• Journal of Microbiology and Biotechnology
• /
• v.26 no.1
• /
• pp.20-27
• /
• 2016

Lactobacillus kefiranofaciens ZW3 was obtained from kefir grains, which have high lactose hydrolytic activity. In this study, a heterodimeric LacLM-type β-galactosidase gene (lacLM) from ZW3 was isolated, which was composed of two overlapping genes, lacL (1,884 bp) and lacM (960 bp) encoding large and small subunits with calculated molecular masses of 73,620 and 35,682 Da, respectively. LacLM, LacL, and LacM were expressed in Escherichia coli BL21(DE3) and these recombinant proteins were purified and characterized. The results showed that, compared with the recombinant holoenzyme, the recombinant large subunit exhibits obviously lower thermostability and hydrolytic activity. Moreover, the optimal temperature and pH of the holoenzyme and large subunit are 60℃ and 7.0, and 50℃ and 8.0, respectively. However, the recombinant small subunit alone has no activity. Interestingly, the activity and thermostability of the large subunit were greatly improved after mixing it with the recombinant small subunit. Therefore, the results suggest that the small subunit might play an important role in maintaining the stability of the structure of the catalytic center located in the large subunit.

• Land and Housing Review
• /
• v.3 no.3
• /
• pp.299-305
• /
• 2012

Various types of bio wood chip for fermentation-extinction of food waste was investigated by comparing their different pore structure with the performance of weight loss rate and microbial activity. The fermentation-extinction of food waste with bio wood chip was examined by adding 700~1,500g of food waste every day during 15 days to the fermentation-extinction reactor with condition of $30{\sim}50^{\circ}C$ temperature and 30~70% humidity, where 1,500g of bio wood chips were existed. The bio wood chips used in this experiment were categorized into 4 different types; microbial-mixing type(A biochip), macro pore type(B biochip) under $2{\mu}m$ of pore size, micro pore type of wood-chips(C biochip) under $0.1{\mu}m$ of pore size, viscous & sticky type(D biochip). As a result, A, B, C, D bio wood chip exhibited 85%, 63%, 92%, 73% weight loss of food waste with fermentation-extinction. The maximum weight loss of food waste was obtained at the fermentation-extinction experiments by using C bio wood chip. On the other hands, the maximum ratio of ATP to COD and TN was obtained from $3.00{\times}10^{-10}$ and $2.31{\times}10^{-11}$ in the case of C bio wood chip, comparing with other types of bio wood chip. Consequently, the performance of weight loss rate was affected with the micro pore structure of bio wood chip which have an advantage of extensive microbial activity space in the fermentation-extinction of food waste.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
• /
• v.17 no.3
• /
• pp.33-37
• /
• 2004

Objective: The aim of this work is to investigate the antibacterial activity of the Sohporaflavanone G isolated from Sophora flavescens (S. flavescens), as the development of microbial resistance to antibiotics make it essential to constantly look for new and active compounds effective against pathogenic bacteria. Method : Sophoraflavanone G was isolated from the dried roots of Sophora flavescens Aiton (Leguminosae) by bioassay?guided fractionation. We investigated the effect of sophoraflavanone G on oral bacterial at various concentrations after incubation of 24 h in strains in the dose?dependent manner. Results: The structure of active compound, Sophoraflavanone G having a lavandulyl group at C?8, was elucidated on the basis of spectral data especially 1H?NMR and I3C?NMR. The antimicrobial activity showed that Sophoraflavanone G exhibited antimicrobial activilies against all the bacteria tested (MICs, 0.39 - 6.25 ㎍／ml). Sophoraflavanone G showed the strong antimicrobial activity against all the facultative bacteria and microaerophilic bacteria (MICs, 0.78 - 1.56 ㎍／ml) and also Sophoraflavanone G showed the strong antimicrobial activity against obligate anaerobic bacteria (MICs, 0.39 - 6.25 ㎍／ml).

