A novel bioprospecting strategy via 13C-based high-throughput probing of active methylotrophs inhabiting oil reservoir surface soil.

Methane-oxidizing bacteria (MOB) have long been considered as a microbial indicator for oil and gas prospecting. However, due to the phylogenetically narrow breath of ecophysiologically distinct MOB, classic culture-dependent approaches could not discriminate MOB population at fine resolution, and accurately reflect the abundance of active MOB in the soil above oil and gas reservoirs. Here, we presented a novel microbial anomaly detection (MAD) strategy to quantitatively identify specific indicator methylotrophs in the surface soils for bioprospecting oil and gas reservoirs by using a combination of 13C-DNA stable isotope probing (SIP), high-throughput sequencing (HTS), quantitative PCR (qPCR) and geostatistical analysis. The Chunguang oilfield of the Junggar Basin was selected as a model system in western China, and type I methanotrophic Methylobacter was most active in the topsoil above the productive oil wells, while type II methanotrophic Methylosinus predominated in the dry well soils, exhibiting clear differences between non- and oil reservoir soils. Similar results were observed by quantification of Methylobacter pmoA genes as a specific bioindicator for the prediction of unknown reservoirs by grid sampling. A microbial anomaly distribution map based on geostatistical analysis further showed that the anomalous zones were highly consistent with petroleum, geological and seismic data, and validated by subsequent drilling. Over seven years, a total of 24 wells have been designed and drilled into the targeted anomaly, and the success rate via the MAD prospecting strategy was 83 %. Our results suggested that molecular techniques are powerful tools for oil and gas prospecting. This study indicates that the exploration efficiency could be significantly improved by integrating multi-disciplinary information in geophysics and geomicrobiology while reducing the drilling risk to a greater extent.

Identifying Active Rather than Total Methanotrophs Inhabiting Surface Soil Is Essential for the Microbial Prospection of Gas Reservoirs.

Methane-oxidizing bacteria (MOB) have long been recognized as an important bioindicator for oil and gas exploration. However, due to their physiological and ecological diversity, the distribution of MOB in different habitats varies widely, making it challenging to authentically reflect the abundance of active MOB in the soil above oil and gas reservoirs using conventional methods. Here, we selected the Puguang gas field of the Sichuan Basin in Southwest China as a model system to study the ecological characteristics of methanotrophs using culture-independent molecular techniques. Initially, by comparing the abundance of the pmoA genes determined by quantitative PCR (qPCR), no significant difference was found between gas well and non-gas well soils, indicating that the abundance of total MOB may not necessarily reflect the distribution of the underlying gas reservoirs. 13C-DNA stable isotope probing (DNA-SIP) in combination with high-throughput sequencing (HTS) furthermore revealed that type II methanotrophic Methylocystis was the absolutely predominant active MOB in the non-gas-field soils, whereas the niche vacated by Methylocystis was gradually filled with type I RPC-2 (rice paddy cluster-2) and Methylosarcina in the surface soils of gas reservoirs after geoscale acclimation to trace- and continuous-methane supply. The sum of the relative abundance of RPC-2 and Methylosarcina was then used as specific biotic index (BI) in the Puguang gas field. A microbial anomaly distribution map based on the BI values showed that the anomalous zones were highly consistent with geological and geophysical data, and known drilling results. Therefore, the active but not total methanotrophs successfully reflected the microseepage intensity of the underlying active hydrocarbon system, and can be used as an essential quantitative index to determine the existence and distribution of reservoirs. Our results suggest that molecular microbial techniques are powerful tools for oil and gas prospecting.

Complete degradation of PET waste using a thermophilic microbe-enzyme system.

