• Title/Summary/Keyword: optimization of the enzyme production

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Optimization of Conditions for the Production of Algin-like Polysaccharide by Polyglucuronic Acid C5-Epimerase (Polyglucuronic Acid C5-Epimerase에 의한 Algin 유사 다당류 생산 조건의 최적화)

  • Cho, Gye-Bong;Chang, Pahn-Shick
    • Korean Journal of Food Science and Technology
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    • v.32 no.1
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    • pp.147-153
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    • 2000
  • We could produce algin-like biomaterial of polyiduronan using polyglucuronic acid C5-epimerase with polyglucuronic acid prepared by specific oxidation of primary alcohol groups of four kinds of polysaccharides(corn starch, rice starch, sweet potato starch, and cellulose). The enzyme activity was determined by the modified Dische carbazole methodology with the isolated crude enzyme from the supernatant centrifuged at $100,000{\times}g$ for 1 hr after grinding fresh bovine liver. And then, the optimal substrate, pH, and temperature for the enzyme reaction of polyglucuronic acid C5-epimerase were determined as the oxidized sweet potato starch, 7.0, and $30^{\circ}C$, respectively. Conclusively, it could be possible to epimerize polyglucuronic acid in the oxidized sweet potato starch to polyiduronic acid. Therefore, we could obtain algin-like polysaccharide using the oxidized sweet potato starch and polyglucuronic acid C5-epimerase isolated from bovine liver.

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Optimization of Bovine Testicular PH-20 hyaluronidase Production in Pichia pastoris (소의 히아론산 분해효소(PH-20)의 Pichia pastoris에서의 생산 최적화)

  • Shin, Hwa Shook;Kim, Eunki
    • Korean Chemical Engineering Research
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    • v.46 no.4
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    • pp.764-768
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    • 2008
  • Bovine testicular hyaluronidase PH-20 was cloned into pPIC9 vector and expressed in Pichia pastoris. Recombinant PH-20 was 75 kDa MW and 7460 units/L activity. Extracellular hyaluronidase activity was two times higher than that of intracellular activity. Non-buffered medium and $30^{\circ}C$ cultivation was favorable for PH-20 production. 1M sorbitol as an osmotic pressure and 0.3% methanol inducer increased cell growth and enzyme activity. 0.4 M arginine augmentation decreased the proteolytic degradation of recombinant hyaluronidase.

Stepwise Addition Technology for the High Yield Production of Chitosanoligosaccharide (키토산올리고당 생산을 위한 다단계 첨가방법)

  • Lee, Ki-Sun;Kim, Seung-Mo;Lim, Hyun-Soo
    • KSBB Journal
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    • v.14 no.6
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    • pp.639-642
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    • 1999
  • Optimization of the chitosanoligosaccharide production was studied with chitosanase. The optimum conditions for the enzymic reaction have been determined. Enzyme stability was maintained above 90% after 6 days at pH 5.0. The optimum initial reaction rate was appeared in 1.0% of chitosan solution. The production yield of chitosanoligosaccharides was over at 0.4%~2.0% of chitosan. At 4.0% of chitosan solution, however, the production yield was decreased to 64.6%. To increase the yield, stepwise addition of substrate into the reactor was investigated. In this case, the yield was increased from 64.6% to 83.2% and the final concentrations of chitosanoligosaccharide was 12.26 mg/mL. By TLC analysis, most of the chitosanoligosaccharides produced were 3-5 mers.

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Isolation and Characterization of Various Strains of Bacillus sp. having Antagonistic Effect Against Phytopathogenic Fungi (식물 병원성 곰팡이에 길항작용을 갖는 다양한 Bacillus sp.의 균주 분리와 특성에 관한 연구)

  • Kim, Hee Sook;Kim, Ji-Youn;Lee, Song Min;Park, Hye-Jung;Lee, Sang-Hyeon;Jang, Jeong Su;Lee, Mun Hyon
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.603-613
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    • 2019
  • This study was carried out to examine the antagonistic effect against phytopathogenic fungi of isolated strains from soil samples collected from Busan, Changwon, and Jeju Island: Botrytis cinerea, Colletotrichum acutatum, Corynespora cassiicola, Fusarium sp., Rhizoctonia solani, Phytophthora capsici, and Sclerotinia sclerotiorum. According to results of our studies, isolated strains showed an antagonistic effect against phytopathogenic fungi. Such an antagonistic effect against phytopathogenic fungi is seen due to the production of siderophores, antibiotic substances, and extracellular amylase, cellulase, protease, and xylanase enzyme activities. Extracellular enzymes produced by isolated strains were significant, given that they inhibited the growth of phytopathogenic fungi by causing bacteriolysis of the cell wall of plant pathogenic fungi. This is essential to break down the cell wall of plant pathogenic fungi and thus help plant growth by converting macromolecules, which cannot be used by the plant for growth, into small molecules. In addition, they are putative candidates as biological agents to promote plant growth and inhibit growth of phytopathogenic fungi through nitrogen fixation, indole-3-acetic acid production, siderophore production, and extracellular enzyme activity. Therefore, this study suggests the possibility of using Bacillus subtilis ANGa5, Bacillus aerius ANGa25, and Bacillus methylotrophicus ANGa27 as new biological agents, and it is considered that further studies are necessary to prove their effect as novel biological agents by standardization of formulation and optimization of selected effective microorganisms, determination of their preservation period, and crop cultivation tests.

