KR101442682B1 - 미생물의 세포 내 대사 프로파일을 위한 추출 및 분석 방법 - Google Patents
미생물의 세포 내 대사 프로파일을 위한 추출 및 분석 방법 Download PDFInfo
- Publication number
- KR101442682B1 KR101442682B1 KR1020120038484A KR20120038484A KR101442682B1 KR 101442682 B1 KR101442682 B1 KR 101442682B1 KR 1020120038484 A KR1020120038484 A KR 1020120038484A KR 20120038484 A KR20120038484 A KR 20120038484A KR 101442682 B1 KR101442682 B1 KR 101442682B1
- Authority
- KR
- South Korea
- Prior art keywords
- phosphate
- metabolites
- intracellular
- biomass
- metabolite
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
- G01N27/622—Ion mobility spectrometry
- G01N27/623—Ion mobility spectrometry combined with mass spectrometry
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/483—Physical analysis of biological material
- G01N33/487—Physical analysis of biological material of liquid biological material
- G01N33/48707—Physical analysis of biological material of liquid biological material by electrical means
- G01N33/48735—Investigating suspensions of cells, e.g. measuring microbe concentration
-
- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- H01J49/00—Particle spectrometers or separator tubes
- H01J49/26—Mass spectrometers or separator tubes
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Pathology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- Biophysics (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
도 2는 (a) ME, (b) BE, (c) MC, (d) HW, (e) PH 및 (f) PA 추출방법에 대한 UPLC/Q-TOF-MS ES-총 이온 크로마토그램을 보여준다.
도 3은 크렙시엘라 옥시토카의 세포내 대사물질의 농도를 나타낸다. 배양 후 10시간에 샘플을 수득하여 각 추출방법[냉장 매탄올(검정색), 고온 에탄올(회색), 메탄올-클로로포름(백색), 열수(사선), 수산화칼륨(가로줄) 및 과염소산(교차줄)]별로 나타낸다.
도 4는 20 대사물질에 대한 추출 효율의 최대값 및 중간값을 보여준다.
Claims (11)
- 크렙시엘라 옥시토카의 세포 내 대사 프로파일(intracellular metabolic profile)을 얻는 방법:
(a) 크렙시엘라 옥시토카의 바이오매스를 -80℃ 내지 -30℃의 100 % 냉장 메탄올(cold methanol)에 현탁한 다음 동결시키는 단계;
(b) 상기 동결된 바이오매스를 해동시키는 단계;
(c) 상기 단계 (a) 및 (b)를 1-10 차례 반복하여 상기 바이오매스로부터 누출된 대사물의 현탁액을 얻는 단계; 및
(d) 상기 대사물의 현탁액에 대하여 매스 스펙트로미트리(mass spectrometry) 분석을 실시하여 상기 크렙시엘라 옥시토카의 세포 내 대사물을 동정하는 단계; 상기 대사물은 3-포스포글리세린산염(3-phosphoglycerate; 3-PG), 아세틴-코엔자임 A(acetyl-coenzyme A; AcCoA), 프럭토즈 6-포스페이트(fructose 6-phosphate; F6P), 프럭토즈 1,6-비스포스페이트(fructose 1,6-bisphosphate; FBP), 글루코즈 1-포스페이트(glucose 1-phosphate; G1P), 글루코오즈 6-포스페이트(glucose 6-phosphate; G6P), 포스포에놀피루브산(phosphoenolpyruvate; PEP), α-케토글루타레이트(α-ketoglutarate; α-KG), 글리세롤 3-포스페이트(glycerol 3-phosphate; G3P), NAD(Nicotinamide Adenine Dinucleotide), NADH(Nicotinamide Adenine Dinucleotide reduced form), NADP(Nicotinamide Adenine Dinucleotide Phosphate), NADPH(Nicotinamide Adenine Dinucleotide Phosphate reduced form), ADP(Adenosine Diphosphate), AMP(Adenosine Monophosphate), ATP(Adenosine Triphosphate), 글리세레이트(glycerate), 시트레이트(citrate), 글루타메이트(glutamate) 및 말레이트(malate)로 구성된 군으로부터 선택되는 하나 이상의 대사물이다.
