• Preventive Nutrition and Food Science
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• 제21권1호
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• pp.52-56
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• 2016

This study was performed to investigate the antifungal substances and the antifungal activity against fungi of lactic acid bacteria (LAB) isolated from kimchi. LAB from kimchi in Imsil showed antifungal activity against Penicillium brevicompactum strain FI02. LAB LI031 was identified as Lactobacillus sakei subsp. Antifungal substances contained in L. sakei subsp. ALI033 culture media were unstable at high pH levels. Both, the control and proteinase K and protease treated samples showed clear zones, suggesting that the antifungal substances produced by ALI033 were non-protein substances unaffected by protesases. Both, the control and catalase showed clear zones, suggesting that the antifungal metabolite was not $H_2O_2$. The molecular weights of the antifungal substances were ${\leq}3,000Da$. The organic acid content of crude antifungal substances produced by L. sakei subsp. ALI033 showed high concentrations of lactic acid (502.47 mg/100 g). Therefore, these results suggest that antifungal substance produced by L. sakei subsp. ALI033 is most likely due to its ability in producing organic acid.

• Parasites, Hosts and Diseases
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• 제31권2호
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• pp.117-128
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• 1993

이 연구는 환자로부터 분리하여 무균 배양된 10개의 질트리코모나스 분리주에 대하여 병원성 여부를 판정하고 단백분해효소 관련 여부를 알아보고자 시도된 것이다. 질트리코모나스 분리주들은 마우스 피내 접종 실험을 통한 병원성 판정에서 약병원성 주, 중등도 병원성 주 및 강 병원성 주 등 3개 그룹으로 나눌 수 있었으며 중성 단백분해효소 및 산성 단백분해효소 활성도는 질트리코모나스 추출물 및 그 배양액에서 약 병원성 주에 비해 강 병원성 주의 활성도가 높게 나타나 피하농양 크기에 따른 병원성과 상기 단백분해효소의 비활성도(specific activi쇼) 사이에 상관관계가 있었음을 알 수 있었다(p < 0.05) 질트리코모나스 단백분해효소는 gelatin을 기질로 하는 SDS-PAGE 전기영동에서 RF치를 달리하는 5가지 분획대가 나타났으며 그 분획양상은 각 분리주 의 병원성에 따라 일정한 양상을 나타내었다. 그리고 여러가지 단백분해효소 억제제를 전기 영동 효소액에 처리했을 경우 antlpaln과 leupeptin 처리군에서는 분획이 전혀 나타나지 않았으며 EUTA 처리군에서는 대조군에 비해 그 활성이 약화된 분획이 관찰되었고, PMSF 처리군에서의 분획들은 대조군과 그 활성의 차이를 볼 수 없어 이들 단백 분해효소는 cystelne 단백분해효소로 추정되었다. 조직 세포에 대한 질트리코모나스 추출물의 세포독성은 병원성에 따라 차이가 있었고 추출물의 단백질 농도 $12.0{\;}\mu\textrm{g}/100{\mu}\ell$ 이상에서 세포 독성에 따른 병원성 구분이 용이하였다. 그리고 질트리코모나스 추출물에 단백분해효소 억제제를 처리한 결과 대조군에 비하여 세포 독성이 낮게 나타났으며, 특히 antipain 처리군에서는 조직 세포에 대한 세포 독성이 현저하게 낮았다. 이상의 결과로 보아 cysteine계로 추정되는 질트리코모나스의 단백분해효소는 특이한 전기영동 활성 분획상을 나타내었는 바 이들은 모두 충체의 병원성 및 세포 독성과 밀접한 관련이 있었다.

