Paenibacillus Illinoisensis Research Articles

Deciphering genome sequence of Paenibacillus illinoisensis strain YWY-3.1: A chitinase, cellulase, and amylase producer

The chitin-degrading Paenibacillus illinoisensis YWY-3.1 was isolated from the lake water at Yok Don National Park, Vietnam. Previous evaluations reported that strain YWY-3.1 had high activities of chitinases, cellulases, and amylases; it also produced remarkable levels of numerous promoting traits for plant growth. This study aimed to sequence and analyze the genome sequence of strain YWY-3.1 for further explorations and applications in agriculture and related fields. The whole genome sequencing revealed the P. illinoisensis YWY-3.1 genome contained 6,451,623 bp, 47.3 % GC, 5786 coding sequences, 88 tRNA, and 1 rRNA. It shared 95.49 % ANI and 87.1 % dDDH to those of P. illinoisensis NBRC 15959 (GCA 004000925). Among the coding sequences, COG deduced 4,991, and KEGG predicted 2819. The genome harbored 338 carbohydrate-active enzymes, including 195 GHs, 44 GTs, 13 PLs, 37 CEs, 3 AAs, and 46 CBMs; among them, 6 GH18 chitinases, 5 GH5 cellulases, 2 GH10 xylanases, 1 GH11 xylanase, and 2 GH13 α-amylases were identified. It contained 49 genes involved in environmental adaptation and plant growth. Moreover, 7 BGCs, with 3 being novel clusters, were identified from the genome. This work provided insight into the genomic information of P. illinoisensis YWY-3.1 and potential gene resources, especially chitinases, cellulases, xylanases, α-amylases, and novel BGCs for further explorations and applications.

Influence of different microbial inoculants on nitrogen retention and diazotroph community succession during cotton straw composting

Inoculation with microorganisms is an effective method for promoting compost maturity and quality. This study investigated the effects of different microbial inoculants on compost maturity, nitrogen retention, and the diazotroph community during cotton straw composting. Results revealed that inoculation improved organic matter degradation and compost maturity, increasing the total nitrogen masses by 6.6–33.0% and the total nitrogen by 84.6–112.1%. Particularly, the microbial inoculants containing Anoxybacillus rupiensis, Paenibacillus illinoisensis, and Bacillus velezensis (1:1:1, T1 treatment) exhibited the highest total nitrogen masses and total nitrogen content. Furthermore, inoculation enhanced nitrogenase activity and nifH abundance, and markedly influenced the succession of the diazotroph community, increasing their sensitivity to environmental variables during composting. Furthermore, the total nitrogen, C/N ratios, and moisture content significantly affected the diazotroph community. Results from a network analysis indicated that inoculation affected the co-occurrence patterns among the diazotroph species and increased complexity between diazotrophs. In conclusion, this study provided three promising microbial inoculants for high-quality compost production and a reference for promoting nitrogen retention in cotton straw composting.

Increased production of chitinase by a Paenibacillus illinoisensis isolated from Brazilian coastal soil when immobilized in alginate beads.

The accumulation of chitin waste from the seafood industry is a serious environmental problem. However, this residue can be degraded by chitinases and its subproducts, such as chitosan, economically exploited. In this study, a chitinase producer bacteria, identified as Paenibacillus illinoisensis, was isolated from the Brazilian coastal city of Terra de Areia - Rio Grande Do Sul (RS) and was immobilized within alginate beads to evaluate its chitinase production. The alginate beads containing cells presented an average size of 4mm, 99% of immobilization efficiency and increased the enzymatic activity in 40.71% compared to the free cells. The biomass during enzymatic production increased 62.01% and the total cells leaked from the alginate beads corresponded to 6.46% after 96h. Immobilized cells were reused in a sequential batch system and remained stable for production for up to four 96-h cycles, decreasing only 21.04% of the initial activity at the end of the fourth cycle. Therefore, the methodology used for cell immobilization resulted in adequate beads to maintain cell viability during the enzymatic production, increasing enzymatic activity, showing low cell leakage from the support and appropriate recyclable capacity.

Update of the list of QPS-recommended biological agents intentionally added to food or feed as notified to EFSA 11: suitability of taxonomic units notified to EFSA until September 2019.

