Production of medium chain length polyhydroxyalkanoate by metabolically engineered Pseudomonas gessardii using acetate-formate carbon sources : 탄소원으로 아세트산과 포름산을 이용하여 대사공학이 적용된 슈도모나스 게사디에서 MCL-PHA 생산

Woo Young Kim 고려대학교 대학원 2024 국내석사

The production medium-chain-length polyhydroxyalkanoates (mcl-PHA) was attempted using Pseudomonas gessardii NIBRBAC000509957. This strain effectively utilizes acetate and formate as carbon sources. This work first evaluated the utilization of acetate as a carbon source, revealing optimal growth at 5 g/L acetate. Then, formate was supplied as a substrate together with acetate to enhance cell growth. After overexpressing formate assimilation pathway enzymes in Methylorubrum extorquens AM1, this strain enabled to grow significantly faster in acetate-formate minimal medium than acetate as a sole carbon source. Furthermore, this work engineered the strain to produce mcl-PHA by overexpressing the tesA from Escherichia coli and phaC gene from Rhodococcus aetherivorans. Finally, this study conducted fed-batch fermentations with acetate-formate minimal medium, resulted 0.35 g/L of mcl-PHA production.

Acetate metabolism in drug resistance and malignancy of cancer

이수진 서울대학교 대학원 2021 국내박사

Acetate metabolism has become a popular topic in cancer research, as cancer metabolic dependence on acetate has been reported in relation to cancer bioenergy, macromolecular synthesis, and epigenetics. However, researches on the effects of acetate metabolism on acquired resistance to cancer drug and the changes in tumor grade are still lacking. Therefore, I investigated the metabolic characteristics of acetate with regards to cisplatin resistance in bladder cancer and the tumor grade of liver cancer, and the clinical significance of these characteristics. First, with the recent recognition of lipid metabolic alterations in bladder cancers, I studied the metabolic implications of cisplatin resistance using cisplatin-sensitive (T24S) and resistant (T24R) bladder cancer cells. Real-time live metabolomics revealed that T24R cells consume more glucose, leading to a higher production of glucose-derived acetate and fatty acids. Along with the activation of general metabolic regulators, enzymes involved in acetate usage (ACSS2) and fatty acid synthesis (ACC), as well as a precursor for fatty acid synthesis (acetyl-CoA), were elevated in T24R cells. Consistently, metabolic analysis with 13C isotope revealed that T24R cells preferred glucose to acetate as the exogenous carbon source for the increased fatty acid synthesis, in contrast to T24S cells. In addition, acetyl-CoA synthetase 2 (ACSS2), rather than the well-established ATP citrate lyase (ACLY), was the key enzyme supplying acetyl-CoA in T24R cells through glucose-derived endogenous acetate. The relevance of ACSS2 in cisplatin resistance was further confirmed by the abrogation of resistance by an ACSS2 inhibitor and, additionally, by the higher expression of ACSS2 in patient tissues with cisplatin resistance than tissues without resistance. Second, the relationships among acetate uptake, metabolic characteristics, and tumor malignancy were comprehensively studied in liver cancer. Among the cell lines used, HepG2 avidly utilized acetate, even in a glucose-sufficient environment, whereas Hep3B