• Title/Summary/Keyword: optimization conditions

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Optimization of Culture Medium for the Production of an Exopolysaccharide (p-CY02) with Cryoprotective Activity by Pseudoalteromonas sp. RosPo-2 from the Antarctic Sea

  • Pilsung Kang;Sung Jin Kim;Ha Ju Park;Il Chan Kim;Se Jong Han;Joung Han Yim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.5
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    • pp.1135-1145
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    • 2024
  • When cells are exposed to freezing temperatures, high concentrations of cryoprotective agents (CPA) prevent ice crystal formation, thus enhancing cell survival. However, high concentrations of CPAs can also cause cell toxicity. Exopolysaccharides (EPSs) from polar marine environments exhibit lower toxicity and display effects similar to traditional CPA. In this study, we sought to address these issues by i) selecting strains that produce EPS with novel cryoprotective activity, and ii) optimizing culture conditions for EPS production. Sixty-six bacteria producing mucous substances were isolated from the Ross Sea (Antarctic Ocean) using solid marine agar plates. Among them, Pseudoalteromonas sp. RosPo-2 was ultimately selected based on the rheological properties of the produced EPS (p-CY02). Cryoprotective activity experiments demonstrated that p-CY02 exhibited significantly cryoprotective activity at a concentration of 0.8% (w/v) on mammalian cells (HaCaT). This activity was further improved when combined with various concentrations of dimethyl sulfoxide (DMSO) compared to using DMSO alone. Moreover, the survival rate of HaCaT cells treated with 5% (v/v) DMSO and 0.8% (w/v) p-CY02 was measured at 87.9 ± 2.8% after freezing treatment. This suggests that p-CY02 may be developed as a more effective, less toxic, and novel non-permeating CPA. To enhance the production of EPS with cryoprotective activity, Response Surface Methodology (RSM) was implemented, resulting in a 1.64-fold increase in production of EPS with cryoprotective activity.

Overproduction of Xanthophyll Pigment in Flavobacterium sp. JSWR-1 under Optimized Culture Conditions

  • Jegadeesh Raman;Young-Joon Ko;Jeong-Seon Kim;Da-Hye Kim;Soo-Jin Kim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.3
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    • pp.710-724
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    • 2024
  • Flavobacterium can synthesize xanthophyll, particularly the pigment zeaxanthin, which has significant economic value in nutrition and pharmaceuticals. Recently, the use of carotenoid biosynthesis by bacteria and yeast fermentation technology has shown to be very efficient and offers significant advantages in large-scale production, cost-effectiveness, and safety. In the present study, JSWR-1 strain capable of producing xanthophyll pigment was isolated from a freshwater reservoir in Wanju-gun, Republic of Korea. Based on the morphological, physiological, and molecular characteristics, JSWR-1 classified as belonging to the Flavobacterium species. The bacterium is strictly aerobic, Gram-negative, rod-shaped, and psychrophilic. The completed genome sequence of the strain Flavobacterium sp. JSWR-1 is predicted to be a single circular 3,425,829-bp chromosome with a G+C content of 35.2% and 2,941 protein-coding genes. The optimization of carotenoid production was achieved by small-scale cultivation, resulting in zeaxanthin being identified as the predominant carotenoid pigment. The enhancement of zeaxanthin biosynthesis by applying different light-irradiation, variations in pH and temperature, and adding carbon and nitrogen supplies to the growth medium. A significant increase in intracellular zeaxanthin concentrations was also recorded during fed-batch fermentation achieving a maximum of 16.69 ± 0.71 mg/l, corresponding to a product yield of 4.05 ± 0.15 mg zeaxanthin per gram cell dry weight. Batch and fed-batch culture extracts exhibit significant antioxidant activity. The results demonstrated that the JSWR-1 strain can potentially serve as a source for zeaxanthin biosynthesis.

