• 한국식품영양과학회지
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• 제45권1호
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• pp.76-83
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• 2016

본 연구는 커피를 추출한 후 발생되는 커피박을 식재료로 활용할 수 있는 방안을 모색하고자 커피박 첨가 형태를 달리하여 머핀을 제조하고 품질 특성을 검토하였다. 커피박 열수추출물(CRE) 머핀의 중량(62.01~62.36 g)은 대조구(61.64 g)에 비해 높았으며, 높이(6.03~6.23 cm)와 부피(186~493 mL)도 대조구(5.87 cm, 185 mL)에 비해 높았다. 수분 함량은 CRE 머핀(29.41~29.92%)과 커피박 분말(CRP) 머핀(30.10~31.11%)이 대조구에 비해 높았다. L(lightness) 값과 b(yellowness) 값은 CRE 및 CRP 첨가량이 증가할수록 감소하였으나 a(redness) 값은 증가하였다. CRE 머핀의 경도는 대조구에 비해 유의적으로 낮았으나, CRP 첨가구는 대조구보다 높았으며 첨가량이 증가할수록 증가하였다. 총 폴리페놀 함량은 CRE 및 CRP 머핀이 대조구보다 높았으며, 첨가량이 증가할수록 유의적으로 증가하였다(P<0.05). CRE 머핀의 DPPH 라디칼 소거능은 15.91~83.25%로 대조구(5.53%)에 비해 유의적으로 높았으며, 농도 증가에 비례하여 증가하였다. ABTS 라디칼 소거능도 CRE 첨가구(1.91~48.09%)가 대조구(25.20%)에 비해 유의적으로 높았으며, 농도가 증가함에 따라 유의적으로 증가하였다(P<0.05). 관능검사 결과 머핀 내부의 외관과 색, 풍미와 맛은 CRE 첨가구가 대조구보다 우수하였다. 맛과 종합적 기호도는 1.0% 첨가구가 가장 우수하였다. CRP 머핀의 외관과 내부 색은 1.0% 첨가구가 가장 우수하였으며, 풍미는 CRP 첨가구가 대조구보다 높았고 맛과 종합적 기호도는 1.0% 첨가구가 가장 우수하였다. 이상의 결과를 종합해 보면 항산화 활성이 다량 잔존하고 있는 커피박은 제과제빵의 항산화 활성 등 기능성 강화 소재로 활용이 가능하며, 머핀 이외에도 다양한 식품 산업에 활용이 가능할 것으로 판단된다.

• Journal of Microbiology and Biotechnology
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• 제11권1호
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• pp.131-137
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• 2001

Previous work has identified that only the catabolite responsive element A (creA; previously called cre-2) out of two potential cre sequences (cre-1: nucleotide +160 to +173 and cre-2: +173 to +186), recognized within the coding region of the xylanaseA gene (xynA) of Bacillus stearothermophilus No.236, was actually, was actually involved in the carbon catabolite repression(CCR) of xynA expression in B. subtilis. However, the level of CCR of xynA expression in the original B.stearothermophilus No.236 strain (70-fold repression). Therefore, to search for an additional cre element in the promoter region, the upstream region of the xynA gene was subcloned by chromosome walking, and as a result, another potential cre element (nucleotide -124∼-137; designated creB) was recognized in this region. The cre-like sequence revealed a high homology to the cre consensus sequence. The xylanase activity of B. subtilis MW15 bearing pWPBR14 (containing creA and creB) cultured in a medium containing xylose as the sole carbon source was about 7.7 times higher than that observed for the same culture containing glucose. B. subtilis MW15 bearing pWPBR23 (containing only creA) produced an activity about 2.4 times higher. This pattern of CCR was confirmed using derivatives of xynA::aprA fusion plasmids. Furthermore, a measurement of the amounts of the xynA transcript showed a similar pattern as that for the production of xylanase. In addition, the synthesis of xylanase in B. subtilis QB7115 [a catabolite control protein A (ccpA) mutant strain] carrying pWPBR14 was almost completely relieved from glucose repression. Together, these results lead to a conclusion that the CCR of the expression of the xynA gene is mediated by CcpA binding at creA and creB sites in B. subtilis.

