RESUMEN

The ocean harbors a variety of bacteria that contain huge protease resources and offer a great potential for industrial and biotechnological applications. Here, we isolated a protease-secreting bacterium Vibrio pomeroyi strain 12613 from Atlantic seawater and purified a protease VP9 from strain 12613. VP9 was identified as a metalloprotease of the M4 family. VP9 could hydrolyze casein and gelatin but not elastin and collagen. With gelatin as the substrate, VP9 showed the highest activity at 40°C and pH 6.0-8.0. It was stable at temperatures of 50°C and less and in the range of pH 5.0-11.0. VP9 also had good tolerance to NaCl, non-ionic detergents, and organic solvent methanol. Unlike other M4 metalloproteases, VP9 has distinct collagen-swelling ability, and its collagen-swelling effect was concentration dependent. The relative expansion volume of collagen increased by approximately eightfold after treatment with 10 µM VP9 at 37°C for 12 h. The collagen-swelling mechanism of VP9 on bovine-insoluble type I collagen was further studied. Atomic force microscopy observation and biochemical analyses showed that VP9 can degrade proteoglycans in collagen fibers, resulting in the release of collagen fibrils from collagen fibers and the swelling of the latter. In addition, VP9 can degrade glycoproteins, a non-collagenous constituent interacting with collagen in the skin. The characteristics of VP9, such as sufficient specificity toward proteoglycans and glycoproteins but no activity toward collagen, suggest its promising potential in the unhairing and fiber-opening processing in leather industry.

RESUMEN

A Gram-stain-negative, pink-pigmented, rod-shaped, non-flagellated, aerobic bacterium, designated strain SM1701T, was isolated from a rotten seaweed collected off Fildes Peninsula, King George Island, West Antarctica. The strain grew at 4-30 °C, pH 6.0-8.0 and with 0.5-5 % (w/v) NaCl. It hydrolysed gelatin and Tweens (40, 60 and 80), but did not reduce nitrates to nitrites. The major cellular fatty acids of strain SM1701T were iso-C15 : 0, iso-C15 : 1G, iso-C16 : 1G, C16 : 0 and iso-C17 : 0 3-OH. Polar lipids included phosphatidylethanolamine, one unidentified aminolipid, two unidentified glycolipids and one unidentified aminoglycolipid. The major respiratory quinone was MK-7. The genomic DNA G+C content of strain SM1701T was 34.1 mol%. It showed high 16S rRNA gene sequence similarities to Crocinitomix algicola (93.8 %) and Crocinitomix catalasitica (92.5 %) and less than 91 % sequence similarities to other known members in the family Crocinitomicaceae. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain SM1701T constituted a distinct lineage within the family Crocinitomicaceae. The phylogenetic trees based on concatenated 261 protein sequences from genome sequences showed that strain SM1701T occupied a branch separated from those of known genera in the family of Crocinitomicaceae, indicating it may belong to a new genus. On the basis of the polyphasic characterization of strain SM1701T in this study, it is considered to represent a novel species in a new genus in the family Crocinitomicaceae, for which the name Putridiphycobacter roseus gen. nov., sp. nov. is proposed. The type strain is SM1701T (=KCTC 62302T=NBRC 113201T=CGMCC 1.16510T).

Asunto(s)

Flavobacteriaceae/clasificación , Filogenia , Algas Marinas/microbiología , Regiones Antárticas , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/genética , Ácidos Grasos/química , Flavobacteriaceae/aislamiento & purificación , Glucolípidos/química , Fosfolípidos/química , Pigmentación , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Vitamina K 2/análogos & derivados , Vitamina K 2/química

RESUMEN

Peptidoglycan is the fundamental structural constituent of the bacterial cell wall. Despite many years of research, the architecture of peptidoglycan is still largely elusive. Here, we report the high-resolution architecture of peptidoglycan from the model Gram-positive bacterium Bacillus subtilis. We provide high-resolution evidence of peptidoglycan architecture remodeling at different growth stages. Side wall peptidoglycan from B. subtilis strain AS1.398 changed from an irregular architecture in exponential growth phase to an ordered cable-like architecture in stationary phase. Thickness of side wall peptidoglycan was found to be related with growth stages, with a slight increase after transition to stationary phase. Septal disks were synthesized progressively toward the center, while the surface features were less clear than those imaged with side walls. Compared with previous studies, our results revealed slight differences in architecture of peptidoglycan from different B. subtilis strains, expanding our knowledge about the architectural features of B. subtilis peptidoglycan.