• KSBB Journal
• /
• 제31권1호
• /
• pp.58-65
• /
• 2016

In this study, Arthrobacter scleromae SYE-3, which was isolated from indigenous plant in a subtropical region, Neigeria, with plant growth promoting activity was evaluated to determine the optimal culture condition. A bacterial strain SYE-3 had the IAA productivity ($89.15{\pm}0.36mg/L$) and ACC deaminase activity ($0.20{\pm}0.06$ at 72 hours). Also, optimal culture conditions such as temperature and pH of strain SYE-3 were $20^{\circ}C$ and 10 in LB medium, respectively. Strain SYE-3 had up to 3% salt tolerance in the LB medium. Plant growth promoting ability of strain SYE-3 using yam (Dioscorea japonica Thunb.) was evaluated. As a result, strain SYE-3 had showed very powerful effect on the increase of the shoot length and root biomass of yam (190.0% and 282.41% increase for 112 days, respectively). These results indicated that Arthrobacter scleromae SYE-3 can serve as a promising microbial resource for the biofertilizers of subtropical crops.

• 생명과학회지
• /
• 제31권2호
• /
• pp.175-182
• /
• 2021

기능성 화장품 소재로서 활용할 수 있는 효모의 분리를 위하여 순창군 베리류 및 과수원 토양에서 분리주 80종을 1차로 선별하였다. 80종의 분리주를 대상으로 항산화 활성 및 tyrosinase 저해 활성을 측정한 결과 DPPH 라디칼 소거능은 51.41%, SOD 활성은 62.23%, tyrosinase 저해 활성 64.75%로 가장 우수한 FT4-4를 최종적으로 선별하였다. 18S rRNA 염기서열 분석을 통해 Saccharomyces cerevisiae FT4-4로 명명하였으며, API ZYM을 이용하여 세포 외 효소 활성을 추가로 측정하였다. 발효 시간에 따른 균체 성장 및 tyrosinase 저해 활성의 변화를 측정한 결과 배양 후 16시간에 최대 균체량인 3.16 g/l와 67.68%의 tyrosinase 저해활성을 나타내었다. 또한, S. cerevisiae FT4-4의 화장품 소재로 활용하기 위한 세포 독성과 melanoma B16F10 세포 멜라닌 억제능을 확인한 결과, 세포독성은 50 mg/ml 이하의 농도에서 100% 이상의 세포 생존율을 보였으며, 시료 10 mg/ml에서의 멜라닌 생합성 저해능은 72.02%로 측정되었다. 향후 FT4-4의 화장품 소재로 활용하기 위해서는 생산 수율을 증가하기 위한 생산조건 확립 이외에도 안전성을 향상시키기 위한 추가적인 독성연구 등 다양한 연구가 수반되어야 하나, 본 연구에서의 항산화 및 미백 효능만으로도 충분히 활용할 가치가 있는 소재로 사료된다.

• 한국식품과학회지
• /
• 제54권1호
• /
• pp.103-108
• /
• 2022

본 연구에서는 조미용 저염된장을 개발하기 위해 정미성과 안전성이 우수한 고초균과 황국균을 이용하여 쌀 및 콩 발효액을 제조하고 숙성시키면서 이에 따른 장류 적용 특성을 분석하고 이를 조미용 저염된장에 적용하고자 하였다. 선별된 균주 3종을 이용하여 쌀과 콩의 발효물을 제조하였고 숙성기간에 따른 품질특성 변화를 확인하였다. 그 결과 쌀, 콩 발효물 모두 숙성기간에 따라 pH는 감소하고 적정산도, AN, 총당 및 환원당은 증가하는 경향을 보였으며 염도와 수분은 유의적인 차이를 보이지 않았다. 이 결과 콩, 쌀발효물의 발효 균주 및 숙성기간은 AN, 총당, 환원당 증가를 보인 균주 및 기간으로 선정하였다(콩: SRCM102487 및 3주, 쌀: SRCM104466 및 6일). 숙성로 선정하였다. 최종 선발된 균주와 대조구의 균주를 이용하여 각 발효물을 제조, 혼합한 후 조미저염된장을 제조한 후 특성을 비교하였다. 된장의 이화학적 특성인 pH와 적정산도, 염도는 유의적인 차이가 없으며, 아미노산성 질소 함량은 선발된 균주를 이용한 된장이 더 높은 함량을 보여주었다. 선발된 균주를 이용한 된장의 유기산은 citric acid와 lactic acid가 주요 유기산으로, 유리당은 glucose가 주요 유리당으로 나타났다. 된장의 유리아미노산 함량은 선발된 균주를 이용한 된장이 지미, 고미, 감미를 나타내는 아미노산 성분의 함량이 더 높은 결과를 보여주었다. 핵산관련물질은 hypoxanthine, inosine, IMP, AMP 순으로 높게 나타났으며, 선발된 균주를 이용한 된장이 IMP를 제외하고 나머지 3가지 물질 중에서 유의적으로 높은 함량을 나타내었다. 이와 같이 쌀과 콩을 선발된 균주로 발효하여 이를 된장에 적용하였을 때 다양한 아미노산과 핵산물질로 인해 된장의 풍미를 증진시키고 산업적으로 적용가능한 조미저염된장 제조가 가능할 것으로 판단되었다.