did not. These characteristics correlated with ACSS2 expression levels in the cells. Metabolomic isotope tracing showed high-ACSS2 HepG2 cells exhibited higher acetate incorporation and enhanced lipid anabolic metabolism than Hep3B cells, which was consistent with the separate gene expression profiles. These metabolic characteristics were confirmed by knockdown and inhibition studies of ACSS2, a master anabolic gene (mTOR), and a lipid catabolic gene (CPT1). Upon in vivo 13C isotopic tracing, orthotopic mouse liver tumors from HepG2 exhibited higher anabolism and less malignancy. Consistently, normal human liver tissue showed higher ACSS2 levels than cancerous tissues. Patients with lower ACSS2 expression, particularly those in the lower ~13th percentile (13VLA group), had notably poorer prognoses in the analysis of two independent large cohorts (total n = 486) than those with higher expression. The 13VLA patients also exhibited decreased lipid anabolic pathways, increased glycolysis, and enhanced hypoxia, which were associated with higher ACSS2 promoter methylation. Finally, positron emission tomography-computed tomography (PET-CT) imaging of liver cancer patients showed that lower-grade cancer had higher 11C-acetate but lower 18F-FDG uptake, and this was reversed in higher-grade cancer. Overall, acetate uptake seems to be independent of nutrient depletion and contributes to lipid anabolic metabolism and better prognosis in liver cancer. This research can help to improve the treatment options for bladder and liver cancers and provide vulnerability targets for customized treatment. 아세테이트 대사는 생물에너지학, 거대 분자 합성, 및 후성 유전학과 관련하여 암의 아세테이트에 대한 대사적 의존성이 보고되면서 암 연구에서 새로운 흐름이 되고 있다. 그러나 아세테이트 대사가 항암제 내성 획득과 종양 등급 변화에 미치는 영향에 대한 연구는 아직 부족하다. 따라서 방광암의 시스플라틴 내성과 간암의 악성도 등급과 관련된 아세테이트의 대사적 특성과 이의 임상적 의의를 조사하였다. 첫째로 최근 방광암에서 지질 대사가 변화되어 있다는 것이 보고되어 시스플라틴 민감성 (T24S) 및 내성 (T24R) 방광암 세포를 사용하여 시스플라틴 저항성의 대사적 의미를 연구하였다. 실시간 라이브 대사체학을 이용하여 시스플라틴 저항성 방광암 세포 (T24R)에서 시스플라틴 민감성 방광암 세포 (T24S) 대비 글루코스를 더 많이 소모하며 글루코스 유래 아세테이트와 지방산을 더 많이 생산하는 것을 확인하였다. T24R 세포에서 일반적인 대사 조절제의 활성화와 더불어 아세테이트 사용 (ACSS2) 및 지방산 합성 (ACC) 효소와 지방 합성을 위한 전구체 (아세틸-CoA)가 상승했다. 13C 동위 원소를 사용한 대사 분석에서도 일관되게 T24R 세포가 T24S 세포와 다르게 지방산 합성 증가를 위한 외인성 탄소 공급원으로 아세테이트보다 글루코스를 선호한다는 것이 나타났다. 또한 T24R 세포에서 글루코스 유래 내인성 아세테이트를 통해 아세틸-CoA를 공급하는 핵심 효소는 잘 확립된 ACLY가 아닌 ACSS2 였다. ACSS2의 상관성은 ACSS2 억제제에 의한 시스플라틴 저항성 상실에 의해 추가로 확인되었으며, 시스플라틴 저항성을 가진 환자 조직에서 ACSS2의 더 높은 발현에 의해 확인되었다. 둘째로 간암에 대한 아세테이트 흡수, 대사적 특성, 및 악성 종양 간의 관계를 종합적으로 연구했다. 세포주 중 HepG2는 글루코스가 충분한 환경에서도 아세테이트를 사용했지만 Hep3B에서는 그렇지 않았고 이는 ACSS2의 발현과 연관이 있었다. 대사체 동위 원소 추적에서 ACSS2가 높은 HepG2는 Hep3B보다 더 높은 아세테이트 유입과 향상된 지질 단백 대사를 나타냈다. 이러한 대사 특성은 ACSS2, 마스터 단백 동화 유전자 (mTOR), 그리고 지질 이화 유전자 (CPT1)에 대한 유전자 발현 억제 및 억제제 연구를 통해 확인되었다. in-vivo 13C 동위 원소 추적에서 HepG2의 동소 (orthotopic) 마우스 간암은 단백 동화가 더 잘 일어나고 악성도가 낮음이 확인되었다. 일관되게 사람의 정상 간 조직은 암 조직보다 더 높은 ACSS2 수준을 나타냈다. ACSS2 발현이 낮은 환자, 특히 13번째 백분위수 이하 (13VLA 그룹)의 환자는 두 개의 독립적인 대규모 코호트에서 현저하게 예후가 나빴다 (총 n = 486). 또한 13VLA 환자는 더 높은 ACSS2 프로모터 메틸화, 감소된 지질 합성 경로, 그리고 증가된 해당 과정 및 강화된 저산소증을 나타냈다. 환자의 PET-CT 영상은 악성도가 낮은 간암이 11C-아세테이트의 흡수가 더 많고 18F-FDG의 흡수가 적다는 것을 보여주었으며 이는 악성도가 높은 간암에서 역전되었다. 전반적으로 아세테이트 흡수는 영양소 고갈과 무관한 것으로 보이며 지질 단백 대사 증가와 간암의 더 나은 예후에 기여한다. 본 연구는 방광암과 간암 환자에 대한 치료 옵션을 개선하고 맞춤형 치료를 위한 환자의 취약성에 기반한 표적 개발에 도움을 줄 수 있다.