Qualified presumption of safety (QPS) was developed to provide a generic safety evaluation for biological agents to support EFSA's Scientific Panels. The taxonomic identity, body of knowledge, safety concerns and antimicrobial resistance are assessed. Safety concerns identified for a taxonomic unit (TU) are where possible to be confirmed at strain or product level, reflected by ‘qualifications’. No new information was found that would change the previously recommended QPS TUs and their qualifications. The list of microorganisms notified to EFSA was updated with 54 biological agents, received between April and September 2019; 23 already had QPS status, 14 were excluded from the QPS exercise (7 filamentous fungi, 6 Escherichia coli, Sphingomonas paucimobilis which was already evaluated). Seventeen, corresponding to 16 TUs, were evaluated for possible QPS status, fourteen of these for the first time, and Protaminobacter rubrum, evaluated previously, was excluded because it is not a valid species. Eight TUs are recommended for QPS status. Lactobacillus parafarraginis and Zygosaccharomyces rouxii are recommended to be included in the QPS list. Parageobacillus thermoglucosidasius and Paenibacillus illinoisensis can be recommended for the QPS list with the qualification ‘for production purposes only’ and absence of toxigenic potential. Bacillus velezensis can be recommended for the QPS list with the qualification ‘absence of toxigenic potential and the absence of aminoglycoside production ability’. Cupriavidus necator, Aurantiochytrium limacinum and Tetraselmis chuii can be recommended for the QPS list with the qualification ‘production purposes only’. Pantoea ananatis is not recommended for the QPS list due to lack of body of knowledge in relation to its pathogenicity potential for plants. Corynebacterium stationis, Hamamotoa singularis, Rhodococcus aetherivorans and Rhodococcus ruber cannot be recommended for the QPS list due to lack of body of knowledge. Kodamaea ohmeri cannot be recommended for the QPS list due to safety concerns.

Isolation and characterization of halotolerant phosphate solubilizing bacteria naturally colonizing the peanut rhizosphere in salt-affected soil

In search of efficient salt-tolerant phosphate solubilizing bacteria (PSB), we conducted a survey of PSB that naturally colonize the rhizosphere of peanuts in Xinjiang, China, a typical inland saline area. A total of 23 PSB were isolated, which included 12 Bacillus strains, three Acinetobacter strains, two Pseudomonas strains, two Brevibacillus, and one strain of Gordonia terrae, Chryseobacterium lathyri, Ensifer sesbaniae, and Paenibacillus illinoisensis. All PSB have high potential for dissolving calcium phosphate [Ca3(PO4)2] within the range of 65–496 mg·L−1, but relatively weak ability to dissolve AlPO4 in amounts ranging from 0.4 to 11.4 mg·L−1, FePO4 ranging from 0.1 to 5.5 mg·L−1, and lecithin ranging from 2.1 to 10.3 mg·L−1. The maximum tolerance for pH is 10, and the maximum tolerance for Na2CO3 is 50 mm. Six isolates that can grow under the condition of 1.5 M NaCl (YMX5, YMX11, TPMX5, TPMX16, TPMX18, and TPMX19) were selected to determine their ability to dissolve Ca3(PO4)2 and tolerate pH changes. This study characterized salt-tolerant PSB isolates that can be used as bioinoculants to protect plants against salt stress. In addition, we describe the phosphate-solubilizing ability of Gordonia terrae at high NaCl concentrations.

Effect of auxin producing and phosphate solubilizing bacteria on mobility of soil phosphorus, growth rate, and P acquisition by wheat plants

Microorganisms capable of mobilizing phosphate promote plant growth, this activity being frequently accompanied by production of plant hormones auxins. However, the extent of contribution of these characteristics to promotion of plant growth remains unclear. Paenibacillus illinoisensis IB 1087 and Pseudomonas extremaustralis IB-Ki-13-1A strains were selected for their capacity to mobilize phosphates and to synthesize auxins in vitro. The effects of inoculating these bacteria on the content of mobile phosphorus in the soil as well as on the phosphorus and hormone content in wheat plants were studied and the observed responses were related to the changes in plant growth. Inoculation of bacteria into the soil increased P concentration in the plants suggesting their increased capacity for the efficient acquisition of phosphorus compounds, while concentration of mobile phosphorus in the soil was increased by its inoculation with bacteria only in the absence of plants. The treatment increased plants mass (to greater extent in the case of P. illinoisensis) in accordance with the increased level of auxins in the treated plant. Increased mass accumulation did not correlate with the potential ability of bacteria strains for production of auxins or phosphate mobilization in vitro. Our data indicate importance of increased auxin content in the plants for the stimulation of root growth and capacity for P uptake as influenced by growth-promoting bacteria.