• 한국식품영양과학회지
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• 제41권3호
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• pp.345-350
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• 2012

본 실험은 CRE 80% 에탄올 추출물을 식이에 0.5%, 1% 혼합하여 5주간 섭취시켰을 때 CRE 추출물이 SHR의 혈압에 미치는 영향을 확인함으로써 CRE 추출물이 고혈압 흰쥐의 지질성분과 혈압에 미치는 영향을 조사함으로써 CRE의 기능성식품으로서의 이용 가능성을 확인하였다. 실험기간 중 실험쥐의 일일평균 체중변화량은 0.5% CRE 보충군(II)은 1.63 g/day로 정상식이 급여군보다 일일평균 체중증가량이 유의적으로 감소하는 결과를 나타내었다. 간, 신장, 비장의 무게 변화를 측정한 결과 유의적인 차이를 나타내지 않았으며, 혈청 내 포도당 농도 또한 정상식이 급여군에 비해서 0.5%, 1% CRE 추가 보충군이 감소하는 경향은 보였으나 유의적인 차이는 나타나지 않았다. 본 실험에서 총 cholesterol 함량은 정상식이 급여군보다 0.5%, 1% CRE 보충 급여군에서 유의적으로 증가하는 경향을 보였지만 HTR을 확인한 결과 정상식이 급여군보다 0.5%, 1% CRE 보충 급여군이 각각 11% 증가한 결과를 나타냈기 때문에 혈중 총 cholesterol 함량의 증가는 HDL-cholesterol의 농도 증가에 의한 것으로 생각된다. HDL-cholesterol 농도는 정상식이 급여군과 0.5%, 1% CRE 보충 급여군 모두에서 유의적으로 증가하는 경향을 나타냈다. LDL-cholesterol 역시 유의적으로 증가하는 경향을 나타냈지만 HDL-cholesterol/LDL-cholesterol의 증가 비율로 봤을 때 HDL-cholesterol이 정상식이 급여군보다 0.5% CRE 보충 급여군이 15%, 1% CRE 보충급여군이 36.5%로 유의적으로 증가하는 결과를 나타냈다. 실험기간 동안 SHR의 혈압 변화는 정상식이 급여군(I)의 경우 거의 일정하게 혈압이 유지되는 경향을 보이다가 5주차에는 다시 증가한 반면 CRE 0.5% 보충군(II)은 초기혈압에서 떨어지는 경향을 보이다가 2주째에 혈압이 다시 상승했다. 하지만 그 후 다시 감소하는 경향을 보였고, CRE 1% 보충군(III)은 일정하게 혈압이 감소하는 경향을 나타냈다. 따라서 CRE 추출물이 SHR의 혈압을 낮추고 지질대사 개선에 영향을 주어 혈관계 장애로 인해 발병되는 고혈압, 당뇨, 뇌혈관질환, 동맥경화증과 같은 질환을 예방하는 기능성식품으로서 이용 가능성이 기대된다.

• 대한본초학회지
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• 제28권5호
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• pp.21-28
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• 2013

Objectives : The aim of this study was to investigate the neuroprotective effects and mechanisms of Cyperi Rhizoma extracts (CRE) using in vitro and in vivo models of Parkinson's disease (PD). Methods : We evaluated the neuroprotective effect of CRE against 1-methyl-4-phenylpyridinium (MPP+) toxicity using tyrosine hydroxylase immunohistochemistry (IHC) in primary rat mesencephalic dopaminergic neurons. In addition, the effect of CRE was evaluated in mice PD model induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). For evaluations, C57bl/6 mice were orally treated with CRE 50 mg/kg for 5 days and were injected intraperitoneally with MPTP (20 mg/kg) at 2 h intervals on the last day. To identify the CRE affects on MPTP-induced neuronal loss of dopaminergic neurons in substantia nigra pars compacta (SNpc) and striatum of mice, the behavioral tests and IHC analysis were carried out. Also, we conducted nitric oxide (NO) and tumor necrosis factor-alpha (TNF-${\alpha}$) assay in dopaminergic neurons and IHC using glial markers in SNpc of mice to assess the anti-inflammation effects. Results : In primary mesencephalic culture system, CRE protected dopaminergic cells against $10{\mu}M$ MPP+-induced toxicity at 0.2 and $1.0{\mu}g/mL$. In the behavior tests, CRE treated group showed improved motor deteriorations than those in the MPTP only treated group. CRE significantly protected striatal dopaminergic damage from MPTP-induced neurotoxicity in mice. Moreover, CRE inhibited productions of NO and TNF-${\alpha}$ in dopaminergic culture system and activation of astrocyte and microglia in SNpc of the mice. Conclusion : We concluded that CRE shows anti-parkinsonian effect by protecting dopaminergic neurons against MPP+/MPTP toxicities through anti-inflammatory actions.