Acetate Yarn을 이용한 소재기획에 관한 연구 : 2002 춘하를 위한 circular knit fabric을 중심으로

정지연 이화여자대학교 대학원 2001 국내석사

본 논문의 목적은 환경친화적 Cellulose계 섬유로 재해석되고 있는 Acetate에 대한 고찰을 통해 Acetate를 Fashion 소재로 더욱 발전시키고 소재정보 수집과 Trend의 이해를 바탕으로 한 Acetate 소재기획 방향을 토대로 2002년 춘하 시즌의 Acetate Circular Knit Fabric을 개발하여 제시하는데 그 목적이 있다. 연구방법은 각종 해외 기술 자료 및 논문, 업체별 카탈로그, 해외 전시회 자료들를 토대로 Acetate 특성과 연구 개발 동향을 고찰하고, 패션정보지, 소재관련회사의 정보지 등을 수집 분석하여 2002년도 춘하 시즌의 소재 Trend 예측 및 기획방향을 설정 Theme별 Acetate Circular Knit Fabric을 제작하여 제시하였다. 연구결과 Acetate는 Fashion 소재로서 필요한 심미적인 성격을 고루 갖추고 있으며 2002년도 춘하 시즌의 감각적이고 인체 친화적 소재군이 부상하며, 신선하고 밝은 컬러의 유행을 예견하고 있는 Trend와도 잘 부합하는 소재이다. Acetate의 가장 큰 특징인 Silk like한 성격과 우수한 Drapery를 이용하여 Sensual, Flash, Crispy의 세가지 Theme를 제안하고 원단을 제작하였다. 첫째, Sensual은 부드럽고 감각적인 피부친화적인 촉감을 위해 저 De' 원사 및 세섬사와 Spandex를 이용하여 Soft한 반발감을 가진 Second Skin Circular Knit를 제작하였다. 둘째, Flash는 Acetate의 Silk like한 고급스럽고 우아한 광택을 위해 Bright Filament Yarn을 이용하여 빛의 율동감을 표현 Color에 의해 재해석 될 수 있는 원단을 제작하였다. 셋째, Crispy는 Natural한 Crispy 감각을 위해 다양한 연사물로 시도한 원단을 제작하였다. 매끄럽고 Soft하면서 Dry한 이중감성을 표현하였다. This thesis starts with the inquiries on Acetate that is regarded as environment-friendly Cellulose textile recently. The purpose of this analysis is not only to promote Acetate for Fashion materials but also to develop Acetate Circular Knit Fabric for 2002 Spring/Summer through collecting material informations and understanding trends in industry. To understand characters and trends of Acetate, I investigate foreign articles, technical informations, catalogues by companies and materials of industrial exhibitions. After these investigation and analysis of forecast and planning direction of fabric trend, I introduced Acetate Circular Knit Fabric by Theme for 2002 Spring/Summer. Estimated that the trends of near future requires biological superiority and fresh and bright color, I concluded that Acetate is pertinent fabric for 2002 Spring/Summer because Acetate contains Aesthestic nature requiring on Fashion fabric. Considering characteristic nature, Silk-like and Drapery, of Acetate, I suggest three Theme Sensual, Flash and Crispy to fabricate Acetate Circular Knit Fabric For Sensual, firstly, I manufactured Second Skin Circular Knit with low De' yarn and Spandex to express soft and skin-friendly feelings. For Flash, secondly, I made fabric with Bright Filament Yarn that contains colors of light fluctuation to express silk-like, elegant, high quality feelings. For Crispy, thirdly, I fabricate with various Twisted yarns that contains dual-senses like slippery and crispy, dry and softy to express natural crispy feelings.