Promotion of iron nutrition and growth on peanut by Paenibacillus illinoisensis and Bacillus sp. strains in calcareous soil.

This study aimed to explore the effects of two siderophore-producing bacterial strains on iron absorption and plant growth of peanut in calcareous soil. Two siderophore-producing bacterial strains, namely, YZ29 and DZ13, isolated from the rhizosphere soil of peanut, were identified as Paenibacillus illinoisensis and Bacillus sp., respectively. In potted experiments, YZ29 and DZ13 enhanced root activity, chlorophyll and active iron content in leaves, total nitrogen, phosphorus and potassium accumulation of plants and increased the quality of peanut kernels and plant biomass over control. In the field trial, the inoculated treatments performed better than the controls, and the pod yields of the three treatments inoculated with YZ29, DZ13, and YZ29 + DZ13 (1:1) increased by 37.05%, 13.80% and 13.57%, respectively, compared with the control. Based on terminal restriction fragment length polymorphism analysis, YZ29 and DZ13 improved the bacterial community richness and species diversity of soil surrounding the peanut roots. Therefore, YZ29 and DZ13 can be used as candidate bacterial strains to relieve chlorosis of peanut and promote peanut growth. The present study is the first to explore the effect of siderophores produced by P. illinoisensis on iron absorption.

Saccharification of Lignocellulosic Materials by Cellulolytic and Xylanolytic <i>Paenibacillus illioisensis </i>CX11

The utilization of lignocellulosic materials to produce a variety of building blocks (e.g. fermentable sugars) is an interesting alternative approach to meeting the growing demand for high value chemicals. Cellulose and hemicellulose can be hydrolyzed by cellulase and xylanase enzymes into their respective building blocks (hexoses and pentoses), which can later be converted into the targeted compounds. The aim of this study was to test the ability of Paenibacillus illinoisensis CX11 to saccharify different lignocellulosic materials, and to determine its ability to produce cellulolytic and xylanolytic enzymes for possible use in converting lignocellulosic materials into their respective fermentable sugars. The ability of P. illinoisensis CX11 to produce CMCase, xylanase, FPase, and avicelase was tested using SSF of corn stalk. Furthermore, the ability of P. illinoisensis CX11 to saccharify lignocellulosic materials was tested using corn stalk, wheat bran, sawdust, and corn cob. The amount of reducing sugars released from the saccharification of lignocellulosic materials was determined by the 3,5-dinitro-salicylic acid (DNS) method. Obtained results showed that P. illinoisensis CX11 can produce CMCase (400.12 ± 1.23 U/L), xylanase (385.57 ± 2.25 U/L), FPase (266.93 ± 2.22 U/L), avicelase (187.85 ± 2.22 U/L) and extracellular protein (4.56 ± 0.14 mg/L). Moreover, P. illinoisensis CX11 showed an ability to saccharify lignocellulosic materials. These findings confirm that P. illinoisensis CX11 has the ability to produce cellulolytic and xylanolytic enzymes, and to hydrolyze different lignocellulosic materials into fermentable sugars. Therefore, this study concludes that P. illinoisensis CX11 can be considered a good source of cellulase and xylanase enzymes to saccharify different lignocellulosic materials.

Cellulase- and Xylanase-Producing Bacterial Isolates with the Ability to Saccharify Wheat Straw and Their Potential Use in the Production of Pharmaceuticals and Chemicals from Lignocellulosic Materials