• 한국식품과학회지
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• 제34권2호
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• pp.311-318
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• 2002

Lactobacillus plantarum에 대해 항균활성을 지니는 유산균을 김치로부터 분리하였다. 이 균은 Leuconostoc mesenteroides으로 동정되었으며 Leuconostoc mesenteroides B7으로 명명하였다. Leuconostoc mesenteroides B7이 생산하는 박테리오신은(박테리오신 B7) pH 및 열 안정성이 뛰어나 pH $2.5{\sim}9.5$ 그리고 $4^{\circ}C{\sim}120^{\circ}C$ 열처리에서도 항균활성을 안정하게 유지하였다. 박테리오신 B7은 proteinase K, trypsin, ${\alpha}-chymotrypsin$, 그리고 protease 처리에 의해 항균활성이 실활되므로 peptide나 단백질로 이루진 구조임을 알 수 있었다. 정제된 박테리오신 B7은 Tricine-SDS-PAGE를 통하여 분자량이 약 3,500 dalton임을 확인하였다. Leuconostoc mesenteroides B7과 이에 감수성인 균주, Lb. plantarum KFRI 464 혹은 Lb. delbruekii KFRI 347와의 혼합배양에 의하여 박테리오신 B7 생산이 증가됨을 확인하였고, 이는 박테리오신 생산 유도물질이(inducing factor) 감수성균주내에 존재함을 시사한다. 감수성균주의 세포분획을 통하여 inducing factor는 감수성균주의 세포벽과 세포내에 존재함을 알았다. 이 inducing factor가 proteinase K처리로 박테리오신 유도활성을 상실함으로써 이는 단백질성물질임을 시사하였다.

• Applied Biological Chemistry
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• 제38권5호
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• pp.382-386
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• 1995

감자의 단백질 분해효소 억제제-II(PI-II)단백질은 카이모트립신 저해부위와 트립신 저해부위로 나뉘어 있는데 PI-II 유전자중의 하나인 PI-IIT 유전자의 염기서열에서 비롯된 아미노산 서열이 폴리펩타이드 카이모트립신 저해제(PCI) 단백질의 아미노산 서열과 84%의 높은 동질성을 가지고 있으므로 감자의 단백질 분해효소 억제제유전자 집단 (family) 중의 하나인 PCI와 homology가 있는 유전자단편을 클로닝하기 위하여 이미 클론된 PI-IIT 유전자로부터 PCR을 행하여 얻어진 DNA 단편을 백터에 클로닝하고 염기서열을 결정하였다. 염기서열을 확인한 유전자 단편을 박테리오파아지 T7 promoter와 terminator를 갖고있는 플라스미드 pET3a에 옮겨 대장균 BL2l(DE3)에서 발현시켰던바 IPTG의 부가에 따라 유기되는 것을 확인 하였으나 발현수준은 기대했던것에 미치지 못하였다.

• 미생물학회지
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• 제7권4호
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• pp.159-166
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• 1969

It is necessry to develop and strengthen the activity of enzymic source which in low applied for maggerley brewing as an amylolytic and proteolytic starter, recently in this country the active and strong enzymic starter is required for the better brewing and to substitute another starch material for the present wheat flour. In this study, manufacturing method the strong enzymic source have been developed and established with use of raw wheat bran plus fungal strains of Rhizopus sp. and Aspergillus usamii the culture of starter. The results on experimental the activities of enzymic sources (stater) are as following ; 1. Method of making the enzymic source (starter) is to cultivate the strains of Asperguillus orzyae, Asp. kawachii, Asp usamii and Rhizopus sp. in the acid treated raw or heatboiled wheat bran. 2. The saccharogenic pwoer (S.P.) of enzymic source which consisted of raw bran plus fungi and cultured in it is generally stronger than those of heat-boiled bran plus fungi, the strongest power was shown in the culture of Rhizopus plus raw bran, and the next other is in mixture of Asp.usamii and Rhizopus on raw wheat bran. 3. The most strong alpha amylase activity was expressed in the plot of Asp.oryzae on heat-boiled wheat bran, the next was in the culture of Rhizopus nad Aspergillus usamii on raw wheatbran. 4. The most vigourous acidic proteinase activity was expressed in the micture of raw bran plus Asp. usamii and Rhizopus those were independentlu cu;tured before mixing for neutral proteinase activity, it was shown in the mixed culture of Asp. usamii and Rhizopus on raw wheat bran, the msot active alkaline proteinase activity of enzymic source was found in the plot of raw bran material. 5. For poly-preptidase activity in pH 6.5 it is found that the culture of Rhizopus and Asp.usamii on raw bran was most active among them of enzymic sources. 6. Generally, it is concluded that culture of fungi on acid treated raw wheat bran is stronger in its activity than those of heat boiled wheat bran, especially the culture of Rhizopus nad Asp.usamii on raw bran exhibited the most vigorous and non-polarized activity for all aspects, so it is considered to be most desirable enzymic stater in Korean Maggerley brewing and this would be able to substitute brewing material for the present wheat flour because of its strong and wide hand activity of amylolytic and proteolytic action.