• Journal of Korean Biological Nursing Science
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• 제25권4호
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• pp.285-294
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• 2023

Purpose: This study aimed to identify the colonization rate of carbapenem-resistant Enterobacteriaceae (CRE), the characteristics of CRE isolates, and risk factors for CRE colonization in patients transferred to the general wards of a small/medium-sized hospital. Methods: This retrospective study was conducted on patients who underwent CRE culture tests within 24 hours of admission among patients transferred to a small/medium-sized hospital. Forty-seven patients confirmed as positive for CRE were classified as belonging to the patient group. For the control group, 235 patients (five times the number of the patient group) were matched by sex, age, and diagnosis, and then selected at random. Data were analyzed using descriptive analysis and multiple logistic regression analysis. Results: The CRE colonization rate was 5% (47 out of 933 patients), and Klebsiella pneumoniae (68.0%) was the most common isolate of CRE. The positivity rate of carbapenemase-producing Enterobacteriaceae was 61.7%. The risk factors for CRE colonization included renal disease (odds ratio [OR]=4.93; 95% confidence interval [CI], 1.49-16.31), heart disease (OR=3.86; 95% CI, 1.35-11.01), indwelling urinary catheters (OR=4.43; 95% CI, 1.59-12.36), and cephalosporin antibiotic use (OR=8.57; 95% CI, 1.23-59.60). Conclusion: Having a comorbid renal or cardiac disease, an indwelling urinary catheter, or a history of exposure to cephalosporin antibiotics could be classified as risk factors for CRE colonization in patients transferred to small and medium-size hospitals. It is necessary to perform active infection control through proactive CRE culture testing of patients with risk factors.

• Journal of Microbiology and Biotechnology
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• 제27권3호
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• pp.514-523
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• 2017

Carbon catabolite repression is a crucial regulation mechanism in microorganisms, but its characteristic in Rhizopus is still unclear. We extracted a carbon regulation gene, cre, that encoded a carbon catabolite repressor protein (CRE) from Rhizopus stolonifer TP-02, and studied the regulation of CRE by real-time qPCR. CRE responded to glucose in a certain range, where it could significantly regulate part of the cellulase genes (eg, bg, and cbh2) without cbh1. In the comparison of the response of cre and four cellulase genes to carboxymethylcellulose sodium and a simple carbon source (lactose), the effect of CRE was only related to the concentration of reducing sugars. By regulating the reducing sugars to range from 0.4% to 0.6%, a glucose-containing medium with lactose as the inducer could effectively induce cellulases without the repression of CRE. This regulation method could potentially reduce the cost of enzymes produced in industries and provide a possible solution to achieve the largescale synthesis of cellulases.

• Journal of Microbiology and Biotechnology
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• 제30권1호
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• pp.109-117
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• 2020

Cre recombinase is widely used to manipulate DNA sequences for both in vitro and in vivo research. Attachment of a trans-activator of transcription (TAT) sequence to Cre allows TAT-Cre to penetrate the cell membrane, and the addition of a nuclear localization signal (NLS) helps the enzyme to translocate into the nucleus. Since the yield of recombinant TAT-Cre is limited by formation of inclusion bodies, we hypothesized that the positively charged arginine-rich TAT sequence causes the inclusion body formation, whereas its neutralization by the addition of a negatively charged sequence improves solubility of the protein. To prove this, we neutralized the positively charged TAT sequence by proximally attaching a negatively charged poly-glutamate (E12) sequence. We found that the E12 tag improved the solubility and yield of E12-TAT-NLS-Cre (E12-TAT-Cre) compared with those of TAT-NLS-Cre (TAT-Cre) when expressed in E. coli. Furthermore, the growth of cells expressing E12-TAT-Cre was increased compared with that of the cells expressing TAT-Cre. Efficacy of the purified TAT-Cre was confirmed by a recombination test on a floxed plasmid in a cell-free system and 293 FT cells. Taken together, our results suggest that attachment of the E12 sequence to TAT-Cre improves its solubility during expression in E. coli (possibly by neutralizing the ionic-charge effects of the TAT sequence) and consequently increases the yield. This method can be applied to the production of transducible proteins for research and therapeutic purposes.