• Asian-Australasian Journal of Animal Sciences
• /
• v.12 no.1
• /
• pp.129-138
• /
• 1999

If rumen bacteria can be manipulated to utilize nutrients (i.e., ammonia and plant cell wall carbohydrates) more completely and efficiently, the need for protein supplementation can be reduced or eliminated and the digestion of fiber in forage or agricultural residue-based diets could be enhanced. However, these approaches require a complete and accurate description of the rumen community, as well as methods for the rapid and accurate detection of microbial density, diversity, phylogeny, and gene expression. Molecular ecology techniques based on small subunit (SSU) rRNA sequences, nucleic acid probes and the polymerase chain reaction (PCR) can potentially provide a complete description of the microbial ecology of the rumen of ruminant animals. The development of these molecular tools will result in greater insights into community structure and activity of gut microbial ecosystems in relation to functional interactions between different bacteria, spatial and temporal relationships between different microorganisms and between microorganisms and reed panicles. Molecular approaches based on SSU rRNA serve to evaluate the presence of specific sequences in the community and provide a link between knowledge obtained from pure cultures and the microbial populations they represent in the rumen. The successful development and application of these methods promises to provide opportunities to link distribution and identity of gastrointestinal microbes in their natural environment with their genetic potential and in situ activities. The use of approaches for assessing pupulation dynamics as well as for assessing community functionality will result in an increased understanding and a complete description of the gastrointestinal communities of production animals fed under different dietary regimes, and lead to new strategies for improving animal growth.

• Archives of Pharmacal Research
• /
• v.28 no.9
• /
• pp.1031-1036
• /
• 2005

[ $(+)-Lyoniresinol-3{\alpha}-O-\beta-D-glucopyranoside$ ] (1) was isolated from an ethyl acetate extract of the root bark from Lycium chinense Miller, and its structure was determined using 1D and 2D NMR spectroscopy including DEPT, HMQC, and HMBC. $(+)-Lyoniresinol-3{\alpha}-O-\beta-D-glucopyranoside$ exhibited potent antimicrobial activity against antibiotic-resistant bacterial strains, methicillin-resistant Staphylococcus aureus (MRSA) isolated from patients, and human pathogenic fungi without having any hemolytic effect on human erythrocytes. In particular, compound 1 induced the accumulation of intracellular trehalose on C. albicans as stress response to the drug, and disrupted the dimorphic transition that forms pseudo-hyphae caused by the pathogenesis. This indicates that $(+)-Lyoniresinol-3{\alpha}-O-\beta-D-glucopyranoside$ has excellent potential as a lead compound for the development of antibiotic agents.

• Journal of Microbiology and Biotechnology
• /
• v.20 no.1
• /
• pp.21-29
• /
• 2010

The phenolic compounds are a major contaminant class often found in industrial wastewaters and the biological treatment is an alternative tool commonly employed for their removal. In this sense, monitoring microbial community dynamics is crucial for a successful wastewater treatment. This work aimed to monitor the structure and activity of the bacterial community during the operation of a laboratory-scale continuous submerged membrane bioreactor (SMBR), using PCR and RT-PCR followed by denaturing gradient gel electrophoresis (DGGE) and 16S rRNA libraries. Multivariate analyses carried out using DGGE profiles showed significant changes in the total and metabolically active dominant community members during the 4-week treatment period, explained mainly by phenol and ammonium input. Gene libraries were assembled using 16S rDNA and 16S rRNA PCR products from the fourth week of treatment. Sequencing and phylogenetic analyses of clones from the 16S rDNA library revealed a high diversity of taxa for the total bacterial community, with predominance of Thauera genus (ca. 50%). On the other hand, a lower diversity was found for metabolically active bacteria, which were mostly represented by members of Betaproteobacteria (Thauera and Comamonas), suggesting that these groups have a relevant role in the phenol degradation during the final phase of the SMBR operation.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.41 no.3
• /
• pp.310-319
• /
• 2012

In order to investigate the biological effects of conformational changes in the folding state of bovine ${\alpha}$-lactalbumin (${\alpha}$-La), the protein was prepared and classified as apo form, microbial transglutaminase (MTGase) added form, or bovine ${\alpha}$-La made lethal to tumor cell (BAMLET) form. Apo ${\alpha}$-La form showed weaker cancer cell inhibitory activity (apoptosis) than native ${\alpha}$-La, which suggests that the metal ion-like $Ca^{2+}$ had a positive effect, whereas BAMLET form showed strong cancer cell apoptotic activity. The BAMLET form seemed to be a molten globule structure that increased hydrophobicity. MTGase added to apo ${\alpha}$-La polymer showed similar anti-cancer activity as native ${\alpha}$-La, and it was well hydrolyzed by digestive enzymes. NMR results showed that BAMLET interacted with oleic acid and produced a complex.

