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Applied and Environmental Microbiology, November 2002, p. 5750-5755, Vol. 68, No. 11
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.11.5750-5755.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Large-Scale Production of Coenzyme F420-5,6 by Using Mycobacterium smegmatis

Dale Isabelle, D. Randall Simpson, and Lacy Daniels*

Department of Microbiology, University of Iowa, Iowa City, Iowa 52242

Received 7 March 2002/ Accepted 30 July 2002


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ABSTRACT
 
Production of coenzyme F420 and its biosynthetic precursor FO was examined with a variety of aerobic actinomycetes to identify an improved source for these materials. Based on fermentation costs, safety, and ease of growth, Mycobacterium smegmatis was the best source for F420-5,6. M. smegmatis produced 1 to 3 µmol of intracellular F420 per liter of culture, which was more than the 0.85 to 1.0 µmol of F420-2 per liter usually obtained with Methanobacterium thermoautotrophicum and ~10-fold higher than what was previously reported for the best aerobic actinomycetes. An improved chromatography system using rapidly flowing quaternary aminoethyl ion-exchange material and Florisil was used to more quickly and easily purify F420 than with previous methods.


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INTRODUCTION
 
F420 is a 7,8-didemethyl-8-hydroxy-5-deazaflavin electron transfer coenzyme first described for methanogenic archaea (10, 11). In the archaea, it is used for a variety of redox reactions that are involved in energy generation (18, 19, 21, 23, 26, 40). F420 is used by Streptomyces species for tetracycline and lincomycin biosynthesis (7, 28, 35) and may be used in mitomycin C biosynthesis (27). In Mycobacterium and Nocardia species, F420 is used by an F420-dependent glucose-6-phosphate dehydrogenase (32, 33). Photolyases from the green alga Scenedesmus species and the cyanobacterium Synechocystis species contain bound F420 (14, 15). Methanobacterium thermoautotrophicum and Methanococcus voltae F420 contains two glutamates (F420-2) (11, 17), and Methanosarcina barkeri contains F420 with four and five glutamates (F420-4,5) (17) while Mycobacterium spp. contain primarily five- and six-glutamate forms (F420-5,6) (3). FO, a 5-deazaflavin that is a biosynthetic intermediate in the final steps of F420 synthesis, is observed in culture supernatants (22, 24, 28, 35). FO is catalytically active in all F420-dependent reactions examined, but its Km is often higher than that of F420 (10, 19, 41). F420 is mostly retained inside cells due to the negative charges on the phosphate and glutamyl groups, but the neutral FO more easily leaks out of the cells. The only measurement of the intracellular and extracellular distribution of both FO and F420 that we are aware of is that reported by Peck for Methanosarcina species (31).

F420 is not commercially available, and thus, those who work with this coenzyme must produce and purify it themselves or obtain it from others. Much of the F420 used in research is made from M. thermoautotrophicum, due to its relatively high F420-2 content per gram of cells and rapid growth rate, although others may use Methanococcus or Methanosarcina species. Most researchers who study F420 have in the past worked with methanogens and thus have been able to grow these organisms. However, as interest has grown in F420-dependent reactions in nonarchaea, scientists not trained in the growth of strictly anaerobic organisms have begun to obtain F420 or FO from Streptomyces species or related actinomycetes. The general observation has been that with even the best producers ~40-fold-less F420 is available from the actinomycetes on a basis of dry weight of cells. About 1.9 µmol of F420 per g (dry weight) of cells is present in M. thermoautotrophicum (10, 11, 17), whereas the highest yields observed for Streptomyces species have been ~0.05 µmol of F420 per g (dry weight) of cells (9, 13, 16). Assuming cell densities of 0.5 and 3 g (dry weight) of cells per liter, respectively, for the methanogen and an actinomycete, this predicts yields of 0.95 and 0.15 µmol of F420 per liter of culture, respectively. However, one paper has described very high levels of FO in the culture media of some actinomycetes (220 to 550 µmol of FO per liter) (24); unfortunately, F420 in the cells or supernatant was not examined. Since the medium used in that study contained very high nutrient levels compared to those for media used in previous studies, we examined F420 and FO levels in cells and in culture supernatants of a range of aerobic actinomycetes grown in rich medium with hope of finding an improved source of F420.