- 삭제
- 삭제
- 삭제
- 삭제
- 제 1 항에 있어서, 상기 단계 (a)의 동결은 액체 질소를 이용하여 실시하는 것을 특징으로 하는 방법.
- 제 1 항에 있어서, 상기 단계 (b)의 해동은 드라이아이스 상에서 실시하는 것을 특징으로 하는 방법.
- 삭제
- 제 1 항에 있어서, 상기 단계 (c) 및 (d) 사이에 상기 대사물의 현탁액을 여과하는 과정을 추가적으로 포함하며 상기 단계 (d)는 상기 여과 과정을 거친 대사물의 현탁액을 이용하여 실시하는 것을 특징으로 하는 방법.
- 삭제
- 제 1 항에 있어서, 상기 방법은 미생물 대사물 중에서 G1P(glucose 1-phosphate) 및 NADPH의 분석을 위하여 상기 미생물의 바이오매스의 과염소산(perchloric acid) 추출 단계를 추가적으로 포함하는 것을 특징으로 하는 방법.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020120038484A KR101442682B1 (ko) | 2012-04-13 | 2012-04-13 | 미생물의 세포 내 대사 프로파일을 위한 추출 및 분석 방법 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020120038484A KR101442682B1 (ko) | 2012-04-13 | 2012-04-13 | 미생물의 세포 내 대사 프로파일을 위한 추출 및 분석 방법 |
Publications (2)
Publication Number | Publication Date |
---|---|
KR20130115815A KR20130115815A (ko) | 2013-10-22 |
KR101442682B1 true KR101442682B1 (ko) | 2014-09-19 |
Family
ID=49635198
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
KR1020120038484A Expired - Fee Related KR101442682B1 (ko) | 2012-04-13 | 2012-04-13 | 미생물의 세포 내 대사 프로파일을 위한 추출 및 분석 방법 |
Country Status (1)
Country | Link |
---|---|
KR (1) | KR101442682B1 (ko) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108102943B (zh) * | 2017-10-11 | 2021-04-23 | 四川大学 | 一种脱氮微生物及其应用 |
-
2012
- 2012-04-13 KR KR1020120038484A patent/KR101442682B1/ko not_active Expired - Fee Related
Non-Patent Citations (1)
Title |
---|
J Microbiol Methods., Vol. 87, No. 1, pages 24-31 (2011.10.) * |
Also Published As
Publication number | Publication date |
---|---|
KR20130115815A (ko) | 2013-10-22 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Mashego et al. | Microbial metabolomics: past, present and future methodologies | |
Gonzalez et al. | A rapid and reliable method for metabolite extraction in yeast using boiling buffered ethanol | |
Ruijter et al. | Determination of intermediary metabolites in Aspergillus niger | |
Wang et al. | Metabolomic basis of laboratory evolution of butanol tolerance in photosynthetic Synechocystis sp. PCC 6803 | |
Amador-Noguez et al. | Metabolome remodeling during the acidogenic-solventogenic transition in Clostridium acetobutylicum | |
Visser et al. | Analysis of in vivo kinetics of glycolysis in aerobic Saccharomyces cerevisiae by application of glucose and ethanol pulses | |
Neubauer et al. | U 13 C cell extract of P ichia pastoris–a powerful tool for evaluation of sample preparation in metabolomics | |
Yee et al. | Evaluation of the bioconversion of genetically modified switchgrass using simultaneous saccharification and fermentation and a consolidated bioprocessing approach | |
Pluskal et al. | Metabolic profiling of the fission yeast S. pombe: quantification of compounds under different temperatures and genetic perturbation | |
US20200032200A1 (en) | Pretreatment method for lc-ms detecting metabolomics of aspergillus flavus | |
Qin et al. | Flux regulation through glycolysis and respiration is balanced by inositol pyrophosphates in yeast | |
Zhu et al. | Metabolomic analysis reveals functional overlapping of three signal transduction proteins in regulating ethanol tolerance in cyanobacterium Synechocystis sp. PCC 6803 | |
Wang et al. | Intracellular metabolic changes of Clostridium acetobutylicum and promotion to butanol tolerance during biobutanol fermentation | |
CN102033114A (zh) | 一种快速简便分析传统酿造用大曲微生物群落结构的方法 | |
CN102174634A (zh) | 检测维生素c二步发酵中细胞内代谢物变化的方法 | |
Huang et al. | The fungus gardens of leaf‐cutter ants undergo a distinct physiological transition during biomass degradation | |
CN112684160A (zh) | 一种高效的血浆、红细胞、淋巴细胞nad+检测方法 | |
