아산화질소($N_2O$)는 기후변화를 야기하는 온실가스임과 동시에 오존층을 파괴하는 가스이다. 하폐수 처리시 생물학적 질소 제거 공정에서 주로 배출되는 아산화질소가 환경에 미치는 영향은 매우 중요하므로 대책 수립이 필요하다. 본 논문에서는 하폐수 처리과정의 아산화질소 배출 관련 최신 연구동향을 종합적으로 고찰함으로써, 아산화질소의 배출량 및 생성에 미치는 주요 인자의 영향을 이해하고, 아산화질소 배출 저감 전략 수립에 필요한 정보를 도출하였다. 하폐수 처리공정에서 아산화질소가 배출되는 주요 경로는 hydroxylamine 산화, nitrifier 탈질 및 종속영양 탈질공정의 3가지로 구분된다. 실험실, 파일럿 및 실 규모 하폐수 처리 공정을 대상으로 아산화질소 배출량을 측정한 결과 아산화질소 배출량의 질소 부하량의 0-95%로 변이가 매우 컸다. 실 규모 하폐수 처리공정에서는 질소 부하량의 0-14.6%가 아산화질소로 배출되고, 평균값과 중간값은 각각 1.95%와 0.2%이었다. 아산화질소 배출량에 영향을 미치는 가장 중요한 운전인자는 용존산소와 아질산염 농도 및 COD/N 비율이었다. 아산화질소 배출 저감을 위해 운전인자를 조절하는 다양한 전략이 보고되고 있다. 또한, 하폐수 처리공정에서 아산화질소 배출 저감하기 위한 새로운 전략으로, 높은 아산화질소 환원효소 활성을 가진 미생물을 활용하거나, 기존의 탈질공정 대신 산소발생 탈질공정(oxygenic denitrification)을 도입하는 것이 제안되고 있다.

Glycogen plays important roles in bacteria. Its structure and storage capability have received more attention recently because of the potential correlations with environmental durability and pathogenicity. However, the low level of intracellular glycogen makes extraction and structure characterization difficult, inhibiting functional studies. Bacteria grown in regular media such as lysogeny broth and tryptic soy broth do no accumulate large amounts of glycogen. Comparative analyses of bacterial media reported in literature for glycogen-related studies revealed that there was no consistency in the recipes reported. Escherichia coli $DH5{\alpha}$ is a convenient model organism for gene manipulation studies with respect to glycogen. Additionally, M9 minimal salts medium is widely used to improve glycogen accumulation, although its composition varies. In this study, we optimized the M9 medium by adjusting the concentrations of itrogen source, tryptone, carbon source, and glucose, in order to achieve a balance between the growth rate and glycogen accumulation. Our result showed that $1{\times}M9$ minimal salts medium containing 0.4% tryptone and 0.8% glucose was a well-balanced nutrient source for enhancing the growth and glycogen storage in bacteria. This result will help future investigations related to bacterial physiology in terms of glycogen function.

Streptococcus thermophilus, Lactobacillus delbrueckii, Lactobacillus fermentum and Lactobacillus casei were successfully isolated from indigenous Indonesian fresh milk based on the general morphological and biochemical classification as described in Bergey's manual. Verification was conducted by sequencing of 16S rRNA after selection using the classification method mentioned in the manual. All isolates exhibited antimicrobial activity against Escherichia coli and Staphylococcus aureus in the well diffusion test. The susceptibilities of the isolated S. thermophilus 24/S1 and L. delbrueckii 94/L4 against 22 different antibiotics were determined by the disc diffusion method and variable susceptibility patterns were observed. Both isolates were susceptible to amoxicillin, the most commonly prescribed antibiotic, and resistant to sulfonamide. The presence of a plasmid was not detected after extraction. S. thermophilus 24/S1 and L. delbrueckii 94/L4 starter cultures were prepared for yogurt production after 9.5 h of incubation and the yogurt was evaluated for its flavor and quality by 30 volunteers. A score of $4.93{\pm}0.45$ out of 7 was obtained as compared to the yogurt prepared using commercial starter cultures which yielded a score of $4.76{\pm}0.30$ out of 7.

