Arsenic (As), which is ubiquitous throughout the environment, represents a major environmental threat at higher concentration and poses a global public health concern in certain geographic areas. Most of the conventional arsenic remediation techniques that are currently in use have certain limitations. This situation necessitates a potential remediation strategy, and in this regard bioremediation technology is increasingly important. Being the oldest representativse of life on Earth, microbes have developed various strategies to cope with hostile environments containing different toxic metals or metalloids including As. Such conditions prompted the evolution of numerous genetic systems that have enabled many microbes to utilize this metalloid in their metabolic activities. Therefore, within a certain scope bacterial isolates could be helpful for sustainable management of As-contamination. Research interest in microbial As(III) oxidation has increased recently, as oxidation of As(III) to less hazardous As(V) is viewed as a strategy to ameliorate its adverse impact. In this review, the novelty of As(III) oxidation is highlighted and the implication of As(III)-oxidizing microbes in environmental management and their prospects are also discussed. Moreover, future exploitation of As(III)-oxidizing bacteria, as potential plant growth-promoting bacteria, may add agronomic importance to their widespread utilization in managing soil quality and yield output of major field crops, in addition to reducing As accumulation and toxicity in crops.

Dairy products are extensively used as carriers of probiotic strains that have potential health benefits. Assessment of the viability of probiotic strains during manufacturing is important to ensure that products meet recommended levels. Hence, the method for accurately quantifying lactic acid bacteria (LAB) in probiotic or dairy products is required. The present study aims to examine the performance of de-Man Rogosa Sharpe (MRS), plate count agar with bromocresol purple (PCA with BCP), and glucose blood liver (BL) agars recommended in the Korea Food Code guidelines for counting LAB. Analysis of the performance of culture media containing 19 lactic acid bacterial species commonly encountered in probiotic and dairy products showed no statistically significant difference between 18 reference strains and three culture media (p > 0.01). Furthermore, the suitability of three culture media was verified for the quantitative assessment of LAB in 25 probiotic and dairy products. The number of LAB in three culture media was determined to be more than 10⁷ colony-forming unit (CFU)/ml for fermented milk products and 10⁸ CFU/ml for condensed fermented milk and probiotic products, indicating that they all satisfied the Korea Food Code guidelines. Moreover, there was no statistically significant difference in the amount of LAB counted in all three culture media, suggesting that they can be used to isolate or enumerate LAB in commercial products. Finally, three culture media will be useful for isolating and enumerating LAB from fermented foods as well as gut microflora.

After random mutagenesis, the mutant Lactobacillus plantarum GNS300 showed improved exopolysaccharide production as determined by the quantification of total sugar. The mutant L. plantarum GNS300 produced 2.82 g/l of exopolysaccharide which showed 79.62% improved exopolysaccharide production compared with the parental strain. When exopolysaccharide of L. plantarum GNS300 was analyzed, the exopolysaccharide is composed of galactose (93.35%) and glucose (6.65%). Through the optimization of fermentation conditions using a bioreactor, 2.93 g/l of exopolysaccharide was produced from 20 g/l of glucose at 35℃, 500 rpm, and 0.1 vvm for 12 h. The mutant L. plantarum GNS300 exhibited 69.18% higher antioxidant activity than that from the parental strain, which might be caused by higher exopolysaccharide production. The concentrated supernatant of the mutant L. plantarum GNS300 inhibited the growth of gram-positive bacteria (Bacillus cereus and Staphylococcus aureus) and gram-negative bacteria (Escherichia coli, Vibrio parahaemolyticus, and Salmonella typhimurium).

A map gene encoding methionyl-specific aminopeptidase (MAP; EC 3.4.11.18) was cloned from Tetragenococcus halophilus CY54. Translated amino acid sequence of CY54 MAP showed high similarities with those from Enterococcus faecalis (83.8%) and Streptococcus salivarius (62.2%) but low similarities with MAPs from Lactobacillus and Lactococcus genera. The map gene was overexpressed in E. coli BL21(DE3) using pET26b(+),pET26b(+), and the recombinant MAP was purified by using an Ni-NTA column. The size of recombinant MAP was 29 kDa as determined by SDS-PAGE. The optimum pH and temperature of CY54 MAP were pH 5.0 and 60℃, respectively. The activity of CY54 MAP was most significantly increased by Co²⁺ ion (159%), and showed the highest activity at 12% NaCl. K_m and V_max were 0.64 ± 0.006 mM and 10.12 ± 0.014 U/mg protein, respectively when met-pNA was used as the substrate. This is the first report on a MAP from Tetragenococcus species.

