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Shengkai Li,Liwen Zhang,Shuhong Gao 한국생물공학회 2023 Biotechnology and Bioprocess Engineering Vol.28 No.5
Lincomycin, a clinically important antibiotic against Gram-positive bacteria, is a sulfur-containing metabolite produced by Streptomyces lincolnensis. The main problem in the fermentation is the high content of lincomycin B, which is a by-product of insufficient methylation by Sadenosylmethionine (SAM) dependent methyltransferase. In this study, the content of lincomycin B was decreased from 4.04% to 0.75% and from 8.20% to 5.78% by the addition of NaCl as an osmotic regulator in flask and 15 L bioreactor fermentation, respectively. By analysis of the sulfur-containing metabolites (cysteine, homocysteine, SAM, and ergothioneine) and high-throughput RNA sequencing, the mechanism of osmotic regulation on lincomycin fermentation was studied. The results showed that the synthesis of methyl donor (SAM) and sulfur donor of lincomycin (ergothioneine and mycothiol) were increased at the metabolite level and transcription level under osmotic stimulation; the transcription of genes involved in sulfur assimilation (tauA, ssuA1B1C1) and lincomycin biosynthesis were also up-regulated significantly. This study provides a cost-effective method to reduce lincomycin B and increase the production of lincomycin A. The mechanisms of osmotic pressure regulation on lincomycin fermentation was also elucidated, which may provide more genetically modified targets for overproduction of lincomycin and other sulfur containing secondary metabolites.
김경자,이재훈,양용준,Kim, Kyoung-Ja,Lee, Jae-Hun,Yang, Yong-Joon The Microbiological Society of Korea 2014 미생물학회지 Vol.50 No.1
Protocatechualdehyde (PA)는 항산화 활성과 항암 활성을 가진 페놀성 물질이다. Streptomyces lincolnensis M-20 균주에서 생산된 PA를 균주 상등액에서 분리, 정제하였다. 항산화 활성을 가진 PA가 구리 이온 존재 하에서는 산화촉진제로 작용하였다. 항산화 활성은 DPPH를 이용한 방법으로 측정하였으며, 구리 이온 존재 하에서 PA의 산화 촉진 작용은 pBR322 플라스미드의 DNA 절단 작용으로 측정하였다. DNA 손상으로 생성되는 활성산소 종의 확인은 활성 산소종의 포집자인 글루타치온에 의해 DNA 절단이 억제되는 것으로 확인하였다. PA와 구리 이온의 복합체 형성은 금속 이온의 킬레이트인 EDTA가 존재할 경우와 존재하지 않을 경우를 자외선/가시광선 분광학적 분석법으로 비교, 확인하였다. Protocatechualdehyde (PA) is phenolic compound having antioxidative and antitumor activities. PA was purified from supernatant of Streptomyces lincolnensis M-20. In the presence of copper ion, PA acted as pro-oxidant. The antioxidant activity was assessed with the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, and the pro-oxidant effect of PA on DNA damage as pBR322 plasmid DNA-cleaving agents in the presence of Cu(II) ions was investigated. The involvement of reactive oxygen species (ROS) in the DNA damage was confirmed by the inhibition of the DNA breakage by using glutathione (GSH), specific scavenger of ROS. When the increase in ROS reaches a certain level (the toxic threshold), it may trigger cell death. The formation of the PA/Cu(II) chelate complex was confirmed by reaction with ethylenediamine-tetraacetic acid (EDTA), a well-known chelating agent for metal ions, by using UV/Vis spectroscopic analysis.
Kim, Kyoung-Ja,Kim, Mi-Ae,Jung, Jee-Hyung 대한약학회 2008 Archives of Pharmacal Research Vol.31 No.12
We characterized the biological functions of protocatechualdehyde (PA) isolated from the butanol extract of culture supernatant from Streptomyces lincolnensis M-20. Following butanol extraction, it was purified by silica gel and Sephadex LH-20 column chromatography. PA was analyzed by Furier Transform Infrared Spectroscopy (FT-IR), Gas chromatograph-Mass Spectrometer (GC-MS), and Nuclear Magnetic Resonance (NMR). PA had potent antioxidant activity, as measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. Antitumor activity against MCF-7 human breast cancer cells was evaluated by the 3-(4,5-dimethylthazol-2-yl)-2,5-diphenyl tetrazolium-bromide (MTT) assay. PA treatment ($0{\sim}150{\mu}M$) dose-dependently blocked apoptosis, as shown by improved cell viability and inter-nucleosomal DNA fragmentation. Our findings suggest that Streptomyces lincolnensis M-20, a lincomycin producer, also produces protocatechualdehyde.