Warqe (Ensete ventricosum) has been traditionally fermented in an earthen pit to yield a carbohydrate-rich food product named kocho, for generations. A fermentation enhancer (gammaa) was added to this fermenting mass to enhance the fermentation process. The objectives of this study were to assess the physicochemical properties and microbiota of the kocho fermentation enhancer culture to reduce losses. Cross-sectional study design was implemented to collect 131 gammaa samples on the first day of fermentation. The samples were further classified into four groups according to the duration of fermentation (14, 21, 30, and 60 days) practised in various households traditionally. The results showed that the fermentation time significantly affected the physicochemical properties and microbial load of gammaa (p < 0.01). As the fermentation progressed from day 1 to 60, the pH decreased and the titratable acidity increased. The total coliform, Enterobacteriaceae, aerobicmesophilic bacteria (AMB), yeast, and mould counts were significantly reduced at the end of fermentation. In contrast, the number of lactic acid bacteria (LAB) increased significantly until day 30 of fermentation, because of the ability of the LAB to grow at low pH. Lactobacillus species from LAB isolates and Enter obacteriaceae from AMB isolates were the most abundant microorganisms in gammaa fermentation. However, the Enterobacteriaceae and Lactobacilli species count showed decreasing and increasing trends, respectively, as the fermentation progressed. These isolates must be investigated further to identify the species and strain, so as to develop gammaa at the commercial scale.

In this exploratory study, eight types of microalgae from different phyla (Chlamydomonas reinhardtii, Chlorella species, Haematococcus pluvialis, Porphyridium purpureum, Porphyridium cruentum, Isochrysis species, Isochrysis galbana, and Pavlova lutheri) were tested for their antibacterial activities against eight target pathogenic bacterial strains. The agar well diffusion method and broth micro dilution assay were conducted to estimate the antibacterial activity. Microalgae cell-free supernatants, exopolysaccharides (EPS), water, and organic solvent extracts were used for inhibition analysis. EPS extracted from P. lutheri showed activity against Bacillus subtilis and Pseudomonas aeruginosa. Inhibition zone diameters of 14-20 mm were recorded on agar plates, while the minimum inhibitory concentrations in the broth micro dilution assay were $0.39-25mg\;ml^{-1}$. During this study, haptophyte microalgae, Isochrysis species, and P. lutheri extracts showed the highest activity against most of the tested pathogenic bacterial strains, while most of the extracts were active against the important foodborne pathogen P. aeruginosa. This study showed promising results regarding important microalgae phyla, which will further aid research related to extracts and exploitation of bioactive metabolic compounds in the food and pharmaceutical industries.

남극해에는 산업적으로 유용한 신규 효소촉매를 생산하는 미생물들이 들어 있다. 우리는 로스해(Ross Sea)로부터 분리한 여러 저온성 박테리아를 조사하였으며, 그 중에서 지방분해 능력이 뛰어난 Croceibacter atlanticus (No. 40-F12)를 찾았다. Shotgun 클로닝 방법으로 리파아제 유전자(lipCA)를 찾았으며 Escherichia coli 균에서 LipCA 효소를 발현하였다. Spain Arreo metagenome alpha/beta hydrolase를 기준으로 LipCA 상동구조모델을 만들어서 분석한 결과, ${\alpha}/{\beta}$ hydrolase fold, Gly-X-Ser-X-Gly motif, 그리고 lid 구조를 갖고 있기 때문에 전형적인 리파아제 효소임이 밝혀졌다. Ammonium sulfate 침전법과 겔여과 크로마토그래피를 통해서 세포추출액으로부터 LipCA 효소를 순수하게 분리한 후, 최적 온도, pH, 안정성, 기질특이성, 유기용매 안정성 등의 효소특성을 규명하였다. LipCA를 cross-linked enzyme aggregate (CLEA) 방법으로 고정화하고 효소특성을 조사, 비교하였다. 고정화를 통해 온도, pH, 유기용매에 대한 안정성이 증가하였고 기질특이성의 변화는 관찰되지 않았다. $LipCA^{CLEA}$는 원심분리 방법으로 쉽게 회수되었고 4번의 재사용 후에 40% 이상의 활성이 잔재하였다. 이 논문은 C. atlanticus 리파아제의 발현, 특성규명, Cross-linked Enzyme Aggregated 고정화를 바탕으로 안정성을 높여 산업적 활용 가능성을 제시한 최초의 보고이다.

