Streptomyces beigongshangae sp. nov., isolated from baijiu fermented grains, could transform ginsenosides of Panax notoginseng.

A Gram-stain-positive actinomycete, designated REN17T, was isolated from fermented grains of Baijiu collected from Sichuan, PR China. It exhibited branched substrate mycelia and a sparse aerial mycelium. The optimal growth conditions for REN17T were determined to be 28 °C and pH 7, with a NaCl concentration of 0 % (w/v). ll-Diaminopimelic acid was the diagnostic amino acid of the cell-wall peptidoglycan and the polar lipids were composed of phosphatidylethanolamine, phosphatidylinositol, an unidentified phospholipid, two unidentified lipids and four unidentified glycolipids. The predominant menaquinone was MK-9 (H2), MK-9 (H4), MK-9 (H6) and MK-9 (H8). The major fatty acids were iso-C16 : 0. The 16S rRNA sequence of REN17T was most closely related to those of Streptomyces apricus SUN 51T (99.8 %), Streptomyces liliiviolaceus BH-SS-21T (99.6 %) and Streptomyces umbirnus JCM 4521T (98.9 %). The digital DNA-DNA hybridization, average nucleotide identity and average amino acid identify values between REN17T and its closest replated strain, of S. apricus SUN 51T, were 35.9, 88.9 and 87.3 %, respectively. Therefore, REN17T represents a novel species within the genus Streptomyces, for which the name Streptomyces beigongshangae sp. nov. is proposed. The type strain is REN17T (=GDMCC 4.193T=JCM 34712T). While exploring the function of the strain, REN17T was found to possess the ability to transform major ginsenosides of Panax notoginseng (Burk.) F.H. Chen (Araliaceae) into minor ginsenoside through HPLC separation, which was due to the presence of ß-glucosidase. The recombinant ß-glucosidase was constructed and purified, which could produce minor ginsenosides of Rg3 and C-K. Finally, the enzymatic properties were characterized.

Bosea beijingensis sp. nov., Telluria beijingensis sp. nov. and Agrococcus beijingensis sp. nov., isolated from baijiu mash.

The baijiu fermentation environment hosts a variety of micro-organisms, some of which still remain uncultured and uncharacterized. In this study, the isolation, cultivation and characterization of three novel aerobic bacterial strains are described. The cells of strain REN20T were Gram-negative, strictly aerobic, motile and grew at 26-37 °C, at pH 6.0-9.0 and in the presence of 0-5.0   % (w/v) NaCl. The cells of strain REN29T were Gram-negative, strictly aerobic, motile and grew at 15-30 °C, at pH 6.0-9.0 and in the presence of 0-10.0   % (w/v) NaCl. The cells of strain REN33T were Gram-positive, strictly aerobic, motile and grew at 15-37 °C, at pH 5.0-10.0 and in the presence of 0-7.0   % (w/v) NaCl. The digital DNA-DNA hybridization and average nucleotide identity by orthology values between type strains in related genera and REN20T (20.3-36.8 % and 79.8-89.9  %), REN29T (20.3-36.8  % and 74.5-88.5  %) and REN33T (22.6-48.6  % and 75.8-84.2  %) were below the standard cut-off criteria for the delineation of bacterial species, respectively. Based on polyphasic taxonomy analysis, we propose three new species, Bosea beijingensis sp. nov. (=REN20T=GDMCC 1.2894T=JCM 35118T), Telluria beijingensis sp. nov. (=REN29T=GDMCC 1.2896T=JCM 35119T) and Agrococcus beijingensis sp. nov. (=REN33T=GDMCC 1.2898T=JCM 35164T), which were recovered during cultivation and isolation from baijiu mash.

Sporosarcina beigongshangi sp. nov., isolated from pit mud of Baijiu.

A Gram-positive, aerobic and short rod-shaped bacterium designated REN13T, was isolated from pit mud of Baijiu, in Sichuan province, China. Strain REN13T could grow at 10-50 â„ƒ, pH 6.0-9.0 and 0-2% (w/v) NaCl, with the optimal growth occurred at 28 â„ƒ, pH 7.0, and 2% (w/v) NaCl. 16S rRNA gene sequence analysis showed that strain REN13T was closely related to Sporosarcina globispora DSM 4T (98.6%). The DNA G + C content of strain REN13T was 41.1 mol %. DDH and ANI value between strain REN13T and S. globispora DSM 4T was 24.4% and 67.6%, respectively. The major fatty acids were iso-C15:0 and antesio-C15:0. The respiratory quinone was MK-7, and the polar lipids were phosphatidylethanolamine, phospholipids, phosphatidylglycerol and diphosphatidylglycerol. Based on the above polyphasic taxonomic analysis, strain REN13T represents a novel species of the genus Sporosarcina, for which the name Sporosarcina beigongshangi sp. nov. is proposed. The type strain is strain REN13T (= JCM 34409T = GDMCC 1.2151T).

Umezawaea beigongshangensis sp. nov., Isolated From the Mash of Baijiu.