In this study, the effect of sodium hypochlorite on biofilm removal was evaluated using three bacterial strains; Aeromonas hydrophila, Streptococcus mutans, and Yersinia enterocolitica. For maximum biofilm removal in 10 min, sodium hypochlorite is required at 1.65, 0.83, and 0.41 g/l for A. hydrophila, S. mutans, and Y. enterocolitica, respectively. Resistance to sodium hypochlorite was increased by the biofilms of all three tested strains, while the change in bactericidal activity according to sodium hypochlorite concentration was strain-specific. Therefore, we aimed to determine the effective concentration of sodium hypochlorite required for hygiene, considering that higher concentrations are needed to remove biofilms than to kill cells.

Neutral and non-essential amino acid, glutamine (Gln), plays an essential role in supplying nitrogen to all the amino acids and nucleotides in the mammalian body. Gln is also the most important carbon source that provides intermediates for gluconeogenesis and fatty acid synthesis and supplements the tricarboxylic acid cycle in fast-growing cancer cells. Among the known 14 Gln transporter genes, soluted carrier family 1 member 5 (SLC1A5) has been reported to be closely associated with cancer cell growth. Three variants (v1, v2, and v3) have been derived from SLC1A5. Here, we established a heterologous gene expression system for the active form of human SLC1A5 variant-2 (hSLC1A5v2) in Escherichia coli. v2 is the smallest variant that has not yet been studied. Four expression systems were investigated: pBAD, pCold, pET, and pQE. We also addressed the problem of codon usage bias. Although pCold and pET overexpressed hSLC1A5v2 in E. coli, they were functionally inactive. hSLC1A5v2 using the pBAD system was able to catalyze the successful transport of Gln, even if it was not highly expressed. Initial activity of hSLC1A5v2 for [¹⁴C] Gln uptake in E. coli reached up to 6.73 μmole·min^-1·gDW^-1 when the cell was induced with 80 mM L-arabinose. In this study, we demonstrated a heterologous expression system for the human membrane protein, SLC1A5, in E. coli. Our results can be used for the functional comparison of SLC1A5 variants (v1, v2, and v3) in future studies, to facilitae the developement of SLC1A5 inhibitors as effective anticancer drugs.

The purpose of this study is to confirm the antioxidant, black hair, and hair growth effects of the N. jatamansi, O. basilicum, and C. sativus mixed extracts that pharmacological efficacy has been verified. Accordingly, four samples (NOC-1 to-4) produced under different extraction conditions were prepared and the results are as follows. First, all samples showed an increase in antioxidant content in a concentration-dependent manner from the results of antioxidant efficacy. In particular, NOC-4 extracted by steaming and ultrasonic methods showed the highest antioxidant effect among the four samples. As a result of analysis of the amount of melanin production in mouse melanoma cells, NOC-4 with concentration of 500 ㎍/ml showed higher melanin production compared to the control group, so the black hair efficacy was the best. Also, in the hair growth test results, it was found that the hair growth was the best at 0.94 ± 0.10 mm at experimental group orally administered with 500 mg/kg of NOC-4. In addition, as a result of cytotoxicity analysis in mouse melanoma cells, the safety of samples was demonstrated by maintaining cell viability of 95% or more at all concentrations. These results suggest that the steaming and ultrasonic extraction method increased the extraction yield of active ingredients for antioxidant, melanin, and hair generation, thereby affecting physiological activity. Based on these results, if the steaming and ultrasonic extraction methods are applied to the mixed extraction of N. jatamansi, O. basilicum, and C. sativus, it is judged that the practical potential as a natural material for black hair and hair growth agents will increase.

Lactococcus lactis HY7803 균주를 대두 발효에 적용하여 맛 성분을 향상시키는 조미료 제조에 대한 이용 가능성을 확인하고자 대두 발효를 진행하였다. 대두발효물의 이화학적 분석 결과에 따르면, L. lactis HY7803 균주를 접종한 대두 발효물은 발효가 진행되면서 아미노태질소 함량, 아미노산 함량이 증가한 반면, 생균수가 감소하였다. 또한 맛성분에 기여하는 아미노산인 글루탐산의 함량을 보면, 발효가 진행됨에 따라 114.99 ± 9.37 pmol/ul에서 138.14 ± 1.76 pmol/ul로 함량이 증가하였고, 글루탐산을 포함한 12개의 아미노산 함량이 음성대조군인 균을 첨가하지 않은 대두발효물에 비해 높았고, Aspergillus oryzae SNU-G를 첨가한 대두발효물과 유사한 함량으로 측정되었다. 따라서 본 연구의 결과는 L. lactis HY7803가 A. oryzae SNU-G의 대체종균으로써 이용 가능하며, 대두 발효를 통해 감칠맛 성분을 생산하는데 기여하는 종균으로써 효과적일 것으로 판단하였다.