• Horticultural Science & Technology
• /
• v.29 no.5
• /
• pp.467-473
• /
• 2011

Flavonoid are large group of the polyphenolic compounds which are distinguished by an aromatic or phenolic ring structure and the phenolic compounds are induced by microbial infection, ultraviolet radiation, temperature and chemical stress. They are known for their antioxidant activity, anti-allergic, anti-inflammatory, anti-microbial and anti-cancer activities. In this study, changes in flavonoid content were investigated using heterologous chalcone isomerase (CHI) expression system. Also, phenolic compounds level was measured to examine the relation between flavonoids and phenols contents. Explants of lettuce (Lactuca sativa L.) were transformed with Agrobacterium tumefaciens LBA 4404 strain containing pFLH-CHI (derived from pPZP2Ha3) vector constructed with CHI gene from Brassica rapa. The putative transgenic plants were confirmed by genomic DNA PCR analysis. Also the transcription levels of the gene were analyzed by semi-quantitative RT-PCR with gene specific primers. The total flavonoid contents were increased at $T_0$ and $T_1$ generations over 1.4 and 4.0 fold, respectively. Total phenol contents also increased at $T_1$ generation. These results indicate that CHI gene plays an important role to regulate the accumulation of flavonoids and its component changes.

• 한국생물공학회:학술대회논문집
• /
• 2002.04a
• /
• pp.457-460
• /
• 2002

In an effort to isolate novel strains expressing a thermostable esterase that hydrolyzed the rac-ketoprofen ethyl ester to ketoprofen in the stereospecific manner, we screened various soils and composts from broad ecological niches in which the activity was expected to be found. Three hundreds of microbial strains were tested to determine their ester-hydrolyzing activity by using an agar plate containing insoluble tributyrin as an indicative substrate, and then further screened by activity on the (R,S)-ketoprofen ethyl ester. Twenty-six strains were screened primarily at high growth and incubation temperature and further compared the ability to ethyl ester-hydrolyzing activity in terms of conversion yield and chiral specificity. Consequently, a strain JYl44 was isolated as a novel strain that produced a (R)-stereospecific esterase with high stability and systematically identified as a Bacillus stearothermophilus JY144. The enzyme indeed stables at a broad range of temperature, upto 65 $^{\circ}C$, and pH ranging from 6.0 to 10.0. The optimal temperature and pH for enzymatic conversion were 50 $^{\circ}C$ and 9.0, respectively. Based on the observations that resulted a poor cell growth, and enzyme expression in wild type strain, we further attempted the gene cloning into a general host Escherichia coli and determined its primary structure, concomitantly resulting a high level expression of the enzyme. The cloned gene had an open reading frame (250 amino acids) with a calculated molecular mass of 27.4 kDa, and its primary structure showed a relative high homology (45-52 %) to the esterases from Streptomyces and Bacillus strains. The recombinant whole cell enzyme could efficiently convert the rac-ketoprofen ethyl ester to (R)-ketoprofen, with optical purity of 99 % and yield of 49 %.

• Journal of Ecology and Environment
• /
• v.33 no.3
• /
• pp.261-270
• /
• 2010

Increasing atmospheric $CO_2$ affects the soil carbon cycle by influencing microbial activity and the carbon pool. In this study, the effects of elevated $CO_2$ on extracellular enzyme activities (EEA; ${\beta}$-glucosidase, N-acetylglucosaminidase, aminopeptidase) in salt marsh sediment vegetated with Suaeda japonica were assessed under ambient atmospheric $CO_2$ concentration (380 ppm) or elevated $CO_2$ concentration (760 ppm) conditions. Additionally, the community structure of sulfate-reducing bacteria (SRB) was analyzed via terminal restriction fragments length polymorphism (T-RFLP). Sediment with S. japonica samples were collected from the Hwangsando intertidal flat in May 2005, and placed in small pots (diameter 6 cm, height 10 cm). The pots were incubated for 60 days in a growth chamber under two different $CO_2$ concentration conditions. Sediment samples for all measurements were subdivided into two parts: surface (0-2 cm) and rhizome (4-6 cm) soils. No significant differences were detected in EEA with different $CO_2$ treatments in the surface and rhizome soils. However, the ratio of ${\beta}$-glucosidase activity to N-acetylglucosaminidase activity in rhizome soil was significantly lower (P < 0.01) at 760 ppm $CO_2$ than at 380 ppm $CO_2$, thereby suggesting that the contribution of fungi to the decomposition of soil organic matter might in some cases prove larger than that of bacteria. Community structures of SRB were separated according to different $CO_2$ treatments, suggesting that elevated $CO_2$ may affect the carbon and sulfur cycle in salt marshes.