Hanamia caeni gen. nov., sp. nov., a Member of the Family Chitinophagaceae Isolated from Activated Sludge in Korea.

A novel Gram-stain-negative, aerobic, yellowish-pigmented, non-motile, rod-shaped bacterial strain, designated strain BO-59T, was isolated from the activated sludge of a wastewater treatment plant in Hanam City, South Korea. Phylogenetic study based on the 16S rRNA gene sequence positioned BO-59T in a distinct lineage in the family Chitinophagaceae, sharing less than 92.8% sequence similarity with members of the closely related genera Ferruginibacter, Flavitalea, Pseudoflavitalea, Flavisolibacter, Niastella, and Terrimonas. Phylogenomic- and genomic relatedness analyses revealed that strain BO-59T is clearly distinguished from other genera in the family Chitinophagaceae by average nucleotide identity < 66.9%) and the genome-to-genome distance (< 29.5%) values. The strain BO-59T contained MK-7 as the predominant quinone, and iso-C15:0, iso-C17:0 3OH, and iso-C15:1 G as major fatty acids (> 10%). The DNA G + C content was 39.1 mol% based on genome sequence analysis. The polar lipids of strain BO-59T were phosphatidylethanolamine, an unidentified aminophospholipid and three unidentified polar lipids. 16S rRNA gene sequence similarity, physiological, and biochemical characteristics indicated that strain BO-59T represents a novel species of a new genus, for which the name Hanamia caeni gen. nov., sp. nov. is proposed. The type strain is BO-59T (= KACC 19646T = LMG 30865 T).

Effects of Light Regulation on Proteome Expression in Rhodobacter sphaeroides 2.4.1.

Light plays an important role in the transcriptional regulation of photosynthetic apparatus. The influence of oxygen and light conditions on the protein expression of Rhodobacter sphaeroides was investigated using a proteomic approach. The R. sphaeroides was grown aerobically under dark cultivation (D24) and light cultivation (L24) for 24 h. An average of 950 distinguishable spots were obtained on 2-D analytic gel for D24 and L24 conditions, of which 48 proteins exhibited significant changes in protein expression levels. Among the 48, 31 proteins were upregulated and 17 proteins were downregulated in L24 when compared with D24. The results depict the comparative protein expression in R. sphaeroides mediated through growth under light or dark conditions. The data suggest that the overexpressed proteins, phosphoribosyl-ATP pyrophosphatase (HisE), in the D24/aerobic culture are involved in the positive regulation of PAC production can be functionally applied in metabolic engineering and industrial processes.

Cell blocking: An enhancement of foodborne pathogen detection by fluorescent signals of recombinant yeasts.

Cell blocking (CB) technique has been widely applied in many studies since the last century. In our research, this technique was mostly used to study the enhancement of the vacuolar response-based system that could detect Shigella sp. and Salmonella sp. investigated in previous studies. The recombinant yeast cells were blocked by mixing with agarose gel on a 96-wells plate, then storing this plate in -80 °C before using. The optimal conditions for the new system, such as agarose concentration, maximum storage time, were also established. Finally, the efficiency of the vacuolar response-based system was improved, and this system could be used as a portable detector for the foodborne pathogen.

Luteimonas granuli sp. nov., Isolated from Granules of the Wastewater Treatment Plant.

A Gram-reaction negative, aerobic, non-motile, light yellow colored, and rod-shaped bacterium (designated Gr-4T) isolated from granules of a wastewater treatment plant, was characterized by a polyphasic approach to clarify its taxonomic position. Strain Gr-4T was observed to grew optimally at 30 ºC and at pH 7.0 on R2A medium. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain Gr-4T belongs to the genus Luteimonas of the family Xanthomonadaceae and was most closely related to Luteimonas padinae CDR SL 15T (99.1%), Luteimonas terricola DSM 22344T (98.5%) and Luteimonas arsenica 26-35T (97.6). The genome comprises 2,917,404 bp with a G+C content of 70.5 mol%. The ANI value between strain Gr-4T and Luteimonas padinae CDR SL 15T was 87.3%. The DNA-DNA relatedness value between strain Gr-4T and Luteimonas padinae CDR SL 15T, Luteimonas terricola DSM 22344T was 36.4 ± 1.3% and 14.2 ± 1.7%, respectively. The predominant quinone was Q-8. The major fatty acids were iso-C15:0, iso-C16:0 and summed feature 9 (comprising iso-C17:1ω9c and/or C16:0 10-methyl) supported the affiliation of strain Gr-4T to the genus Luteimonas. Moreover, the physiological, biochemical results, and low level of ANI and DNA-DNA relatedness value allowed the phenotypic and genotypic differentiation of strains Gr-4T from other Luteimonas species with validly published names. The novel isolate therefore represents a novel species, for which the name Luteimonas granuli sp. nov. is proposed, with the type strain Gr-4T (=KACC 16614T = JCM 18203T).