The efficient use of lignocellulosic materials for pharmaceutical and chemical production relies on the hydrolysis of their components into their building blocks (e.g. hexoses and pentoses), which can be converted later into chemicals of interest. This study aimed to isolate cellulase- and xylanase-producing bacteria for the bioconversion of lignocellulosic materials into their respective monomeric sugars. Bacterial isolates were screened using CMC- and Xylan–Trypan blue agar, and then cellulase and xylanase activities were evaluated by the 3,5-dinitro-salicylic acid (DNS) method. Furthermore, bacterial ability to saccharify wheat straw was tested. Ten bacterial isolates were found to have the ability to saccharify wheat straw, and to produce cellulase and xylanase enzymes simultaneously. The bacterial isolates were identified at molecular level using 16S rRNA gene sequencing and phylogenetic analysis. Bacterial isolates were identified as Cellulomonas sp. CX4, Cellulomonas sp. CX5, Bacillus sp. CX6, Bacillus sp. CX10, Paenibacillus illinoisensis CX11, Paenibacillus sp. CX14, Bacillus cereus CX15, and Bacillus sp. CX16, Paenibacillus barcinonensis CX17 and Cellulomonas sp. CX20. Among all the isolates, Bacillus sp. CX6 showed the highest ability to produce total reducing sugar (6.03 and 6.16 mg/ml), while the lowest ability to saccharify wheat straw was found with Cellulomonas sp. CX5 (2.01 and 2.12 mg/ml). This study presents cellulase- and xylanase- producing bacterial isolates for their potential to saccharify lignocellulosic materials for possible use in the production of pharmaceuticals and chemicals.

English

 Paenibacillus strain, UKCH21, isolated from Uttarakhand State, north western Indian Himalayas was found to produce high levels of extracellular chitinases. The 16s rRNA gene sequence showed 100% homology with P. illinoisensis (Accession No. KR856190) available in the public domain and further phylogenetic study also verified the species identity. The culture supernatants have a maximum chitinase activity of 110.8 U/ml after 3 days of culturing. The isolate showed strong antifungal activity manifested in the form of progressive mycelia degradation in dual culture plates. The pathogenicity was observed as structural deformities like uneven thickening of mycelia as a result of direct degradation of chitin. The optimum pH and temperature of UKCH21 chitinases was found to be 5 and 50°C, respectively. Partial characterization of chitinase gene also confirms the family 18 status of glycosyl hydrolase with substantial variability presented here with. Above all, percent inhibition of growth and the rapid degradation of mycelia of tested plant pathogenic fungi (Rhizoctonia solani, Fusarium solani and Sclerotium rolfsi) in bacteria seeded medium suggest its utilization as potent antifungal biocontrol agent.   Key words: Paenibacillus illinoisensis, UKCH21, chitinase, antifungal, Indian Himalayas.

ISOLATION OF MODERATELY HALOPHILIC LIPASE PRODUCING BACTERIA FROM SPONGES IN PAHANG COASTAL WATER, MALAYSIA

Sponges (Porifera) harbour diverse microorganisms which can be the potential source for microbial enzymes such as lipase. In this study, moderately halophilic lipase producing bacteria were isolated from sponges tissues collected near Balok, at Pahang coastal water. Out of 70 isolates that grew on tributyrin agar plate, only 7 isolates had produced clear zones surrounding their colonies. Out of these, 5 isolates appeared to be gram-positive rod; meanwhile, the other 2 isolates were gram-negative rod in morphologies. These isolates were subjected to several biochemical tests i.e., oxidase, gelatin hydrolysis, lactose fermentation, citrate and motility test, and 16S rRNA gene amplification and sequencing. The results from 16S rRNA sequencing showed that 2 isolates (NHTH 6B and NHTH 28A) were highly similar (>97%) with Paenibacillus illinoisensis; isolate NHTH 26A with Stenotrophomonas pavanii; and isolate NHTH 29A with Enterobacter aerogenes. Phylogenetic analysis on selected isolates (NHTH 6B, NHTH 26A, NHTH 28A and NHTH 29A) with other species from the database showed high bootstrap values of above 50%. This showed that diverse phyla of lipase producing bacteria were isolated from the sponge collected from Pahang coastal water. In the isolation of industrial important species, the presence of pathogenic group of microorganism in this sponge could indicate issues on water quality and safety in this area.

Paenibacillus kyungheensis sp. nov., isolated from flowers of magnolia.