Screening New Antihypertensive Angiotensin I-Converting Enzyme Inhibitor -Producing Yeast and Optimization of Production Condition (항고혈압성 안지오텐신 전환효소 저해제를 생산하는 새로운 효모의 선별 및 저해물질 최적 생산조건)

  • Kang, Min-Gu;Kim, Ha-Kun;Yi, Sung-Hun;Lim, Sung-Il;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.39 no.3
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    • pp.194-197
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    • 2011
  • Forty eight strains of yeast were cultured in potato dextorse(PD) broth at $30^{\circ}C$ for 24 hr and centrifuged with 12,000 rpm for 20 min. After concentrated the cultures, antihypertensive angiotensin I-converting enzyme(ACE) inhibitory activities of its concentrates were investigated. Among them, the concentrates from Saccharomyces cerevisiae Y183-3 showed the highest ACE inhibitory activity of 71.8%. The ACE inhibitor from Saccharomyces cerevisiae Y183-3 was maximally produced when Saccharomyces cerevisiae Y183-3 cultured in PD broth at $30^{\circ}C$ for 36 hr.

Optimization of Xylanase Production from Paenibacillus sp. DG-22 (Paenibacillus sp. DG-22로부터 xylanase 생산의 최적화)

  • Lee, Yong-Eok
    • Journal of Life Science
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    • v.13 no.5
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    • pp.618-625
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    • 2003
  • Investigations were carried out to optimize the culture conditions for the production of xylanase by Paenibacillus sp. DG-22, a moderately thermophilic bacterium isolated from timber yard soil. Xylanase production showed a cell growth associated profile. Xylanase activity was found only in the culture supernatant, while $\beta-xylosidase$ activity was mainly associated with the cells. The formation of xylanase activity was induced by xylan and repressed by glucose and xylose. The production profile of xylanase was examined with various commercial xylan and maximum yield was achieved with 0.1∼ 0.5% birchwood xylan. Among various nitrogen sources tested, yeast extract was optimal for the production of xylanase. The xylanase activity was inhibited by $Co^{2+},\; Cu^{2+},\; Fe^{3+},\; Hg^{2+}\;$ and$\;Mn^{2+}$ ions while $Ca^{2+},\; Mg^{2+},\; Ni^{2+},\; Zn^{2+}$ions and DTT stimulated xylanase activity Mercury (II) ion at 5 mM concentration abolished all the xylanase activity. The predominant products of xylan-hydrolysate were xylobiose, xylotriose, and higher xylooligo-saccharides, indicating that the enzyme was an endoxylanase.

Optimization of Production of Trehaolse from Maltose using Recombinant Trehalose Synthase from Thermus caldophilus GK24 (재조합 트레할로스 합성효소에 의한 맥아당으로부터 트레할로스 생산 최적화)

  • 조연정;고석훈;이대실;신현재
    • KSBB Journal
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    • v.18 no.1
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    • pp.8-13
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    • 2003
  • Recombinant trehalose synthase from Thermus caldophilus GK24 showed an ability to produce trehalose from maltose. The activity of the partially purified enzyme was not influenced by most metal ions at 1 mM but was inhibited by 10 mM $Co^{2+}$, $Mn^{2+}$, and $Fe^{2+}$. Enzyme activity varied during prolonged reaction due to changes in the environmental conditions. Thus, the reaction was carried out for an extended time with optimized conditions of $45^{\circ}C$ and pH 7.0. An yield of 32.9% was reached at $60^{\circ}C$ after reaction for 22 h, and, maximum trehalose conversion (69.2%) was attained at $25^{\circ}C$. The yields obtained using enzyme dosages of 10, 25, and 50 U/g were 62.3, 62.3 and 59.0 %, respectively, though the initial conversion rate was higher when the higher dose was used. Similar profiles of trehalose production yields were observed with reaction working volumes of 10 ml to 2,000 ml.