Characterization of Cellobiohydrolases from Schizophyllum commune KMJ820.

A novel cellobiohydrolase (CBH)-generating fungi have been isolated and categorized as Schizophyllum commune KMJ820 based on morphology and rDNA gene sequence. Cellulose powder was used as carbon source, the total enzyme activity was 11.51 U/ml is noted; which is among the highest amounts of CBH-generating microbes studied. CBH have been purified to homogenize, with pursual of serial chromatography using S. commune supernatants and two different CBHs were found; CBH 1 and 2. The filtered CBHs showed greater activity (V max = 51.4 and 20.8 U/mg) in contrast to CBHs from earlier studies. The MW (molecular weights) of S. commune CBH 1 and 2 were verified to be approximately 50 kDa and 150 kDa, respectively, by size exclusion chromatography. Even though CBHs have been evaluated from other sources, but S. commune CBH is prominent in comparison to other CBHs by its high enzyme activity.

Vacuolar targeting of aldehyde dehydrogenase 6 tagging with signal peptide of proteinase A.

Vacuoles are useful materials with antimicrobial and anticancerous properties. Vacuolar proteins can discompose macromolecules from the outside of yeast cells. The objective of this study was to determine the function of a protein transported into a vacuole. Specifically, cytosolic protein aldehyde dehydrogenase 6 (ALD6) was used for the delivery to the vacuole. To transport cytosolic protein to the vacuole in this study, a transfer vector including a signal peptide sequence isolated from vacuolar protein proteinase A was designed. A signal peptide is an amino acid sequence in front of the transported protein. Signal peptides have various delivery pathways according to the kind of signal sequence they contain. They play important roles in transporting proteins to organelles, in cellular mechanisms, and the transfer of protein outside and inside eukaryotes. Thus, we focused on the design of a transfer vector containing a signal peptide sequence isolated from the DNA sequence of proteinase A (PEP4). In addition, this study evaluated the expression level of cytosolic ALD6 after being transported into the yeast vacuole. Our results showed that the developed transfer vector was useful for delivering proteins to vacuole by using signal peptide sequence. Therefore, this transfer vector might be used as a tool to deliver target proteins to organelles of interest in eukaryotes.

Ochrobactrum soli sp. nov., Isolated from a Korean Cattle Farm.

A Gram stain negative, motile, non-spore-forming, rod-shaped, strictly aerobic, beige-pigmented bacterium, designated strain BO-7T, was isolated from soil of cattle farm, in Seosan, Republic of Korea. On the basis of 16S rRNA gene sequencing, strain BO-7T clustered with species of the genus Ochrobactrum and appeared closely related to O. haematophilum CCUG 38531T (98.9%), O. daejeonense KCTC 22458T (98.1%), O. rhizosphaerae DSM 19824T (98.1%), O. pituitosum DSM 22207T (98.0%), and O. pecoris DSM 23868T (98.0%). The digital DNA-DNA hybridization and average nucleotide identity between strain BO-7T and the closely related strains were 21.9-39.1%, 78.5-89.5%, respectively, indicating that BO-7T is a novel species of the genus Ochrobactrum. The DNA G + C content of the genomic DNA was 57.1 mol%, and ubiquinone Q-10 was the predominant respiratory quinone. The polar lipids consisted of phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylmonomethyl-ethanolamine, di-phosphatidylglycerol, the major polyamines were spermidine, putrescine, and sym-homospermidine. The major cellular fatty acids (> 5%) were C16:0, C19:0 cycle ω7c, and C18:1ω7c and/or C18:1ω6c (summed feature 8). ANI calculation, digital DNA-DNA hybridization, physiological and biochemical characteristics indicated that strain BO-7T represents a novel species of the genus Ochrobactrum, for which the name Ochrobactrum soli sp. nov. is proposed. The type strain is BO-7T (= KACC 19676T = LMG 30809T).

Cellular Lysosomes' Activity for Melanin Reduction on Artificial Skin Tissue.