Influence of bornyl acetate on blood pressure and aortic strip's contractility of the rat

기영우 조선대학교 2003 국내석사

본 연구는 bornyl acetate가 정상혈압 쥐에서 혈압반응과 적출혈관 평활근의 수축반응에 대한 bornyl acetate 의 효과를 검색하고 작용기전을 규명하고자 수행하였다. Phenylephrine(α₁-아드레날린 수용체 효능약)과 고농도 칼륨(막탈분극약)은 적출 대동맥 편에서 각각 현저한 수축반응을 일으켰다. 이 같은 고농도칼륨 (5.6×10^-2M)에 의한 수축반응은 bornyl acetate (2.5∼20 ㎍/ml) 존재 하에서 용량-의존적으로 억제되었다. 또한 Phenylephrine (10^-5M)에 의한 수축반응도 높은 농도의 bornyl acetate (10-20 ㎍/ml) 존재 하에서는 억제되었으나 낮든 농도(2.5-5.0 ㎍/ml에서는 영향을 받지 않았다. 더욱이, bornyl acetate (3 mg/kg/30min)을 정상혈압의 urethane 마취흰쥐의 정맥 내로 30분간 주입시 norepinephrine에 의한 승압반응을 현저히 억제하였다. 이상과 같은 연구결과를 종합하여 보면, 마취흰쥐에서 정맥 내로 투여한 bornyl acetate는 적어도 아드레날린 α₁-수용체를 차단함으로써 혈압하강을 나타내는 것으로 생각되며 또한 bornyl acetate는 기전 미상의 직접작용 외에 α₁-아드레날린 수용체의 차단작용에 의한 혈관이완작용을 나타내는 것으로 사료된다. The extract of magnolia bark has been shown to have depressant actions on the central nervous system (Watanabe et al., 1973). Recently, it has been reported that the crude extract of magnolia bark, an herbal drug, inhibited the secretion of catecholamines from bovine adrenal chromaffin cells stimulated by acetylcholine in a concentration-dependent manner (Tachikawa et al., 2000). Therefore, the present study was conducted to investigate the effects of bornyl acetate on arterial blood pressure and vascular contractile responses in the normotensve rats and to establish the mechanism of action. Phenylephrine (an adrenergial-receptor agonist) and high potassium (a membrane-depolarizing agent) caused greatly contractile responses in the isolated aortic strips, respectively. These phenylephrine (10^-5 M)-induced contractile responses were depressed in the presence of high concentrations of bornyl acetate (10 ∼ 20 ㎍/㎖), but not affected in low concentration of bornyl acetate (2.5 ∼ 5 ㎍/㎖). Also, high potassium (5.6 × 10^-2 M)-induced contractile responses were greatly inhibited in the presence of bornyl acetate (2.5 ∼ 20 ㎍/㎖) in a dose-dependent fashion. Bornyl acetate (1 ∼ 10 ㎎/㎏) given into a femoral vein of the normotensive rat produced a dose-dependent depressor response, which is transient (data not shown). Interestingly, the infusion of a moderate dose of bornyl acetate (3 ㎎/㎏/30 min) made a significant reduction in pressor responses induced by intravenous norepinephrine. Collectively, these results obtained from the present study demonstrate that intravenous bornyl acetate causes a dose-dependent depressor action in the anesthetized rat at least partly through the blockade of adrenergic α₁-receptors. bornyl acetate also causes vascular relaxation in the isolated aortic strips of the rat via the blockade of adrenergic α₁-receptors, in addition to the unknown mechanism of the direct vasorelaxation.

