• 농업과학연구
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• 제50권2호
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• pp.323-336
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• 2023

As a byproduct obtained from cheese manufacture, whey protein was developed as a functional food that contains multi-functional proteins. In this study, the biochemical activity of fermented milk prepared by fortifying whey protein with excellent physiological activity was investigated. Immunoglobulin (IgG) content was higher in 10% fortified whey protein fermented milk than in the control. The viable cell counts were 20% higher in the fermented milk with 10% fortified whey protein than in the control group. The antibacterial effect of 10% fortified whey protein fermented milk compared to the control group was shown to be effective against four pathogenic microorganisms, Escherichia coli (KCTC1039), Pseudomonas aeruginosa 530, Salmonela Typhimurium (KCTC3216), and Staphylococcus aureus (KCTC1621). The antioxidant effect by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities wasincreased two-fold in 10% fortified whey protein fermented milk compared to the control. The 10% fortified whey protein fermented milk inhibited the expression of the inflammatory cytokines (interleukin [IL]-6, tumor necrosis factor [TNF]-α, and induced nitric oxide synthase [iNOS]) in a concentration-dependent manner. In a piglets feeding test, the weight gain with 10% fortified whey protein fermented milk was increased by 18% compared to the control group, and no diarrhea symptoms appeared. Our results clearly demonstrated that 10% fortified whey protein fermented milk could be a useful functional ingredient for improving health.

• Pediatric Infection and Vaccine
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• 제24권1호
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• pp.1-6
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• 2017

목적: 본 연구에서는 delta neutrophil index (DNI)가 신생아 균혈증을 예측하는 지표로서의 효용성을 다른 지표들과의 비교를 통해 알아보고자 하였다. 방법: 원주세브란스 기독병원 신생아 중환자실에 발열을 주소로 입원한 환아들과 입원 중 발열이 있었던 생후 31일 미만 환아 146명을 대상으로 혈액배양검사와 동시에 시행한 총 백혈구 수, 절대호중구수, DNI, 혈소판 수, C-반응단백(C-reactive protein, CRP)에 대하여 분석하였다. 결과: 균혈증이 있었던 환아 77명의 평균 재태주수는 38.74주, 출생 체중은 3.20 kg였다. 대조군의 평균 재태주수는 33.34주, 출생 체중은 2.20 kg였다. 균혈증의 원인은 Staphylococcus aureus (22명), Staphylococcus epidermidis (18명), Streptococcus agalactiae (8명) 등이었다. DNI와 CRP만이 재태주수와 출생 체중 보정 후 균혈증과 연관성을 보여 area under the ROC curve를 조사하였고 DNI 0.70, CRP 0.68이었다. 결론: DNI는 신생아 균혈증을 예측하는 데 효과적인 지표이다. 다른 인자들과 함께 고려한다면 균혈증을 예측하는 데 더 도움이 될 것이다.

• 생명과학회지
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• 제16권2호
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• pp.245-252
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• 2006

피부 트러블을 가진 남,여 40명의 피부에서 분리한 collagenase를 생산하는 균주를 분리, 동정한 결과 Staphylococcus aureus로 판명되었으며 이를 S. aureus JJ-11이라 명명하였다. S. aurells JJ-11 균주의 collagenase의 최적 생산 조건은 1.5%(w/v) gelatin, 1%(w/v) yeast extract, 0.4% (w/v) $K_2HPO_4$, 0.005%(w/v) $NiSO_4{\cdot}6H_2O$를 함유한 배지 (pH 7.0)에서 $37^{\circ}C$, 200 rpm으로 18시간 동안 배양하는 것이다. 분리 균주가 생산하는 collagenase를 정제하기 위해서 amberlite IRA-900과 sephacryl S-300 HR columns를 이용하였고, 6.66-folds로 정제되었다. S. allreus JJ-11 균주가 생산하는 collagenase를 정제한 결과 분자량은 약 62 kDa이었으며, pH 7.0과 $37^{\circ}C$에서 각각 최대의 활성을 가졌고, pH와 온도에 대한 안정성은 pH 4.0-8.0, $40^{\circ}C$까지 100%의 활성이 있었다. 금속이온에 대해서는 $Fe^{2+},\;Co^{2+},\;Ba^{2+}$ 존재 하에서는 5 mM 농도에서도 활성을 유지하였다. $Sr^{2+},\;Hg^{2+}$에서는 30% 이상이 저해를 받는 것으로 확인되었다. 또한 EDTA와 O-phenanthroline에 의해 65% 이상이 저해되는 일 반적인 collagenase의 특정인 metalloproteinase의 특정을 보였으며, 그리고, 여러 가지 기절에 대해 효소활성을 비교한 결과, insoluble collagen (type I)에 대해 효소 활성이 가장 높았다.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1994년도 제2회 추계심포지움
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• pp.143-145
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• 1994