• 미생물학회지
• /
• 제50권3호
• /
• pp.233-238
• /
• 2014

전통주 양조에 적합한 종균 선발의 일환으로 에탄올 저항성 및 산 생성능이 낮은 한 유산균주를 전통 메주에서 분리하였고, 생화학적 동정 및 16S rRNA 유전자 염기 서열의 분석 결과 유산균인 Lactobacillus brevis로 동정되었다. 8% (v/v) 에탄올을 농도 별로 첨가한 MRS 배지에 이 균주를 $30^{\circ}C$. 96시간 배양한 결과 잘 생육하였으며, 배양 후 최종 pH는 4.5까지 감소하였다. 이 균주는 또한 식품 부패 및 병원성 균들인 Escherichia coli, Bacillus cereus, Listeria monocytogenes, Stapylococcus aureus, Micrococcus luteus, Pseudomonas aeruginosa에 대해 증식 억제능을 나타냈다. 이 결과들로부터 L. brevis JBE 30은 전통주 양조에 적합한 종균으로 사용 될 수 있음을 보였다.

• Journal of Microbiology and Biotechnology
• /
• 제19권11호
• /
• pp.1456-1463
• /
• 2009

Vibrio parahaemolyticus possessing urease-positive property is relatively rare, but such strains consistently exhibit the TDH-related hemolysin (TRH) gene. In this study, we examined the effects of incubation temperature on urease activity expression, using the TH3996 and AQ4673 strains where the enzyme activity is known to be temperature-dependent and -independent, respectively. In the TH3996 strain, $\beta$-galactosidase activity was 4.4-fold lower after $30^{\circ}C$ cultivation than after $37^{\circ}C$ in a ureR-lacZ fusion strain, but temperature dependency was not found in ureD- or nikA-lacZ fusion strains. However, ureR-, ureD-, and nikA-lacZ fusions of the AQ4673 strain was not influenced by incubation temperature. We compared the promoter sequences of ureR between the above two strains. Intriguingly, we detected mismatches of two nucleotides between the two strains located at positions -66 and -108 upstream of the methionine initiation codon for UreR. Additionally, urease activity was not affected by culture temperature at either $30^{\circ}C$ or $37^{\circ}C$ by allelic introduction of the AQ4673 ureR gene into the TH3996 ureR deletion mutant. Taken together, our study demonstrates that the transcriptional factor UreR is involved in the temperature dependency of urease activity, and two nucleotides within the ureR promoter region are of particular importance for the urease activity dependency of V. parahaemolyticus.

• Journal of Microbiology
• /
• 제43권4호
• /
• pp.325-330
• /
• 2005

Rhodococcus sp. strain YU6 was isolated from soil for the ability to grow on o-xylene as the sole carbon and energy source. Unlike most other o-xylene-degrading bacteria, YU6 is able to grow on p-xylene. Numerous growth substrate range experiments, in addition to the ring-cleavage enzyme assay data, suggest that YU6 initially metabolizes 0- and p-xylene by direct aromatic ring oxidation. This leads to the formation of dimethylcatechols, which was further degraded largely through meta-cleavage path-way. The gene encoding meta-cleavage dioxygenase enzyme was PCR cloned from genomic YU6 DNA using previously known gene sequence data from the o-xylene-degrading Rhodococcus sp. strain DK17. Subsequent sequencing of the 918-bp PCR product revealed a $98\%$ identity to the gene, encoding meth-ylcatechol 2,3-dioxygenase from DK17. PFGE analysis followed by Southern hybridization with the catechol 2,3-dioxygenase gene demonstrated that the gene is located on an approximately 560-kb megaplasmid, designated pJY J1

• Mycobiology
• /
• 제40권2호
• /
• pp.138-141
• /
• 2012

We measured physiological functionalities, including antihypertensive angiotensin I-converting enzyme inhibitory activity and immun-stimulating ${\beta}$-glucan content for sixty kinds of Makgeolli that is commercially available from the market. As a result, we selected R-12 commercial raw Makgeolli, with a high content of immuno-stimulating ${\beta}$-glucan, and R-14 commercial raw Makgeolli, exhibiting high antihypertensive activity. Due to the similarities in their overall physicochemical properties and raw materials used for fermentation, we compared the microbial flora in order to investigate the reason for the differences in their functionalities. Nested PCR and denaturing gradient gel electrophoresis for yeasts and bacteria were performed for analysis of microbial diversity of two different kinds of Makgeolli (i.e., R-12, R-14), which showed immuno-stimulating ${\beta}$-glucan content and exhibited a very high level of antihypertensive activity, respectively. Analysis of the 18S rDNA amplicon revealed a major presence of the yeast strain Pichia burtonii in every Makgeolli sample. Analysis of the 16S rDNA amplicon revealed a predominance of lactic acid bacteria, and the most frequent lactic acid bacteria were Lactobacillus ingluviei, L. fermentum, and L. harbinensis, and Lactobacillus sp. Among these, L. harbinensis was detected only in R-12 and L. ingluviei was found only in R-14. Different functionalities from the individual commercially available Makgeolli may be attributed to actions of different microbial flora during fermentation.

