• Journal of Applied Biological Chemistry
• /
• v.65 no.1
• /
• pp.33-41
• /
• 2022

Esculetin (also known as 6, 7-dihydroxycoumarin) a type of coumarin, has been exhibited anti-inflammatory and anti-aging effects. Biorenovation is the microbe-mediated enhancement of biological efficacies and structurally diversified compounds relative to their substrate compounds. The production of different kinds of esculetin derivatives using Bacillus sp. JD3-7 and their effects on lipopolysaccharide (LPS)-triggered inflammatory response in RAW 26.7 cells were assessed. One of the biorenovation products, identified as esculetin 6-O-phosphate (ESP), at concentrations of 1.25, 2.5, and 5 μM inhibited the LPS-stimulated production of inflammation markers of nitric oxide synthase 2 and cyclooxygenase 2 as well as their respective enzymatic reaction products of nitric oxide and prostaglandin E2 in the order of increasing concentrations (1.25, 2.5, and 5 μM). Additionally, ESP treatment suppressed the LPS-stimulated secretion of pro-inflammatory cytokines of interleukin (IL)-1β, IL-6, and tumor necrosis factor- α. Furthermore, these anti-inflammatory effect of ESP was associated with the downregulation of mitogen-activated protein kinase signaling, that is, extracellular signal-regulated kinase, c-Jun NH2-terminal kinase, and p38 mitogen-activated protein kinase signaling pathways. This study would therefore provide interesting insights into the biorenovation-assisted generation of a novel anti-inflammatory compound. ESP may be used to develop treatments for inflammatory disorders.

• Research in Plant Disease
• /
• v.17 no.1
• /
• pp.38-43
• /
• 2011

Almost 10~30% of vegetables were discarded by the spoilage from farms to tables. After harvest, vegetables are often spoiled by a wide variety of microorganisms including many bacterial and fungal species. This investigation was conducted to extent the knowledge of relationship the spoilage of vegetables and the diversity of microbes. The total aerobic bacterial numbers in fresh lettuce, perilla leaf, and chicory were $2.6{\sim}2.7{\times}10^6$, $4.6{\times}10^5$, $1.2{\times}10^6\;CFU/g$ of fresh weight, respectively. The most common bacterial species were Pseudomonas spp., Alysiella spp., and Burkholderia spp., and other 18 more genera were involved in. After one week of incubation of those vegetables at $28^{\circ}C$, the microbial diversity had been changed. The total aerobic bacterial numbers increased to $1.1{\sim}4.6{\times}10^8$, $4.9{\times}10^7$, and $7.6{\times}10^8\;CFU/g$ of fresh weight for lettuce, perilla leaf, and chicory that is about $10^2$ times increased bacterial numbers than that before spoilage. However, the diversity of microbes isolated had been simplified and fewer bacterial species had been isolated. The most bacterial population (~48%) was taken up by Pseudomonas spp., and followed by Arthrobacter spp. and Bacillus spp. The spoilage activity of individual bacterial isolates had been tested using axenic lettuce plants. Among tested isolates, Pseudomonas fluorescence and Pantoea agglomerans caused severe spoilage on lettuce.

• Applied Biological Chemistry
• /
• v.15 no.2
• /
• pp.111-142
• /
• 1972

An experimental Chung-Kook-Jang was prepared using the strain Bacillus subtilis sp. isolated by the author Samples were taken in 12 hrs interval during the fermentation and the oligopeptides were separated by the method of molecular sieving using the ion exchange resin column of Dowex-50. Only the X-16 fraction of oligopeptides was taken and the components of oligopeptides were developed in two dimensional thin layer chromatograms. The each peptide spot was eluted and each peptide was isolated. The pattern and kinds of amino acids, and N and C-terminal amino acids were studied. Fourteen different oligopeptides could be detected by the two dimensional thin layer chromatography, all of which were consisted of $4{\sim}9$ kinds of amino acids. No dipeptides and no tripeptides could be found. The N and C-terminal amino acids and the residual component amino acids of all these 14 peptides could be summarized as the follows. [P]-I. Pro (Cys Ala Asp Trp Ile Val) Glu [P]-II. Val (His Arg Glu Thr Ala Met) Asp [P]-III. Glu (Cys Lys Asp Thr Met) Ala [P]-IV. Glu(His Ser Ala) Met) [P]-V. Ile (Cys Asp Arg Gly Pro T.p Phe) His [P]-VI. Gly(Asp ser) Lys [P]-VII. Thr(Pro Tyr Phe) Asp [P]-VIII. Phe(Tyr Leu Ile) Val [P]-IX. Trp (Phelle) Thr [P]-X. Ile (Arg Leu) Phe [P]-XI. Asp (Lys His Ser Gly Glu Pro) Ala [P]-XII. Glu (Cys Asp Gly) Ser [P]-XIII. Ala (Arg Tyr) Glu [P]-XIV. Met (Glu Ala) His It appears that the protease of the Bacillus subtilis K-27 syrain has rather wider range of specificity than proteases of Aspergoillus soya, pepsin, chymotrypsin, and trypsin.

