Paenibacillus

Heyndrickx, M. Vandemeulebroecke, K. Hoste, B. Janssen, P. Kersters, K. Devos, R Logan, N. A. Ali, N. Berkeley, R. C. W. Reclassification of Paenibacillus (Formerly Bacillus) Pulvifaciens (Nakamura 1984) Ash et al. 1994, a later subjective synonym of Paenibacillus (Formerly Bacillus) larvae (White 1906) Ash et al. 1994, as a subspecies of Paenibacillus-larvae, with emended descriptions of Paenibacillus-larvae as Paenibacillus-larvae subsp larvae and Paenibacillus-larvae subsp pulvifaciens. Int. J. Syst. Bacteriol. 1996, 46, 270-279. 

The data reported identifies sulfur substrates tested for growth as sole sulfur source for the various strains. The strains may metabolize other sulfur compounds (not listed). A complete name of listed strains in Table 3 comprises Rhodococcus sp. SY1 Rhodococ-cus sp. H-2 Rhodococcus sp. D-l Rhodococcus ECRD-1 Gordona CYKS1 Nocar-dia sp. CYKS2 Paenibacillus All-2 Mycobacterium sp. WU-F1 Mycobacterium sp. WU-0103 Mycobacterium phlei sp. GTIS10 and Agrobacterium MC501. 

Figure 6. The postulated pathway of benzothiophene desulfurization by Paenibacillus sp. Strain A-112 [31].

In addition to DBT and BT, strain A11-2 could utilize methyl, dimethyl, and trimethyl DBTs as sulfur sources. The desulfurization of asymmetric alkylated DBTs was assessed to understand the sulfur specificity of this organism. It was shown to desulfurize several asymmetric alkyl DBTs up to C3-DBTs. It was shown that the rates of desulfurization depended on not only the position of alkyl substitution but also the number and length of alkyl substitution. An attempt was made to co-relate the data based on a molecular shape parameter. Selectivity of this organism was compared with R. erythropolis KA2-5-1 and, although clear differences were observed, the parameter fitting was not perfect. Two Paenibacillus strains, Paenibacillus sp. A11-1 and All-2, were patented [87] and were deposited as PERM BP-6025 and PERM BP-6026 in 1996 [122,123],... 

Rhodococcus sp. Strain WU-K2R A Rhodococcus strain capable of sulfur-specific desulfurization of benzothiophene, naphthothiophene (NT), and some of their alkyl derivatives was reported [35]. The metabolites of BT desulfurization were BT sulfone, benzo[c][l,2]oxanthiin S-oxide, benzo[c][l,2]oxanthiin S,S-dioxide, o-hydroxystyrene, 2,(2 -hydroxyphenyl)ethan-l-al, and benzofuran. The NT metabolites were NT sulfone, 2 -hydroxynaphthyl ethene, and naphtho[2,l-b]furan [35], The exact biochemical pathway was not determined, however, part of the pathway for BT desulfurization was speculated to be similar to Paenibacillus All-2. 

Rhodococcus sp. Strain KT462 This was the first strain discovered to be capable of BT and DBT desulfurization. The pathway of BT desulfurization is similar to that in Paenibacillus All-2 and Sinorhizobium sp. KT55 and that for DBT is the 4S pathway [37], The substrate specificity of this organism is different from R. erythropolis KA2-5-1 and Paenibacillus sp. All-2. It is able to desulfurize 2-methyl BT, 3-methyl BT, 5-methyl BT, 7-methyl BT, 1-methyl DBT, 3-methyl DBT, and 4-methyl DBT in addition to BT and DBT. 

The desulfurization pathway was proposed to be BT -> BT sulfone -> benzo[e][l,2]oxanthiin S-oxide -> o-hydroxystyrene. Additionally, formation of the intermediate benzo(e)(l,2)oxathiin S,S dioxide was inferred to a side pathway resulting in formation of benzofuran as shown in Fig. 7. This pathway is similar to that reported for Sinorhizobium KT55, Paenibacillus sp. strain All-2 and R. erythropolis KT462. 

Dsz enzymes from R. erythropolis strains IGTS8, KA2-5-1, and D-l as well as the desulfurization enzymes from Paenibacillus All-2 and B. subtilis WU-S2B have been purified and characterized. These enzymes have been reported to be associated with the soluble fraction of the cell extract, indicating that these are soluble enzymes. The characteristics of the enzymes purified from various organisms and their comparison is given below (Table 6). 