Enzymatic degradation has been proposed as a suitable solution for addressing PET pollution, but approximately 10 % of PET is left as nonbiodegradable. Microbes can completely degrade PET at the gram level per year. Based on the complementary benefits of microbes and enzymes, a microbe-enzyme system was created to completely degrade PET. Here, a thermophilic microbe-enzyme (TME) system composed of Bacillus thermoamylovorans JQ3 and leaf-branch compost cutinase variant (ICCG) was used to demonstrate the synergistic degradation of PET, enabling 100 % degradation of PET waste at a high PET loading level (360 g/L). Six endogenous PET hydrolases of strain JQ3 were discovered by employing an ester bond hydrolysis function-first genome mining (EGM) strategy and first successfully expressed in E. coli. These hydrolases could release TPA as the final product from PET and preferentially degraded BHET instead of MHET. Of these, carboxylesterase 39_5 and ICCG could degrade PET in a synergistic manner to generate 50 µM of TPA, which was greater than the sum of the individual treatments. Finally, the degradation pathway of the TME system was speculated to include biofilm formation, PET degradation and utilization. The successful implementation of this study rendered a scale-up degradation feasible of PET at a lower cost.

Current Fungal Taxonomy and Developments in the Identification System.

A functional identification system is the core and basis of fungal taxonomy, which provides sufficient diagnostic characteristics for species delimitation. Phenotype-based identification systems have exhibited significant drawbacks, such as being laborious and time-consuming. Thus, a molecular-based identification system (rDNA, DNA fingerprint, etc.) is proposed for application to fungi that lack reliable morphological characteristics. High Throughput Sequencing also makes great contributions to fungal taxonomy. However, the formal naming of nonculturable fungi from environmental sequencing is a significant challenge. Biochemical profile-based identification systems have outstanding value in fungal taxonomy and can occasionally be indispensable. This method utilizes biomarker metabolites and proteins that are expected to be unequivocal and stable. Of these, Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry has become the method of choice for chemotaxonomy. In sum, these described identification systems cannot solve all problems of species delimitation, and considerable attention to the updating of fungal nomenclature, standardization of techniques, knowledge sharing, and dissemination will be necessary.

Multiple approaches of loop region modification for thermostability improvement of 4,6-α-glucanotransferase from Limosilactobacillus fermentum NCC 3057.

4,6-α-glucanotransferase (4,6-α-GT), as a member of the glycoside hydrolase 70 (GH70) family, converts starch/maltooligosaccharides into α,1-6 bond-containing α-glucan and possesses potential applications in food, medical and related industries but does not satisfy the high-temperature requirement due to its poor thermostability. In this study, a 4,6-α-GT (ΔGtfB) from Limosilactobacillus fermentum NCC 3057 was used as a model enzyme to improve its thermostability. The loops of ΔGtfB as the target region were optimized using directed evolution, sequence alignment, and computer-aided design. A total of 11 positive mutants were obtained and iteratively combined to obtain a combined mutant CM9, with high resistance to temperature (50 °C). The activity of mutant CM9 was 2.08-fold the activity of the wild type, accompanied by a 5 °C higher optimal temperature, a 5.76 °C higher melting point (Tm, 59.46 °C), and an 11.95-fold longer half-life time (t1/2). The results showed that most of the polar residues in the loop region of ΔGtfB are mutated into rigid proline residues. Molecular dynamics simulation demonstrated that the root mean square fluctuation of CM9 significantly decreased by "Breathing" movement reduction of the loop region. This study provides a new strategy for improving the thermostability of 4,6-α-GT through rational loop region modification.

Synergism Between Multi-Pseudomonas and Cutinase for Biodegradation of Crude Oil-Based Derivatives.

