Cellulosimicrobium Cellulans Research Articles

Plant growth promotion by a hexavalent chromium reducing bacterial strain, Cellulosimicrobium cellulans KUCr3

This article reports on the isolation and characterization of a Cr(VI) resistant bacterial strain, having plant growth promoting properties to improve general growth of plant in chromium-contaminated soil through rhizosphere colonization. The strain was isolated from the sludge of waste canal carrying industrial effluents. The minimum inhibitory concentration of chromium to this strain was found to be 450 and 400 mM in complex and minimal media, respectively. The strain also showed varied degree of resistance to Cd, Co, As, Ni and Zn. It exhibited potential Cr(VI) reducing ability under aerobic culture conditions, and the factors affecting Cr(VI) reduction by this strain were evaluated. The optimum pH and temperature required to achieve maximum Cr(VI) reduction values were 7 and 35°C, respectively. Higher concentration of Cr(VI) slowed down the reduction, but with longer incubation time it reduced nearly all detectable amount of Cr(VI). The strain showed positive response to IAA production and phosphate solubilization. It promoted the growth of chilli plants in waste-fed soil with or without additional Cr through its establishment in rhizosphere. The successful establishment of KUCr3 in the rhizosphere of chilli plants helped to reduce Cr uptake by the test plant. This strain shows a promise that the multifarious role of this strain would be useful in the Cr-contaminated rhizosphere soil as a good bioremediation and plant growth promoting agent as well. Through biochemical characterization and 16S rDNA sequence analysis, the strain KUCr3, as the name given to it, was identified as a strain of Cellulosimicrobium cellulans.

Elucidação parcial da estrutura de aminoglucanooligossacarídeos (AGO's) produzidos enzimaticamente

O presente trabalho visou a aplicação da enzima quitinolítica purificada da linhagem Cellulosimicrobium cellulans 191 e da preparação comercial de papaína na hidrólise da quitina coloidal. A quitina coloidal foi caracterizada quanto ao grau de desacetilação e apresentou GD de 14% por espectroscopia na região do infravermelho. A quitinase purificada hidrolisou a quitina coloidal liberando di-N-acetilquitobiose, enquanto que a preparação comercial de papaína atuando sobre o mesmo substrato formou di-N-acetilquitobiose e tri-N-acetilquitotriose. A estrutura química dos aminoglucanooligossacarídeos foi elucidada por espectrometria de massa.

Isolation of cellulose-hydrolytic bacteria and applications of the cellulolytic enzymes for cellulosic biohydrogen production

Nine cellulolytic bacterial strains were isolated from soil sample taken in southern Taiwan. Through 16S rRNA sequence matching; eight of those isolates belong to Cellulomonas sp., while the other one belongs to Cellulosimicrobium cellulans. The activity of cellulolytic enzymes (cellulases and xylanase) produced from those strains was mainly present extracellularly and the enzyme production was dependent on cellulosic substrates (xylan, rice husk and rice straw) used for growth. HPLC analysis confirmed the bacterial hydrolysis of these cellulosic substrates for soluble sugars production. The efficiency of fermentative H 2 production from the enzymatically hydrolyzed rice husk was examined with seven H 2-producing pure bacterial isolates. With an initial reducing sugar concentration of 0.36 g l −1, only Clostridium butyricum CGS5 exhibited efficient H 2 production from the rice husk hydrolysates with a cumulative H 2 production and H 2 yield of 88.1 ml l −1 and 19.15 mmol H 2 (g reducing sugar) −1 (or 17.24 mmol H 2 (g cellulose) −1), respectively.