Optimization of Solid-State Fermentation Condition Using Distiller's Dried Grain (주정박을 이용한 고체발효 조건의 최적화)

  • Choi, Gi-Wook;Moon, Se-Kwon;Kim, Yule;Jang, Byung-Wook;Kim, Young-Ran;Chung, Bong-Woo
    • KSBB Journal
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    • v.23 no.4
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    • pp.345-349
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    • 2008
  • To enhance the value as a feedstuff of distiller's dried grain (DDG) and develop fermented feedstuff, we investigated the effects of the culture conditions affecting glucoamylase activity, such as pH in submerged culture and moisture content in solid-state culture. Also, we investigated the optimal mixing ratio of DDG and wheat bran for the production of fermented feedstuff containing high content of amino acids. In culture conditions for high fermented activity, pH and moisture were optimum at pH 4 and 60%, respectively. In the case of mixing ratio, the glucoamylase activity was decreased with increase of DDG content. On the other hand, the content of crude protein was increased slowly. For the development of fermented feedstuff, the optimal mixing ratio of DDG and wheat bran was 1 to 4. Finally, we could produce approximately 1 ton (dry matter) of trial product in incubator of pilot-scale. The glucoamylase activity and the crude protein content were 1,024 U/g and 33.6%, respectively.

Development of Loop-mediated Isothermal Amplification (LAMP) for Detection of Vibrio alginolyticus (Loop-mediated isothermal amplification (LAMP)법을 이용한 Vibrio alginolyticus의 신속 진단법 개발)

  • Hong, Seung-hyun;Heo, Moon-Soo
    • Journal of Life Science
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    • v.25 no.8
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    • pp.903-909
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    • 2015
  • Loop-mediated isothermal amplification (LAMP), a PCR-based diagnostic method, is based on autocycling strand displacement DNA synthesis in the presence of exonuclease-negative Bst DNA polymerase under isothermal conditions. With the help of four specific primers that recognize six different sequences of a target DNA, LAMP has high specificity in pathogenic identification in a short time. Hence, in the present study, LAMP is used as a diagnostic tool in the identification of the most dreadful aquatic pathogenic species, Vibrio alginolyticus, and to develop species-specific LAMP primers and optimization of LAMP reaction conditions such as annealing temperature, elongation time, and other PCR chemical concentrations, including MgSO4, dNTPs, Betaine, and Bst polymerase. The optimized LAMP primers were also checked for specificity with other Vibrio species, which showed that the designed primers were very specific to V. alginolyticus After the first introduction of a species name like this one, the first part (“Vibrio” in this case) should be abbreviated to only the first letter.only. These are usually the most harmful pathogens of the Vibrio species that appear in shrimp and crabs. The results also revealed that the LAMP assay could be 10-fold more sensitive than conventional PCR in detecting V. alginolyticus. This could be the first report on using a rapid and highly sensitive technique, the LAMP assay, in the effective diagnosis of the pathogenic bacteria V. alginolyticus, which could help in the early detection of diseases, particularly in aquaculture.

Characterization and Culture Optimization of an Glucosidase Inhibitor-producing Bacteria, Gluconobactor oxydans CK-2165 (α-Glucosidase 저해제 생산 균주, Gluconobacter oxydans CK-2165의 특성 및 배양 최적화)

  • Kim, Byoung-Kook;Suh, Min-Jung;Park, Ji-Su;Park, Jang-Woo;Suh, Jung-Woo;Kim, Jin-Yong;Lee, Sun-Young;Choi, Jongkeun;Suh, Joo-Won;Lee, In-Ae
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.13 no.11
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    • pp.5179-5186
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    • 2012
  • Miglitol, a well-known therapeutic intervention agents for diabetes, exhibits competitive inhibitory activity against ${\alpha}$-glucosidase and it is usually produced through three sequential steps including chemical and bioconversion processes. Gluconobactor oxydans (G. oxydans) belonging to acetic acid bacteria biologically, converts 1-deoxy-1-(2-hydroxyethylamino)-D-glucitol (P1) into a key intermidiate, 6-(2-hydroxyetyl) amino-6-deoxy-${\alpha}$-L-sorbofuranose (P2) by incomplete oxidation. In this study, we identified and optimized fermentation conditions of CK-2165, that was selected in soil samples by comparing the bioconversion yield. CK-2165 strain was found to be closely related to G. oxydans based on the result of phylogenetic analysis using 16S rDNA sequence. Utilization of API 20 kits revealed that this strain could use glucose, mannose, inositol, sorbitol, rhamnose, sucrose, melibiose, amygdalin and arabinose as carbon sources. The culture conditions were optimized for industrial production and several important factors affecting bioconversion rate were also tested using mycelial cake. Cell harvested at the late-stationary phase showed the highest bioconversion yield and $MgSO_4$ was critically required for the catalytic activity.