• Journal of Microbiology and Biotechnology
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• 제8권1호
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• pp.21-27
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• 1998

The xylA gene of Bacillus stearothermophilus encoding the major ${\beta}$-xylosidase was previously cloned and sequenced. In the present study we examined the regulation of the cloned xylA gene expression in Bauillus subtilis MW15 carrying the xylA::aprA fusion plasmids. The induction of the fused xylA gene expression remained uninfluenced by any of the carbon sources tested but the gene expression was repressed about 2-3 fold in the presence of glucose. Two CRE-like sequences (CRE-1: nucleotides ＋ 124 to ＋136 and CRE-2: ＋247 to ＋259) were recognized within the reading frame region of the xylA gene. The deletion experiments showed that the CRE-2 sequence had a role in catabolite repression (CR) as a true CRE of the xylA gene, but the CRE-1 had no effect on CR of the xylA gene expression. Surprisingly, the deletion of the CRE- 1 sequence reduced about 2~3 fold of the expression of the xylA fused gene. The repression ratios of the xylA gene expression were estimated to be about 0.4 from the assay of subtilisin activity, and about 0.3 at the level of transcription by determining the amounts of xylA transcripts in B. subtilis. While, the level of CR of the xylA gene was assessed to be about l0-fold in previous work when the relative amounts of the xylA transcripts were measured in B. stearothermophilus.

• 한국임상약학회지
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• 제30권2호
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• pp.120-126
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• 2020

Objective: The rising number of carbapenemase-resistant Enterobacteriaceae (CRE) cases has become a concern worldwidely. This study investigated patient characteristics with CRE and analyzed the risk factors associated with its acquisition. Methods: A retrospective review of the electronic medical records of the Kangbuk Samsung Medical Center from May 2016 to April 2019 was performed. The inclusion criterion was hospitalized patients aged ≥18 years with confirmed CRE acquisition. Patients were divided by CRE acquired and non-required patients. CRE acquired patients were those with CRE confirmed by their active surveillance cultures, while non-acquired patients were those with carbapenemase-sensitive Enterobacteriaceae (CSE). If CRE was isolated more than once during hospitalization, only the first isolation was used for data analysis. Patient characteristics, antibiotic used, and the duration of use were compared between two groups using univariate analysis, and the risk factors associated with CRE were analyzed using multiple logistic regression analysis. Results: Among the 73 CRE acquired patients, 44 (60.3%) were positive for carbapenemase-producing Enterobacteriaceae (CPE). Infection from Klebsiella pneumonia (42 cases, 57.5%), Escherichia coli (17 cases, 23.3%), and Enterobacter cloacae (5 cases, 6.8%). The risk of CRE acquisition was significantly increased by 4.99 times [confidence interval (CI), 1.40-17.78; p=0.013] with mechanical ventilation, 3.86 times (CI, 1.59-9.36; p=0.003) with penicillin administration, and 21.19 times (CI, 6.53-68.70; p<0.001) with carbapenem administration. Conclusions: Proper antibiotic use including the selection, frequency, and duration, and patients on mechanical ventilators need close monitoring.

• BMB Reports
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• 제34권4호
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• pp.322-328
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• 2001

Excision and amplification of pre-determined, large genomic segments (taken directly from the genome of a natural host, which provides an alternative to conventional cloning in foreign vectors and hosts) was explored in human cells. In this approach, we devised a procedure for excising a large segment of human genomic DNA, the iNOS gene, by using the Cre/loxP system of bacteriophage P1 and amplifying the excised circles with the EBNA-1/oriP system of the Epstein-Barr virus. Two loxP sequences, each of which serves as a recognition site for recombinase Cre, were integrated unidirectionally into the 5'-UTR and 3'-UTR regions of the iNOS gene, together with an oriP sequence for conditional replication. The traps-acting genes cre and EBNA-1, which were under the control of a tetracycline responsive $P_{hcmv^*-1}$ promoter, were also inserted into the 5'-UTR and 3'-UTR regions of the iNOS gene, respectively, by homologous recombination. The strain carrying the inserted elements was stably maintained until the excision and amplification functions were triggered by the induction of cre and EBNA-1. Upon induction by doxycycline, Cre excised the iNOS gene that was flanked by two ZoxP sites and circularized it. The circularized iNOS gene was then amplified by the EBNA-1/oriP-system. With this procedure, approximately a 45.8-kb iNOS genomic fragment of human chromosome 17 was excised and successfully amplified in human cells. Our procedure can be used effectively for the sequencing of unclonable genes, the functional analysis of unknown genes, and gene therapy.