Enhancement of intestinal permeability of leuprolide acetate using permeation enhancers

노시원 중앙대학교 대학원 2023 국내석사

자동화장치를 이용한 PET 영상용 표지 화합물 [11C]acetate의 합성 및 임상 사용을 위한 최적화에 관한 연구

장화연 전남대학교 2015 국내석사

[11C]Acetate, a radiotracer for PET imaging, is a promising radiopharmaceutical for overcoming the limitation of 2-deoxy-2-[18F]fluoro-D-glucose in a number of cancers. Here, the optimized automatic synthesis of [11C]acetate using an in-house-developed module under different conditions has been reported for routine production. [11C] CO2 was produced in a 16.4MeV PETtrace cyclotron, and methyl magnesium chloride was used for synthesis. For product purification, ion-exchange solid-phase extraction cartridges were used, connected in series. Highperformance liquid chromatography and gas chromatography were used to measure radiochemical and chemical purity. The Limulus amebocyte lysate test and the fluid thioglycollate medium test were performed for quality control of [11C]acetate. The total reaction time of [11C] acetate was within 15min, and the overall decay-corrected radiochemical yield was 84.33±8.85%. Radiochemical purity was greater than 98% when evaluated on an analytical high-performance liquid chromatography system. No endotoxins or anaerobic bacteria were seen on quality control checks. Optimized production of [11C]acetate was achieved by the in-house module. Radiochemical and biological properties of the [11C]acetate produced were appropriate for clinical PET study. [11C]Acetate는 여러 암 종류의 진단에 사용되던 2-deoxy-2-[18F]fluoro-d-glucose([18F]FDG)의 단점을 보완하기 위한 유망한 방사성의약품 중 하나이다. 본 연구는 기존에 보고 되었던 [11C]acetate 생산 방법의 장점을 극대화 하고 단점을 극복 할 수 있는 새로운 합성장치를 자체적으로 개발하고 임상사용을 위한 최적화 조건을 확립하는 것을 목표로 하였다. [11C]CO2는 16.4Mev의 PETtrace cyclotron 에서 생성하고 이를methyl magnesium chloride와 반응시켜 합성하였다. [11C]Acetate의 정제를 위해 이온 교환 고상 추출 카트리지를 사용하였고, 방사화학적 순도와 화학적 순도를 측정하기 위해 고성능 액체 크로마토그래피와 가스 크로마토그래피를 사용하였다. [11C]Acetate의 생물학적 품질관리를 위해 limulus amebocyte lysate 와 fluid thioglycollate medium 검사를 진행하였다. [11C]Acetate 의 생산시간은 총 15분으로 반감기가 20분인 [11C]CO2보다 짧았고 반감기를 고려한 방사화학적 수율은 84.33 ± 8.85%였다. 방사화학적 순도는 고성능 액체 크로마토그래피에서 98%이상임이 확인되었다. 또한 독소 또는 혐기성 박테리아는 품질관리 검사를 통해 음성임을 확인하였다. [11C]Acetate 는 자체 개발한 모듈로 최적의 조건을 확립하여 간단하고 효율적으로 합성하였고 방사화학적 및 생물학적 특성이 임상사용에 적합함을 확인하였다.

Retinyl acetate가 白鼠 乳房癌 發生에 미치는 影響에 對한 硏究

김동구 忠南大學校 1988 국내박사

An experimental study was performed to study the effects of retinyl acetate on N-methyl-N-nitrosourea induced mammary carcinogenesis in Sprague-Dawley rats. Female Sprague Dawley rats were given an intravenuous injection of 5 mg of N-methyl-N-nitrosourea per 100 gm body weight at 60 and 67 days of age, respectively. After carcinogenic treatment, feeding of a diet supplemented 1/2 mmol (164 mg) or 1mmol (328 mg) retinyl acetate per kilogram diet was initated from the day of carcinogenic treatment. During the experiment the animals were checked biweekly for the detection of mammary tumors. Thus, the incidence of tumors, the trend and numbers of tumor occurrence, changes of body weight, and the size of tumors were evaluated. The results were follows: 1.Tumor incidence in the NMU treated group with control diet was 46.7%, 33.3% in the group with 1/2 mmol retinyl acetate, and 13.3% in the group with 1 mmol retinyl acetate supplied group compared with the NMU-only treated group. 2.Tumors occurred between 14th and 16th week after injection of NMU. The number of tumors was not significantly different between the NMU-only treated group and the 1/2 mmol retinyl acetate supplied group. There was, however, a significant difference in the 1 mmol retinyl acetate supplemented group compared to the NMU-only treated group. 3.Body weights of the animals gradually increased during the experiments and there was no statistical difference among the groups. 4.The average size of the tumors was 3.9cm in the NMU-only treated group, 3. 93cm in the 1/2 mmol retinyl acetate group, and 4.63cm in the 1 mmol retinyl acetate group. Histologic features of the tumors were identical in the experimental groups.