Polyethylene terephthalate (PET) as one of the main crude oil-based derivatives, produces a significant amount of waste that is difficult to degrade. Currently, microbial degradation of PET is an eco-friendly, efficient, and economical method. This study was conducted to propose a novel screening strategy for PET-degrading bacteria, and evaluate their degradation efficiency of PET. Two strains, Pseudomonas nitroreducens S8 and Pseudomonas monteilii S17, were isolated and could utilize PET as a carbon source by co-culture. The combined use of both bacteria gave a synergistic effect on the disruption of the PET surface through colonization behavior, which could enhance the subsequent degradation of PET. Its time of reaching a peak value of PET degradation rate (94.5% at 6 d) was 2 days earlier than these of single bacteria. A similar synergistic effect was also observed in the metabolization of PET monomers, and the metabolic rate was expressed as 82.4% of bis (2-hydroxyethyl) terephthalate (BHET), 64.0% of mono (2-hydroxyethyl) terephthalate (MHET), and 20.0% of terephthalic acid (TPA), respectively. This study is novel in showing the degradation of PET waste by combinations of bacterial pretreatment and enzymatic treatment, which can be a promising method.

Directional-path modification strategy enhances PET hydrolase catalysis of plastic degradation.

Poly (ethylene terephthalate) (PET) is a widely used type of general plastic that produces a significant amount of waste due to its non-degradable properties. We propose a novel directional-path modification (DPM) strategy, involving positive charge amino acid introduction and binding groove remodeling, and apply it to Thermobifida fusca cutinase to enhance PET degradation. The highest value of PET degradation (90%) was achieved in variant 4Mz (H184S/Q92G/F209I/I213K), exhibiting values almost 30-fold that of the wild-type. We employed molecular docking, molecular dynamics simulations, and QM/MM MD for the degradation process of PET, accompanied by acylation and deacylation. We found that the distance of nucleophilic attack was reduced from about 4.6 Å in the wild type to 3.8 Å in 4Mz, and the free energy barrier of 4Mz dropped from 14.3 kcal/mol to 7.1 kcal/mol at the acylation which was the rate-limiting step. Subsequently, the high efficiency and universality of the DPM strategy were successfully demonstrated in LCC, Est119, and BhrPETase enhancing the degradation activity of PET. Finally, the highest degradation rate of the pretreated commercial plastic bottles had reached to 73%. The present study provides insight into the molecular binding mechanism of PET into the PET hydrolases structure and proposes a novel DPM strategy that will be useful for the engineering of more efficient enzymes for PET degradation.

Genetic and Chemical Diversity of Edible Mushroom Pleurotus Species.

The genus Pleurotus is one of the most widely cultivated and edible mushrooms with various cultivators. Three molecular characteristics were used to evaluate the genetic diversity of 132 tested samples. Phylogenetic analysis showed five clades for tested samples of the genus Pleurotus by the combined ITS and LSU sequences with strong bootstraps and Bayesian posterior probability supports. A total of 94 polymorphic fragments ranging from 10 to 100 bp were observed by using an intersimple sequence repeat (ISSR) marker. The DNA fragment pattern showed that P. ostreatus cultivator (strain P9) was clearly distinguished from wild strain based on their clear banding profiles produced. DNA GC content of the genus Pleurotus varied from 55.6 mol% to 43.3 mol%. Their chemical composition was also determined, including sugar, amino acid, polar lipid, mycolic acid, quinone, and fatty acid, which presented some high homogeneity. Most of the tested samples contained mycolic acid; glucose and arabinose as the main sugars; aspartic acid, arginine, lysine, tyrosine, and alanine as the main amino acids; and C16:0, C18:0, C18:2 cis-9,12, anteiso-C14:0, and summed feature 8 as the main fatty acids. In addition, their polar lipid profiles were investigated for the first time, which significantly varied among Pleurotus species. The genus Pleurotus contained menaquinone-6 as the sole respiratory quinone, which showed a significant difference with that of its closely related genera. These results of this study demonstrated that the combined method above could efficiently differentiate each Pleurotus species and thus be considered an efficient tool for surveying the genetic diversity of the genus Pleurotus.

Accelerated biodegradation of polyethylene terephthalate by Thermobifida fusca cutinase mediated by Stenotrophomonas pavanii.

