 from food waste may have the potential to degrade protein and carbohydrate in food waste.

Isolation of Bacteria with Protease Activity from Cheonggukjang and Purification of Fibrinolytic Enzyme (청국장으로부터 혈전용해 활성이 우수한 균주 분리 및 혈전용해효소정제)

  • Choi, Yeon Hee;Lee, Jun Seung;Bae, So Young;Yang, Keun Jae;Yeom, Kyu Won;Jo, Dong Hyeok;Kang, Ock Hwa;Baik, Hyung Suk
    • Journal of Life Science
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    • v.23 no.2
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    • pp.259-266
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    • 2013
  • To isolate the fibrinolytic enzyme, 268 strains from 21 samples were morphologically isolated from Cheonggukjang collected from Korea and Japan. Among the 268 strains, protease-producing bacteria were isolated in nutrient agar medium including 1% skimmed milk. As a result of this, 22 strains were isolated. Apiweb site was used to identify these strains based on their biochemical properties. In addition, 16S rRNA sequencing was performed to identify the strain. Most of the identified strains were Bacillus subtilis and B. amyloliquefaciens. Fibrinolytic enzyme activity was measured with the fibrin plate method. Five strains were finally selected: A2-14, A2-20, C1-05, C1-09, and F2-01. Of those five strains, the A2-20 strain, which is close to B. amyloliquefaciens, showed the strongest fibrinolytic activity. The fibrinolytic enzyme produced by the A2-20 strain was partially purified from culture supernatant by gel filtration and ion exchange chromatography. The optimal pH and temperature values of the partially purified enzyme were 7.0 and $35^{\circ}C$, respectively. Purified protein analysis was carried out with SDS-PAGE and zymography. A genetic analysis was also conducted by PCR based on the consensus sequence of fibrinolytic enzyme. Corresponding genes with a partial sequence of the A2-20 strain were identified.

Screening of conjugated linoleic acid (CLA) producing Lactobacillus plantarum and production of CLA on soy-powder milk by these stains (공액리놀레산 생성 Lactobacillus plantarum 선발 및 이를 이용한 콩-분말 두유에서 공액리놀레산 생산)

  • Kim, Baolo;Lee, Byong Won;Hwang, Chung Eun;Lee, Yu-Young;Lee, Choonwo;Kim, Byung Joo;Park, Ji-Yong;Sim, Eun-Yeong;Haque, Md. Azizul;Lee, Dong Hoon;Lee, Jin Hwan;Ahn, Min Ju;Lee, Hee Yul;Ko, Jong Min;Kim, Hyun Tae;Cho, Kye Man
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.231-240
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    • 2015
  • In this study, a total of 16 conjugated linoleic acid (CLA) producing lactic acid bacteria (LAB) were isolated from fermented foods. Among those strains, the S48 and P1201 strains were capable of producing higher CLA contents than other LABs. The two strains were classified as Lactobacillus plantarum based on morphological, physiological, chemotaxonomic, and molecular-genetic properties. The survival rates of these strain appeared to be 59.57% and 62.22% under artificial gastric conditions after 4 h at pH 2.5, respectively. These strains produced the cis-9, trans-11, and trans-10, cis-12 CLA isomers from 8% skim milk medium supplemented with the different free LA concentration at $37^{\circ}C$ for 48 h and the production of two CLA isomers constantly increased in the growth until 48 h of incubation. After 48 h of fermentation, the levels of CLA appeared highest in steamed soy-powder milk than fresh and roasted soy-powder milks. In particular, the CLA contents were produced $183.57{\mu}g/ml$ and $198.72{\mu}g/ml$ from steamed soy-powder milk after fermentation (48 h) with S48 and P1201 strains, respectively.