• Korean Journal of Food Science and Technology
• /
• v.4 no.4
• /
• pp.276-284
• /
• 1972

A strain of Bacillus sp. which has powerful enzymatic activity and sets free a little amount of ammonia nitrogen and large amount of amino nitrogen was separated from Meju, Denzang, Chungkukjang, and paddy straw to make the soy-sauce rapidly and conveniently in the various mixing ratio of steamed soy-bean and wheat parched or steamed. Total nitrogen and amino nitrogen were increased during the maturing. The acidity of soy-sauce was increased to $1.15{\sim}1.98%$ than ordinary soy-sauce. pH were ranged in $4.6{\sim}6.0%$. The fermenting time takes 30 days while it takes at least 4 monthes in ordinary method. The best results were obtained with the mixed ratio of 1 : 1 or 1 : 2 (wheat : soy-bean).

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.35 no.1
• /
• pp.35-41
• /
• 2006

Antibacterial activities of the trace elements in combination with the food additives were measured against 6 kinds of food-borne microorganisms such as Escherichia coli, Vibrio parahaemolyticus, Staphylococcus aureus, Bacillus cereus, Bacillus subtilis and Pseudomonas fluorescens. The difference of antibacterial activity was not shown among the kinds of food additives, such as dextrin, gelatin and collagen. Zn and Ge in combination with food additives had strong antibacterial effect. Especially, $1\%$ zinc acetate in combination with $1\%$ gelatin was more effective against P. fluorescens and S. aureus than against Bacillus sp., E. coli and V. parahaemolyticus. Minimum inhibitory concentration of zinc acetate in combination with $1\%$ gelatin appeared to be 0.5 mg/mL on S. aureus and P. fluorescens, and 1.0 mg/mL on E. coli, V. parahaemolyticus, B. cereus and B. subtilis. Minimum bactericidal concentration of zinc acetate in combination with $1\%$ gelatin appeared to be 0.5 mg/mL on P. fluorescens and 1.0 mg/mL on E. coli, V. parahaemolyticus, S. aureus, B. cereus and B. subtilis. The zinc acetate in combination with gelatin showed stronger inhibitory effect in acidic range below pH 6.0, and remained active even after heat treatment at $121^{\circ}C$ for 15 min. In comparison with control, the viable cell counts of fish pastes, which were coated with the solution containing both $1\%$ zinc acetate and $3\%$ gelatin, were decreased by more than 100-fold until the storage of 7 days at $10^{\circ}C$. The results indicate that the combined use of zinc acetate and some food additives could prolong the shelf life of fish pastes by 8 days or more at $10^{\circ}C$.

• 한국균학회소식:학술대회논문집
• /
• 2015.05a
• /
• pp.45-46
• /
• 2015

Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

• Korean Journal of Microbiology
• /
• v.49 no.4
• /
• pp.343-352
• /
• 2013

In this study, we have evaluated the effect of efficient microorganisms on odor-removal efficiency and early stabilization of the burial sites. We have developed an efficient microorganism designated as 'KEM' which have the ability to degrade organic compounds and remove odor effectively. Other efficient microorganisms already used on site, such as EM and Bacillus sp., were also compared. We preceded these experiment using lab-scale reactors under three conditions (control, only media and only body) and comparing the effect of with or without the application of tree efficient microorganisms separately. Analysis was focused on eight components (ammonia, TMA, $H_2S$, methyl mercaptan, dimethyl sulfide, dimethyl disulfide, $CO_2$ and $CH_4$), and as a result, efficient microorganisms were shown efficiency in the removal of ammonia and methyl mercaptan. The applied KEM decayed up to 71.2% of the buried meat. We were unable to observe significant differences in microbial communities between efficient microorganisms-treated and non-treated reactors due to the large presence of microorganisms in both soil and carcasses. However, it was possible to observe the effect on odor control and decay rate through the application of efficient microorganisms.