김경자,김미애,정지형 대한약학회 2008 Archives of Pharmacal Research Vol.31 No.12
We characterized the biological functions of protocatechualdehyde (PA) isolated from the butanol extract of culture supernatant from Streptomyces lincolnensis M-20. Following butanol extraction, it was purified by silica gel and Sephadex LH-20 column chromatography. PA was analyzed by Furier Transform Infrared Spectroscopy (FT-IR), Gas chromatograph-Mass Spectrometer (GC-MS), and Nuclear Magnetic Resonance (NMR). PA had potent antioxidant activity, as measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. Antitumor activity against MCF-7 human breast cancer cells was evaluated by the 3-(4,5-dimethylthazol-2-yl)-2,5-diphenyl tetrazolium-bromide (MTT) assay. PA treatment (0~150 μM) dose-dependently blocked apoptosis, as shown by improved cell viability and inter-nucleosomal DNA fragmentation. Our findings suggest that Streptomyces lincolnensis M-20, a lincomycin producer, also produces protocatechualdehyde.
이유리,이미자,최윤이,전계택,정용섭 한국생물공학회 2014 Biotechnology and Bioprocess Engineering Vol.19 No.6
Production of lincomycin by Streptomyceslincolnensis was studied by varying medium compositionand environmental conditions. With the medium compositionoptimized by statistical experimental design (45 g/L solublestarch, 15 g/L sugar cane molasses, 13.33 g/L peptonewater, 6.67 g/L NaNO3 and 4.0 g/L CaCO3), lincomycinproduction increased by 2 ~ 3 fold compared to thatobtained with un-optimized basal medium. Lincomycinproduction was further improved by optimizing cultureconditions such as agitation speed, impeller type and pHunder the optimized medium condition. The highest titer of350 mg/L lincomycin was achieved from 240 h bioreactorculture. These results demonstrate that fermentation conditionsfor maximal lincomycin production by Streptomyceslincolnensis were optimized via biotechnological processes.
Ki-An Cho,Hoon Cho 대한환경위생공학회 2008 대한환경위생공학회지 Vol.23 No.2
Using crude soybean oil as the sole carbon source, the lincomycin production from Streptomyces lincolnensis LC 345 was investigated in the air lift bioreactor. When 30 g/L of crude soybean oil was used, the maximum lincomycin concentration reached 0.89 g/L, after 5 days of culture. When CSL concentration was increased from 10 to 30 g/L, Lincomycin concentration was increased from 0.6 to 1.2. On the other hand, when CSL concentration was increased from 40 to 60 g/L, it was decreased from 1.15 to 0.7 g/L. Using these results, fed batch cultures for comparing the use of crude soybean oil and glucose as a conventional carbon source were carried out in a 5 L air lift bioreactor. When crude soybean oil was used as the sole carbon source, the maximum lincomycin concentration was 2.0 g/L, which was about 2.0 fold higher than that of glucose medium after 7 day of culture. The product yield from olive oil was 0.042 gig consumed carbon source, which was about 3.8 fold higher than that of glucose.
Cho, Ki-An,Cho, Hoon Korean Society for Environmental Sanitary Engineer 2008 대한환경위생공학회지 Vol.13 No.2
Using crude soybean oil as the sole carbon source, the lincomycin production from Streptomyces lincolnensis LC 345 was investigated in the air lift bioreactor. When 30 g/L of crude soybean oil was used, the maximum lincomycin concentration reached 0.89 g/L, after 5 days of culture. When CSL concentration was increased from 10 to 30 g/L, Lincomycin concentration was increased from 0.6 to 1.2. On the other hand, when CSL concentration was increased from 40 to 60 g/L, it was decreased from 1.15 to 0.7 g/L. Using these results, fed batch cultures for comparing the use of crude soybean oil and glucose as a conventional carbon source were carried out in a 5 L air lift bioreactor. When crude soybean oil was used as the sole carbon source, the maximum lincomycin concentration was 2.0 g/L, which was about 2.0 fold higher than that of glucose medium after 7 day of culture. The product yield from olive oil was 0.042 g/g consumed carbon source, which was about 3.8 fold higher than that of glucose.