All eukaryotes have lysosomes which contain hydrolytic enzymes such as protease to degrade waste materials and cellular fragments. As a cellular organelle, lysosomes function as the digestive system of the cell, serving both to degrade material taken up from outside the cell and to digest obsolete components of the cell itself. Conversely, melanin has photochemical functions to protect tissue from the harmful effects of ultraviolet rays. However, too much of melanin leads to problems such as hyperpigmentation, requiring materials to maintain and control the amount of melanin. In this study, we found evidence of correlation between lysosome and melanin in a new eco-friendly material, MelanoDerm, a reconstituted 3D human skin model containing normal melanocytes and keratinocytes. Melanin content assay and cell viability were measured, using 2% kojic acid as positive control, while MelanoDerm was exposed to various concentrations of lysosome. Our results indicate that lysosome may be a useful cosmetic agent for the treatment of hyperpigmentation.

Hydrogenophaga borbori sp. nov., isolated from activated sludge.

A novel Gram-stain-negative, yellowish-pigmented bacterial strain, designated LA-38T, was isolated from activated sludge of wastewater treatment plants in Hanam city, South Korea. Cell of LA-38T were rod-shaped, aerobic, motile and non-spore-forming. In phylogenetic analyses based on 16S rRNA genes, LA-38T clustered with species of the genus Hydrogenophaga and appeared closely related to Hydrogenophaga intermedia DSM 5680T (99.2 % similarity), Hydrogenophaga palleronii DSM 63T (98.2 %), Hydrogrenophaga laconesensis KCTC 42478T (98.1 %), Hydrogenophaga. atypica DSM 15342T (98.1 %), Hydrogenophaga defluvii DSM 15341T (98.0 %) and Hydrogenophaga taeniospiralis DSM 2082T (97.2 %). The average nucleotide identities between LA-38T and the closely related strains were 79.3-88.5 %, indicating that LA-38T represents a novel species of the genus Hydrogenophaga. The DNA G+C content of the genomic DNA was 69.9 mol% and ubiquinone Q-8 was the predominant respiratory quinone. The major cellular fatty acids (>5 %) were C16 : 0, cyclo-C19 : 0, C16 : 1ω7c and/or C16 : 1ω6c (summed feature 3), and C18 : 1ω7c and/or C18 : 1ω6c (summed feature 8). The major polar lipids consisted of phosphatidylglycerol, diphosphatidylglycerol and phosphatidylethanolamine, the major polyamines were 2-hydroxyputrescine and putrescine. ANI calculation, physiological and biochemical characteristics indicated that LA-38T represents a novel species of the genus Hydrogenophaga, for which the name Hydrogenophaga borbori sp. nov. is proposed. The type strain is LA-38T (=KACC 19730T=LMG 30805T).

Encapsulation of daunorubicin into Saccharomyces cerevisiae-derived lysosome as drug delivery vehicles for acute myeloid leukemia (AML) treatment.

Lysosome, an intracellular organelle with an acid interior, contains acidic hydrolases and specific membrane proteins. Saccharomyces cerevisiae contains vacuoles (corresponding to lysosomes) that have similar lipid composition membrane to mammalian cell membrane. However, yeast vacuoles do not cause significant immune stimulation in vivo. Taking advantage of these structural similarities and bio-derived strengths, the present study describes encapsulation of daunorubicin into lysosome derived from S. cerevisiae as drug delivery vehicles for acute myeloid leukemia (AML) treatment. Daunorubicin is a chemotherapy medication used to treat cancer, specifically for AML. In this study, recombinant S. cerevisiae that could keep the small size of lysosomal vacuoles was constructed. Appropriate time and concentration to encapsulate the drug were then identified. In addition, release profile and anticancer effect of the drug in lysosome carriers were confirmed. According to this study, a more accurate encapsulation condition into lysosome can be optimized and potential application of S. cerevisiae derived lysosomes as drug carriers is confirmed.

Simplicispira hankyongi sp. nov., a novel denitrifying bacterium isolated from sludge.

A Gram-stain-negative, facultatively anaerobic, motile, light-yellow, and rod-shaped bacterium, designated as NY-02T, was isolated from the sludge of a wastewater treatment plant in Hanam City, Republic of Korea. The comparison of 16S rRNA gene sequences revealed that the strain formed a distinct lineage within the genus Simplicispira and was most closely related to S. suum SC1-8T (99.0%), S. limi EMB325T (98.3%), S. psychrophila LMG 5408T (98.2%), and S. piscis RSG39T (97.4%). Both average nucleotide identity (ANI) and DNA-DNA hybridization (DDH) values between strain NY-02T, and its closes type strains [S. suum SC1-8T, S. limi EMB325T, S. psychrophila LMG 5408T, and S. piscis RSG39T] were lower than the cut-off (≥ 95-96% for ANI and ≥ 70% for DDH) to define a bacterial species. The genome comprises of 3,709,074 bp with a G + C content of 64.2 mol%. Ubiquinone 8 (Q-8) was the predominant quinone. The major fatty acids were C16:0 and summed feature 3 (C16:1ω7c and/or C16:1ω6c), and the major polar lipids were phosphatidylglycerol, diphosphatidylglycerol, and phosphatidylethanolamine. The results of the physiological, biochemical, and taxonomic analyses in addition to low ANI and DNA-DNA relatedness values (82.2% and < 34.0%, respectively) indicate that the strain NY-02T represents a novel species of the genus Simplicispira. The name proposed for strain NY-02T (= KACC 19731T = LMG 31165T) is Simplicispira hankyongi sp. nov.

