Production of siderophore from L-glutamic acid as both carbon and nitrogen sole sources in Acinetobacter sp. B-W

국문 초록 (Abstract)

포도당과 글루탐산을 함유한 배지에서 시드로포어인 2, 3-dihydroxybenzoic acid (DHB)를 생산하는 Acinetobacter sp. B-W 균주를 글루탐산을 유일한 탄소원과 질소원으로 함유한 배지에 배양한 결과, 상등액에서 2, 3-DHB가 아닌 카테콜 형의 시드로포어를 생산하는 것으로 조사되었다. 글루탐산의 농도는 3%에서 시드로포어 생산이 최고로 나타났으며, 3% 보다 높은 농도에서는 감소하는 것으로 조사되었다. 글루탐산을 유일한 탄소원과 질소원으로 함유한 배지에서 자란 균주 B-W는 배양 온도 $28^{\circ}C$에서는 시드로포어를 생산하지만 $36^{\circ}C$에서는 생산하지 않았다. 또한 $10{\mu}M\;FeCl_3$를첨가한 배지에서는 시드로포어 생산이 완전히 억제되었다. 글루탐산 배지에서 생산된 균주 B-W의 시드로포어는 TLC 전개 용매 butanol: acetic acid: water =12:3:5에서 Rf치가 0.32로 나타나 Rf치가 0.82인 2, 3-DHB와는 다른 것으로 조사되었으며, 또한 항산화 활성도 없는 것으로 나타나 항산화 활성을 가진 2, 3-DHB와는 다른 시드로포어인 것으로 추정되었다.

다국어 초록 (Multilingual Abstract)

Catechol type siderophore different from 2, 3-dihydroxybenzoic acid (DHB) was produced from Acinetobacter sp. B-W grown in medium containing L-glutamic acid as both carbon and nitrogen sole sources at $28^{\circ}C$. Optimal concentration of glutamic acid for siderophore production was 3% and production of siderophore was decreased above 3% glutamic acid. In previous report, siderophore, 2, 3-DHB was produced from strain B-W grown in medium containing glucose as carbon source and glutamic acid as nitrogen source. Rf value of siderophore produced from strain B-W grown in medium glutamic acid as both carbon and nitrogen sole sources at $28^{\circ}C$ was 0.32, while 2, 3-DHB was 0.84 in butanol-acetic acid-water (12:3:5) as developing solvent. Antioxidative activity of 2, 3-DHB was not detected in that siderophore produced from glutamic acid. Catechol nature of siderophore was detected by Arnow test. Although in iron-limited media optimal cell growth was identified at $36^{\circ}C$, significant quantities of siderophore were produced only at $28^{\circ}C$. Biosynthesis of siderophore was strongly inhibited by growth at $36^{\circ}C$. Production of siderophore was completely inhibited by $10{\mu}M\;FeCl_3$.

참고문헌 (Reference)

1 Schwyn, R, "Universal chemical assay for detection and determination of siderophores" 160 : 47-56, 1987

2 Skaar, E. P, "The battle for iron between bacterial pathogens and their vertebrate hosts" 6 : e1000949-, 2010

3 Cogswell, R.L, "Temperature restriction of iron acquisition in Proteus vulgaris" 15 : 69-71, 1980

4 김경자, "Temperature dependent 2,3-dihydroxybenzoic acid production in Acinetobacter sp. B-W" 한국미생물학회 51 (51): 249-255, 2015

5 Jalal, M, "Structure of anguibactin, a unique plasmid related bacterial siderophore from the fish pathogen Vibrio anguillarum" 111 : 292-296, 1989

6 Miethke, M, "Siderophore-based iron acquisition and pathogen control" 71 : 413-451, 2007

7 Meyer, J. M, "Siderophore production by Pyoverdin is essential for virulence of Pseudomonas aeruginosa" 64 : 518-523, 1996

8 Gillam, A.H, "Quantitative determination of hydroxamic acids" 5 : 841-844, 1981

9 Walsh, C. T, "Molecular studies on enzymes in chorismate metabolism and enterobactin biosynthetic pathway" 90 : 1105-1129, 1990

10 Payne, S, "Methods in Enzymology" Academic Press 1994

11 Neilands, J. B, "Methodology of siderophores" 58 : 1-24, 1984

12 Wencewicz, T, "Is drug release necessary for antimicrobial activity of siderophore-drug conjugates? Syntheses and biological studies of the naturally occurring salmycin “Trojan Horse” antibiotics and synthetic desferridanoxamine antibiotic conjugates" 22 : 633-648, 2009

13 Rogers, H. J, "Iron-binding catechols and virulence in Escherichia coli" 7 : 445-456, 1973

14 Raymond, K, "Iron Transport in Bacteria" ASM Press 1-16, 2004

15 Hoefte, M, "Iron Chelation in Plants and Soil Microorganisms" Academic Press 1993

16 Garibaldi, J.A, "Influence of temperature on the biosynthesis of iron transport compounds by Salmonella typhimurium" 110 : 262-265, 1972

17 Earhart, C.F, "Escherichia coli and Salmonella: cellular mechanisms and molecular biology" ASM Press 472-482, 1996

18 Worsham, P.L, "Effect of growth temperature on the acquisition of iron by Salmonella typhimurium and Escherichia coli" 158 : 163-168, 1984

19 Milagres, A. M. F, "Detection of siderophore production from several fungi and bacteria by a modification of chrome azurol S (CAS) agar plate assay" 37 : 1-6, 1999

20 Sonenshein, A. L, "Control of key metabolic intersections in Bacillus subtilis" 5 : 917-927, 2007

21 Gunka, K, "Control of glutamate homeostasis in Bacillus subtilis: a complex interplay between ammonium assimilation, glutamate biosynthesis and degradation" 85 : 213-224, 2012

22 Arnow, L.E, "Colorimetric determination of the components of 3, 4-dihydroxy phenylalanine tyrosine mixtures" 118 : 531-537, 1937

23 Carrillo-Castaneda, G, "Characterization of siderophore-mediated iron transport from Rhizobium leguminosarum by Phaseoli" 23 : 1669-1683, 2000

24 Loehr, J, "Characterization of anguibactin, a novel siderophore from Vibrio anguillarum 775 (pJM1)" 167 : 57-65, 1986

25 O’Brien, I. G, "Biologically active compounds containing 2,3-dihydroxybenzoic acid and serine formed by Escherichia coli" 20 : 453-460, 1970

26 Wandersman, C, "Bacterial iron sources: from siderophores to hemophores" 58 : 611-647, 2004

27 Csaky, T.Z, "An estimation of bound hydroxylamine in biological materials" 2 : 450-454, 1948

28 Magasanik, B, "Ammonia assimilation by Saccharomyces cerevisiae. Eukaryot" 2 : 827-829, 2003

29 Yokoyama, S, "A modified ninhydrin reagent using ascorbic acid instead of potassium cyanide" 95 : 204-205, 2003

30 Abe, N, "1.1-Diphenyl-2-picrylhydrazylradical scavengers, bisorbicillin and demethyltrichodimerol, from a fungus" 62 : 661-662, 1998