Bacterial communities of tyre monofill sites: growth on tyre shreds and leachate

To investigate bacterial communities of tyre monofill sites, colonization of tyre material by bacteria and the effect of tyre leachate on bacteria. Culturable bacteria were isolated from buried tyre shreds and identified using fatty acid methyl ester analysis. Isolates belonged to taxonomic groups such as Bacilli, Actinobacteria, Clostridia, Flavobacteria, beta and gamma-proteobacteria. For tyre material colonization experiments, Bacillus megatarium, Bacillus cereus, Hydrogenophaga flava, Janthinobacterium lividum, Cellulosimicrobium cellulans, Arthrobacter globiformis (isolated from tyre shreds or leachate at the study site); Escherichia coli and Acidithiobacillus ferrooxidans were used. Beakers containing tyre shreds and artificial rain water were inoculated with a given bacterial culture, incubated at room temperature and sampled at regular intervals. 4',6-diamidino-2-phenylindole (DAPI) staining followed by epifluorescent microscopy was used to enumerate bacteria in samples. Of the bacteria tested, B. megatarium, J. lividum, E. coli, C. cellulans and A. globiformis exhibited the most extensive colonization of the tyre shreds. However, the extent of colonization varied among bacteria. Response to tyre leachate was also examined using B. cereus and J. lividum. Both bacteria increased in abundance due to the addition of leachate. Bacteria associated with buried tyre shreds were identified and found to include typical soil and freshwater organisms. The majority of indigenous isolates grew on tyre material (or leachate) suggesting that they play an active role in the ecology of these sites and that their potential role in tyre degradation should be explored. This study provides information on bacterial communities of tyre-waste disposal sites, explores the interaction between tyre material and bacteria and identifies bacteria that could be involved in or employed for recycling tyre-waste.

Production of alkaline protease from Cellulosimicrobium cellulans

Cellulosimicrobium cellulans is one of the microorganisms that produces a wide variety of yeast cell wall-degrading enzymes, β-1,3-glucanase, protease and chitinase. Dried cells of Saccharomyces cerevisiae were used as carbon and nitrogen source for cell growth and protease production. The medium components KH2PO4, KOH and dried yeast cells showed a significant effect (p<0.05) on the factorial fractional design. A second design was prepared using two factors: pH and percentage of dried yeast cells. The results showed that the culture medium for the maximum production of protease was 0.2 g/l of MgSO4.7H2O, 2.0 g/l of (NH4)2SO4 and 8% of dried yeast cells in 0.15M phosphate buffer at pH 8.0. The maximum alkaline protease production was 7.0 ± 0.27 U/ml over the center point. Crude protease showed best activity at 50ºC and pH 7.0-8.0, and was stable at 50ºC.

Production, purification and application of extracellular chitinase from Cellulosimicrobium cellulans 191

This study concerned the production, purification and application of extracellular chitinase from Cellulosimicrobium cellulans strain 191. In shaken flasks the maximum yield of chitinase was 6.9 U/mL after 72 h of cultivation at 25°C and 200 rpm. In a 5 L fermenter with 1.5 vvm aeration, the highest yield obtained was 4.19 U/mL after 168 h of fermentation at 25°C and 200 rpm, and using 3 vvm, it was 4.38 U/mL after 144 h of fermentation. The chitinase (61 KDa) was purified about 6.65 times by Sepharose CL 4B 200 gel filtration with a yield of 46.61%. The purified enzyme was able to lyse the cell walls of some fungi and to form protoplasts.

Comparative Study on the in vitro Antibacterial Efficacy of Aqueous and Methanolic Extracts of Quercus infectoria Gall`s Against Cellulosimicrobium cellulans

The in vitro antibacterial efficacy of aqueous and methanolic extract of Quercus infectoria Olivier (Fagaceae) galls was tested against Cellulosimicrobium cellulans using extract concentration ranging from 0.25 to 4 mg mL-1. Both types of extract showed significant inhibition of C. cellulans growth with strong correlation between extract concentrations and degrees of antibacterial activity for concentrations ranging from 0.5 to 4 mg mL-1. Although, slight reduction of average diameter of inhibition zones after 24 h of incubation for aqueous extract (0.96 ± 0.148 cm) compared to methanolic extract (1.00 ± 0.182 cm), both extracts still attained the MIC value beginning at a concentration of 0.5 mg mL-1 but established higher concentration for the MBC at 2 mg mL-1. The antibacterial activity of methanolic extract was also significantly affected by the temperature with an optimum inhibition zone being obtained at 30 °C (1.38 ± 0.05 cm) and this was reduced to approximately 20% at temperatures of above 50 °C.

Isolation of Bacteria which Produce Yeast Cell Wall-Lytic Enzymes and Their Characterization

To isolate bacteria capable of producing enzymes that lyse cell wall, we screened soil samples from Rokko Mountain where is situated in west Japan. The isolated strains KH1, KH2 and KH3 that secrete yeast cell wall lytic enzymes are described herein. The activity of the enzymes, after isolation from culture supernatants, were examined. The lytic enzyme produced by strain KH3 was the most active in both cell wall degradation and beta-1,3 glucanase activity in comparison with commercially available enzymes. This enzyme also had the highest thermo- and pH-stability. On the baisis of partial 16S rDNA sequence analyses, KH1 and KH3 were identified as Cellulosimicrobium sp. and KH2 as Bacillus sp. KH3 was tested by DNA-DNA hybridization and GC content in an attempt to precisely classify it, because the 16S rDNA sequence similarities to other strains were very high at 99.4%. These chemotaxonomic experiments indicate that KH3 belongs to Cellulosimicrobium cellulans.