Protein disulfide isomerase A3 (PDIA3) is a major member of the protein disulfide isomerase (PDI) family. PDI proteins commonly reside in the endoplasmic reticulum and mediate important thiol-disulfide interchanges during post-translational protein folding. Unlike other PDI family members, PDIA3 is ubiquitous in various organ systems. However, its physiological activity varies in other tissues. PDIA3 has been associated with cancer, airway inflammation, neurodegenerative diseases, and metabolic diseases. However, the mechanisms of the association of PDIA3 with these pathological conditions remain unclear. Recombinant PDIA3 (rPDIA3) is needed to clarify the interactions between PDIA3 and certain physiological phenomena. In the present study, we aimed to produce highly purified rPDIA3 for use in pathological experiments. We expressed rPDIA3 with a histidine-enriched elongated peptide tag in Escherichia coli and obtained rPDIA3 at 97.8% purity using consecutive His-tag and reverse-phase chromatography. Elongated peptide tags screened from artificially designated library had dual functions for protein expression and simple purification.

유용미생물은 임업과 축산 분야에 활용될 뿐만 아니라 병해충 방제와 작물 생육 증진 등의 용도로 농업에서 널리 이용되고 있다. 하지만 유용미생물의 토양에서의 생존율과 정착율에 대한 연구는 미미한 형편이다. 본 연구에서는 마이크로코즘을 이용해 바실러스 3 균주를 토양에 처리한 후, 이들의 토양 내 생존능을 정량 PCR을 이용하여 13일 동안 정량적으로 분석하였다. 또한 Illumina MiSeq 플랫폼을 이용하여 바실러스 3 균주 처리구와 대조구의 토양미생물 군집 분포를 비교 및 분석하였다. 바실러스 3 균주의 처리 직후 토양 내 밀도는 건조토양 1 그람당 평균 $4.4{\times}10^6$ 유전자수로 대조구에 비해 1,000배 이상 높았다. 바실러스 균주의 토양 내 밀도는 처리 후 약 일주일 간 유지되었고 그 후부터는 유의성 있게 감소하였지만 여전히 대조구보다 100배 이상 높았다. 바실러스 균주 처리 후 토양 내 미생물 군집 구조 분석 결과, 대조구와 처리구 모두 Acidobacteria 문($26.3{\pm}0.9%$), Proteobacteria 문($24.2{\pm}0.5%$), Chloroflexi 문($11.1{\pm}0.4%$), Actinobacteria 문($9.7{\pm}2.5%$)에 속하는 세균이 우점하였다. 대조구 대비 처리구에서 Actinobacteria 문의 비율은 뚜렷하게 감소하였지만 Bacteroidetes 문과 Firmicutes 문의 비율은 증가하는 경향이었다. 속 수준에서 바실러스 3 균주를 처리함에 따라 일부 세균 군집의 종 풍부도를 변화되었고, 결국 전체 토착 미생물 군집 구조가 변화되었음을 확인할 수 있었다. 본 연구에서 수행한 유용한 바실러스의 토양 접종 후 이들의 토양 내 생존능 분석 및 토착 세균 군집의 변화는 유용미생물을 생물적 제제로 시설재배지에 사용할 때 중요한 정보를 제공할 것으로 판단된다.

Differences in host ethnicities and geographical distributions may influence the genetic variation and pathogenesis of Helicobacter pylori strains, particularly with respect to those with a high risk of gastric cancer and in Asian Enigma regions. We simultaneously identified H. pylori virulence-associated genes involved in inflammation and cell damage in Thai and Korean dyspeptic patients. The virulence-associated gene cagA, cagA genotypes (East Asian and Western type cagA), vacA genotypes (s- and m-), oipA, and sabA were detected in Thai and Korean dyspeptic patients by polymerase chain reaction (PCR), real-time PCR, and DNA sequence analysis. Comparisons between the two regions showed that cagA, East Asian type cagA, and vacA s1/m1 in Korean dyspeptic patients occurred at rates of 100%, 86.67%, and 88.89%, respectively (p < 0.05). The oipA- and sabA-positive samples were significantly more predominant in the Korean population (95.56%, 91.11%) than in the Thai population (32%, 34%). DNA sequence analysis revealed differences in the patterns of cytosine-thymine dinucleotide repeats of oipA and sabA among the two populations of dyspeptic patients. Our results indicate that the H. pylori strains detected in the two regions were divergent, and strains colonizing the Korean dyspeptic patients may be more virulent than those in the Thai population. Our data may help explain H. pylori pathogenesis in Asian Enigma areas with a low gastric cancer incidence. However, other factors involving H. pylori infection in these two regions should be further analyzed.