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Growth of actinomycetes.
 
We obtained cultures of Actinomadura kijaniata (ATCC 31588), Actinoplanes missouriensis (ATCC 14538), Streptomyces avermitilis (ATCC 31267), Streptomyces flocculus (ATCC 13257), Streptomyces coelicolor (ATCC 19894), Mycobacterium phlei (ATCC 11758), Nocardioides simplex (ATCC 6946), and Rhodococcus rhodochrous (ATCC 13808) from the American Type Culture Collection (Manassas, Va.). Mycobacterium smegmatis mc2 155 was a gift of W. Jacobs, Jr. (Albert Einstein College of Medicine, New York, N.Y.). Rhodococcus opacus Rb1 was a gift of R. Blasco (Universidad de Córdoba, Cordoba, Spain). Aerobic actinomycetes were routinely grown in flasks with shaking at 28, 30, or 37°C, depending on their optimal temperature. Inoculum medium contained (in grams per liter) soluble starch (25), glucose (5), soy peptone (10), yeast extract (5), ammonium sulfate (2), and KH2PO4 (0.3). MSR production medium contained (in grams per liter) glucose (40), yeast extract (15), soy peptone (15), NaH2PO4 · H2O (1.75), and ferric ammonium citrate (0.04). MSR medium pH was adjusted to 7.0 before autoclaving. Flasks were autoclaved with all components present, but for fermentors, glucose was sterilized separately. Cells were grown in inoculum medium for 1 to 3 days. For flask growth, a 10% inoculation was made into MSR medium, which was incubated for 2 to 6 days, until late log or stationary phase. For fermentor growth, a 5% inoculation was made into MSR medium, and cells were grown for about 4 days.

Fermentor growth was conducted in a New Brunswick Scientific Bioflo II 5-liter-working-volume fermentor, with 4 liters of medium. The vessel height was 30 cm, and the internal diameter was 16 cm. The liquid level was 19 cm above the vessel bottom. One Rushton-type six-bladed 7.8-cm-diameter impeller was positioned 7.5 cm from the hemispheric bottom of the vessel. A second impeller was positioned above the liquid surface of the medium, 23.5 cm above the bottom of the vessel; this top impeller helped reduce foam problems. Agitation was initially set at 200 rpm, and the gas rate was set at 1.5 liters/min. After about 20 h of growth, agitation was increased to 400 or 500 rpm. Small amounts (~0.1 ml) of polyethylene glycol 1000 were added several times after the first day of growth to control foam.


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HPLC analysis.
 
F420 and FO were measured by C18 high-pressure liquid chromatography (HPLC) using a fluorescence detector (excitation, 400 nm; emission, 470 nm) (3). Deazaflavin was estimated with a standard curve of known F420 concentrations based on the {varepsilon}400 of 25.7 mM-1 cm-1. M. thermoautotrophicum strain Marburg (grown in a 100-liter fermentor on H2-CO2 as described previously [8]) was used as a source of standard F420-2. M. smegmatis was used as an F420-5 and F420-6 standard (3). FO purified from the supernatant of M. smegmatis was used as a routine FO standard, the identity of which was confirmed by coelution with synthetic FO (2). For analytical purposes, supernatant was separated from cells by centrifugation of culture samples at 3,500 x g for 10 min, followed by decanting of the supernatant into another tube and frozen storage until direct analysis by appropriate dilution and HPLC. Cells (from 10 or 35 ml of culture) were extracted twice by being boiled in 1 ml of 25 mM Na acetate (pH 4.7) for 15 min, followed by centrifugation, and the two supernatants were combined, diluted, and analyzed. Pellets from this extraction were dried at 95°C for >2 days and weighed to determine dry weights. Extraction with 50 to 70% ethanol without boiling was not as effective.


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Predominant F420 forms made by aerobic actinomycetes.
 
HPLC analysis of the actinomycete cell extracts revealed two major peaks which corresponded to F420-5 and F420-6 from M. smegmatis. Very little F420-2 was present. Although earlier work had presented evidence that the F420 in some Streptomyces species had more than two glutamyl groups (based on thin-layer chromatography or thin-layer electrophoresis data [12, 25]), our analysis provides good evidence that specifically the F420-5 and F420-6 forms predominate in many aerobic actinomycetes. FO was also identified in cells and the medium.