Polyethylene terephthalate (PET) is a general plastic that produces a significant amount of waste due to its non-biodagradable properties. We obtained four bacteria (Stenotrophomonas pavanii JWG-G1, Comamonas thiooxydans CG-1, Comamonas koreensis CG-2 and Fulvimonas soli GM-1) that utilize PET as a sole carbon source through a novel stepwise screening and verification strategy. PET films pretreated with S. pavanii JWG-G1 exhibited weight loss of 91.4% following subsequent degradation by Thermobifida fusca cutinase (TfC). S. pavanii JWG-G1 was able to colonize the PET surface and maintain high cell viability (over 50%) in biofilm, accelerating PET degradation. Compared with PET films with no pretreatment, pretreatment with S. pavanii JWG-G1 caused the PET surface to be significantly rougher with greater hydrophilicity (contact angle of 86.3 ± 2° vs. 96.6 ± 2°), providing better opportunities for TfC to contact and act on PET. Our study indicates that S. pavanii JWG-G1 could be used as a novel pretreatment for efficiently accelerating PET biodegradation by TfC.

Synergistic biodegradation of poly(ethylene terephthalate) using Microbacterium oleivorans and Thermobifida fusca cutinase.

Poly(ethylene terephthalate) (PET) is a major source of plastic pollution. Biodegradation technologies are of paramount interest in reducing or recycling PET waste. In particular, a synergistic microbe-enzyme treatment may prove to be a promising approach. In this study, a synergistic system composed of Microbacterium oleivorans JWG-G2 and Thermobifida fusca cutinase (referred to as TfC) was employed to degrade bis(hydroxyethyl) terephthalate (BHET) oligomers and a high crystalline PET film. A novel degradation product that was obtained by M. oleivorans JWG-G2 treatment alone was identified as ethylene glycol terephthalate (EGT). With the addition of TfC as a second biocatalyst, the highest synergy degrees for BHET oligomers and PET film degradation were 2.79 and 2.26, respectively. The largest amounts of terephthalic acid (TPA) and mono(2-hydroxyethyl) terephthalate (MHET) (47 nM and 330 nM, respectively) were detected after combined treatment of PET film with M. oleivorans JWG-G2 at 5 × 103 µL/cm2 and TfC at 120 µg/cm2, and the degree of PET film surface destruction was more significant than those produced by each treatment alone. The presence of extracellular PET hydrolases in M. oleivorans JWG-G2, including three carboxylesterases, an esterase and a lipase, was predicted by whole genome sequencing analysis, and a predicted PET degradation pathway was proposed for the synergistic microbe-enzyme treatment. The results indicated that synergistic microbe-enzyme treatment may serve as a potentially promising tool for the future development of effective PET degradation. KEY POINTS: • An ecofriendly synergistic microbe-enzyme PET degradation system operating at room temperature was first introduced for degrading PET. • A novel product (EGT) was first identified during PET degradation. • Potential PET hydrolases in M. oleivorans JWG-G2 were predicted by whole genome sequencing analysis.

A dual-functional aminopeptidase from Streptomyces canus T20 and its application in the preparation of small rice peptides.

An aminopeptidase that derived from Streptomyces canus T20 (ScAP) was successfully expressed and characterized in Escherichia coli BL21 (DE3) for first time. The specific activity was 6000 U/mg, which is highest in Streptomyces aminopeptidases. Its optimal conditions were 60 °C and pH 8.0, respectively. ScAP exhibited excellent thermal and alkaline pH stability, retained 80.0% maximal activity at 50 °C for 200 h or at pH 9.0 for 24 h. Its activity observed to be complete inhibited by 0.1 mM EDTA and enhanced by Ca2+ and Co2+ to 115.4% and 104.0% respectively. ScAP also has exhibited high specificity towards rice protein on preparation of small peptides. The yield of small rice peptides achieved 66.5%, which is highest by far. Besides, ScAP have significant debittering effect on rice peptides. Results showed that bitter intensity score decreased by 49.0% with optimum condition (0.048% ScAP at 50 °C for 6 h). Therefore, ScAP as dual functional aminopeptidase of hydrolytic and debittering might have a potential application in the production of high yield and low bitterness of small rice peptides.