Rabies virus (RABV)-specific antibodies in animals and humans are measured using standard methods such as fluorescent antibody virus neutralization (FAVN) tests and rapid fluorescent focus inhibition tests, which are based on cell culture systems. An alternative assay that is safe and easy to perform is required for rapid sero-surveillance following mass vaccination of animals. Two purified monoclonal antibodies (4G36 and B2H17) against RABV were selected as capture and detection antibodies, respectively. A genetically modified RABV, the ERAGS strain, was propagated and concentrated by polyethylene glycol precipitation. Optimal conditions for the RABV antigen, antibodies, and serum dilution for a blocking enzymelinked immune sorbent assay (B-ELISA) were established. We evaluated the sensitivity, specificity, and accuracy of the B-ELISA using serum samples from 138 dogs, 71 raccoon dogs, and 25 cats. The B-ELISA showed a diagnostic sensitivity of 95.8-96.3%, specificity of 91.3-100%, and accuracy of 96.0-97.2% compared to the FAVN test. These results suggest that the B-ELISA is useful for sero-surveillance of RABV in dogs, raccoon dogs, and cats.

Current reports suggest that obesity is a serious global health issue. Emerging evidence has predicted strong links between obesity and the human gut microbiota. However, only a few such studies have been conducted in Asia, and the gut microbiota of lean and obese adult Asians remains largely unexplored. Here, we investigated the potential relationship between gut microbiota, body massindex (BMI), and metabolic parameters in adults from Thailand, where obesity is increasing rapidly. Fecal and blood samples were collected from 42 volunteers who were allocated into lean, overweight, and obese groups. The fecal microbiota was examined by quantitative PCR analysis. Bacteroidetes, Firmicutes, and Staphylococcus spp. and methanogens were most abundant in lean volunteers. Overweight volunteers majorly harbored Christensenella minuta and Akkermansia muciniphila, ${\gamma}-Proteobacteria$, and bacteria belonging to the genus Ruminococcus. Methanogens and bacteria belonging to the phylum Bacteroidetes were negatively correlated with adiposity markers (BMI and waist circumference), but positive correlated with high-density lipoprotein, suggesting that they can be used as leanness markers. While some of our results agree with those of previous reports, results regarding the contributions of specific taxa to obesity were inconsistent. This is the first study to report the adult gut microbiota in Southeast Asian populations using molecular techniques and biochemical markers and provides a foundation for future studies in this field.

Food bio preservation is of major interest in the food industry. Many types of antimicrobial compounds can be produced by lactic acid bacteria (LAB), including bacteriocins. Bacteriocins increase the shelf-life of food by decreasing some food-borne diseases. In this study, a multi-coding sequence of bacteriocin genes was used for primer design to produce bacteriocin genes in Enterococcus faecium AH2 strain and Pediococcus pentosaceus AH1. Multi-coding sequences were aligned to detect conserved sequences in the bacteriocin gene. Eight genes encoding proteins involved in bacteriocin production were isolated and sequenced, including six from E. faecium AH2 (entA, entI, entF, entR, orfA2, orfA3) and two from P. pentoceseus AH1 (papA, pedB), and all gene sequences were deposited in the Gen Bank database under accession numbers LC064146-LC064151, LC101300, and LC101789, respectively. P. pentosaceus AH1 and E. faecium AH2 strains displayed bacteriocin activities of $2610AU\;mL^{-1}$ and $690AU\;mL^{-1}$ and inhibition zones of 26 mm and 19 mm, respectively. Overexpression of entA in E. faecium AH2 increased the bacteriocin and antimicrobial activities.

Whole-genome sequencing of the wood-rotting fungus, Flammulina fennae, was carried out to identify carbohydrate-active enzymes (CAZymes). De novo genome assembly (31 kmer) of short reads by next-generation sequencing revealed a total genome length of 32,423,623 base pairs (39% GC). A total of 11,591 gene models in the assembled genome sequence of F. fennae were predicted by ab initio gene prediction using the AUGUSTUS tool. In a genome-wide comparison, 6,715 orthologous groups shared at least one gene with F. fennae and 10,667 (92%) of 11,591 genes for F. fennae proteins had orthologs among the Dikarya. Additionally, F. fennae contained 23 species-specific genes, of which 16 were paralogous. CAZyme identification and annotation revealed 513 CAZymes, including 82 auxiliary activities, 220 glycoside hydrolases, 85 glycosyltransferases, 20 polysaccharide lyases, 57 carbohydrate esterases, and 45 carbohydrate binding-modules in the F. fennae genome. The genome information of F. fennae increases the understanding of this basidiomycete fungus. CAZyme gene information will be useful for detailed studies of lignocellulosic biomass degradation for biotechnological and industrial applications.