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Production levels of F420 and FO in flask experiments.
 
The best producers of F420 in flasks were S. flocculus, M. smegmatis, and M. phlei, and most actinomycetes examined contained ~0.1 to 0.6 µmol per g (dry weight) of cells (Table 1). FO accounted for about 2 to 10% of the cellular deazaflavin. FO generally predominated in the supernatant and represented a significant amount of total deazaflavin for potential recovery. F420 yields here were higher than those that we previously reported for several actinomycetes grown in weaker medium (~0.02 to 0.06 µmol per g [dry weight] of cells) (9). These high yields could be due to the increased medium strength in the present work (two- and threefold-higher levels of carbohydrate and peptone, respectively, plus other nutrients, compared to medium that we had previously used [9]). The M. thermoautotrophicum cellular F420 yield in term of micromoles per liter was 1.5- to 5-fold lower than those of M. smegmatis and S. flocculus, although on a per-gram basis the methanogen was higher; this reflects the much higher actinomycete cell densities (Table 2). Volumetric production was highest with S. flocculus. Volumetric production by flask-grown M. smegmatis was 7.5- to 23-fold higher than the levels produced by S. coelicolor and Streptomyces griseus, which until the present work had been the best F420-producing actinomycetes (Table 1).


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  		TABLE 1. F420 and FO levels in cells and culture supernatantsa


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  		TABLE 2. Comparison of factors involved in choosing an organism for producing F420

Due to the very high FO levels reported in the medium of actinomycetes by Kuo et al. (24) and the fact that we did not see such levels in our work (Table 1), we were concerned that a special medium was required to obtain such high FO levels. Thus, we examined F420 and FO production by S. avermitilis, A. kijaniata, M. smegmatis, and A. missouriensis in a molasses-cottonseed medium that was essentially identical to their medium and which contained (in grams per liter) glucose (15), blackstrap molasses (20), soluble starch (40), cottonseed hydrolysate (25), CaCO3 (8), and K2SO4 (2). In our initial experiments, for F420 and FO estimation we used quaternary aminoethyl (QAE) ion-exchange chromatography followed by conventional bench top non-HPLC C18 chromatography of cell extract and supernatant samples while monitoring the effluent with spectrofluorimetry, followed by deazaflavin measurement by UV-visible light spectral analysis, not by HPLC. The molasses-cottonseed medium was problematic because much dark material bound permanently to the QAE column material, limiting QAE recycle options, but repeatable 5-deazaflavin estimates were obtained. We varied growth temperature and agitation speed and took samples during growth of several of the higher-yielding species (data not shown). In all cases, using this partial purification and absorbance measurement approach, we observed less than 1 µmol of FO per liter (data not shown), which was significantly less than that reported by Kuo et al. (24). We then used HPLC without prior chromatography to directly examine cultures of S. flocculus, S. avermitilis, and M. smegmatis grown on molasses-cottonseed medium. These yielded, respectively, 1.7, 1.5, and 2.1 µmol of F420 per liter, and FO levels in the spent medium ranged from not detectable to 0.75 µmol/liter. We conclude that similar results were obtained with both of our methods but that the lower values that arose from the absorbance approach were due to both chromatography processing losses and the inherent low sensitivity of absorbance and that the higher values resulting from HPLC are a result of the higher specificity of fluorescence and little loss from chromatographic steps, since the sample was only diluted and injected. Also, our HPLC measurements with M. thermoautotrophicum (Table 1) are consistent with yields reported by several other laboratories using different methods (10, 11, 17), giving us confidence in our methodology.

