Complete sequence and organization of the Serratia marcescens biotin operon

Sakurai, Naoki; Akatsuka, Hiroyuki; Kawai, Eri; Imai, Yuji; Komatsubara, Saburo

doi:10.1099/13500872-142-11-3295

Cited by 5 publications

(3 citation statements)

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“…1) (6,7,15,16,23,26). Many microorganisms, including Mesorhizobium loti, have biotin synthesis genes organized in operons (1,18,19,21,28,35), but neither the organization of bio genes in S. meliloti nor the molecular basis of this organism's dependence on external biotin has been defined.The present study therefore was initiated to answer two questions. First, do new genomic data (10; http://sequence.toulouse .inra.fr/rhime/public/Access/RhimeFormRA.html) support the concept that S. meliloti is a biotin auxotroph?…”

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Functional Analysis of Sinorhizobium meliloti Genes Involved in Biotin Synthesis and Transport

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Phillips

Streit

2002

Appl Environ Microbiol

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External biotin greatly stimulates bacterial growth and alfalfa root colonization by Sinorhizobium meliloti strain 1021. Several genes involved in responses to plant-derived biotin have been identified in this bacterium, but no genes required for biotin transport are known, and not all loci required for biotin synthesis have been assigned. Searches of the S. meliloti genome database in combination with complementation tests of Escherichia coli biotin auxotrophs indicate that biotin synthesis probably is limited in S. meliloti 1021 by the poor functioning or complete absence of several key genes. Although several open reading frames with significant similarities to genes required for synthesis of biotin in gram-positive and gram-negative bacteria were found, only bioB, bioF, and bioH were demonstrably functional in complementation tests with known E. coli mutants. No sequence or complementation evidence was found for bioA, bioC, bioD, or bioZ. In contrast to other microorganisms, the S. meliloti bioB and bioF genes are not localized in a biotin synthesis operon, but bioB is cotranscribed with two genes coding for ABC transporter-like proteins, designated here bioM and bioN. Mutations in bioM and bioN eliminated growth on alfalfa roots and reduced bacterial capacity to maintain normal intracellular levels of biotin. Taken together, these data suggest that S. meliloti normally grows on exogenous biotin using bioM and bioN to conserve biotin assimilated from external sources.Microorganisms from the gram-negative genera Rhizobium, Sinorhizobium, Bradyrhizobium, Mesorhizobium, and Azorhizobium, collectively termed rhizobia, are well known for their capacity to establish N 2 -fixing symbioses with legume plants (2). Although the molecular basis of rhizobial N 2 reduction is defined (17) and a foundation has been constructed for understanding other bacterial genes expressed in the plant (25), our knowledge of how rhizobia grow on plant roots is less complete. Good growth of S. meliloti strain 1021 (Rm1021) on alfalfa roots requires external biotin (33), often supplied by plant roots (27), which regulates a gene, bioS, that helps S. meliloti compete under such conditions (13, 34). Whether biotin is essential or simply stimulatory for rhizobial growth has been long debated (36-38), but clearly, cell densities of Rm1021 and many other rhizobia under biotin-limiting conditions are increased greatly by small amounts of biotin (8). Growing S. meliloti serially under biotin-limited conditions produces several physiological and metabolic changes, including the accumulation of polyhydroxybutyrate and a significant reduction in cell size (8,14). Biotin-dependent enzymes such as pyruvate carboxylase are also affected under biotin-limited conditions, and several tricarboxylic acid cycle auxiliary enzymes show decreased activities (4, 5).Biotin is formed in bacteria by a well-defined pathway ( Fig. 1) (6,7,15,16,23,26). Many microorganisms, including Mesorhizobium loti, have biotin synthesis genes organized in operons (1,18,19,21,2...