Chimeric fusion proteins involving IgG have proven valuable in studying protein-protein interactions and may possess therapeutic applications as well. For example, three receptor subtypes for the natriuretic peptides, when fused to the Fc portion of human IgG ${\gamma}$ chain, were quantitatively and qualitatively indistinguishable from the native receptor, thus allowing detailed structure-function studies of the receptor. In an attempt to block human immunodeficiency virus infectivity with soluble derivatives of CD4, a CD4/IgG Fc chimeric molecule was shown to increase the plasma half life of soluble CD4 and possessed the added advantage of IgG Fc-mediated placental transfer. In the case of the KGFR, this approach provided a framework for dissection of its ligand binding domains and made it possible to demonstrate that high affinity binding sites for two ligands, aFGF and KGF, reside within different receptor Ig-like domains. Chimeric molecules fused to immunoglobulins would have the advantages of secretion from transfected cells as well as detection and purification from medium utilizing Staphylococcus aureus Protein A. In addition, where highly related receptors make their discrimination very hard due to the difficulties in generating specific immunochemical probes, IgG fusion protein with tailor-made specificities confers particular advantages to elucidate patterns of receptor distribution and expression. The approach described here may have general applications in defining ligand-receptor interactions as well as searching for specific agonists and antagonists of receptor function.

• Biomolecules & Therapeutics
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• 제9권2호
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• pp.79-87
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• 2001

Heat shock proteins serve as chaperone by preventing the aggregation of denatured proteins and promote survival of pathogens in harsh environments. In this study, heat shock gene encoding a 84-kDa (p84) protein, which is one of the three major heat shock proteins in S. pneumoniae, was cloned and characterized. PCR with a forward primer derived from N-terminal amino acid sequence of the p84 and a reverse primer derived from the conserved second ATP-binding region of Clp family was used for amplification of the gene encoding the p84 and subsequently the PCR product was used for sequence determination. Sequence analysis of the p84 gene demonstrated that it is a member of ClpL. The deduced amino acid sequence of pneumococcal ClpL shows homology with other members of the Clp family, and particularly, even in variable leader region, with bovine Clp-like protein and L. lactis ClpL. S. pneumoniae clpL is the smallest clop member (701 amono acids) containing the two conserved ATP-binding regions, and hydrophilic N-terminal variable region of pneu-mococcal Clp ATPase is much shorter than any known Clp ATPases. Histidine tagged ClpL was overexpressed and purified from E. coli. Immunoblot analysis employing antisera raised against pneumococcus p84 demonstrated no cross-reactivity with Clp analog in Eschericha coli, Staphylococcus aureus and human HeLa cells. Preimmunization of mice with ClpL extended mice life partially but did not protect them from death.

• 한국임상수의학회지
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• 제26권2호
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• pp.176-180
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• 2009

2년령의 암컷 miniature schnauzer가 침울, 복통 및 식욕부진으로 내원하였다. 신체검사를 통해 빈맥, 발열과 복통을 확인하였다. 총혈구계수 검사에서 호중구수의 증가를 동반한 백혈구증가증이 관찰되었다. 방사선 검사에서 하행결장의 변위와 척추와 평행하게 방사선투과가 감소한 부위가 관찰되었다. 초음파검사시 방광내 결석과 슬러지, 우측 신장의 수신증 및 근위뇨관의 확장이 관찰되었다. 뇨검사결과 단백뇨와 혈뇨가 나타났으며 뇨침사검사에서 다수의 상피세포, calcium oxalate 결정, struvite 결정, 무정형 결정과 과립원주, 백혈구 및 구균이 관찰되었다. 상기의 결과와 배설성 요로 조영술을 통해 신우신염과 편측성 뇨관 완전폐색증으로 진단하였다. 신장절제술을 위한 개복술시 후복막강에 풍선형의 구조물을 발견하였다. 구조물로부터 채취한 시료의 세포검사와 배양검사결과 다수의 퇴행성 호중구와 탐식된 구균을 관찰하였다. 결론적으로 뇨관폐색과 방광결석을 동반한 Staphylococcus intermedius에 의한 복막후농양으로 최종 진단하였고 우측신장의 절제를 실시하여 치료하였다.