We could not find conditions that led to production of the very high FO levels (>200 µmol/liter) reported previously (24). The differences between our results and those of Kuo et al. do not arise from strain differences, since at least in the case of A. kijaniata and S. avermitilis the same American Type Culture Collection strains were used (D. A. Yurek, personal communication). We do not know the reason for the difference between our data and those reported previously (24), but we noted that the molasses-cottonseed medium showed several large HPLC peaks which eluted near FO, and that may interfere with HPLC analysis. The lack of agreement concerning the quantities of 5-deazaflavin produced in our study and the previous study does not diminish the importance of the two major conclusions of Kuo et al. that 5-deazaflavin is required for lincomycin production and that many aerobic actinomycetes produce 5-deazaflavin (7, 24). Furthermore, the earlier work led us to try a much stronger medium, which allowed us to produce levels of F420 per liter that are >10-fold higher than previously reported for the best-producing actinomycetes.

We conclude that S. flocculus, M. smegmatis, and M. phlei produce more cellular F420 and more supernatant FO than are produced by the four strains previously described as very high FO producers (A. kijaniata, S. avermitilis, A. missouriensis, and S. coelicolor). These findings are important for those wishing to make F420 for their own use, because it corrects the impression that ~100 µmol of F420 per liter might be produced by these organisms and identifies several organisms which produce ~2 µmol of F420 per liter and which can serve as good sources of F420 compared to the methanogens and most aerobic actinomycetes.


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F420 production in a fermentor.
 
We examined deazaflavin production by two of our best F420 producers in a fermentor. After 70 to 90 h, six separate fermentor runs with M. smegmatis made 1 to 3 µmol of cellular F420 per liter and 1 to 6 µmol of FO per liter of supernatant. Little 5-deazaflavin was present before 50 h. Impeller speed (400 versus 500 rpm) did not affect yield. Two fermentor runs with M. phlei gave similar results but created more wall growth than with M. smegmatis, making M. phlei cultures more difficult to monitor and harvest.


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Identification of the best organisms for F420 production.
 
Based on volumetric F420 production (micromoles per liter of culture) and on several factors listed in Table 2, the best actinomycetes are more attractive than methanogens for F420 production, if it is not important to have F420-2 instead of F420-5,6. Growth of methanogens requires extensive experience with anaerobic technique and requires a variety of tools for the task, many of which are not commercially available (8, 37). M. thermoautotrophicum is grown in a fermentor that contains pressurized hydrogen gas, which along with the methane produced creates a flammability hazard. Most fermentors require modification prior to use for this purpose. Methanosarcina barkeri, Methanobacterium formicicum, and Methanococcus species (e.g., Methanococcus voltae or Methanococcus vannielii) can be grown without the use of hydrogen gas, with the use instead of methanol or formate as growth substrate and nitrogen gas to maintain anaerobic conditions (8, 36, 37). While this simplifies growth of these methanogens, specialized equipment and experience with anaerobic growth are still required. Methanosarcina barkeri in particular is prone to contamination (although this can be avoided by appropriate use of antibiotics), and obtaining colonies of any methanogen on petri plates requires a great deal of experience and generally is done with an anaerobic glove bag and anaerobic jar. In contrast, aerobic actinomycetes are easy to grow, and their growth period is more predictable. The cell yields of the better actinomycetes are about 10-fold higher than those of the methanogens. This offsets the lower actinomycete internal F420 yields per gram (dry weight) of cells, since from a cost perspective the yield of F420 per liter of fermentor broth is the most important under most fermentation facility cost structures. Nonetheless, for kinetic reasons, work with a methanogen enzyme may be best done with F420-2, rather than the mix of F420-5 and F420-6 that predominates in Mycobacterium species. Also, if appropriate skilled personnel are available in a financially subsidized facility, F420-2 purification from a methanogen would require processing much less cell mass and would thus save on the costs of purification that have not been estimated here. Thus, in these instances, it is worth exploring F420 production by a methanogen grown on formate in a pH-controlled fermentor (20, 36) and the more traditional use of M. thermoautotrophicum (8).