• 한국산업보건학회지
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• 제17권3호
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• pp.235-244
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• 2007

By comparing the proteins from the workers exposed to styrene with the ones from controls, it may be possible to identify proteins that play a role in the occurrence and progress of occupational disease and thus to study the molecular mechanisms of occupational disease. In order to find the biomarkers for assessing the styrene effects early, before clinical symptoms develop and to understand the mechanisms of adverse health effects, we surveyed 134 employees, among whom 52 workers(30 male and 22 female) were chronically exposed to styrene in 10 glass-reinforced plastic boat manufacturing factories in Korea and 82 controls had never been occupationally exposed to hazardous chemicals including styrene. The age and drinking habits and serum biochemistry such as total protein, BUN and serum creatinine in both groups were significantly different. Exposed workers were divided into three groups according to exposure levels of styrene(G1, below 1/2 TLV; G2, 1/2 TLV to TLV; G3, above TLV). The mean concentration of airborne styrene in G1 group was $10.93{\pm}11.33ppm$, and those of urinary mandelic acid(MA) and phenylglyoxylic acid(PGA) were $0.17{\pm}0.21$ and $0.13{\pm}0.11g/g$ creatinine, respectively. The mean concentration of airborne styrene in G2 and G3 groups were $47.54{\pm}22.43$ and $65.33{\pm}33.47ppm$, respectively, and levels of urinary metabolites such as MA and PGA increased considerably as expected with the increase in exposure level of styrene. The airborne styrene concentration were significantly correlated to the urinary concentration of MA(r=0.784, p=0.000) and PGA(r=0.626, p<0.001). In the 2D electrophoresis, the concentration of five proteins including complement C3 precursor, alpha-1-antitrypsin(AAT), vitamin D binding protein precursor(DBP), alpha-1-B-glycoprotein(A1BG) and inter alpha trypsin inhibitor(ITI) heavy chain-related protein were significantly altered in workers exposed to styrene compared with controls. While expression of complement C3 precursor and AAT increased by exposure to styrene, expression of DBP, A1BG and ITI heavy chain-related protein decreased. These results suggest that the exposure of styrene might affects levels of plasma proteinase, carriers of endogenous substances and immune system. In particular, increasing of AAT with the increase in exposure level of styrene can explain the tissue damage and inflammation by the imbalance of proteinase/antiproteinase and decrease of DBP, A1BG and ITI heavy chain-related protein in workers exposed to styrene is associated with dysfunction and/or declination in immune system and signal transduction

• Asian-Australasian Journal of Animal Sciences
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• 제33권1호
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• pp.100-110
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• 2020

Objective: The objective of this study was to isolate proteolytic microorganisms and evaluate their effects on proteolysis in total mixed ration (TMR) silages of soybean curd residue. Methods: TMRs were formulated with soybean curd residue, alfalfa or Leymus chinensis hay, corn meal, soybean meal, a vitamin-mineral supplement, and salt in a ratio of 25.0: 40.0:30.0:4.0:0.5:0.5, respectively, on a basis of dry matter. The microbial proteinases during ensiling were characterized, the dominate strains associated with proteolysis were identified, and their enzymatic characterization were evaluated in alfalfa (A-TMR) and Leymus chinensis (L-TMR) TMR silages containing soybean curd residue. Results: Both A-TMR and L-TMR silages were well preserved, with low pH and high lactic acid concentrations. The aerobic bacteria and yeast counts in both TMR silages decreased to about 10⁵ cfu/g fresh matter (FM) and below the detection limit, respectively. The lactic acid bacteria count increased to 10⁹ cfu/g FM. The total microbial proteinases activities reached their maximums during the early ensiling stage and then reduced in both TMR silages with fermentation prolonged. Metalloproteinase was the main proteinase when the total proteinases activities reached their maximums, and when ensiling terminated, metallo and serine proteinases played equally important parts in proteolysis in both TMR silages. Strains in the genera Curtobacterium and Paenibacillus were identified as the most dominant proteolytic bacteria in A-TMR and L-TMR, respectively, and both their proteinases were mainly with metalloproteinase characteristics. In the latter ensiling phase, Enterococcus faecium strains became the major sources of proteolytic enzymes in both TMR silages. Their proteinases were mainly of metallo and serine proteinases classes in this experiment. Conclusion: Proteolytic aerobic bacteria were substituted by proteolytic lactic acid bacteria during ensiling, and the microbial serine and metallo proteinases in these strains played leading roles in proteolysis in TMR silages.