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Functional Analysis of Sinorhizobium meliloti Genes Involved in Biotin Synthesis and Transport

Entcheva

Phillips

Streit

2002

Appl Environ Microbiol

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“…Dies liegt zum Teil an einer hohen Instabilität der zur Verfügung stehenden biotinüberproduzierenden Bakterienstämme und an den hohen Produktionskosten. Problematisch sind toxische, inhibitorische Effekte sowie eine hohe Plasmidinstabilität bei Biotinüberproduktion (Ifuku et al, 1995;Sakurai et al, 1996;.…”

Section: B D-mannoseunclassified

“…3A). Eine Vielzahl von Bakterien sind in der Lage Biotin zu produzieren, wobei die Produktionsraten solcher Stämme zwischen 15 µg/l (Shaw et al, 1999) und 200 mg/l liegen (Sakurai et al, 1996). Diese Raten sind zu niedrig, um ein biotechnologisches Verfahren zu entwickeln.…”

Section: Introductionunclassified

Umwelt-Genomik als Quelle für die Isolierung von neuen Operons und Genclustern aus mikrobiellen Konsortien

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American type cilture colection b Basen BCIP 5-Brom-4-chlor-3-indolyl-phosphat bidest. zweifach destilliert bp Basenpaar(e) BSA Rinderserumalbumin C. Corynebacterium ºC Grad Celsius C-Quelle Kohlenstoff-Quelle ca. circa CS Casamonosäuren cfu Klon bildende Einheiten (engl. colony forming units) Cm Chloramphenicol DIG Digoxigenin DMF Dimethylformamid DMSO Dimethylsulfoxide DNA Desoxyribonukleinsäure DNase Desoxyribonuklease DSMZ Deutsche Sammlung von Mikroorganismen und Zellkulturen DTB Dethiobiotin DTT Dithiothreitol E. Escherichia, Erwinia E-Cup Eppendorf-Reaktionsgefäß EDTA Ethylendiamintetraacetat et al. et alii (und andere) ERG Eppendorf Reaktionsgefäss g Gramm G. Gluconobacter Gram (+) Grampositives Bakterium Gram (-) Gramnegatives Bakterium i. d. R. in der Regel IPTG Isopropyl-ß-d-thiogalactopyranosid K. Klebsiella kb Kilobasen K d Dissoziationskonstante Km Kanamycin l Liter L. Lactobacillus LB Luria-Bertani (ein Nährmedium) M molar MCS Multi-Cloning-Site mg Milligramm µg Mikrogramm min Minute bioA bioD bioB B. flavum C. glutamicum M. leprae bioA bioF bioD C3_227 bioB bioA bioD orf1 orf2 bioF bioB bioH orf3 Kurthia sp. bioC bioHII bioFII A. E. coli S. marcescens S. tiphimurium E. herbicola orf1 bioA bioB bioF bioC bioD bioH bioB bioF bioH bioC bioD bioA M. flagelatum Gram (-) Mikroorganismen bioB bioF bioD bioA bioZ M. loti bioB bioF bioD bioA bioZ A. tumefaciens C58 B.In allen Habitaten wie z.B. Boden, Meerwasser, Biofilmen, Teichen oder Intestinaltrakten, sind mikrobielle Biotin-Produzenten zu finden. Die Biotinbiosynthesegene liegen in der Regel in Operons vor (Serebriiskii et al., 1996; Abb. 4). Biotinbiosynthese ist bereits in Escherichia coli und Bacillus subtilis detailliert untersucht worden (Bower et al., 1996a;Shiuan and Campbell, 1988). An Abb. 4: Biotinbiosynthesegene verschiedener Mikroorganismen. (A) Die Organisation der Biotinbiosynthesegene in verschiedenen Gram-positiven Bakterien; (B) in verschiedenen Gram-negativen Bakterien. Die Pfeile zeigen die Transkriptionsrichtungen der bio-Gene. Abb. 5: Isolierung von Biotin überproduzierenden Mikroorganismen aus Umweltproben/ Umwelt DNA-Bänken. II. Material und Methoden 9 II. Material und Methoden II.1. Organismen, Plasmide und Primer Die in dieser Arbeit verwendeten Bakterienstämme, Plasmide, Cosmide und Primer sind mit ihren relevanten phäno-oder genotypischen Eigenschaften sowie der Herkunft in den Tabellen 1-3 aufgeführt. Tab. 1: In dieser Arbeit verwendete Stämme.

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Metabolites, Primary and Secondary

Demain

1999

Encyclopedia of Bioprocess Technology

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Complete sequence and organization of the Serratia marcescens biotin operon

Cited by 5 publications

References 37 publications

Functional Analysis of Sinorhizobium meliloti Genes Involved in Biotin Synthesis and Transport

Functional Analysis of Sinorhizobium meliloti Genes Involved in Biotin Synthesis and Transport

Umwelt-Genomik als Quelle für die Isolierung von neuen Operons und Genclustern aus mikrobiellen Konsortien

Metabolites, Primary and Secondary

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