A Gram-positive, aerobic and non-motile actinobacterial strain, designated REN6T, was isolated from the mash of Baijiu (Chinese spirits, a kind of distilling liquor is made by cooking, saccharification, fermentation and distillation) collected from Sichuan Province Region, China, and characterized using a polyphasic approach. Morphological and chemotaxonomic properties of strain REN6T were consistent with the description of the genus Umezawaea, such as the spore arrangement, the abundant aerial mycelium and the fragmented substrate mycelium. The diamino acid of peptidoglycan is meso-diaminopimelic acid. The diagnostic phospholipids were DPG (diphosphatidylglycerol), PG (phosphatidylglycerol), PI (phosphatidylinositol), PE (phosphatidylethanolamine). The major fatty acids were C16:0, C17:0, Iso-C16:0, Iso-C16:1 H, C17:1ω6c and C17:1 ω8c. Menaquinone-9 (MK-9) (H4) was the predominant menaquinones. The genomic DNA G + C contents were 72.7 mol%. Phylogenetic analysis based on 16S rDNA gene sequences demonstrated that strain REN6T should also be classified in the genus Umezawaea, with U. tangerina (98.7%) and U. endophytica (98.7%). However, it can be distinguished from the closest strains U. tangerina JCM 10302T based on the low levels of DNA-DNA hybridization 22.1%. Based upon the morphological, physiological, chemotaxonomic and molecular characteristics differences from other members of the genus, a novel species, Umezawaea beigongshangensis sp. nov., is proposed, with REN6T (= JCM 33954T = CGMCC 19205T) as the type strain.

Pseudoxanthomonas beigongshangi sp. nov., a novel bacteria with predicted nitrite and nitrate reduce ability isolated from pit mud of Baijiu.

A Gram-negative and rod-shape bacterium designated REN9T, was isolated from pit mud of Baijiu in Sichuan, China. The 16S rRNA sequence of strain REN9T had a high similarity to Pseudoxanthomonas indica P15T (99.21%), P. mexicana AMX 26BT (97.74%) and P. japonensis 12-3T (97.43%). Phylogenetic analysis showed that REN9T belongs to the genus Pseudoxanthomonas and formed distinct cluster. The ANI and DDH values between strains REN9T and P15T were 80.94% and 24%, respectively. Strain REN9T grew optimally at 37 °C, pH 7.0 and 2% NaCl. PE (phosphatidylethanolamine), PG (phosphatidylglycerol) and DPG (diphosphatidylglycerol) were the major polar lipids of REN9T. Ubiquinone 8 (Q-8) was the predominant quinone, and Iso-C15:0 (64.32%), anteiso-C15:0 (12.04%) and Iso-C14:0 (4.56%) were the majority fatty acids. The DNA G + C content of strain REN9T was 67.3 mol%. Genomic analysis showed that strain REN9T had two secondary metabolite biosynthesis gene clusters. Moreover, strain REN9T had 43 glycoside hydrolases, 41 glycosyl transferases and 41 carbohydrate esterases. Based on the polyphasic taxonomic analysis, strain REN9T is recommended as a novel species within the genus Pseudoxanthomonas, for which the name Pseudoxanthomonas beigongshangi is proposed. The type stain is REN9T (= JCM 33961T = GDMCC 1.2210T).

Brevibacterium renqingii sp. nov., isolated from the Daqu of Baijiu.

Two bacterial strains, designated REN4T and REN4-1, were isolated from daqu sample collected from baijiu factory located in Shanxi, China. The two strains shared highly similar 16S rRNA gene sequences (99.67% identities) and formed a monophyletic clade within the Brevibacterium 16S rRNA gene tree, showing 97.56-97.85% 16S rRNA gene sequence identities with type strains Brevibacterium permense VKM Ac-2280 T, Brevibacterium sediminis FXJ8.269 T, Brevibacterium oceani BBH7T and Brevibacterium epidermidis NCIMB 702286 T. They contained MK-8(H2) as the most predominant menaquinone, antesio-C15:0, antesio-C17:0, Iso-C16:0 and Iso-C17:0 as the major cellular fatty acids, DPG (diphosphatidylglycerol), PG (phosphatidylglycerol), PGL (phosphatidylglycerollipids), and PL (phospholipids) as the main polar lipids. The genomic DNA G + C content of strains REN4 and REN4-1 were 64.35, 65.82 mol%. Moreover, the low DNA-DNA relatedness values, physiological and biochemical characteristics, and taxonomic analysis allowed the differentiation of strains REN4T and REN4-1 from the other recognized species of the genus Brevibacterium. Therefore, strain REN4T represents a novel species of the genus Brevibacterium, for which the name Brevibacterium renqingii sp. nov. is proposed, with the type strain REN4T (= JCM 33953 T = KCTC 49366 T).

Pontibacter beigongshangensis sp. nov., Isolated from the Mash of Wine.

A Gram-negative, aerobic, oval-shaped, and light red pigmented bacterium, designated T6-1T, was isolated from the mash of wine collected from a wine-making laboratory simulated fermenter located in Beijing, China. The optimal growth of T6-1T occurred at 30 °C, pH 7.0 with 1% NaCl. The sole respiratory quinone was menaquinone-7 (MK-7). The principal cellular fatty acids (>5%) were iso-C15:0, iso-C17:0 3OH, C16:1 ω5c, and iso-C17:0. The major polar lipids were PE (phosphatidylethanolamine), PL (unidentified phospholipid), and L1-2 (unidentified lipids). 16S rRNA phylogenetic analysis indicated that strain T6-1T belonged to the genus Pontibacter. The 16S rRNA gene sequence of strain T6-1T was most similar to Pontibacter amylolyticus 9-2T (95.92%). The genomic DNA G+C content of strain T6-1 was 50.34 mol%. The digital DNA-DNA relatedness and average nucleotide identity value between T6-1T and 9-2T was 20.20% and 74.18%, respectively. Polyphasic taxonomy analysis indicated that strain T6-1T represents a novel species of the genus Pontibacter, for which the name Pontibacter beigongshangensis sp. nov. is proposed, with the type strain T6-1T (= CGMCC 1.17104T = KCTC 72413T).