Of the top five F420 producers in Table 1, we are most confident in the safety of working on a large scale with the two Mycobacterium species. S. flocculus presents no infection hazard, but some strains produce streptonigrin, a toxic antitumor antibiotic, which could be a hazard (4). A similar objection could be made to using A. kijaniata and S. avermitilis, which produce the toxic compounds kijanimicin and avermectin, respectively (5, 6). Thus, we prefer to not use these organisms for F420 production. M. smegmatis is not normally considered pathogenic, but more than 20 human infections with this organism have been documented, mostly due to accidental or surgical trauma (29, 39, 42). Two cases of aspiration pneumonia have been reported, but the patients already had serious pulmonary disease. Only three instances of M. phlei infections have been reported, one each in the foot, knee, and peritoneal cavity (1, 30, 38). Since fermentor growth involves the risk of scraping and puncture wounds, when M. smegmatis and M. phlei are being grown we recommend that latex or plastic gloves be worn when working with the system and that puncture-resistant leather gloves be used when inoculating the culture by needle. All wounds that may have introduced mycobacteria should be cleaned thoroughly, and if appropriate (e.g., for penetrating wounds), medical attention should be sought. These precautions are no greater than those required for safe growth of a conventional Escherichia coli strain.

As shown in Table 2, consideration of fermentor costs, and cellular yield per liter, leads to estimates of $27/µmol of F420 with M. thermoautotrophicum and $3.40 to 7.50 with S. flocculus, M. smegmatis, or M. phlei. Although FO is catalytically active, and it may be useful for some experiments, most researchers need F420 more than FO. This makes the cost per micromole of F420 the most important consideration for most situations. Thus, based on ease of growth, fewer hazards, and lower costs, M. smegmatis is the best source for F420 out of the organisms that we have examined.


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Purification of F420 from actinomycetes.
 
A procedure for purifying F420 from cells was developed by using Macro-Prep High Q QAE (Bio-Rad) and Florisil (Sigma; F-9127) for column chromatography. As an example, fermentor-grown M. smegmatis cells (600 g [wet weight]), which based on HPLC analysis contained 44.2 µmol of F420 and 7.5 µmol of FO, were processed by this procedure. Cells were mixed with 600 ml of 25 mM NaPO4 buffer (pH 7.0), autoclaved at 121°C for 10 min, and centrifuged. The procedure was repeated three more times (but with 450 ml of buffer), and the pooled supernatant was adjusted to pH 7.0. The supernatant (2.1 liters) was applied to a QAE column (5 by 20 cm) that had been equilibrated with 25 mM NaPO4 (pH 7.0), and the column was washed with 2 liters of 100 mM NaCl in the buffer. The FO was eluted with 4.0 liters of 250 mM NaCl in buffer, and F420 was eluted in four 2.0-liter volumes of 400 mM NaCl in buffer. QAE chromatography yielded 34.8 µmol of F420 (79% recovery, by HPLC). Florisil was cleaned with 6 N HCl, water, and acetone and then dried at 90°C prior to use. The F420 fractions (8 liters) were pooled and adjusted to pH 4.7, then applied to a 5- by 20-cm Florisil column equilibrated with 25 mM Na acetate (pH 4.5), and washed with 2 liters of 400 mM NaCl in buffer. The F420 was eluted by a series of 2.0-liter solutions: 100 mM NaCl in buffer, Na acetate buffer alone, water at pH 4.7, and finally twice with water at pH 7. F420 (31.5 µmol, by HPLC) eluted in the pH 7 water (95% recovery for the column). Material from this large-scale preparation still contained impurities as indicated by its slightly browner color than that of pure F420, but after rotary evaporation to 500 ml, HPLC indicated that it was about 70% F420-5,6 and 25% F420-2,3,4, with no other significant fluorescent peaks. Omission of the wash or elution steps prior to pH 7 water reduced the purity of the final product, and reduction of the pH 7 water volume reduced the yield. The F420 produced by this method can be used effectively for routine enzyme assays and other purposes, but it can then be purified further by gradient chromatography at a more convenient scale, and by HPLC, as needed. The High Q column material had an improved flow rate and good binding capacity compared to the more conventional QAE Sepharose or Sephadex. The Florisil was more efficient at binding the deazaflavins and had a better flow rate than that of the C18 material that we have used in the past (32). It is likely that FO purification from a fermentor supernatant by similar QAE and Florisil techniques would also have advantages over previously used methods.


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ACKNOWLEDGMENTS
 
We thank William R. Jacobs, Jr. (Albert Einstein College of Medicine), for providing us with a culture of M. smegmatis mc2 155, R. Blasco (Universidad de Córdoba) for giving us a culture of R. opacus Rb1, and David Gibson (University of Iowa) for use of his fermentor.