Evaluation of the Flux According to Membrane Distillation Module Structure and Operating Conditions Using PVDF Hollow Fiber Membrane (PVDF 중공사 분리막을 이용한 MD 모듈 구조 및 운전 조건에 따른 플럭스 영향 평가)

  • Min, Ji Hee;Lee, Seul ki;Gil, Nam Seok;Park, Min Soo;Kim, Jin Ho
    • Membrane Journal
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    • v.28 no.2
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    • pp.83-89
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    • 2018
  • Hydrophobic porous PVDF hollow fiber membranes for Membrane Distillation (MD) were fabricated by a combination of thermally induced phase separation (TIPS) and stretching. The purpose of this study is to investigate the shape and operating conditions of the module and the effect of piping size on parallel connection. In the optimization experiment of the vacuum membrane distillation module, the flux decreased as the packing density and length of the membrane in the module increased. When the module was connected vertically, it was confirmed that the nearest to the inlet of the vacuum port was the highest flux. In selecting the size of the header pipe of the module, it was confirmed that the maximum flux is shown when the inner diameter area of the hollow fiber membrane and the inner diameter area of the header pipe are the same. Also, it is necessary to find the optimal linear velocity because the higher the linear velocity in the module, the higher the flux, but the pressure acting on the module also increases proportionally.

Case Studies for SMR Natural Gas Liquefaction Plant by Capacity in Small Scale Gas Wells through Cost Analysis (소규모 가스전 규모에 따른 SMR 천연가스 액화 플랜트 용량별 비용 분석 사례연구)

  • Lee, Inkyu;Cho, Seungsik;Lee, Seungjun;Moon, Il
    • Journal of the Korean Institute of Gas
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    • v.20 no.3
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    • pp.46-51
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    • 2016
  • Natural gas liquefaction process which spends a huge amount energy is operated under cryogenic conditions. Thus, many researchers have studied on minimizing energy consumption of LNG plant. However, a few studied for cost optimization have performed. This study focused on the cost analysis for the single mixed refrigerant (SMR) process, one of the simplest natural gas liquefaction process, which has different capacity. The process capacity is increased from 1 million ton per annum (MTPA) to 2.5 MTPA by 0.5 MTPA steps. According to the increase of plant size, only flow rate of natural gas and mixed refrigerant are increased and other operating conditions are fixed. Aspen Economic Evaluator(v.8.7) is used for the cost analysis and six tenths factor rule is applied to obtain multi stream heat exchanger cost data which is not supplied by Aspen Economic Evaluator. Moreover, the optimal plant sizes for different sizes of gas wells are found as the result of applying plant cost to small scale gas wells, 20 million ton (MT), 40 MT, and 80 MT. Through this cost analysis, the foundation is built to optimize LNG plant in terms of the cost.

Establishment of the Optimum Culture Conditions for Mozzarella Cheese manufacturing by Streptococcus macedonicus LC743 with Immunomodulating Activity (면역 활성능을 가진 Streptococcus macedonicus LC743을 이용한 모짜렐라 치즈 제조의 최적 배양조건 확립)