The DBT sulfone monooxygenase (TdsA) from Paenibacillus sp. strain All-2 has been isolated and characterized by Konishi et al [162], This enzyme was compared with the mesophilic enzyme dszA from R. erythropolis and found to have very little sequence homology however, had similar properties. The molecular mass of the enzyme TdsA was calculated by gel filtration and found to be 120 KDa, whereas with a subunit molecular mass calculated by SDS-PAGE to be 48 KDa. 

Table 7. Properties of the desulfinase enzyme from different strains R. erythropolis IGTS8, R. erythropolis KA2-5-1, and Paenibacillus sp. All-2...

The thermophilic enzyme DszD from Paenibacillus All-2 has been cloned into E. coli and characterized [172], The sequence of this enzyme showed 30% similarity to the major flavin reductase of Vibrio fischeri. The optimum activity was reported to be at 45°C in resting cell cultures and 55°C in cell-free extracts. 

Desulfurization of diesel oils by thermophilic bacteria has also been demonstrated. A LGO with 800ppm sulfur was biotreated with Paenibacillus All-2 at 50°C in a... 

Recently, several thermophilic organisms have been reported to be capable of sulfur-specific biodesulfurization. These include the Paenibacillus [87,151], Mycobacterium [30,31,85,94,294,295], etc. The ability to desulfurize sulfur compounds other than DBT derivatives, including benzothiophene, naphthothiophene, and benzonaphthothio-phene derivatives has also been demonstrated, thus widening the substrate specificity of the biodesulfurization process. Second, the thermophilic ability of the organisms offers temperature and operational advantages to further improve the commercialization potential of the BDS process. 

Konishi, J. Onaka, T. Ishii, Y., and Suzuki, M., Demonstration of the Carbon-Sulfur Bond Targeted Desulfurization of Benzothiophene by Thermophilic Paenibacillus Sp Strain Al 1-2 Capable of Desulfurizing Dibenzothiophene. Ferns Microbiology letters, 2000. 187(2) pp. 151-154. 

Konishi, J. Ishii, Y. Onaka, T., and Maruhashi, K., Purification and Characterization of the Monooxygenase Catalyzing Sulfur-Atom Specific Oxidation of Dibenzothiophene and Benzothiophene From the Thermophilic Bacterium Paenibacillus Sp Strain All-2. Applied Microbiology and Biotechnology, 2002. 60(1-2) pp. 128-133. 

Konishi, J., and Maruhashi, K., 2-(2 -hydroxyphenyl)benzene sulfinate desulfinase from the thermophilic desulfurizing bacterium Paenibacillus sp strain All-2 Purification and Characterization. Applied Microbiology and Biotechnology, 2003. 62(4) pp. 356-361. 

Ohshiro, T. Aoi, Y. Torii, K., and Izumi, Y., Flavin Reductase Coupling With Two Monooxygenases Involved in Dibenzothiophene Desulfurization Purification and Characterization From a Non-Desulfurizing Bacterium, Paenibacillus Polymyxa A-l. Applied Microbiology, and Biotechnology, 2002. 59(6) pp. 649-657. 

Ishii, Y. Konishi, 1 Suzuki, M., and Maruhashi, K., Cloning and Expression of the Gene Encoding the Thermophilic NAD(P)H-FMN Oxidoreductase Coupling With the Desulfurization Enzymes From Paenibacillus Sp All-2. Journal of Bio science and Bioengineering, 2000. 90(6) pp. 591-599. 

Ishii, Y. Ohshiro, T. Aoi, Y., et al., Identification of the Gene Encoding a NAD(P)H-Flavin Oxidoreductase Coupling With Dibenzothiophene (DBT)-Desulfurizing Enzymes From the DBT-Nondesulfurizing Bacterium Paenibacillus Polymyxa A-l. Journal of Bioscience and Bioengineering, 2000. 90(2) pp. 220-222. 

Paenibacillus polymyxa, Micrococcus luteus Reactive Violet 5R The bacterial consortium showed complete decolorization in 36 h [76]... 

Girardin, H., Albagnac, C., Dargaignaratz, C., Nguyen-The, C. and Carlin, F., Antimicrobial activity of foodborne Paenibacillus and Bacillus spp. against Clostridium botulinum, J. Pood Prot., 65, 806-813, 2002. 

Bertaux, J., Schmid, M., Chemidlin Prevost-Boure, N. et at. (2003). In situ identification of intracellular bacteria related to Paenibacillus spp. in the mycelium of the ectomycorrhizal fungus Laccaria bicolor S238N. Applied and Environmental Microbiology, 69, 4243-8. 

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