This work was supported by National Institutes of Health grant GM56177 and U.S. Department of Agriculture grant 4132008 to L.D.


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FOOTNOTES
 
* Corresponding author. Mailing address: Department of Microbiology, University of Iowa, Iowa City, IA 52242. Phone: (319) 335-7780. Fax: (319) 335-9006. E-mail: lacy-daniels{at}uiowa.edu. Back


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REFERENCES
 
    1
  1. Aguilar, J. L., E. E. Sanchez, C. Carrillo, G. S. Alarcon, and A. Silicani. 1989. Septic arthritis due to Mycobacterium phlei presenting as infantile Reiter's syndrome. J. Rheumatol. 16:1377-1378.[Medline]
    2. 2
  2. Ashton, W. T., R. D. Brown, F. S. Jacobson, and C. Walsh. 1979. Synthesis of 7-8-didemethyl-8-hydroxy-5-deazariboflavin and confirmation of its identity with the deazaisoalloxazine chromophore of Methanobacterium redox coenzyme F420. J. Am. Chem. Soc. 101:4419-4420.[CrossRef]
    3. 3
  3. Bair, T. B., D. W. Isabelle, and L. Daniels. 2001. Structures of coenzyme F420 in Mycobacterium species. Arch. Microbiol. 176:37-43.[CrossRef][Medline]
    4. 4
  4. Bolzan, A. D., and M. S. Bianchi. 2001. Genotoxicity of streptonigrin: a review. Mutat. Res. 488:25-37.[CrossRef][Medline]
    5. 5
  5. Bradner, W. T., C. A. Claridge, and J. B. Huftalen. 1983. Antitumor activity of kijanimicin. J. Antibiot. (Tokyo) 36:1078-1079.[Medline]
    6. 6
  6. Chung, K., C. C. Yang, M. L. Wu, J. F. Deng, and W. J. Tsai. 1999. Agricultural avermectins: an uncommon but potentially fatal cause of pesticide poisoning. Ann. Emerg. Med. 34:51-57.[CrossRef][Medline]
    7. 7
  7. Coats, J. H., G. P. Li, M. S. Kuo, and D. A. Yurek. 1989. Discovery, production, and biological assay of an unusual flavenoid cofactor involved in lincomycin biosynthesis. J. Antibiot. (Tokyo) 42:472-474.[Medline]
    8. 8
  8. Daniels, L. 1995. Large-scale culturing of methanogenic bacteria, p. 63-74. In F. T. Robb, A. R. Place, K. R. Sowers, H. J. Schreier, S. DasSarma, and E. M. Fleischmann (ed.), Archaea: a laboratory manual. Cold Spring Harbor Laboratory Press, Plainview, N.Y.
    9. 9
  9. Daniels, L., N. Bakhiet, and K. Harmon. 1985. Widespread distribution of a 5-deazaflavin cofactor in actinomycetes and related bacteria. Syst. Appl. Microbiol. 6:12-17.
    10. 10
  10. Eirich, L. D., G. D. Vogels, and R. S. Wolfe. 1979. Distribution of coenzyme F420 and properties of its hydrolytic fragments. J. Bacteriol. 140:20-27.[Abstract/Free Full Text]
    11. 11
  11. Eirich, L. D., G. D. Vogels, and R. S. Wolfe. 1978. Proposed structure for coenzyme F420 from Methanobacterium. Biochemistry 17:4583-4593.[CrossRef][Medline]
    12. 12
  12. Eker, A. P. 1980. Photoreactivating enzyme from Streptomyces griseus. II. Evidence for the presence of an intrinsic chromophore. Photochem. Photobiol. 32:593-600.[Medline]
    13. 13
  13. Eker, A. P., J. K. Hessels, and R. Meerwaldt. 1989. Characterization of an 8-hydroxy-5-deazaflavin:NADPH oxidoreductase from Streptomyces griseus. Biochim. Biophys. Acta 990:80-86.[Medline]
    14. 14
  14. Eker, A. P., J. K. C. Hessels, and J. van de Velde. 1988. Photoreactivating enzyme from the green alga Scenedesmus acutus. Evidence for the presence of two different flavin chromophores. Biochemistry 27:1758-1765.[CrossRef]
    15. 15