A Gram-staining-positive, catalase-positive, oxidase-negative, facultatively anaerobic, rod-shaped bacterium designated strain DCY88T, was isolated from flowers of magnolia. Phylogenetic analysis based on 16S rRNA gene sequence comparison revealed that the strain formed a distinct lineage within the genus Paenibacillus that was closely related to Paenibacillus hordei RH-N24T (97.8 %). The other most closely related species were Paenibacillus illinoisensis NRRL NRS-1356T (94.3 %), Paenibacillus hunanensis DSM 22170T (94.2 %), Paenibacillus peoriae DSM 8320T (93.9 %), Paenibacillus kribbensis Am49T (93.8 %) and the type species of the genus, Paenibacillus polymyxa ATCC 842T (93.3 %). Cells of the strain were endospore-forming and motile by peritrichous flagella. Strain DCY88T formed pink-pigmented colonies on trypticase soy agar and R2A agar medium. Growth of strain DCY88T occurs at temperatures 5-37 °C, at pH 4-9 and 0.5-5.5 % NaCl (w/v). The menaquinone was MK-7.The cell wall peptidoglycan of strain DCY88T contained meso-diaminopimelic acid. The major fatty acids were anteiso-C15 : 0 (61.0 %) and C16 : 0 (11.0 %). The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and an unidentified polar lipid. The strain DCY88T contained spermidine as the major polyamine. The DNA G+C content was 51.6 mol%. The DNA-DNA hybridization relatedness between strain DCY88T and P. hordei RH-N24T was 48 ± 2 %. The phenotypic, phylogenetic and chemotaxonomic results indicate that the strain DCY88T represents a novel species of the genus Paenibacillus, for which the name Paenibacillus kyungheensis sp. nov. is proposed. The type strain is DCY88T ( = JCM 19886T = KCTC 33429T).

Bioefficacy of Rhizobacterial Isolates against Root Infecting Fungal Pathogens of Chickpea (Cicer arietinum L.)

Chickpea is considered to be food for the poor in Pakistan. Its yield is much lower than expected due to infestation of a number of fungal pathogens. The present study was designed to determine the effect of rhizobacterial isolates against fungal pathogens infecting chickpea roots. RH-31, RH-32 and RH-33 were isolated from groundnut rhizosphere. Antifungal activities of these isolates were tested by seed treatment and soil application methods against three root fungal pathogens. Data on disease incidence, bio-control efficiency and root biomass was recorded. Phylogenetic analysis indicated that sequences of RH-31, RH-32 and RH-33 showed >99% identity with Paenibacillus illinoisensis, Bacillus subtilis, and Pseudomonas psychrotolerans respectively. RH-33 was effective against Fusarium oxysporum and Macrophomina phaseolina with highest levels of inhibition, whereas RH-32 inhibited Fusarium solani. However, RH-31 showed best activity against F. oxysporum. Disease incidence and bio-control efficiency revealed that all isolates reduced disease severity and increased overall plant biomass as compared to control treatment. Present findings show potential of bacterial isolates from rhizosphere of Pakistan. Application of selected rhizobacteria through seed treatment method might be a promising strategy to lower damage caused by root pathogens in chickpea. This could be efficient, economical, environment friendly and might serve as a biocontrol agent.

Isolation and characterization of methane utilizing bacteria from wetland paddy ecosystem

Methylotrophic bacteria which are known to utilize C1 compounds including methane. Research during past few decades increased the interest in finding out novel genera of methane degrading bacteria to efficiently utilize methane to decrease global warming effect. Moreover, evaluation of certain known plant growth promoting strains for their methane degrading potential may open up a new direction for multiple utility of such cultures. In this study, efficient methylotrophic cultures were isolated from wetland paddy fields of Gujarat. From the overall morphological, biochemical and molecular characterization studies, the isolates were identified and designated as Bacillus aerius AAU M 8; Rhizobium sp. AAU M 10; B. subtilis AAU M 14; Paenibacillus illinoisensis AAU M 17 and B. megaterium AAU M 29. Gene specific PCR analysis of the isolates, P. illinoisensis, B. aerius, Rhizobium sp. and B. subtilis showed presence of pmoA gene encoding α subunit particulate methane monooxygenase cluster. B. megaterium, P. illinoisensis, Rhizobium sp. and Methylobacterium extrorquens showed presence of mmoX gene encoding α subunit of the hydroxylase component of the soluble methane monooxygenase cluster. P. illinoisensis and Rhizobium sp. showed presence mxaF gene encoding α subunit region of methanol dehydrogenase gene cluster showing that both isolates are efficient utilizers of methane. To the best of our knowledge, this is the first time report showing presence of methane degradation enzymes and genes within the known PGPB group of organisms from wet land paddy agro-ecosystem, which is considered as one of the leading methane producer.