  • Park, Sun-Young;Han, Noori;Lim, Sang-Dong
    • Journal of Dairy Science and Biotechnology
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    • v.33 no.3
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    • pp.215-221
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    • 2015
  • This study aimed to establish the optimum culture conditions for Mozzarella cheese by using Streptococcus macedonicus LC743, a strain selected for its immunomodulatory activity. For process optimization, 1.0% and 2.0% strain was inoculated and incubated at $32^{\circ}C$ and $35^{\circ}C$, respectively. The general components, bacterial count, total solids, yields, and immunomodulatory activity of the Mozzarella cheese were investigated. When the strain was inoculated at 2.0% and incubated at $32^{\circ}C$, the product quality and immunomodulatory activity was optimal and required minimal processing time. Therefore, 2.0% of S. macedonicus LC743 starter culture was added to milk at $32^{\circ}C$, after pasteurization at $63^{\circ}C$ for 30 min, and agitated for 4~5 min after addition of $230{\mu}L/kg$ of rennet. Curd was made by setting the milk 25~35 min after addition of 0.01~0.02% calcium chloride. The curd was cut at 0.1~0.12% acidity (81 min later) and after heating the cheese to up to $43^{\circ}C$. Whey was removed at an acidity of 0.17~0.18% by agitation for 53 min. Next, cheddaring for 210 min up to an acidity of 0.6~0.65%, stretching at $72{\sim}75^{\circ}C$, and molding at $65{\sim}70^{\circ}C$ were performed, and the product was allowed to cool down to $5^{\circ}C$. Salting was done with a solution of $18{\sim}20^{\circ}B{\acute{e}}$ at $12^{\circ}C$ for 20 min and drying occurred at 80~90% relative humidity at $10^{\circ}C$ for 2~3 days.

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Optimization of Production Conditions of Biosurfactant from Bacillus sp. and its Purification (Bacillus sp.에 의한 생물계면활성제의 생산 및 그의 성질)

  • Kim, Jin-Sook;Song, Hee-Sang;Chung, Nam-Hyun;Bang, Won-Gi
    • Applied Biological Chemistry
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    • v.48 no.2
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    • pp.109-114
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    • 2005
  • A bacterium capable of emulsifying hydrocarbon, n-hexadecane, and decreasing surface tension of the culture media using oil collapsing method was isolated. The bacterium was partially identified as Bacillus sp. and named BJS-51. n-Hexadecane was the most effective carbon source for production of biosurfactant. Surface tension was decreased from 76 dyne/cm to 31 dyne/cm and CMD (critical micelle dilution) had the highest value of 5.7 at 3% n-hexadecane. Ammonium phosphate was the most effective nitrogen source, when C/N ratio was 60, surface tension and CMD were 29 dyne/cm and 9.2, respectively. Optimum pH and temperature were 7.2 and $30^{\circ}C$, respectively. Produced biosurfactant was extracted and purified using organic solvent extraction method and preparative HPLC systems. After analysis by various color reaction, this biosurfactant was identified as lipopolysaccharide. Surface tension and CMC (critical micelle concentration) of purified biosurfactant were 27 dyne/cm and 0.08 g/l, repectively. CMD was 9.2, so the yield of biosurfactant was about 0.74 g/l at the optimal conditions. The biosurfactant was very stable at wide range of $pH\;2{\sim}12$ with surface tension $29{\sim}31\;dyne/cm$ and showed $29{\sim}30\;dyne/cm$ of surface tension after heat treatment at $100^{\circ}C$ for 60 min.

The Optimization of Mevinolin Production by Medium Composition of Penicillium citrinum (Penicillium citrinum 의 배지 조성에 의한 Mevinolin 생산 최적화 연구)

  • 차월석;신성의;권규혁;김선일;이동병;이태범
    • KSBB Journal
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    • v.15 no.3
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    • pp.232-237
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    • 2000
  • Theses studies were made on the mevinolin production from Penicillium citrinum Thom (KCTC 6990) Culture conditions pH temperature carbon sources nitrogen sources mineral sources surfactants and glucose concentration were optimized. The results of glucose concentration and maximum mevinolin production according to incubating time in the flask nearly disappeared after 5 days and appeared after 7 days respectively. temperature and pH conditions of maximum mevinolin production were $24^{\circ}C$ and 3.7 pH respectively. The results of maximum mevinolin production according to the kind of nutrients were as follows. Glucose of carbon sources were 3.5 mg/L. Peptone of nitrogen sources were 3.5 mg/L TEX>$K_2HP0_4$ of mineral sources was 3.8 mg/L Tween 20 of surfactants were 4.5 mg/L Maximum mevinolin productioni of glucose con-centration was 4.0mg/L of glucose 100 g/L In the batch culture Maximum mevinolin concentration was 10.3 mg/L after 8 days. maximum mevinolin specific production rate 0.016 mg/g-hr. These results need to be studied more than ever about temperature pH 야ㅕㅡ and treatment of by-product oil in the batch culture and must do the fad batch from now to increase mevinolin productivity.

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