  15. Eker, A. P., P. Kooiman, J. K. Hessels, and A. Yasui. 1990. DNA photoreactivating enzyme from the cyanobacterium Anacystis nidulans. J. Biol. Chem. 265:8009-8015.[Abstract/Free Full Text]
    16. 16
  16. Eker, A. P., A. Pol, P. van der Meyden, and G. D. Vogels. 1980. Purification and properties of 8-hydroxy-5-deazaflavin derivatives from Streptomyces griseus. FEMS Microbiol. Lett. 8:161-165.
    17. 17
  17. Gorris, L. G., and C. van der Drift. 1994. Cofactor contents of methanogenic bacteria reviewed. Biofactors 4:139-145.[Medline]
    18. 18
  18. Hartzell, P. L., G. Zvilius, J. C. Escalante-Semerena, and M. I. Donnelly. 1985. Coenzyme F420 dependence of the methylenetetrahydromethanopterin dehydrogenase of Methanobacterium thermoautotrophicum. Biochem. Biophys. Res. Commun. 133:884-890.[CrossRef][Medline]
    19. 19
  19. Jacobson, F. S., L. Daniels, J. A. Fox, C. T. Walsh, and W. H. Orme-Johnson. 1982. Purification and properties of an 8-hydroxy-5-deazaflavin-reducing hydrogenase from Methanobacterium thermoautotrophicum. J. Biol. Chem. 257:3385-3388.[Abstract/Free Full Text]
    20. 20
  20. Jones, J. B., and T. C. Stadtman. 1977. Methanococcus vannielii: culture and effects of selenium and tungsten on growth. J. Bacteriol. 130:1404-1406.[Abstract/Free Full Text]
    21. 21
  21. Jones, J. B., and T. C. Stadtman. 1980. Reconstitution of a formate-NADP+ oxidoreductase from formate dehydrogenase and a 5-deazaflavin-linked NADP+ reductase isolated from Methanococcus vannielii. J. Biol. Chem. 255:1049-1053.[Abstract/Free Full Text]
    22. 22
  22. Kern, R., P. J. Keller, G. Schmidt, and A. Bacher. 1983. Isolation and structural identification of a chromophoric coenzyme F420 fragment from culture fluid of Methanobacterium thermoautotrophicum. Arch. Microbiol. 136:191-193.[CrossRef]
    23. 23
  23. Kunow, J., D. Linder, K. O. Stetter, and R. K. Thauer. 1994. F420H2: quinone oxidoreductase from Archaeoglobus fulgidus. Characterization of a membrane-bound multisubunit complex containing FAD and iron-sulfur clusters. Eur. J. Biochem. 223:503-511.[Medline]
    24. 24
  24. Kuo, M. S., D. A. Yurek, J. H. Coats, and G. P. Li. 1989. Isolation and identification of 7,8-didemethyl-8-hydroxy-5-deazariboflavin, an unusual cosynthetic factor in streptomycetes, from Streptomyces lincolnensis. J. Antibiot. (Tokyo) 42:475-478.[Medline]
    25. 25
  25. Lin, X. L., and R. H. White. 1986. Occurrence of coenzyme F420 and its {gamma}-monoglutamyl derivative in nonmethanogenic archaebacteria. J. Bacteriol. 168:444-448.[Abstract/Free Full Text]
    26. 26
  26. Ma, K., and R. K. Thauer. 1990. Purification and properties of N5, N10-methylene-tetrahydromethanopterin reductase from Methanobacterium thermoautotrophicum (strain Marburg). Eur. J. Biochem. 191:187-193.[Medline]
    27. 27
  27. Mao, Y., M. Varoglu, and D. H. Sherman. 1999. Molecular characterization and analysis of the biosynthetic gene cluster for the antitumor antibiotic mitomycin C from Streptomyces lavendulae NRRL 2564. Chem. Biol. 6:251-263.[CrossRef][Medline]
    28. 28
  28. McCormick, J. R. D., and G. O. Morton. 1982. Identity of cosynthetic factor 1 of Streptomyces aureofaciens and fragment FO from coenzyme F420 of Methanobacterium sp. J. Am. Chem. Soc. 104:4014-4015.[CrossRef]
    29. 29