β-cyclodextrin production by the cyclodextrin glucanotransferase from Paenibacillus illinoisensis ZY-08: cloning, purification, and properties

The gene encoding the cyclodextrin glucanotransferase (CGTase, EC2.4.1.19) of Paenibacillus illinoisensis was isolated, cloned, sequenced and expressed in Escherichia coli. Sequence analysis showed that the mature enzyme (684 amino acids) was preceded by a signal peptide of 34-residues. The deduced amino acid sequence of the CGTase from P. illinoisensis ZY-08 exhibited highest identity (99 %) to the CGTase sequence from Bacillus licheniformis (P14014). The four consensus regions of carbohydrate converting domain and Ca(2+) binding domain could be identified in the sequence. The CGTase was purified by using cold expression vector, pCold I, and His-tag affinity chromatography. The molecular weight of the purified enzyme was about 74 kDa. The optimum temperature and pH of the enzyme were 40 °C and pH 7.4, respectively. The enzyme activity was increased by the addition of Ca(2+) and inhibited by Ba(2+), Cu(2+), and Hg(2+). The K m and V max values calculated were 0.48 mg/ml and 51.38 mg of β-cyclodextrin/ml/min. The ZY-08 and recombinant readily converted soluble starch to β-cyclodextrin but ZY-08 did not convert king oyster mushroom powder and enoki mushroom powder. However the recombinant CGTase converted king oyster mushroom powder and enoki mushroom powder to β-cyclodextrin.

Paenibacillus hordei sp. nov., isolated from naked barley in Korea

A Gram-staining positive, facultative aerobic bacterium, designated strain RH-N24(T), was isolated from naked barley in South Korea. Cells of the isolate were observed to be motile rods by means of peritrichous flagella and showed catalase-positive and oxidase-negative reactions. Growth of strain RH-N24(T) was observed at 4-40°C (optimum: 35-37°C) and at pH 5.0-9.0 (optimum: pH 6.0-7.0). Chemotaxonomic data (major isoprenoid quinone: MK-7; DNA G+C content: 53.5mol%; cell wall type: A1γ-meso-diaminopimelic acid; major fatty acids: anteiso-CB(15:0) and CB(16:0B)) supported the affiliation of the isolate to the genus Paenibacillus. The major cellular polar lipids were identified as phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol and two unidentified polar lipids. Phylogenetic analysis based on 16S rRNA gene sequences also supported the conclusion that strain RH-N24(T) belonged to the genus Paenibacillus. Comparative 16S rRNA gene sequence analysis showed that strain RH-N24(T) was most closely related to Paenibacillus hunanensis FeL05(T) and Paenibacillus illinoisensis NRRL NRS-1356(T) with similarities of 94.64 and 94.54%, respectively. On the basis of phenotypic and molecular properties, strain RH-N24(T) represents a novel species within the genus Paenibacillus for which the name Paenibacillus hordei sp. nov. is proposed. The type strain is RH-N24(T) (=KACC 15511(T)=JCM 17570(T)).

Immobilization of chitinases from Streptomyces griseus and Paenibacillus illinoisensis on chitosan beads

To investigate the enzyme activity of the chitosan beads immobilized with chitinases this study was carried out. Protein content of the Streptomyces griseus enzyme in supernatant was not detected in test tubes including chitosan beads treated with GA 1, 2, 3, 4, 5, and 10% at 6 h after the reaction except for glutaraldehyde (GA) 0%. Also, chitinase activity of the supernatant was highest (2.68 unit/mL), in GA 0%. Regarding enzyme stability of S. griseus, total chitinase activities were 1.4, 0.4, and 0.3 units/2 mL in GA 0%, 3%, and 5%, respectively, at the initial reaction time. The supernatant protein from Paenibacillus illinoisensis KJA-424 was bound at the highest level of 42% in chitosan beads treated with GA 10% after reaction at 4

Isolation and Purification of Thermostable β-Mannanase from Paenibacillus illinoisensis ZY-08

A bacterium, ZY-08 that produced extracellular mannanase, was isolated and identified as Paenibacillus illinoisensis (P. illinoisensis) on the basis of 16S rDNA phylogenetic analysis. The enzyme was purified to apparent homogeneity by ultrafiltration, DEAE-Sepharose chromatography, and Sephadex G-50 chromatography procedures. The β-mannanase was purified 3.5 fold to homogeneity with a final recovery of 22% and specificity of 9.3 U/mg protein as judged by SDS-polyacrylamide gel electrophoresis. The molecular mass was about 60 kDa. It was active at 60°C and pH 6.0 and it’s retained about 40% of activity at 70°, with a broad pH range of 3.0-6.0. The mannanase was highly specific towards glucomannan and galactomannan, but exhibited very low activity towards starch, carboxy-methyl cellulose, and birchwood xylan. Result of thin-layer chromatography analysis showed that the enzyme produced various kinds of oligosacchrides. These unique properties of the thermostable mannanase and broad substrate specificity (low activity towards cellulosic substrates) from P. illinoisensis ZY-08 make this enzyme attractive for biotechnological applications.