Mesorhizobium denitrificans sp. nov., a novel denitrifying bacterium isolated from sludge.

A Gram-stain-negative, non-spore-forming, facultative, rod-shaped bacterium (designated LA-28T) was isolated from a sludge sample from a wastewater treatment plant in Hanam city, Republic of Korea. On the basis of 16S rRNA gene sequencing, strain LA-28T clustered with species of the genus Mesorhizobium and appeared closely related to M. jarvisii LMG 28313T (96.8%), M. waimense ICMP 19557T (96.7%), and M. huakuii LMG 14107T (96.7%). Growth occurs at 18-40°C on R2A medium in the presence of 1-4% NaCl (w/v) and at pH 6-8. The DNA G+C content was 61.2 mol%, and the predominant quinone was ubiquinone-10 (Q-10). The major cellular fatty acids (> 5%) were C16:0, C19:0ω8c cyclo, C18:1ω7c 11-methyl, and C18:1ω7c and/or C18:1ω6c (summed feature 8). Major polar lipids were phosphatidylglycerol (PG), phosphatidylethanolamine (PE), phosphatidyl-N-methylethanolamine (PME), and phosphatidylcholine (PC). Physiological and biochemical characteristics indicated that strain LA-28T represents a novel species of the genus Mesorhizobium, for which the name Mesorhizobium denitrificans sp. nov. is proposed. The type strain is LA-28T (= KACC 19675T = LMG 30806T).

Co-digestion of Biowastes to Enhance Biological Hydrogen Process by Defined Mixed Bacterial Cultures.

Co-digestion of biowastes for hydrogen (H2) production using defined mixed cultures can overcome the high risk of failure due to contamination and imbalanced nutrient status. H2 production from biowastes-pea-shells, potato peels (PP), onion peels (OP) and apple pomace, either individually or in various combinations was evaluated by hydrolyzing with defined hydrolytic mixed bacterial culture (MHC5) and subjecting the hydrolysate to mixture of defined H2 producers (MMC6). Co-digestion of OP and PP hydrolysate supplemented at H2 production stage with GM-2 and M-9 media resulted in 95 and 102 l H2/kg of Total solids (TS), respectively compared to 84 l H2/kg of TS in control. Upscaling the process by digesting 4.0 l slurry (16-fold) resulted in 88.5 and 95 l H2/kg of TS, respectively compared to 72 l H2/kg of TS in control. Thus, H2 production by co-digestion of biowastes could be improved through the supplementation with very dilute medium (0.1 ×) and selection of suitable biowastes under unsterile conditions. The overall efficiency can be further enhanced by integrating it with bioprocesses for biopolymers such as polyhydroxyalkanoates and or biofuels like methane production.

Brevibacterium anseongense sp. nov., isolated from soil of ginseng field.

Gram-positive, aerobic, non-motile, pale-yellow, and rodshaped bacterium, designated as Gsoil 188T, was isolated from the soil of a ginseng field in Pocheon, South Korea. A phylogenetic analysis based on 16S rRNA gene sequence comparison revealed that the strain formed a distinct lineage within the genus Brevibacterium and was most closely related to B. epidermidis NBRC 14811T (98.4%), B. sediminis FXJ8.269T (98.2%), B. avium NCFB 3055T (98.1%), and B. oceani BBH7T (98.1%), while it shared less than 98.1% identity with the other species of this genus. The DNA G + C content was 68.1 mol%. The predominant quinone was MK-8(H2). The major fatty acids were anteiso-C15:0 and anteiso-C17:0. The cell wall peptidoglycan of strain Gsoil 188T contained meso-diaminopimelic acid. The major polar lipids were phosphatidylglycerol, diphosphatidylglycerol, and an unidentified aminolipid. The physiological and biochemical characteristics, low DNA-DNA relatedness values, and taxonomic analysis allowed the differentiation of strain Gsoil 188T from the other recognized species of the genus Brevibacterium. Therefore, strain Gsoil 188T represents a novel species of the genus Brevibacterium, for which the name Brevibacterium anseongense sp. nov. is proposed, with the type strain Gsoil 188T (= KACC 19439T = LMG 30331T).