  29. Newton, J. A., Jr., and P. J. Weiss. 1994. Aspiration pneumonia caused by Mycobacterium smegmatis. Mayo Clin. Proc. 69:297-298.
    30. 30
  30. Paul, E., and P. Devarajan. 1998. Mycobacterium phlei peritonitis: a rare complication of chronic peritoneal dialysis. Pediatr. Nephrol. 12:67-68.[CrossRef][Medline]
    31. 31
  31. Peck, M. W. 1989. Changes in concentrations of coenzyme F420 analogs during batch growth of Methanosarcina barkeri and Methanosarcina mazei. Appl. Environ. Microbiol. 55:940-945.[Abstract/Free Full Text]
    32. 32
  32. Purwantini, E., and L. Daniels. 1996. Purification of a novel coenzyme F420-dependent glucose-6-phosphate dehydrogenase from Mycobacterium smegmatis. J. Bacteriol. 178:2861-2866.[Abstract/Free Full Text]
    33. 33
  33. Purwantini, E., T. Gillis, and L. Daniels. 1997. Presence of F420-dependent glucose-6-phosphate dehydrogenase in Mycobacterium and Nocardia species, but absence in Streptomyces and Corynebacterium species and methanogenic Archaea. FEMS Microbiol. Lett. 146:129-134.[CrossRef][Medline]
    34. 34
  34. Reuke, B., S. Korn, W. Eisenreich, and A. Bacher. 1992. Biosynthetic precursors of deazaflavins. J. Bacteriol. 174:4042-4049.[Abstract/Free Full Text]
    35. 35
  35. Rhodes, P. M., N. Winskill, E. J. Friend, and M. Warren. 1981. Biochemical and genetic comparison of Streptomyces rimosus mutants impaired in oxytetracycline biosynthesis. J. Gen. Microbiol. 124:329-338.
    36. 36
  36. Sowers, K. R. 1995. Large-scale growth of methanogens that utilize acetate and formate in a pH auxostat, p. 75-77. In F. T. Robb, A. R. Place, K. R. Sowers, H. J. Schreier, S. DasSarma, and E. M. Fleischmann (ed.), Archaea: a laboratory manual. Cold Spring Harbor Laboratory Press, Plainview, N.Y.
    37. 37
  37. Sowers, K. R., and K. M. Noll. 1995. Techniques for anaerobic growth, p. 15-47. In F. T. Robb, A. R. Place, K. R. Sowers, H. J. Schreier, S. DasSarma, and E. M. Fleischmann (ed.), Archaea: a laboratory manual. Cold Spring Harbor Laboratory Press, Plainview, N.Y.
    38. 38
  38. Spiegl, P. V., and C. M. Feiner. 1994. Mycobacterium phlei infection of the foot. Foot Ankle Int. 15:680-683.[Medline]
    39. 39
  39. Wallace, R. J., D. R. Nash, M. Tsukamura, Z. M. Blacklock, and V. A. Silcox. 1988. Human disease due to Mycobacterium smegmatis. J. Infect. Dis. 158:52-59.[Medline]
    40. 40
  40. Widdel, F., and R. S. Wolfe. 1989. Expression of secondary alcohol dehydrogenase in methanogenic bacteria and purification of the F420-specific enzyme from Methanogenium thermophilum strain TCI. Arch. Microbiol. 152:322-328.[CrossRef]
    41. 41
  41. Yamazaki, S., L. Tsai, and T. C. Stadtman. 1982. Analogues of 8-hydroxy-5-deazaflavin cofactor: relative activity as substrates for 8-hydroxy-5-deazaflavin-dependent NADP+ reductase from Methanococcus vannielii. Biochemistry 21:934-939.[CrossRef][Medline]
    42. 42
  42. Young, L. S., C. B. Inderlied, O. G. Berlin, and M. S. Gottleib. 1986. Mycobacterial infections in AIDS patients, with an emphasis on the Mycobacterium avium complex. Rev. Infect. Dis. 8:1024-1033.[Medline]

Applied and Environmental Microbiology, November 2002, p. 5750-5755, Vol. 68, No. 11
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.11.5750-5755.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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