Cyclodextrinase from Thermococcus sp expressed in Bacillus subtilis and its application in the preparation of maltoheptaose.

BACKGROUND: Maltoheptaose as malto-oligosaccharides with specific degree of polymerization, has wide applications in food, medicine and cosmetics industries. Currently, cyclodextrinase have been applied as prepared enzyme to prepare maltoheptaose. However, the yield and proportion of maltoheptaose was lower, which is due to limited substrate and product specificity of cyclodextrinase (CDase). To achieve higher maltoheptaose yield, cyclodextrinase with high substrate and product specificity should be obtained. RESULTS: In this study, cyclodextrinase derived from Thermococcus sp B1001 (TsCDase) was successfully expressed and characterized in Bacillus subtilis for the first time. The specific activity of TsCDase was 637.95 U/mg under optimal conditions of 90 °C and pH 5.5, which exhibited high substrate specificity for cyclodextrins (CDs). When the concentration of ß-CD was 8%, the yield of maltoheptaose achieved by TsCDase was 82.33% across all reaction products, which exceeded the yields of maltoheptaose in other recent reports. Among malto-oligosaccharides generated as reaction products, maltoheptaose was present in the highest proportion, about 94.55%. CONCLUSIONS: This study provides high substrate and product specificity of TsCDase. TsCDase is able to prepare higher yield of maltoheptaose through conversion of ß-CD in the food industry.

Arthrobacter sedimenti sp. nov., isolated from river sediment in Yuantouzhu park, China.

A Gram-stain positive, motile, aerobic and rod-shaped strain (MIC A30T) was isolated from river sediment in Yuantouzhu park, Wuxi City, China. Growth occurred at 20-40 °C, at pH 6.0-9.0 and at 0-5.0% NaCl. Strain MIC A30T was moderately related to Arthrobacter liuii CGMCC 1.12778T (97.9%), Arthrobacter pokkaliiT (97.9%) and Arthrobacter globiformis NBRC 12137T (96.7%) by 16S rRNA analysis. The DNA-DNA relatedness values between strain MIC A30T and these reference strains were below 30%. The DNA G+C content was 63.1 mol%. Average nucleotide identity (ANI) and genome-to-genome distance (GGD) values between strain MIC A30T and A. liuii CGMCC 1.12778T were 60.34% and 29.39%, respectively. Quinone was identified as MK-9(H2). Major polar lipids were diphosphatidylglycerol and phosphatidylglycerol. Major fatty acids were iso-C15:0, anteiso-C15:0 and anteiso-C17:0. Whole-cell sugars were galactose, mannose and rhamnose. The cell wall peptidoglycan contained A4α peptidoglycan type with lysine as the diagnostic diamino acid. Based on several taxonomic results, strain MIC A30T is identified as a novel species in genus Arthrobacter, whose name is proposed as Arthrobacter sedimenti sp. nov. The type strain is MIC A30T (= KACC 19599T = CGMCC 1.13474T).

Luteimonas cellulosilyticus sp. nov., Cellulose-Degrading Bacterium Isolated from Soil in Changguangxi National Wetland Park, China.