• Journal of Microbiology and Biotechnology
• /
• 제24권2호
• /
• pp.197-208
• /
• 2014

A novel protease gene from Bacillus gibsonii, aprBG, was cloned, expressed in B. subtilis, and characterized. High-level expression of aprBG was achieved in the recombinant strain when a junction was present between the promoter and the target gene. The purified recombinant enzyme exhibited similar N-terminal sequences and catalytic properties to the native enzyme, including high affinity and hydrolytic efficiency toward various substrates and a superior performance when exposed to various metal ions, surfactants, oxidants, and commercial detergents. AprBG was remarkably stable in 50% organic solvents and retained 100% activity and stability in 0-4 M NaCl, which is better than the characteristics of previously reported proteases. AprBG was most closely related to the high-alkaline proteases of the subtilisin family with a 57-68% identity. The secretion and maturation mechanism of AprBG was dependent on the enzyme activity, as analyzed by site-directed mutagenesis. Thus, when taken together, the results revealed that the halo-solvent-tolerant protease AprBG displays significant activity and stability under various extreme conditions, indicating its potential for use in many biotechnology applications.

• Plant Biotechnology Reports
• /
• 제5권3호
• /
• pp.245-254
• /
• 2011

This study describes an efficient protocol for Agrobacterium tumefaciens-mediated transformation of two subgroups of genotype AAA bananas (Musa acuminata cv. Pei Chiao and Musa acuminata cv. Gros Michel). Instead of using suspension cells, cauliflower-like bud clumps, also known as multiple bud clumps (MBC), were induced from sucker buds on MS medium containing $N^6$-Benzylaminopurine (BA), Thidiazuron (TDZ), and Paclobutrazol (PP333). Bud slices were co-cultivated with A. tumefaciens C58C1 or EHA105 that carry a plasmid containing Arabidopsis root-type ferredoxin gene (Atfd3) and a plant ferredoxin-like protein (pflp) gene, respectively. These two strains showed differences in transformation efficiency. The EHA105 strain was more sensitive in Pei Chiao, 51.3% bud slices were pflp-transformed, and 12.6% slices were Atfd3-transformed. Gros Michel was susceptible to C58C1 and the transformation efficiency is 4.4% for pflp and 13.1% for Atfd3. Additionally, gene integration of the putative pflp was confirmed by Southern blot. Resulting from the pathogen inoculation assay, we found that the pflp transgenic banana exhibited resistance to Fusarium oxysporum f. sp. cubense tropical race 4. This protocol is highly advantageous to banana cultivars that have difficulties in setting up suspension cultures for the purpose of quality improvement through genetic transformation. In addition, this protocol would save at least 6 months in obtaining explants for transformation and reduce labor for weekly subculture in embryogenic cell suspension culture systems.

• The Plant Pathology Journal
• /
• 제32권2호
• /
• pp.136-144
• /
• 2016

Pseudomonas fluorescens pc78 is an effective biocontrol agent for soil-borne fungal diseases. We previously constructed a P43-gfp tagged biocontrol bacteria P. fluorescens pc78-48 to investigate bacterial traits in natural ecosystem and the environmental risk of genetically modified biocontrol bacteria in tomato rhizosphere. Fluctuation of culturable bacteria profile, microbial community structure, and potential horizontal gene transfer was investigated over time after the bacteria treatment to the tomato rhizosphere. Tagged gene transfer to other organisms such as tomato plants and bacteria cultured on various media was examined by polymerase chain reaction, using gene specific primers. Transfer of chromosomally integrated P43-gfp from pc78 to other organisms was not apparent. Population and colony types of culturable bacteria were not significantly affected by the introduction of P. fluorescens pc78 or pc78-48 into tomato rhizosphere. Additionally, terminal restriction fragment length polymorphism profiles were investigated to estimate the influence on the microbial community structure in tomato rhizosphere between non-treated and pc78-48-treated samples. Interestingly, rhizosphere soil treated with strain pc78-48 exhibited a significantly different bacterial community structure compared to that of non-treated rhizosphere soil. Our results suggest that biocontrol bacteria treatment influences microbial community in tomato rhizosphere, while the chromosomally modified biocontrol bacteria may not pose any specific environmental risk in terms of gene transfer.