Biological pretreatment of sugarcane trash for its conversion to fermentable sugars

Pretreatment of lignocellulosic biomasses, the first step in their conversion to utilizable molecules requires very high energy (steam and electricity), corrosion resistant high-pressure reactors and high temperatures. These severe conditions not only add to the cost component of the entire process but also lead to the loss of sugars to the side reactions. Microbial pretreatments have been reported to be associated with reducing the cost factors as well as the severities of the reactions. Eight bioagents, including fungi and bacteria, were screened for their pretreatment effects on sugarcane trash. They narrowed down the C:N ratio of trash from 108:1 to a varying range of approximately 42:1 to 60:1.The maximum drop in C:N ratio of 61% was observed using Aspergillus terreus followed by Cellulomonas uda (52%) and Trichoderma reesei and Zymomonas mobilis (49%). The bioagents helped in degradation of sugarcane trash by production of cellulases, the maximum being produced by A. terreus, (12 fold) followed by C. uda (10 fold), Cellulomonas cartae (9 fold) and Bacillus macerans (8 fold). The microbial pretreatment of trash rendered the easy accessibility of sugars for enzymatic hydrolysis, which can be directed for production of alcohol.

Diffusion characteristics and controlled release of bacterial fertilizers from modified calcium alginate capsules

An indigenous Cellulosimicrobium cellulans GS6 isolate able to solubilize insoluble phosphate complexes in soil is a potential bacterial fertilizer. Enclosure of the phosphate-solubilizing bacterium (PSB) in biodegradable capsules may protect the PSB cells inoculated into soil and, in the meantime, enable the control of cell release that confers long-term fertilizing effects. In this study, calcium alginate (CA) was used as the core matrix to encapsulate cells of C. cellulans GS6. The cell-liberating properties of the CA-based capsules were modified by blending with a variety of supplemental materials (SM), including chitin, cellulose, olive oil, and gelatin. The experimental results showed that the maximum cell-release percentage (MCR%) of the capsules decreased in the order of CA-cellulose > CA-olive oil > CA-chitin > CA-gelatin > CA. Furthermore, a mass transport model was developed to accurately describe the kinetics of cell release results for each capsule. The diffusion coefficient ( D e) of each capsule was also determined from the model simulation. We found that the estimated D e values are positively correlated to the release rate with rare exceptions. Lastly, as our results underscored the crucial roles that the type of capsules plays in the rate and amount of cell release, controlled release of the bacterial fertilizer ( C. cellulans GS6 cells) may be achieved via the design of capsule materials.

Characterization of clinical isolates previously identified as Oerskovia turbata: proposal of Cellulosimicrobium funkei sp. nov. and emended description of the genus Cellulosimicrobium

Taxonomic studies were performed on 13 clinical isolates (ten of which were epidemiologically related) that had been previously identified as Oerskovia turbata. Comparative phylogenetic analysis, based on 16S rRNA gene sequences, indicated that the isolates are closely related to Cellulosimicrobium cellulans with sequence similarity values ranging from 99.5 to 99.8 %. Chemotaxonomic results (fatty acid profiles and menaquinones) supported the inclusion of these isolates in the genus Cellulosimicrobium. The DNA G+C content was 74.5 mol%. The results of DNA-DNA reassociation, whole-cell sugars (with galactose as the characteristic whole sugar) and phenotypic properties, including antimicrobial resistance, indicated that these isolates are representatives of a novel species of the genus Cellulosimicrobium. The name Cellulosimicrobium funkei sp. nov. is proposed for the novel strains, with strain W6122T (=ATCC BAA-886T = DSM 16025T = CCUG 50705T) as the type strain. The definition of this novel Cellulosimicrobium species will assist in the understanding of the epidemiology and clinical significance of these micro-organisms.