A Gram-negative, motile, aerobic, and rod-shaped strain (MIC 1.5T) was isolated from soil in Changguangxi national wetland park. Growth occurred at 20-45 °C, at pH 6.0-8.0, and at 0-4.0% NaCl. Based on 16S rRNA gene sequence analysis, strain MIC 1.5T was related to were identified as Luteimonas dalianensis CGMCC 1.12191T (95.3%), Luteimonas padinae DSM 101536T (94.5%), Luteimonas huabeiensis DSM 26429T (94.1%), and Luteimonas mephitis DSM 12574T (92.5%). The DNA-DNA relatedness values between strain MIC 1.5T , and these strains were well below 31%. The polar lipids were phosphatidylethanolamine, phosphatidylglycerol, and diphosphatidylglycerol. The DNA G+C content of strain MIC 1.5T was 66.3 mol%. Average nucleotide identity (ANI) and genome-to-genome distance (GGD) values between strain MIC 1.5T and L. dalianensis CGMCC 1.12191T were 65.39% and 29.52%, respectively. The quinone was identified as Q-8. The major fatty acids were iso-C15:0, iso-C15:0 3OH, and iso-C17:0 3OH and summed feature 3 (C16:1ω7c and/or iso-C15:0 2-OH). Based on the phylogenetic, physiological, and chemotaxonomic results, strain MIC 1.5T represents a novel species of the genus Luteimonas, for which the name Luteimonas cellulosilyticus sp. nov. is proposed. The type strain is MIC 1.5T (= KACC 19469T = CCTCC AB 2017256T).

Cellulomonas aurantiaca sp. nov., isolated from a soil sample from a tangerine field.

A Gram-stain positive, facultatively aerobic, motile and rod-shaped bacterial strain, designated THG-SMD2.3T, was isolated from a soil sample collected in a tangerine field, Republic of Korea. According to the 16S rRNA gene sequence comparisons, the isolate was identified as a member of the genus Cellulomonas and to be closely related to Cellulomonas fimi ATCC 484T (98.5%), Cellulomonas biazotea DSM 20112T (98.3%), Cellulomonas chitinilytica X.bu-bT (98.0%), Cellulomonas xylanilytica XIL11T (97.2%), Cellulomonas humilata ATCC 25174T (97.1%) and Cellulomonas composti TR7-06T (97.0%). The 16S rRNA gene sequence similarities with other current species of the genus Cellulomonas were in the range 95.4-96.6%. Catalase and oxidase tests were found to be positive. The DNA G+C content was determined to be 73.0 mol%. DNA-DNA hybridization values between strain THG-SMD2.3T and C. fimi ATCC 484T, C. biazotea DSM 20112T, C. chitinilytica X.bu-bT, C. xylanilytica XIL11T, C. humilata ATCC 25174T and C. composti TR7-06T were 58.1 ± 1.6%, 56.7 ± 0.8%, 30.3 ± 1.6%, 22.8 ± 1.6%, 19.9 ± 1.6%, and 13.5 ± 3.0%, respectively. Strain THG-SMD2.3T was also found to be able to grow at 20-42 °C, at 0-3% NaCl and at pH 5.5-10. The major fatty acids were identified as anteiso-C15:0, iso-C15:0, anteiso-C17:0 and iso-C14:0. The predominant menaquinone was identified as tetrahydrogenated menaquinones with nine isoprene units [MK-9(H4)]. The polar lipids were found to be diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, two unidentified aminolipids and two unidentified phospholipids. Based on these phenotypic, genotypic and phylogenetic characterisations strain THG-SMD2.3T (= KACC 19341T = CGMCC 1.16303T) is concluded to represent a novel species of the genus Cellulomonas, for which the name Cellulomonas aurantiaca sp. nov. is proposed.

Chitinophaga aurantiaca sp. nov., isolated from a soil sample from a tangerine field.

A Gram-stain-negative, facultatively anaerobic, non-motile and rod-shaped bacterial strain, designated THG-SD5.5T, was isolated from a soil sample collected in a tangerine field, Republic of Korea. According to the 16S rRNA gene sequence comparisons, the isolate was identified as a member of the genus Chitinophaga and to be closely related to Chitinophaga ginsengihumi KACC 17604T (97.9%) and Chitinophaga rupis KACC 14521T (97.5%). The 16S rRNA gene sequence similarities with other species of the genus Chitinophaga were in the range 92.8-95.5%. Catalase test was positive. Oxidase test was negative. The DNA G + C content was determined to be 46.1 mol%. DNA-DNA hybridization values between strain THG-SD5.5T and C. ginsengihumi KACC 17604T and C. rupis KACC 14521T were 45.1% and 15.6%, respectively. Strain THG-SD5.5T was also found to be able to grow at 24-33 °C, at 0-5% NaCl and at pH 5.5-9.0. The major fatty acids were identified as anteiso-C15:0, C16:0, anteiso-C17:0 and C18:0. The dominant respiratory quinone was identified as menaquinone-7 (MK-7). The polar lipids were found to be diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, an unidentified aminolipid, two unidentified phospholipids and three unidentified lipids. Based on these phenotypic, genotypic and phylogenetic characterisations, strain THG-SD5.5T (= KACC 19338T = CGMCC 1.16304T) is concluded to represent a novel species of the genus Chitinophaga, for which the name Chitinophaga aurantiaca sp. nov. is proposed.