CN104391060A (zh) | Gc-ms研究灰葡萄孢菌代谢组的样品前处理及检测方法 | |
KR101442682B1 (ko) | 미생물의 세포 내 대사 프로파일을 위한 추출 및 분석 방법 | |
Du et al. | Metabolic profiling of Pleurotus tuoliensis during mycelium physiological maturation and exploration on a potential indicator of mycelial maturation | |
Tian et al. | Comprehensive analysis of spatial heterogeneity reveals the important role of the upper-layer fermented grains in the fermentation and flavor formation of Qingxiangxing baijiu | |
US11988651B2 (en) | Metabolome sampling and analysis method for analyzing metabolome during synthetic gas fermentation of synthetic gas fermentation microorganisms | |
Kapoore | Mass spectrometry based hyphenated techniques for microalgal and mammalian metabolomics. | |
Chen et al. | Matrix-assisted laser desorption ionization mass spectrometry based quantitative analysis of cordycepin from Cordyceps militaris | |
Kumar et al. | Unraveling the role of intra-cellular metabolites in the lactic acid production by novel Bacillus amyloliquefaciens using sugarcane molasses as a substratum |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A201 | Request for examination | ||
PA0109 | Patent application |
Patent event code: PA01091R01D Comment text: Patent Application Patent event date: 20120413 |
|
PA0201 | Request for examination | ||
E902 | Notification of reason for refusal | ||
PE0902 | Notice of grounds for rejection |
Comment text: Notification of reason for refusal Patent event date: 20130924 Patent event code: PE09021S01D |
|
PG1501 | Laying open of application | ||
AMND | Amendment | ||
E601 | Decision to refuse application | ||
PE0601 | Decision on rejection of patent |
Patent event date: 20140325 Comment text: Decision to Refuse Application Patent event code: PE06012S01D Patent event date: 20130924 Comment text: Notification of reason for refusal Patent event code: PE06011S01I |
|
AMND | Amendment | ||
PX0901 | Re-examination |
Patent event code: PX09011S01I Patent event date: 20140325 Comment text: Decision to Refuse Application Patent event code: PX09012R01I Patent event date: 20131125 Comment text: Amendment to Specification, etc. |
|
E902 | Notification of reason for refusal | ||
PE0902 | Notice of grounds for rejection |
Comment text: Notification of reason for refusal Patent event date: 20140626 Patent event code: PE09021S01D |
|
AMND | Amendment | ||
PX0701 | Decision of registration after re-examination |
Patent event date: 20140828 Comment text: Decision to Grant Registration Patent event code: PX07013S01D Patent event date: 20140703 Comment text: Amendment to Specification, etc. Patent event code: PX07012R01I Patent event date: 20140526 Comment text: Amendment to Specification, etc. Patent event code: PX07012R01I Patent event date: 20140325 Comment text: Decision to Refuse Application Patent event code: PX07011S01I Patent event date: 20131125 Comment text: Amendment to Specification, etc. Patent event code: PX07012R01I |
|
X701 | Decision to grant (after re-examination) | ||
GRNT | Written decision to grant | ||
PR0701 | Registration of establishment |
Comment text: Registration of Establishment Patent event date: 20140915 Patent event code: PR07011E01D |
|
PR1002 | Payment of registration fee |
Payment date: 20140916 End annual number: 3 Start annual number: 1 |
|
PG1601 | Publication of registration | ||
FPAY | Annual fee payment |
Payment date: 20170901 Year of fee payment: 4 |
|
PR1001 | Payment of annual fee |
Payment date: 20170901 Start annual number: 4 End annual number: 4 |
|
PC1903 | Unpaid annual fee |
Termination category: Default of registration fee Termination date: 20200626 |