In vivo evaluation of hot water extract of Acorus gramineus root against benign prostatic hyperplasia.

BACKGROUND: Acorus gramineus has been reported to exhibit various pharmacological effects including inhibition of cholesterol synthesis, enhancement of lipid metabolism, prevention of dementia and inhibition of mast cell growth. According to the Chinese compendium of materia media, it has been reported that Acorus spp. is effective for sedation, dementia prevention as well as diuretic effect. In addition, it showed more than equivalent activity compared to furosoemide, a drug known to be effective in diuretic action in animal model study. However, their effectiveness against benign prostatic hyperplasia (BPH) of Acorus gramineus has not been reported. This study was designed to evaluate the effect of Acorus gramineus root hot water extract (AG) against BPH in vivo. METHODS: Male rats, 10 weeks of age and weighing 405 g ± 10 g, were used for this study. Biomarkers were evaluated including prostate weight, prostate weight ratio, hormonal changes, 5-α reductase type II androgen receptor (AR) of the prostate gland and anti-oxidant activation factors related to BPH. These biomarkers were measured in vivo test. RESULTS: AG showed significant effect at the 250 and 500 mg/kg/day in rats. Groups treated with AG displayed significantly lower levels of prostate gland weight (0.79 g) compared to the BPH induced group (1.19 g). Also, dihydrotestosterone (DHT) level was decreased from 61.8 to 100% and androgen receptor expression level was decreased from 111 to 658%. Any hematological toxicity of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) level wasn't observed. CONCLUSION: This study indicated that AG was effective for reducing BPH symptoms. TRIAL REGISTRATION: Not applicable.

Effects of Alginate Oligosaccharide Mixture on the Bioavailability of Lysozyme as an Antimicrobial Agent.

In this study, we report an oral drug delivery system without any additional process using pH-sensitive biopolymer, alginate, and alginate oligosaccharide with lysozyme as a model drug. These biopolymers exhibited pH-sensitive characteristics such as shrinking at acidic pH and eroding with dissolution at alkaline pH. The incorporation of lysozyme and biopolymers was performed an artificial intestinal juice (pH 6.8). The immobilization efficiency and lysozyme stability in gastric juice (pH 1.2) has been tested by E coil antimicrobial activity. The lysozyme without biopolymer immobilization lost approximately 80-90% of antimicrobial activity than that of pure lysozyme. However, the pH-sensitive biopolymer-controlled lysozyme maintained similar antimicrobial activity compared to that of pure lysozyme (50-90% of cell mortality). Therefore, this simple, easy, and rapid system can be effectively and practically applied for pathogen treatment for in vivo oral drug delivery.

Purple sweet potato anthocyanins attenuate hepatic lipid accumulation through activating adenosine monophosphate-activated protein kinase in human HepG2 cells and obese mice.

Purple sweet potato is a functional food rich in anthocyanins that possess disease-preventive properties. Anthocyanins are known to possess potent antidiabetic properties. However, the effect of the anthocyanin fraction (AF) from purple sweet potato on hepatic lipid metabolism remains unclear. Our hypothesis is that AF inhibits hepatic lipid accumulation through the activation of adenosine monophosphate-activated protein kinase (AMPK) signaling pathways in vitro and in vivo. In this study, we evaluated body weight, liver histology, and hepatic lipid content in high-fat diet (HFD)-fed ICR mice treated with AF. In addition, we characterized the underlying mechanism of AF's effects in HepG2 hepatocytes through Western blot analysis. Anthocyanin fraction (200 mg/kg per day) reduced weight gain and hepatic triglyceride accumulation and improved serum lipid parameters in mice fed an HFD for 4 weeks. Anthocyanin fraction significantly increased the phosphorylation of AMPK and acetyl-coenzyme A carboxylase (ACC) in the liver and HepG2 hepatocytes. In addition, AF down-regulated the levels of sterol regulatory element-binding protein 1 and its target genes including ACC and fatty acid synthase (FAS). The specific AMPK inhibitor compound C attenuated the effects of AF on the expression of lipid metabolism-related proteins such as SREBP-1 and FAS in HepG2 hepatocytes. The beneficial effects of AF on HFD-induced hepatic lipid accumulation are thus mediated through AMPK signaling pathways, suggesting a potential target for the prevention of obesity.