Glaciecola amylolytica sp. nov., an amylase-producing bacterium isolated from seawater.

A Gram-stain-negative, aerobic, non-motile and coccus-shaped bacterium (THG-3.7T) was isolated from seawater. Growth occurred at 10-30 °C (optimum 25 °C), at pH 6-8 (optimum 7) and in the presence of 1-8 % (w/v) NaCl (optimum 4 %). Based on 16S rRNA gene sequence analysis, the nearest phylogenetic neighbours of strain THG-3.7T were identified as Paraglaciecola mesophila DSM 15026T (95.3 % similarity), Glaciecola pallidula DSM 14239T (95.2 %), Paraglaciecola aquimarina KCTC 32108T (95.1 %), Paraglaciecola arctica KACC 14537T (94.9 %), Glaciecola nitratireducens KCTC 12276T (94.7 %) and Paraglaciecola psychrophila CGMCC 1.6130T (94.7 %). 16S rRNA gene sequence similarities among strain THG-3.7T and other species were lower than 94.7 %. The polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, one unidentified lipid and one unidentified aminolipid. The quinone system was composed of Q-8. The major fatty acids were C16 : 0, C18 : 1ω7c and summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH). The DNA G+C content of strain THG-3.7T was 47.9 mol%. On the basis of the data presented, strain THG-3.7T represents a novel species of the genus Glaciecola, for which the name Glaciecola amylolytica sp. nov. is proposed. The type strain is THG-3.7T (=KACC 19478T=CCTCC AB 2017258T).

Rubus idaeus L. (red raspberry) blocks UVB-induced MMP production and promotes type I procollagen synthesis via inhibition of MAPK/AP-1, NF-κß and stimulation of TGF-ß/Smad, Nrf2 in normal human dermal fibroblasts.

Chronic exposure to ultraviolet (UV) radiation causes photo-oxidation, which in turn results in the upregulation of matrix metalloproteinases (MMPs) and loss of collagen. Rubus idaeus L. (RI), also called red raspberry, is an important cash crop that contains abundant antioxidant compounds. Sanguiin H-6 and lambertianin C are the major ingredients presented in the extracts. Here, we studied the protective effect of RI on UVB-induced photoaging in normal human dermal fibroblasts (NHDFs). We found that RI notably reduced UVB-induced MMPs secretion and pro-inflammatory mediators production, and significantly suppressed UVB-induced activation of mitogen-activated protein kinase (MAPK), nuclear factor-κß, as well as activator protein 1. Additionally, treatment of NHDFs with the ERK inhibitor (PD98059) and JNK inhibitor (SP600125) resulted in the reduction of UVB-induced MMP-1 and IL-6 expressions, which demonstrated that the inhibition of MMP-1 and IL-6 by RI is associated with the MAPK pathway. Furthermore, we also found that RI accelerated procollagen type I synthesis by activating the transforming growth factor-ß/Smad pathway and enhanced the expression of cytoprotective antioxidants such as heme oxygenase-1 and NHD(P)H quinone oxidoreductase 1 by promoting nuclear factor E2-related factor 2 nuclear transfer. Overall, these findings demonstrated that RI was potentially effective in preventing UVB induced skin photoaging.