Revisiting the Cellulosimicrobium cellulans yeast-lytic β-1,3-glucanases toolbox: A review

Cellulosimicrobium cellulans (also known with the synonyms Cellulomonas cellulans, Oerskovia xanthineolytica, and Arthrobacter luteus) is an actinomycete that excretes yeast cell wall lytic enzyme complexes containing endo-β-1,3-glucanases [EC 3.2.1.39 and 3.2.1.6] as key constituents. Three genes encoding endo-β-1,3-glucanases from two C. cellulans strains have been cloned and characterised over the past years. The βglII and βglIIA genes from strain DSM 10297 (also known as O. xanthineolytica LL G109) encoded proteins of 40.8 and 28.6 kDa, respectively, whereas the β-1,3-glucanase gene from strain ATCC 21606 (also known as A. luteus 73–14) encoded a 54.5 kDa protein. Alignment of their deduced amino acid sequences reveal that βglII and βglIIA have catalytic domains assigned to family 16 of glycosyl hydrolases, whereas the catalytic domain from the 54.5 kDa glucanase belongs to family 64. Notably, both βglII and the 54.5 kDa β-1,3-glucanase are multidomain proteins, having a lectin-like C-terminal domain that has been assigned to family 13 of carbohydrate binding modules, and that confers to β-1,3-glucanases the ability to lyse viable yeast cells. Furthermore, βglII may also undergo posttranslational proteolytic processing of its C-terminal domain, resulting in a truncated enzyme retaining its glucanase activity but with very low yeast-lytic activity. In this review, the diversity in terms of structural and functional characteristics of the C. cellulans β-1,3-glucanases has been compiled and compared.

Potential misidentification of a new Exiguobacterium sp. as Oerskovia xanthineolytica isolated from blood culture

Potential misidentification of a new Exiguobacterium sp. as Oerskovia xanthineolytica isolated from blood culture

Equine abortion and premature birth associated with Cellulosimicrobium cellulans infection.

During the 2002 and 2003 foaling seasons, Cellulosimicrobium (Cellumonas) cellulans (formerly Oerskovia xanthineolytica) was the principal microorganism isolated from fetal tissues or placentas from cases of equine abortion, premature birth, and term pregnancies. Significant pathologic findings included chronic placentitis and pyogranulomatous pneumonia. In addition, microscopic and macroscopic alterations in the allantochorion from 4 of 7 cases of placentitis were similar to those caused by Crossiella equi and other nocardioform bacteria. This report confirms a causative role of C. cellulans infection in equine abortion.

Reclassification of Cellulosimicrobium variabile Bakalidou et al. 2002 as Isoptericola variabilis gen. nov., comb. nov.

As already depicted in the original publication, the type strain of the species Cellulosimicrobium variabile Bakalidou et al. 2002, DSM 10177(T), does not cluster unambiguously with the type species, Cellulosimicrobium cellulans DSM 43879(T), in phylogenetic analysis. Strain DSM 10177(T) is moderately related to the recently described species Xylanimonas cellulosilytica, Promicromonospora pachnodae and Xylanibacterium ulmi, forming a lineage that branches between C. cellulans and members of the genus Promicromonospora in most dendrograms generated on the basis of different algorithms and reference strains. The type strains of the two Cellulosimicrobium species resemble each other in morphology, composition of fatty acids, DNA G+C content, phospholipids and the presence of lysine in position 3 of the peptide subunit of peptidoglycan. However, the two strains differ from each other in cell-wall sugars and in the amino acid composition of the A4alpha-type peptidoglycan, which contains serine and aspartic acid in C. cellulans, whereas only aspartic acid is present in the interpeptide bridge of C. variabile. This type is also present in Xylanimonas cellulosilytica XIL07(T), but not in the neighbouring species P. pachnodae DSM 12657(T), which exhibits the L-lys-L-ser-D-Glu type. On the basis of distinct phylogenetic position and the amino acid composition of peptidoglycan, a novel genus and combination for C. variabile, Isoptericola variabilis gen. nov., comb. nov., is proposed.

Effective extracellular trehalose production by Cellulosimicrobium cellulans

A bacterium isolated from a petal of Casa Blanca Lily (ST26 strain) produced a marked amount of extracellular trehalose (alpha- d-glucopyranosyl-[1,1]-alpha- d-glucopyranose) in culture medium containing glucose. 16S rDNA-based phylogeny showed that ST26 belongs to, or is related to, Cellulosimicrobium cellulans, a close relative of Cellulomonas spp. Various Cellulomonas strains obtained from culture collections also showed extracellular trehalose productivity, suggesting that trehalose production is a common property of this bacterial genus. ST26 accumulated trehalose in medium supplied with glucose but not with sucrose, glycerol or maltose. Effective extracellular trehalose production by ST26 was achieved by supplying 0.5-1% ammonium sulfate and 0.5-1% CaCO(3). The addition of CaCO(3) adjusted the pH of the culture to around 5.0. The optimized culture conditions yielded trehalose from glucose at a conversion rate of 61%. The addition of ammonium sulfate greatly reduced the dry cell weight of ST26 and intracellular content of trehalose, which suggests that the addition of ammonium sulfate makes ST26 cells leak trehalose into the medium. ST26 effectively propagated in minimal medium containing trehalose as a sole carbon source, which suggests that trehalose serves as a carbohydrate reserve of this organism.

