• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.378-381
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• 2001

Motierella alpina DSA-12 의 회 분식 발효에서 $MnSO_4$ 첨가로 작은 pellet을 형성하게 하여 유가식 배양으로의 적용 가능성을 보았으나, $KH_2PO_4_4$의 경우에는 균체증식에는 효과가 있었지만, 균체가 풀어져 자라 점도가 증가하여 ARA 의 합성이 억제되었다. 유가식 배양에 14 % 암모니아 용액을 사용할 때 균체증식과 총지질의 함량에 영향이 있음을 확인했으며, 62.1 g/L의 균체량에 60% 이상의 지질 함량으로 12.0 g/L 의 아라키돈산을 생산할 수 있었다.

• 한국미생물·생명공학회지
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• 제23권1호
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• pp.6-11
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• 1995

In order to screen microorganism producing lactic acid with high productivity from nature, we used a medium containing 100 g/l glucose and selected several microorganisms producing more than 80 g/l L-lactic acid. We investigated their physiological characteristics and compared them. The best microorganism was identified as Lactobacillus casei subsp. rhamnosus. The optimum pH for growth and production of lactic acid was 6.0 and this strain showed the highest growth rate at around 30$\circ$C , but the optimum temperature for lactic acid production was 45$\circ$C . The growth was inhibited proportionally from 50 g/l to 300 g/l of glucose and the maximal cell mass increased according to increasing the concentration of corn steep liquor (CSL) protein up to 30 g/l. In batch fermentation for lactic acid production, we produced 128 g/l L-lactic acid with 20 g/l CSL protein and 150 g/l glucose in 35 hours. In pH-stat fed-batch fermentation, we were able to produce 183 g/l L-lactic acid.

• 한국식품과학회지
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• 제24권4호
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• pp.326-329
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• 1992

${\gamma}-linolenic$ acid를 함유한 유지를 생산하기 위해 포도당을 주기적으로 첨가하면서 Mucor sp. KCTC 8405P를 유가배양하였다. 배양 시작 후 2일까지 효모 모양으로 배양한 후 배지의 pH를 높여 균사체 모양으로 유도하여 배양하면, 지방질은 포도당이 고갈되는 배양 7일째까지 축적되었다. 본 배양조건에 의하여 건조 균체량 99.3 g/l, 총지방질 38.0 g/l와 3.5 g/l의 ${\gamma}-linolenic$ acid를 얻을 수가 있었다.

• 미생물학회지
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• 제37권2호
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• pp.158-163
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• 2001

Avermectin B1a의 생산성 향상을 위하여 무기인의 영향을 조사하고, 주요 유기질소원의 최적농도를 response surface methodology를 적용하여 구하였다. 1.5 g/ι의 농도로 무기인을 아버멕틴 생산배지에 첨가하였을 때 B1b 성분의 구성비가 5.8%에서 3%로 감소하였으며 B1a 생성은 영향을 받지 않았다. 배지의 주요성분인 대두분, 면실분, 효모추출물 중 아버멕틴 생성에 가장 큰 영향을 끼치는 유기질소원은 대두분이었다. 실험실 규모 발효조에서 Streptomyces avermifilis YA99-40의 유가식 회분배양에 의해 아버멕틴 생산성을 회분배양에 비해 44.8% 증가시킬 수 있었다. 발효조 배양시작 후 136, 206 시간에 각각 30, 20 g/ι의 당을 추가하는 유가식 회분배양을 실시하였을 때 B1a 성분의 최대 생산성은 회분배양 대비 86.3% 증가하였으며 생성된 총 아버멕틴 중 B1a의 구성비율도 화분배양에 비해 38%에서 45%로 향상되었다. 이 같은 결과는 산업적인 규모로 아버멕틴의 생산성을 향상시키는데 유용하게 적용시킬 수 있다.

• 한국미생물·생명공학회지
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• 제31권1호
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• pp.46-50
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• 2003

젓갈에서 분리된 Lactococcus lactis SA72균주로부터 유용 박테리오신인 lacticin SA72의 생산을 최적화하기 위해 배양용 배지 및 배양조건을 검토하였다. 유산균의 대표적인 배양 배지인 BHI, MRS, Ml7배지에 대해서 검토한 결과, 이중 MRS배지가 최적 배지로 결정되었다. 이 MRS배지에 0.5% glucose를 첨가한 경우, 더 높은 활성을 얻을 수 있었다. pH의 영향을 검토하였을 때는, 초기 pH에서는 pH 7이었고, 발효조에서의 배양기간 동안 pH 6으로 조절할 때가 높은 활성을 보였다. 배양온도는 $32^{\circ}C$로 유지할 때가 활성이 우수하였다. 회분식 배양에서는 최대 3,200AU/m1을 얻을 수 있었으며, 유가식 배양으로 10g/1 glucose를 주기적으로 공급하였을 때 , 약 6,400 AU/ml로 가장 높은 활성을 얻을 수 있었다.

