A novel Gram-stain-positive, aerobic bacterium, designated strain DHS C021(T), was isolated from a limestone sample collected from the Puding Karst Ecosystem Research Station of Guizhou Province, southwest China. This strain developed branched vegetative mycelia, and its aerial mycelia fragmented into rod-shaped spores. The cell-wall peptidoglycan contained meso-diaminopimelic acid and the whole-cell sugars comprised galactose, ribose and mannose. The respiratory quinone was identified as menaquinone MK-9(H-4). The major cellular fatty acids were iso-C-14:0 and iso-C-16:0. The phospholipids detected were diphosphatidylglycerol, phosphatidylethanolamine, hydroxyl-phosphatidyethanolamine, phosphatidylinositol, phosphotidylinositolmannosides and one unidentified phospholipid. The genomic DNA G+C content was 69.8 mol% and 16S rRNA gene sequence analysis showed that the strain belonged to the genus Lentzea and shared highest sequence similarity with Lentzea albida CGMCC 4.1727(T) (98.8 %) and Lentzea waywayandensis CGMCC 4.1646(T) (98.5 %). However, it could be distinguished from these reference strains based on the low levels of DNA-DNA relatedness (54.5 +/- 2.7 and 41.7 +/- 3.2 %, respectively). On the basis of morphological, chemotaxonomic and phylogenetic characteristics, and DNA-DNA hybridization data, strain DHS C021(T) represents a novel species of the genus Lentzea, for which the name Lentzea pudingi sp. nov. is proposed. The type strain is DHS C021T (=CGMCC4.7319(T) =KCTC 39694(T)).