Cellulosimicrobium variabile sp. nov., a cellulolytic bacterium from the hindgut of the termite Mastotermes darwiniensis.

A novel cellulolytic and xylanolytic bacterium, strain MX5T, was isolated from the hindgut contents of the Australian termite Mastotermes darwiniensis (Froggatt). The isolate was a facultative anaerobe and had a Gram-positive cell-wall profile. The rod-shaped bacterium formed irregular coryneform and coccoid cells during growth. Phylogenetic analysis of the 16S rDNA provided evidence that the organism was closely related to the as-yet undescribed cellulolytic strain SR272 and the non-validly described species 'Cellulomonas pachnodae' as well as Promicromonospora citrea and Promicromonospora sukumoe. Strain MX5T was assigned to the genus Cellulosimicrobium on the basis of phylogenetic and chemotaxonomic criteria. The murein of strain MX5T contained the diamino acid lysine. N-Glycolylmuramic acid, mycolic acids and hydroxy fatty acids were absent. The major neutral sugar in the cell wall was galactose and the major quinone was menaquinone MK-9(H4). The predominant fatty acids were ai-C15:0, i-C15:0, i-C16:0 and C16:0. The G+C content of the DNA was in a range 70-72 mol%. On the basis of 16S rDNA sequence similarities and chemotaxonomic features, MX5T was clearly different from Cellulosimicrobium cellulans and other validly described species within this phylogenetic group. For this reason, a novel species is described, for which the name Cellulosimicrobium variabile sp. nov. is proposed.

Overproduction, Purification, and Characterization of β-1,3-Glucanase Type II in Escherichia coli

An Escherichia coli recombinant system produced soluble and full-length beta-1,3-glucanase type II (BglII) cloned from the yeast-lytic actinomycete Oerskovia xanthineolytica. The expression system was designed to produce recombinant BglII with a six-histidine peptide fused to the carboxy end of the protein. The expression level was optimized to produce 30% of total protein of E. coli as the recombinant protein, releasing 75% to the extracellular space. The 43-kDa recombinant protein was purified by IMAC to homogeneity and its molecular and biochemical characteristics were studied, showing that there are no important functional differences with those properties described for the BglII purified from O. xanthineolytica.

Cloning and expression of an Oerskovia xanthineolytica β-1,3-glucanase in Escherichia coli

An Escherichia coli recombinant system produced a soluble β-1,3-glucanase (BglII) cloned from Oerskovia xanthineolytica. The protein was obtained in a truncated form derived from the complete polypeptide. Cell fractionation studies show that 80% of the glucanase was retained in the periplasmic space after 20 h of induction. If cells were grown with glycine, 60% of the glucanase was released from the periplasm

Production of new extracellular glycolipids by a strain ofCellulomonas cellulans(Oerskovia xanthineolytica) and their structural characterization

Glycolipid-producing bacteria were isolated from soil samples. One of the strains, identified as Cellulomonas cellulans (Oerskovia xanthineolytica), was found to produce significant amounts of unusual extracellular glycolipids, which were shown to be composed of at least 11 individual compounds. Hydrolysis of the glycolipid mixture and gas chromatography - mass spectrometry analysis revealed the presence of fatty acids and hydroxy fatty acids ranging from C10to C18, 16 of which were identified. The glycidic moiety consisted of glucose, rhamnose, and ribose. The same sugars were found to be present in the cell wall of Cellulomonas cellulans, which also contained polar lipids including glycolipids. During strain cultivation, glycolipid excretion was stimulated when nitrogen was exhausted from the culture medium. In these conditions, the production in fermenters on glycerol, expressed in glucose equivalents, reached 8.9 g/L. Cell hydrophobicity, which rose to 95% during the growth phase, decreased to 50% during the production phase. The overall results show that the bacterial cell wall is involved in the synthesis of these new extracellular glycolipids.Key words: glycolipid, excretion, Cellulomonas cellulans, Oerskovia xanthineolytica, cell wall.