• 미생물학회지
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• 제51권3호
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• pp.271-279
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• 2015

락토페린(LF)는 철이온과 결합하는 당 단백질로서 항균, 항바이러스, 항진균 등의 기능을 가지고 있으며, 생체의 각종 체액으로부터 분비되는 다기능성 단백질이다. 본 연구에서는 사람의 락토페린(hLF)으로부터 유래된 N-lobe의 유전자를 분리하고 산업용 균주로서 많이 사용되는 메탄올자화 효모인 Pichia pastoris에서 발현시켰다. 재조합 사람 락토페린 N-lobe (rhLF-N)는 배양액으로 분비 발현되었으며, 3L 발효조에서 약 $458{\mu}g/ml$이 수준으로 생성되었다. rhLF-N을 정제한 다음 SDS-PAGE와 western blot으로 분석하여 분자량 35 kDa 단백질을 확인하였으며, hLF에 대한 항체를 이용하여 면역확산법으로 면역성을 확인하였다. rhLF-N의 mRNA 발현양상을 qRT-PCR로 분석한 결과 메탄올 첨가에 의한 발현 유도 후 2-3일째에 발현율이 가장 높았으며, 4일째에는 점차적으로 감소하였다. 정제한 rhLF-N을 이용하여 항균활성을 조사한 결과 Staphylococcus aureus, E. coli, Pseudomonas aeruginosa, Burkholderia cepacia, Salmonella typhimurium과 같은 병원성 균에 대해 광범위한 항균활성을 보였으나, LF유래 항균 peptide들과 항균활성을 비교하였을 때, 항균력이 상대적으로 매우 떨어지는 것으로 나타났다. 비록 본 연구에서 발현한 rhLF-N은 항균력은 떨어지나, hLF에 비해 그 크기가 작고 배양조건 연구로 P. pastoris에서 대량 생산이 가능하며, 배양액으로 분비시킬 수 있기 때문에 정제 비용 등을 고려 할 때 산업적 응용에는 보다 유리할 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제33권12호
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• pp.1692-1697
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• 2023

Staphylococcus aureus integrated with mecA gene, which codes for penicillin-binding protein 2a, is resistant to all penicillins and other beta-lactam antibiotics, resulting in poor treatment expectations in skin and soft tissue infections. The development of a simple, sensitive and portable biosensor for mecA gene analysis in S. aureus is urgently needed. Herein, we propose a dual-toehold-probe (sensing probe)-mediated exonuclease-III (Exo-III)-assisted signal recycling for portable detection of the mecA gene in S. aureus. When the target mecA gene is present, it hybridizes with the sensing probe, initiating Exo III-assisted dual signal recycles, which in turn release numerous "3" sequences. The released "3" sequences initiate catalytic hairpin amplification, resulting in the fixation of a sucrase-labeled H2 probe on the surface of magnetic beads (MBs). After magnet-based enrichment of an MB-H1-H2-sucrase complex and removal of a liquid supernatant containing free sucrase, the complex is then used to catalyze sucrose to glucose, which can be quantitatively detected by a personal glucose meter. With a limit of detection of 4.36 fM for mecA gene, the developed strategy exhibits high sensitivity. In addition, good selectivity and anti-interference capability were also attained with this method, making it promising for antibiotic tolerance analysis at the point-of-care.

• BMB Reports
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• 제43권3호
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• pp.176-181
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• 2010

The primary sigma factor ($\sigma^{A}$) of Staphylococcus aureus, a potential drug target, was little investigated at the structural level. Using an N-terminal histidine-tagged $\sigma^{A}$ (His-$\sigma^{A}$), here we have demonstrated that it exits as a monomer in solution, possesses multiple domains, harbors primarily $\alpha$-helix and efficiently binds to a S. aureus promoter DNA in the presence of core RNA polymerase. While both N- and C-terminal ends of His-$\sigma^{A}$ are flexible in nature, two Trp residues in its DNA binding region are buried. Upon increasing the incubation temperature from 25$^{\circ}$ to 40$^{\circ}C$, $\sim$60% of the input His-$\sigma^{A}$ was cleaved by thermolysin. Aggregation of His-$\sigma^{A}$ was also initiated rapidly at 45$^{\circ}C$. From the equilibrium unfolding experiment, the Gibbs free energy of stabilization of His-$\sigma^{A}$ was estimated to be +0.70 kcal $mol^{-1}$. The data together suggest that primary sigma factor of S. aureus is an unstable protein. Core RNA polymerase however stabilized $\sigma^{A}$ appreciably.

• Preventive Nutrition and Food Science
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• 제22권3호
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• pp.191-194
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• 2017

Ocimum gratissimum is a common plant in the tropics and has been used in food and medicine. Its usage in food and medicine could be attributed to its phtyochemical and antimicrobial properties. In this study we investigated the proximate, phytochemical, and antimicrobial attributes of air dried leaves of O. gratissimum. The aqueous extract was found to contain phtyochemicals with alkaloid and saponin present in appreciable amounts. The proximate analysis (crude protein and crude fibre content were 15.075% and 17.365%, respectively) showed that the leaf could be a good source of protein and fibre. The aqueous ethanolic extract of the leaf exhibited activity against a wider range of organisms when compared to the aqueous extract at the investigated concentrations. Aqueous ethanolic extracts of O. gratissimum leaf was active against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus cereus and the aqueous extract of the leaf was active against P. aeruginosa.