Scopulibacillus cellulosilyticus sp. nov., a cellulose-degrading bacterium isolated from tea.

A Gram-stain positive, aerobic, non-motile, endospore-forming and rod-shaped strain (THG-NT9T) was isolated from a green tea sample. Growth occurred at 20-45 °C (optimum 28-35 °C), at pH 6.0-8.0 (optimum 7.0) and at 0-2.0% NaCl (optimum 0%). Based on 16S rRNA gene sequence analysis, the near phylogenetic neighbours of strain THG-NT9T were identified as Scopulibacillus daqui DSM 28236T (98.6%), Scopulibacillus darangshiensis DSM 19377T (97.4%), Pullulanibacillus pueri CGMCC 1.12777T (96.7%) and Pullulanibacillus camelliae CGMCC 1.15371T (96.3%). The DNA G + C content of strain THG-NT9T was determined to be 47.5 mol %. DNA-DNA hybridization values between strain THG-NT9T and S. daqui DSM 28236T, S. darangshiensis DSM 19377T, P. pueri CGMCC 1.12777T, P. camelliae CGMCC 1.15371T and Pullulanibacillus naganoensis DSM 10191T were 41.3 ± 0.1 (39.4 ± 0.4% reciprocal analysis), 39.1 ± 0.1 (37.3 ± 0.1%), 21.4 ± 0.7 (20.1 ± 0.3%), 20.7 ± 0.1 (20.1 ± 0.4%) and 12.1 ± 0.2% (8.3 ± 0.2%). The polar lipids were identified as diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and three unidentified lipids. The quinone was identified as MK-7. The major fatty acids were C18:3 ω7c, iso-C15:0, iso-C16:0, iso-C17:0 and anteiso-C17:0. The cell wall type was determined to be A1γ peptidoglycan with meso-diaminopimelic acid as the diagnostic diamino acid plus alanine and glutamic acid and glucose as the cell wall sugar. On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics, and DNA-DNA hybridization data, strain THG-NT9T represents a novel species of the genus Scopulibacillus, for which the name Scopulibacillus cellulosilyticus sp. nov. is proposed. The type strain is THG-NT9T (= KCTC 33918T = CGMCC 1.16305T).

Camelliibacillus cellulosilyticus gen. nov., sp. nov., a cellulose-degrading bacterium isolated from tea.

A Gram-stain-positive, oxidase- and catalase-positive, endospore-forming, aerobic, non-motile and rod-shaped bacterium (THG-YT1T) was isolated from green tea. Growth occurred at 10-40 °C (optimum, 25-30 °C), at pH 6-8 (optimum, 7) and at 0-2 % NaCl (optimum, 0 %). Based on 16S rRNA gene sequences, phylogenetic analyses showed that strain THG-YT1T formed a distinct lineage with respect to closely related genera in the family Bacillaceae. Strain THG-YT1T was most closely related to the genera within the families Pullulanibacillus, Scopulibacillus, Tuberibacillus and Caenibacillu, with levels of 16S rRNA gene sequence similarity to the type species of members of these genera of less than 95.0 %. The menaquinone was MK-7. The polar lipids were phosphatidylethanolamine, two unidentified aminophospholipids, two unidentified aminolipids and two unidentified glycolipids. The major fatty acids of strain THG-YT1T were C18 : 3ω7c and anteiso-C17 : 0. The cell-wall peptidoglycan type was A1γ with meso-diaminopimelic acid as the diagnostic diamino acid plus alanine and glutamic acid. The cell-wall sugar was glucose. The DNA G+C content of strain THG-YT1T was determined to be 53.5 mol%. Based on the data presented here, strain THG-YT1T represents a novel species of a new genus of the family Bacillaceae, for which the name Camelliibacillus cellulosilyticus gen. nov., sp. nov. is proposed. The type strain is Camelliibacillus cellulosilyticus THG-YT1T(=KACC 19471T=CGMCC 1.16306T).