• 한국미생물·생명공학회지
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• 제19권5호
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• pp.491-496
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• 1991

Aminoglycoside-3'-phosphotransferase(APH(3'))를 생산하는 균주인 E.coli ATCC 21990을 산업적으로 이용하기 위해서 자외선 조사 및 NTG를 처리하고 고농도의 kanamycin B에 내성을 갖는 변이주인 E.coli M1과 M2를 선별하였다. E.coli M1은 단위 균체당 효소 생산성은 높으나 생육속도가 낮아 실용적이질 못했고 주 질소원인 yeast extract를 사용했을때 E.coli M1보다 E.coli M2가 생육속도가 훨씬 빨랐으며 약 2배의 APH(3')을 얻을 수 있었고 산소 가스를 사용하였을 경우는 약 2.5배의 APH(3')을 얻었다.

• Journal of Microbiology and Biotechnology
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• 제32권9호
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• pp.1178-1185
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• 2022

Steroids are a class of compounds with cyclopentane polyhydrophenanthrene as the parent nucleus, and they usually have unique biological and pharmacological activities. Most of the biosynthesis of steroids is completed by a series of enzymatic reactions starting from cholesterol. Synthetic biology can be used to synthesize cholesterol in engineered microorganisms, but the production of cholesterol is too low to further produce other high-value steroids from cholesterol as the raw material and precursor. In this work, combinational strategies were established to increase the production of cholesterol in engineered Saccharomyces cerevisiae RH6829. The basic medium for high cholesterol production was selected by screening 8 kinds of culture media. Single-factor optimization of the carbon and nitrogen sources of the culture medium, and the addition of calcium ions, zinc ions and citric acid, further increased the cholesterol production to 192.53 mg/l. In the 5-L bioreactor, through the establishment of strategies for glucose and citric acid feeding and dissolved oxygen regulation, the cholesterol production was further increased to 339.87 mg/l, which was 734% higher than that in the original medium. This is the highest titer of cholesterol produced by microorganisms currently reported. The fermentation program has also been conducted in a 50-L bioreactor to prove its stability and feasibility.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.219-222
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• 2000

Physiological changes of Escherichia coli during the fed-batch fermentation process were characterized in this study. Overall cellular protein samples prepared at the different stage of fermentation were separated by 2-dimensional gel electrophoresis (2-DE), and differently expressed 15 proteins, Phosphotransferase enzyme I, GroEL, Trigger factor, ${\beta}$ subunit of ATP synthase, Transcriptional regulator KDGR, Phosphoglycerate mutase 1, Inorganic pyrophosphatase, Serine Hydroxymethyl-transferase, ${\alpha}$ subunit of RNA polymerase, Elongation factor Tu, Elongation factor Ts, Tyrosine-tRNA ligase, DnaK suppressor protein, Transcriptional elongation factor, 30S ribosomal protein S6 were identified using matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS). When bacterial cells grow to high cell density, and IPTG-inducible heterologous protein is produced, expression level of overall cellular proteins was decreased. According to their functions in the cell, identified proteins were classified into three groups, proteins involved in transport process, small-molecule metabolism, and synthesis and modification of macromolecules.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.845-848
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• 2001

For understanding physiology and metabolism under various culture conditions, combined analysis of transcriptome and proteome is attractable way. We have manufactured DNA microarray containing 2,850 genes including all functionally known and putative ones. In this study, we report analysis of transcriptome and proteome during the high cell density culture of E. coli by using DNA microarray and 2-DE. Fed-batch fermentation of E. coli was carried out by exponential feeding of nutrients until the maximum cell density reached 74 g dry cell weight/L (g DCW/L). Changes in transcriptome and proteome during the HCDC are analyzed qualitatively and quantitatively to provide their physiological and metabolic meanings.

• Journal of Microbiology and Biotechnology
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• 제25권6호
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• pp.845-855
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• 2015

The present study describes the gene cloning and high-level expression of an alkaline and thermostable lipase gene from Trichosporon coremiiforme V3. Nucleotide analysis revealed that this lipase gene has an open reading frame of 1,692 bp without any introns, encoding a protein of 563 amino acid residues. The lipase gene without its signal sequence was cloned into plasmid pPICZαA and overexpressed in Pichia pastoris X33. The maximum lipase activity of recombinant lipase was 5,000 U/ml, which was obtained in fed-batch cultivation after 168 h induction with methanol in a 50 L bioreactor. The purified lipase showed high temperature tolerance, and being stable at 60℃ and kept 45% enzyme activity after 1 h incubation at 70℃. The stability, effects of metal ions and other reagents were also determined. The chain length specificity of the recombinant lipase showed high activity toward triolein (C18:1) and tripalmitin (C16:0).