• The Korean Journal of Food And Nutrition
• /
• v.10 no.4
• /
• pp.497-502
• /
• 1997

For the purpose of enzymatic production of galactooligosaccharides from soybean arabinogalactan (SAG) hydrolysis, the $\beta$-1, 4-D-arabinogalactanase($\beta$-1, 4-galactanase) from Bacillus sp. HJ-12 was used. The soybean galactooligosaccharides(SOS) were optimally produced in SAG 1%(w/v), pH 8.0, 5$0^{\circ}C$, $\beta$-1, 4-galactanase 20unit/g SAG and 24-40 hour reaction conditions. The produced galactooligosaccharides had visocity of 11,000 cp at 75%(w/v), $25^{\circ}C$. The viscosity of galactooligosaccharides was 80 fold increasing value than that of sucrose solution. Temperature dependence of viscosity of SOS was 4.6 fold higher value than surose solution below than 5$0^{\circ}C$. Less than 50 Brix, the viscosity of SOS was similar with sucrose solution(20-40 cp), but increasing of concentration, the difference of viscosity between SOS and sucrose solution was increased. And, SOS was very stable at pH and temperature.

• Journal of Applied Biological Chemistry
• /
• v.51 no.6
• /
• pp.272-276
• /
• 2008

Purification and some properties of Xylogone sphaerospora ${\beta}$-mannanase were reprevious previous paper. Locust bean gum galactomannan was hydrolyzed by the purified ${\beta}$-mannanase, and then the hydrolysates was separated by activated carbon column chromatography. The main hydrolysates were composed of D.P. (Degree of Polymerization) 4 and 6 galactosyl mannooligosaccharides. For elucidate the structure of D.P 4 and 6 galactosyl mannooligosaccharides, sequential enzymatic action was performed. D.P 4 and 6 were identified as ${Gal^2}{Man_3}\;(6^2-mono-O-{\alpha}-D-galactopyranosyl-4-O-{\beta}-D-mannotriose)$ and ${Gal^2}{Man_5}\;(6^2-mono-O-{\alpha}-D-galacto- pyranosyl-4-O-{\beta}-D-mannopentaose)$. To investigate the effects of locust bean gum galactosyl mannooligosaccharides on in vitro growth of Bifidobacterium longum, B. bifidum, B. infantis, B. adolescentis, B. animalis, B. auglutum and B. breve. Bifidobacterium spp. were cultivated individually on the modified-MRS medium containing carbon source such as D.P. 4 and D.P. 6 galactosyl mannooligosaccharides, respectively. B. longum and B. bifidum grew up to-fold and 6.6-fold more effectively by the treatment of D.P. 6 galactosyl mannooligosaccharides, compared to those of standard MRS medium. Especially, D.P. 6 was more effective than D.P. 4 galactosyl mannooligosaccharide on the growth of Bifidobacterium spp.

• Journal of Microbiology and Biotechnology
• /
• v.25 no.2
• /
• pp.227-233
• /
• 2015

Two recombinant arabinosyl hydrolases, α-_L-arabinofuranosidase from Geobacillus sp. KCTC 3012 (GAFase) and endo-(1,5)-α-_L-arabinanase from Bacillus licheniformis DSM13 (BlABNase), were overexpressed in Escherichia coli, and their synergistic modes of action against sugar beet (branched) arabinan were investigated. Whereas GAFase hydrolyzed 35.9% of _L-arabinose residues from sugar beet (branched) arabinan, endo-action of BlABNase released only 0.5% of _L-arabinose owing to its extremely low accessibility towards branched arabinan. Interestingly, the simultaneous treatment of GAFase and BlABNase could liberate approximately 91.2% of _L-arabinose from arabinan, which was significantly higher than any single exo-enzyme treatment (35.9%) or even stepwise exo- after endo-enzyme treatment (75.5%). Based on their unique modes of action, both exo- and endo-arabinosyl hydrolases can work in concert to catalyze the hydrolysis of arabinan to _L-arabinose. At the early stage in arabinan degradation, exo-acting GAFase could remove the terminal arabinose branches to generate debranched arabinan, which could be successively hydrolyzed into arabinooligosaccharides via the endo-action of BlABNase. At the final stage, the simultaneous actions of exo- and endo-hydrolases could synergistically accelerate the _L-arabinose production with high conversion yield.