• Title/Summary/Keyword: recombinant fermentation

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Rapid Purification of Recombinant Human Lipocortin-I Secreted from Saccharomyces cerevisiae

  • Chung, Bong-Hyun;Nam, Soo-Wan
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.4
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    • pp.242-246
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    • 2000
  • Human lipocortin-I was expressed as a secretory product by Saccharomyces cerevisiae harboring an expression system consisting of GAL10 promoter, inulinase signal sequence and lipocortin-I terminator. Fed-batch fermentation was carried out to overproduce recombinant human lipocortin-I. The culture medium was desalted and concentrated by ultrafiltration, and then subjected to hydroxyapatite column chromatography. The lipocortin-I was purified to >98% purity by single-step hydroxyapatite column chromato-graphy. However, it was found that the purified lipocortin-I was a proteolytically-cleaved form which was cleaved immediately after the basic amino acid Lys26.

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Scale-up of Recombinant Hirudin Production from Saccharomyces cerevisiae

  • Kim, Chul-Ho;K. Jagannadha Rao;Youn, Duk-Joong;Rhee, Sang-Ki
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.5
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    • pp.303-305
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    • 2003
  • Scale-up of hirudin production from Saccharomyces cerevisiae from bench-scale to pilot-scale was carried out based on constant volumetric oxygen transfer coefficient (K$\sub$L/a). Fed-batch mode of cultivation using step-wise feeding strategy of galactose was employed for the production of hirudin in a 30-L and a 300-L pilot-scale fermentor. The final hirudin concentrations were achieved 390 mg/L and 286.1 mg/L, and the volumetric productivities were 80.4% and 90.7% with the 30-L and 300-L fermentors, respectively, compared to the productivity of the 5-L bench-scale fermentor.

Monitoring of Biological Processes by 2-dimensional Fluorescence Sensor (2차원 형광센서에 의한 생물공정의 모니터링)

  • 이종일;정상윤;서국화
    • KSBB Journal
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    • v.16 no.5
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    • pp.493-499
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    • 2001
  • This work presented the monitoring technique of biological processes by a 2-dimensional fluorescence sensor. The 2-dimensional fluorescence sensor can be used to monitor some important variable during cultivation processes simultaneously. In this study we have monitored fermentation processes of a few microorganisms such as recombinant E.coli, A. terreus and T. vulgaris. and investigated the change of the fluorescence spectra in the fermentation processes qualitatively. The 2-dimensional fluorescence sensor can be also used to monitor biochemical reactions and separation processes and applied for the optimization of biological processes.

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Fructan Biosynthesis by Yeast Cell Factories

  • Hyunjun Ko;Bong Hyun Sung;Mi-Jin Kim;Jung-Hoon Sohn;Jung-Hoon Bae
    • Journal of Microbiology and Biotechnology
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    • v.32 no.11
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    • pp.1373-1381
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    • 2022
  • Fructan is a polysaccharide composed of fructose and can be classified into several types, such as inulin, levan, and fructo-oligosaccharides, based on their linkage patterns and degree of polymerization. Owing to its structural and functional diversity, fructan has been used in various fields including prebiotics, foods and beverages, cosmetics, and pharmaceutical applications. With increasing interest in fructans, efficient and straightforward production methods have been explored. Since the 1990s, yeast cells have been employed as producers of recombinant enzymes for enzymatic conversion of fructans including fructosyltransferases derived from various microbes and plants. More recently, yeast cell factories are highlighted as efficient workhorses for fructan production by direct fermentation. In this review, recent advances and strategies for fructan biosynthesis by yeast cell factories are discussed.

Plasmid Stabilitly of a Recombinant Escherichia coli and Optimization of Fermentation Processes (유전자 재조합 균주에서 플라스미드의 안정성과 발효 공정의 최적화)

  • 박성훈
    • Journal of Life Science
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    • v.2 no.1
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    • pp.26-34
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    • 1992
  • 유전자 재조합 균주의 발효에 있어서 플라스미드의 안정성과 발효공정의 최적화에 대해 개략적으로 서술하였다. 클론된 DNA의 발현은 플라스미드의 안정성을 크게 저해하며, 저하된 플라스미드의 안정성은 재조합 균주의 생산성을 많이 떨어뜨린다. 최적 발효 조건은 각각의 숙주-벡터 시스템, 사용한 배지, 생성물 등에 따라 크게 변한다. 동일한 숙주-벡터 시스템의 경우도, 사용하는 배지에 따라 온도, 희석률 또는 성장속도에 의존하는 정도가 달라지고 또 최적값도 다 변한다. 또한 발효조건의 최적화가 균체 내 플라스미드의 자기복제,mRNA로의 전사, 단백질로의 translation, 더 나아가 미생물 전체의 생리와 밀접하게 관련이 있다.