Enzymatic Synthesis of Ethyl Butyrate Using Ester Synthetase Derived from Banana Peel and Pineapple Peel (바나나 껍질과 파인애플 껍질 Ester Synthetase를 이용한 Ethyl Butyrate의 효소적 합성)

  • Yoon, Ki-Hong;Kim, Kee-huck;Lee, Gyu-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.9
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    • pp.1122-1127
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    • 2017
  • Currently, the consumer trends are increasing towards "natural" in all food systems. Therefore, in the flavor industry, the production of flavor esters by "natural" methods are needed. On the other hand, "natural flavor" is expensive to produce because of the limited natural source. Recently, the flavor obtained from the enzyme or microbial could be represented as "natural flavor". Ethyl butyrate is used most frequently as a fruity aroma in drinks and the processed food industry. In this study, ethyl butyrate was synthesized enzymatically using the ester synthetase obtained from the waste of pineapple and banana peel. The ethyl butyrate production optimization was analyzed using a response surface methodology. The enzyme reaction variances were composed of the ethanol content, butyric acid content, and reaction time. As a result, in ester synthetase obtained from banana peel, the maximum predicted production amounts were 45.8199 mM at an ethanol content of 38.7050 mM, butyric acid content of 50.9019 mM, and reaction time of 4.3662 h. In ester synthetase obtained from pineapple peel, the maximum predicted production was 65.1087 mM at an ethanol content of 54.6502 mM, butyric acid content of 58.7638 mM, and reaction time of 4.7436 h. In conclusion, ethyl butyrate production was shown the more useful using the ester synthetase obtained from pineapple peel than that from banana peel.

Optimization of Combined Process of Enzymatic Hydrolysis and Solvent Extraction for Production of Lycopene from Elaeagnus umbellata (보리수 나무 열매로부터 라이코펜 생산을 위한 효소 분해 및 유기용매 추출 복합 공정의 최적화)

  • Oh, Yun Hye;Lee, Ju Mi;Chae, Hee Jeong
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.4
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    • pp.293-299
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    • 2020
  • This study was undertaken to optimize combining the processes of enzymatic hydrolysis and extraction for lycopene production from autumn olive berry. The autumn olive berry was pulverized and suspended in water, followed by treatment with various hydrolytic enzymes including Ceremix, Celluclast, AMG, Viscozyme, Pectinex, Promozyme, Ultraflo and Tunicase. Reaction solutions were subjected to extraction by applying different organic solvents including acetone, ethyl acetate, hexane and chloroform. Highest yields of lycopene extraction were obtained with the Ceremix (hydrolysis enzyme) and chloroform (extraction solvent) combination. Subsequently, using this ideal combination, enzymatic hydrolysis conditions, including enzyme concentration, pH and temperature, were statistically optimized to 0.58%, 5.5 and 54.4℃, respectively, by applying the response surface method. The lycopene extraction yield increased 2.3-fold (22.6 mg/100g) by using the selected combined process. We propose that these results could be used for the future development of bioactive materials required for bio-health care products.

Optimization of Glycosyl Aesculin Synthesis by Thermotoga neapolitana β-Glucosidase Using Response-surface Methodology (반응표면분석법을 이용한 Thermotoga neapolitana β-glucosidase의 당전이 활성을 통한 glycosyl aesculin 합성 최적화)

  • Park, Hyunsu;Park, Young-Don;Cha, Jaeho
    • Journal of Life Science
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    • v.27 no.1
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    • pp.38-43
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    • 2017
  • Glycosyl aesculin, a potent anti-inflammatory agent, was synthesized by transglycosylation reaction, catalyzed by Thermotoga neapolitana ${\beta}-glucosidase$, with aesculin as an acceptor. The key reaction parameters were optimized using response-surface methodology (RSM) and $2{\mu}g$ of the enzyme. As shown by a statistical analysis, a second-order polynomial model fitted well to the data (p<0.05). The response surface curve for the interaction between aesculin and other parameters revealed that the aesculin concentration and reaction time were the primary factors that affected the yield of glycosyl aesculin. Among the tested factors, the optimum values for glycosyl aesculin production were as follows: aesculin concentration of 9.5 g/l, temperature of $84^{\circ}C$, reaction time of 81 min, and pH of 8.2. Under these conditions, 61.7% of glycosyl aesculin was obtained, with a predicted yield of 5.86 g/l. The maximum amount of glycosyl aesculin was 6.02 g/l. Good agreement between the predicted and experimental results confirmed the validity of the RSM. The optimization of reaction conditions by RSM resulted in a 1.6-fold increase in the production of glycosyl aesculin as compared to the yield before optimization. These results indicate that RSM can be effectively used for process optimization in the synthesis of a variety of biologically active glycosides using bacterial glycosidases.