Xylanase is an industrially relevant enzyme used for the production of xylobiose and xylose. Various methods are used to enhance the microbial yield of xylanase. In the present study, co-culturing of Bacillus subtilis and Bacillus pumilus were investigated using submerged fermentation for xylanase production, which was markedly increased when sal, sagwan, newspaper, wheat bran, and xylan were used as single carbon sources. Maximum xylanase production was reported after 5 days of incubation in optimized media at pH 7.0 and 37℃, resulting in 2.69 ± 0.25 µmol/min by coculture. The 1:1 ratio of sal and sagwan in optimized production media was shown to be suitable for xylanase synthesis in submerged fermentation (SMF). In comparison to mono-culture using B. pumilus and B. subtilis, co-culturing resulted in an overall 3.8-fold and 2.15-fold increase in xylanase production, respectively.

Nitrilases are a hydrolase group of enzymes that catalyzes nitrile compounds and produce industrially important organic acids. The current objective is to optimize nitrilase production using statistical methods assisted with artificial intelligence (AI) tool from novel nitrile degrading isolate. A nitrile hydrolyzing bacteria Bacillus subtilis AGAB-2 (GenBank Ascension number- MW857547) was isolated from industrial effluent waste through an enrichment culture technique. The culture conditions were optimized by creating an orthogonal design with 7 variables to investigate the effect of the significant factors on nitrilase activity. On the basis of obtained data, an AI-driven support vector machine was used for the fitted regression, which yielded new sets of predicted responses with zero mean error and reduced root mean square error. The results of the above global optimization were regarded as the theoretical optimal function conditions. Nitrilase activity of 9832 ± 15.3 U/ml was obtained under optimized conditions, which is a 5.3-fold increase in compared to unoptimized (1822 ± 18.42 U/ml). The statistical optimization method involving Plackett Burman Design and Response surface methodology in combination with an AI tool created a better response prediction model with a significant improvement in enzyme production.

본 연구에서는 한국형 프로바이오틱스와 볶은 쓴메밀을 배양하여 쓴메밀의 대표 성분인 rutin에서 quercetin으로 생물전환을 확인하였다. 프로바이오틱스 17종의 유전체 분석 결과 CBT BG7, LC5, LR5, LP3, LA1과 LGA1 등 6종에서 rutin에서 isoquercetin 전환에 관련된 α-rhamnosidase 유전자가 확인되었다. Isoquercetin에서 quercetin 전환에 관련된 β-glucosidase 유전자는 17종의 프로바이오틱스에서 모두 확인되었다. 17종의 프로바이오틱스 중 CBT BG7, LR5, LP3, LA1, LGA1과 ST3 등 6종의 균주가 배양 7일 후 rutin에서 quercetin으로 최대 21.5 ± 0.3%까지 생물전환 하였다. 생물전환 시간 단축을 위하여 프로바이오틱스와 복합 효소 Cellulase KN^®을 함께 배양하였다. 그 결과, 효소 첨가 없이 가장 높은 생물전환율 21.5 ± 0.3%를 보였던 CBT LA1 균주는 효소와 함께 배양 1일 후 84.6 ± 0.5%의 생물전환율을 보였다. 특히 CBT ST3 (96.2 ± 0.4%), SL6 (90.1 ± 1.4%) 그리고 LP3 (90.0 ± 0.4%) 균주는 quercetin으로 90% 이상의 높은 전환율을 보였다. 추가로 프로바이오틱스 생물전환능이 우수하여 quercetin 함량이 높은 배양여액 동결건조물은 RAW264.7 세포에서 LPS에 의해 유도된 NO 생성 억제 효과를 보였다. 본 연구를 통해 한국형 프로바이오틱스와 효소를 조합하여 볶은 쓴메밀과 함께 배양하였을 때 rutin으로부터 항염증, 항산화, 항비만, 항당뇨 활성을 가지는 생리활성 물질 quercetin으로 생물전환이 증가하고 전환 시간이 단축됨을 확인하였다.

우리나라 4개 지역에서 상업적으로 제조한 8개 누룩을 구입하여 Bacillus 속, Staphylococcus 속 및 유산균을 선택적으로 검출할 수 있는 배지를 이용하여 세균을 분리하고, 최근 확립된 분류기준에 따라 동정하여 세균의 그룹 별 다양성 및 우점종을 검토하였다. 모든 시료로부터 Bacillus 속이 검출되었지만, 3개 시료에서는 Staphylococcus 속 또는 유산균이 검출되지 않았다. 유산균이 가장 많이 검출된 1개 시료를 제외한 7개 시료에서 Bacillus 속이 가장 많이 계수되었고, Staphylococcus 속은 가장 적은 수로 검출되었다. Bacillus 속 6개 종의 존재가 확인되었고, B. subtilis, B. velezensis, B. licheniformis가 우점으로 검출되었다. Staphylococcus 속 9개 종의 존재가 확인되었고, coagulase 음성 S. pseudoxylosus와 S. saprophyticus의 우점을 확인했다. 유산균은 Enterococcus 속, Lactobacillus 및 근연속, Pediococcus 속, Weissella 속 10종의 존재가 확인되었고, P. pentosaceus가 우점종으로 확인되었다.