• Fisheries and Aquatic Sciences
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• 제19권10호
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• pp.42.1-42.10
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• 2016

The objectives of this study were to identify the histamine-forming bacteria and bacteriocin- producing lactic acid bacteria (LAB) isolated from Myeolchi-jeot according to sequence analysis of the 16S rRNA gene, to evaluate the inhibitory effects of the bacteriocin on the growth and histamine accumulation of histamine-forming bacteria, and to assess the physico-chemical properties of the bacteriocin. Based on 16S rRNA gene sequences, histamine-forming bacteria were identified as Bacillus licheniformis MCH01, Serratia marcescens MCH02, Staphylococcus xylosus MCH03, Aeromonas hydrophila MCH04, and Morganella morganii MCH05. The five LAB strains identified as Pediococcus acidilactici MCL11, Leuconostoc mesenteroides MCL12, Enterococcus faecium MCL13, Lactobacillus sakei MCL14, and Lactobacillus acidophilus MCL15 were found to produce an antibacterial compound with inhibitory activity against the tested histamine-producing bacteria. The inhibitory activity of these bacteriocins obtained from the five LAB remained stable after incubation at pH 4.0-8.0 and heating for 10 min at $80^{\circ}C$; however, the bacteriocin activity was destroyed after treatment with papain, pepsin, proteinase K, ${\alpha}$-chymotrypsin, or trypsin. Meanwhile, these bacteriocins produced by the tested LAB strains also exhibited histamine-degradation ability. Therefore, these antimicrobial substances may play a role in inhibiting histamine formation in the fermented fish products and preventing seafood-related food-borne disease caused by bacterially generated histamine.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.88.3-89
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• 2003

Acanthoic acid (AA) is a pimaradiene diterpene isolated from the Korean medicinal plant, Acanthopanax koreanum (Araliaceae), which has been traditionally used as a tonic and sedative as well as in the treatment of rheumatism and diabetes in korea. Proteinase-activated receptor-2 (PAR-2) agonist trypsin plays a role in inflammation, and human leukemic mast cells (HMC-l) express PAR-2. In the present study, the effect of acanthoic acid on production of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and tryptase in trypsin-stimulated HMC-1 was examined. (omitted)

• 한국식품영양과학회지
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• 제26권6호
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• pp.1228-1236
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• 1997

Lactobacillus strains were isolated from volunteer's feces, including from newly-born infants and adults in their 20's, by using differential MRS-BPB plates. Total 56 presumptive Lactobacillus strains were isolated and the bacteriocin productions by the isolates were examined by agar diffusion method. Six bacteriocin-producing strains were confirmed. Among them, two isolates, HU-1 and H22-3, showed the most outstanding antimicrobial activities, which were not affected by pH adjustments or catalase treatments of culture. HU-1 was originated from a two-years old boy and H22-3 was originated from a newly-born infant. HU-1 and H22-3 had the same morphology(short rod) when examined by scanning electron microscope, and the same biochemical traits including growth temperature range, salt tolerance and sugar-fermenting abilities. But the growth-inhibition spectrum and plasmid profiles of HU-1 and H22-3 were different. Both strains inhibited the growth of various Gram (+) microorganisms including Listeria monocytogenes. Micrococcus luteus, and Staphylococcus aureus in addition to many species of lactic acid bacteria, indicating the production of broad-spectrum bacteriocins. Bacteriocins produced by HU-1 and H22-3 were stable up to 90℃, 15 min heat treatments. Their activities were not affected by pepsin or trypsin treatments but destroyed by proteinaseK or pronase treatments.