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Bacillus cereus에 의한 Phospholipase C (PLC) 생산

  • Seo, Guk-Hwa;Lee, Jong-Il;Bornscheuer, Uwe T.
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.232-234
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    • 2002
  • Bacillus cereus secretes a nonspecific phospholipase C (PLC) that catalyzes the hydrolysis of phospholipids to yield diacylglycerol and a phosphate monoester. This study focuses on the production of PLC by B. cereus and recombinant E. coli with fusion protein gene (plc::gfp). Fermentation processes have been monitored by a 2-dimensional fluorescence sensor.

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Genome Shuffling of Mangrove Endophytic Aspergillus luchuensis MERV10 for Improving the Cholesterol-Lowering Agent Lovastatin under Solid State Fermentation

  • El-Gendy, Mervat Morsy Abbas Ahmed;Al-Zahrani, Hind A.A.;El-Bondkly, Ahmed Mohamed Ahmed
    • Mycobiology
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    • v.44 no.3
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    • pp.171-179
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    • 2016
  • In the screening of marine mangrove derived fungi for lovastatin productivity, endophytic Aspergillus luchuensis MERV10 exhibited the highest lovastatin productivity (9.5 mg/gds) in solid state fermentation (SSF) using rice bran. Aspergillus luchuensis MERV10 was used as the parental strain in which to induce genetic variabilities after application of different mixtures as well as doses of mutagens followed by three successive rounds of genome shuffling. Four potent mutants, UN6, UN28, NE11, and NE23, with lovastatin productivity equal to 2.0-, 2.11-, 1.95-, and 2.11-fold higher than the parental strain, respectively, were applied for three rounds of genome shuffling as the initial mutants. Four hereditarily stable recombinants (F3/3, F3/7, F3/9, and F3/13) were obtained with lovastatin productivity equal to 50.8, 57.0, 49.7, and 51.0 mg/gds, respectively. Recombinant strain F3/7 yielded 57.0 mg/gds of lovastatin, which is 6-fold and 2.85-fold higher, respectively, than the initial parental strain and the highest mutants UN28 and NE23. It was therefore selected for the optimization of lovastatin production through improvement of SSF parameters. Lovastatin productivity was increased 32-fold through strain improvement methods, including mutations and three successive rounds of genome shuffling followed by optimizing SSF factors.

Extractive fermentation of Monascus purpureus promotes the production of oxidized congeners of the pigment azaphilone

  • Lim, Yoon Ji;Lee, Doh Won;Park, Si-Hyung;Kwon, Hyung-Jin
    • Journal of Applied Biological Chemistry
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    • v.61 no.4
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    • pp.327-334
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    • 2018
  • Monascus is a source of food colorant with high productivity of the pigment azaphilone. Monascus azaphilone (MAz) is biosynthesized through a single non-reducing polyketide pathway, the major components of which are ankaflavin (1), monascin (2), rubropunctatin (3) and monascorubrin (4); valuable biological activities have been reported for these compounds. Thus, various culture conditions were explored to reduce the cost of culture ingredients, enhance productivity and modulate compound composition. In the present study, we examined an extractive fermentation (EF) method with Diaion HP-20 resin (HP20) in direct comparison to a previously explored method involving Triton X-100 (TX100) to explore the modulated production of the major MAzs. We employed wild-type Monascus purpureus as well as two derivative recombinant strains (${\Delta}mppG$ and ${\Delta}mppE$) that are known to have differential MAz profiles as that of the wild-type strain. The HP20 resin was capable of modulating the MAz profile in favor of orange MAzs 3 and 4, oxidized congeners in this class, as was TX100-a phenomenon not previously observed for TX100 EF with Monascus anka. These finding substantiate that HP20 can be employed for the selective production of oxidized MAz and for diversifying the culture conditions used for Az production.