Saccharification of Lignocellulosic Materials by Cellulolytic and Xylanolytic Paenibacillus illioisensis 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.

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).

Paenibacillus panacisoli sp. nov., a xylanolytic bacterium isolated from soil in a ginseng field in South Korea

A Gram-positive, facultatively anaerobic, motile, spore-forming bacterium, designated Gsoil 1411T, was isolated from soil of a ginseng field in Pocheon Province (South Korea) and was characterized using a polyphasic approach. Comparative analysis of 16S rRNA gene sequences revealed that strain Gsoil 1411T belongs to the family Paenibacillaceae, with closest sequence similarity to the type strains of Paenibacillus xylanilyticus (95.7 %), Paenibacillus illinoisensis (95.2 %) and Paenibacillus pabuli (94.8 %). Strain Gsoil 1411T showed less than 94 % sequence similarity to the type strains of other recognized members of the genus Paenibacillus. In addition, the presence of MK-7 as the major menaquinone, anteiso-C15 : 0 as a major fatty acid (44.8 %) and the presence of PAEN513F and PAEN862F signature sequences suggest that it is affiliated to the genus Paenibacillus. The G+C content of the genomic DNA was 53.9 mol%. On the basis of its phenotypic characteristics and phylogenetic distinctiveness, strain Gsoil 1411T is suggested to represent a novel species within the genus Paenibacillus, for which the name Paenibacillus panacisoli sp. nov. is proposed. The type strain is Gsoil 1411T (=KCTC 13020T=LMG 23405T).