Kallotenue papyrolyticum gen. nov., sp. nov., a cellulolytic and filamentous thermophile that represents a novel lineage (Kallotenuales ord. nov., Kallotenuaceae fam. nov.) within the class Chloroflexia.
Several closely related, thermophilic and cellulolytic bacterial strains, designated JKG1(T), JKG2, JKG3, JKG4 and JKG5, were isolated from a cellulolytic enrichment (corn stover) incubated in the water column of Great Boiling Spring, NV. Strain JKG1(T) had cells of diameter 0.7-0.9 µm and length ~2.0 µm that formed non-branched, multicellular filaments reaching >300 µm. Spores were not formed and dense liquid cultures were red. The temperature range for growth was 45-65 °C, with an optimum of 55 °C. The pH range for growth was pH 5.6-9.0, with an optimum of pH 7.5. JKG1(T) grew as an aerobic heterotroph, utilizing glucose, sucrose, xylose, arabinose, cellobiose, CM-cellulose, filter paper, microcrystalline cellulose, xylan, starch, Casamino acids, tryptone, peptone, yeast extract, acetate, citrate, lactate, pyruvate and glycerol as sole carbon sources, and was not observed to photosynthesize. The cells stained Gram-negative. Phylogenetic analysis using 16S rRNA gene sequences placed the new isolates in the class Chloroflexia, but distant from other cultivated members, with the highest sequence identity of 82.5 % to Roseiflexus castenholzii. The major quinone was menaquinone-9; no ubiquinones were detected. The major cellular fatty acids (>5 %) were C18 : 0, anteiso-C17 : 0, iso-C18 : 0, iso-C17 : 0, C16 : 0, iso-C16 : 0 and C17 : 0. The peptidoglycan amino acids were alanine, ornithine, glutamic acid, serine and asparagine. Whole-cell sugars included mannose, rhamnose, glucose, galactose, ribose, arabinose and xylose. Morphological, phylogenetic and chemotaxonomic results suggest that JKG1(T) is representative of a new lineage within the class Chloroflexia, which we propose to designate Kallotenue papyrolyticum gen. nov., sp. nov., Kallotenuaceae fam. nov., Kallotenuales ord. nov. The type strain of Kallotenue papyrolyticum gen. nov., sp. nov. is JKG1(T) ( = DSM 26889(T) = JCM 19132(T)).
School of Life Sciences, University of Nevada, Las Vegas, 4505 S. Maryland Parkway, Las Vegas, Nevada 89154, USA.
Jessica K
JK
Cole
School of Life Sciences, University of Nevada, Las Vegas, 4505 S. Maryland Parkway, Las Vegas, Nevada 89154, USA.
Brandon A
BA
Gieler
School of Life Sciences, University of Nevada, Las Vegas, 4505 S. Maryland Parkway, Las Vegas, Nevada 89154, USA.
Devon L
DL
Heisler
School of Life Sciences, University of Nevada, Las Vegas, 4505 S. Maryland Parkway, Las Vegas, Nevada 89154, USA.
Maryknoll M
MM
Palisoc
School of Life Sciences, University of Nevada, Las Vegas, 4505 S. Maryland Parkway, Las Vegas, Nevada 89154, USA.
Amanda J
AJ
Williams
Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, Washington 99352, USA.
Alice C
AC
Dohnalkova
Yunnan Institute of Microbiology, School of Life Sciences, Yunnan University, Kunming, 650091, PR China.
Hong
H
Ming
Yunnan Institute of Microbiology, School of Life Sciences, Yunnan University, Kunming, 650091, PR China.
Tian Tian
TT
Yu
School of Life Sciences, University of Nevada, Las Vegas, 4505 S. Maryland Parkway, Las Vegas, Nevada 89154, USA.
Jeremy A
JA
Dodsworth
Yunnan Institute of Microbiology, School of Life Sciences, Yunnan University, Kunming, 650091, PR China.
Wen-Jun
WJ
Li
School of Life Sciences, University of Nevada, Las Vegas, 4505 S. Maryland Parkway, Las Vegas, Nevada 89154, USA.
Brian P
BP
Hedlund
D002482:Cellulose / Q000378:metabolism*; D041862:Chloroflexi / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000235:genetics; D005227:Fatty Acids / Q000737:chemistry; D045482:Hot Springs / Q000382:microbiology*; D008969:Molecular Sequence Data; D009505:Nevada; D010457:Peptidoglycan / Q000737:chemistry; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D024482:Vitamin K 2 / Q000031:analogs & derivatives / Q000737:chemistry
D004269:DNA, Bacterial; D005227:Fatty Acids; D010457:Peptidoglycan; D012336:RNA, Ribosomal, 16S; D024482:Vitamin K 2; C073355:menaquinone 9; D002482:Cellulose