대기 입자상물질(particulate matter, PM) 시료의 곰팡이 메타게놈 분석을 위해 DNA 추출 및 유전자 증폭 시 여러 문제가 발생한다. 본 연구에서는 PM 시료로부터 DNA를 추출하는 방법과 polymerase chain reaction (PCR)을 위한 프라이머 및 온도 조건의 최적화를 위하여 다양한 조건으로 실험하였다. 여러 조건에서 DNA 추출 여부를 비교 평가한 결과, bufffer와 proteinase K를 이용하여 20분 동안 화학적 세포 용해 처리와 bead beating 처리를 한 후 상용 DNA 추출 kit를 사용하면 DNA를 효율적으로 추출할 수 있었다. PCR 조건을 최적화하기 위해 ITS2 유전자 영역을 증폭할 수 있는 10개 조합의 프라이머를 이용하여 PCR을 수행한 결과, ITS3tagmix3/ITS4 조합의 프라이머로 annealing 온도 58℃로 하였을 때 증폭된 PCR 산물의 농도가 상대적으로 높았다. 이 조건에서도 PCR 산물의 농도가 낮은 경우에는 1차 PCR 산물을 주형 DNA로 사용하여 nested PCR을 수행하면 만족스러운 농도로 ITS2 유전자를 증폭할 수 있었다. 본 연구에서 도출한 조건으로 서울 대기 PM2.5를 포집한 필터 시료 15종을 대상으로 DNA 추출과 PCR을 수행한 결과 성공적으로 ITS2 유전자 증폭이 가능하였다. 본 연구에서 최적화한 방법은 대기 PM 시료의 곰팡이 메타게놈을 분석하고 해석하는 연구에 활용 가능하다.

Preterm birth (PTB) is defined as giving birth prior to the 37th week of pregnancy and is a major cause of infant mortality. Studies have indicated that the vaginal microbiota's composition and its dysbiosis, particularly during pregnancy, may play a major role in PTB. While previous research work concentrated on well-studied microorganisms such as Lactobacillus, Prevotella, Gardnerella, various other microbes, and their significance in the vaginal microbiota's stability remain unknown. Moreover, current studies have focused primarily on the relative abundances of the microbes found, without considering their interactions with other members of the vaginal microbiota. In this work, we developed a novel computational approach and performed taxonomic classification of vaginal microbiota samples stratified longitudinally (Term/PTB) to observe compositional disparities and find underexamined microbes that may be contributing to PTB. Furthermore, we carried out a correlational analysis to build a microbial co-interaction network and investigated the functional implications of the genes present in both Term and PTB samples. The co-occurrence network revealed that Lactobacillus acts in solidarity to maintain the stability of the vaginal microbiota and did not have strong co-interactions with any of the other microbes. Similarly, microbes with strong interactions with Atopobium, a well-known marker microbe of PTB, were also observed. Additionally, several genes such as PTXA, FANCM, GPX, and DUSP were found to be playing an important role in the occurrence of PTB. This study provides a novel conceptual framework revealing distinct vaginal microbiota signatures that could be potential therapeutic targets for the prevention of PTB.

Among certain animals, gut microbiomes demonstrate species-specific patterns of beta diversity. This host-specificity is a potent driver of exogenous microbial exclusion. To overcome persistent translational limitations, translational microbiome research and therapeutic development must account for host-specific patterns of microbial engraftment. This commentary seeks to highlight the important implications of host-specificity for microbial ecology, Fecal Microbiota Transplantation (FMT), next-generation probiotics, and translational microbiota research.

Here we report the draft genome sequence of Weissella koreensis strain HJ and genomic analysis of its key features. The genome consists of 1,427,571 bp with a GC content of 35.5%, and comprises 1,376 coding genes. In silico analysis revealed the absence of pathogenic factors within the genome. The genome harbors several genes that play an important role in the survival of the gastrointestinal tract. In addition, a type III polyketide synthase cluster was identified. Pangenome analysis identified 68 unique genes in W. koreensis strain HJ. The genome information of this strain provides the basis for understanding its probiotic properties.

Clonal complex (CC) 398 community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has emerged worldwide in a variety of livestock animals and humans. We report complete genome sequences of Panton-Valentine leucocidin (PVL) and immune evasion cluster (IEC) gene-positive CC398 MRSA strains isolated from patients in Korea.