Paenibacillus taichungensis sp. nov., from soil in Taiwan

Among a large collection of Taiwanese soil isolates, a novel Gram-variable, rod-shaped, motile, endospore-forming bacterial strain, strain V10537(T), was subjected to a polyphasic study including 16S rRNA and gyrB gene sequence analysis, DNA-DNA hybridization experiments, cell wall peptidoglycan type, cellular fatty acid composition analysis and comparative phenotypic characterization. 16S rRNA gene sequence analysis indicated that the organism belonged to the genus Paenibacillus. Strain V10537(T) possessed meso-diaminopimelic acid as the diagnostic diamino acid of the peptidoglycan. It contained menaquinone MK-7 as the predominant isoprenoid quinone and anteiso-C(15 : 0) (53.6 %) and C(16 : 0) (19.0 %) as the major fatty acids. Phylogenetically, the most closely related species to strain V10537(T) were Paenibacillus pabuli, Paenibacillus xylanilyticus, Paenibacillus amylolyticus, Paenibacillus barcinonensis and Paenibacillus illinoisensis, with 16S rRNA gene sequence similarities of 99.5, 98.8, 98.3, 98.2 and 98.1 % to the respective type strains. The gyrB gene sequence similarities between strain V10537(T) and these strains were 76.9-85.0 %. DNA-DNA hybridization experiments showed levels of relatedness of 8.5-45.6 % between strain V10537(T) and these strains. The DNA G+C content of strain V10537(T) was 46.7 mol%. Strain V10537(T) was clearly distinguishable from other Paenibacillus species and thus represents a novel species of the genus Paenibacillus, for which the name Paenibacillus taichungensis sp. nov. is proposed. The type strain is V10537(T) (=BCRC 17757(T) =DSM 19942(T)).

Paenibacillus provencensis sp. nov., isolated from human cerebrospinal fluid, and Paenibacillus urinalis sp. nov., isolated from human urine

Gram-negative, spore-forming rods were isolated from a human urine sample (strain 5402403(T)) and a human cerebrospinal fluid sample (strain 4401170(T)). Based on genotypic characteristics, these strains belonged to the genus Paenibacillus and were closely related. Phylogenetic analysis based on 16S rRNA gene sequence comparison showed that they clustered with Paenibacillus massiliensis 2301065(T) (95.9 and 94.3 % 16S rRNA gene sequence similarity, respectively, for strains 5402403(T) and 4401170(T)), Paenibacillus illinoisensis NRRL NRS-61356(T) (90.6 and 93.8 %), Paenibacillus xylanilyticus XIL14(T) (95.3 and 95.4 %), Paenibacillus barcinonensis BP-23(T) (94.3 and 94.0 %), Paenibacillus pabuli NCIMB 12781(T) (89.1 and 92.3 %) and Paenibacillus amylolyticus NRRL NRS-290(T) (94.2 and 93.8 %). The predominant fatty acids were 15 : 0 anteiso (49.0 and 55.3 %, respectively, for strains 5402403(T) and 4401170(T)), 16 : 0 iso (15.4 and 13.5 %), 16 : 0 (7.6 and 3.6 %), 15 : 0 (6.3 and 2.8 %), 17 : 0 anteiso (5.7 and 7.5 %), 14 : 0 iso (4.1 and 2.7 %) and 15 : 0 iso (4.1 and 3.4 %). 16S rRNA gene sequence similarity between strain 4401170(T) and strain 5402403(T) was 98.4 %, but the DNA-DNA reassociation rate between the two strains was 53.2 %. So, considering the recommendations of the ad hoc committee, they do not belong to the same species. On the basis of phenotypic data and genotypic inference, it is proposed that the strains should be assigned to the novel species Paenibacillus urinalis sp. nov. (type strain 5402403(T) =CIP 109357(T) =CCUG 53521(T)) and Paenibacillus provencensis sp. nov. (type strain 4401170(T) =CIP 109358(T) =CCUG 53519(T)).