Ethyl acetate를 利用한 二中乳劑의 蛋白質 微粒子 製造工程의 開發

李賢泰 대구가톨릭대학교 2002 국내석사

The objective of this study was to develop an ethyl acetate-based double emulsion microencapsulation process to load proteins into poly(d,l-lactide-co-glycolide) 75:25 microspheres. Instead of methylene chloride known as a confirmed carcinogen, ethyl acetate was used as an alternative dispersed solvent. A solvent extraction/quenching technique was used to load a model protein lysozyme into the microspheres. This study was then aimed at investigating the effect of process parameters upon the characteristics of the microspheres and the encapsulation efficiency of lysozyme into the microspheres. Among various process parameters, the timing of the onset of ethyl acetate extraction/quenching critically was found to be a critical parameter affecting the encapsulation efficiency of lysozyme: its efficiency decreased with retardation in the onset of ethyl acetate quenching. For instance, 12.5 ± 2.9 % of lysozyme appeared in the aqueous continuous phase, right after the primary water-in-oil emulsion stirred for 5 min was quenched. By contrast, 51.9 ± 5.3 % of lysozyme diffused to the aqueous continuous phase, right after quenching was performed on the primary emulsion stirred for 45 min. As a result, the highest encapsulation efficiency was attained with the microspheres prepared by carrying out the solvent quenching step after 5-min stirring. It was found that during the ethyl acetate-based microencapsulation process, lysozyme in the inner aqueous phase continued to diffuse across the ethyl acetate phase to the aqueous continuous phase. This was a reason why the degree of the encapsulation efficiency of lysozyme declined as a function of stirring time. The embryonic water-in-oil emulsion droplets were also broken into smaller ones until they became solidified by the solvent quenching step. Such dynamic changes in the size distribution of emulsion droplets also influenced the leaching of lysozyme into the aqueous continuous phase during the microencapsulation process. Increases in the amount of poly(d,l-lactide-co-glycolide) dissolved in the ethyl acetate phase contributed to inhibiting the diffusion of lysozyme to the aqueous continuous phase. Therefore, the onset of ethyl acetate quenching and poly (d,l-lactide-co-glycolide)concentration should be optimized to improve the encapsulation efficiency of lysozyme.

Silver Nanoparticle을 포함하는 Cellulose acetate와 Curcumin의 composite fiber 에 대한 연구 : Study of Cellulose acetate and Curcumin Composite fibers containing Silver Nanoparticles

바다르모니르자이디 한양대학교 2009 국내석사

Cellulose acetate is the acetate ester of cellulose, the primary structural component of the cell wall of green plants and is one of the most common biopolymers on earth. Electrospun cellulose acetate mats have been used as affinity membranes, antimicrobial membranes, and drug-delivery membranes. Silver nanoparticles have unique antimicrobial properties and have been used with a number of different polymers. Curcumin is a naturally occurring compound in the curcuma longa plant mostly used as a food item. It has anti-oxidant, anti tumor and anti-inflammatory properties. Previously cellulose acetate has been electrospun with curcumin but they do not contain antibacterial property. This research focuses on the antibacterial property of the Cellulose acetate / curcumin composite. Silver nanoparticles have been introduced in the cellulose acetate / curcumin composite by direct electrospinning them. The results show that the cellulose acetate / curcumin / silver nanoparticles composite possess antibacterial properties as well as the antioxidant, anti-tumor properties of curcumin. This composite has potential applications in the biomedical field.

Glucose-acetate를 利用한 glutamic acid 生産菌株의 分離 및 同定

정대주 全南大學校 大學院 1983 국내석사

L-Glutamic acid producing bacteria from using glucose - acetate as a carbon source was isolated and identified. The results are summarized as follows. 1. In the media containing glucose-acetate as the carbon source, 214 strains of glucose-acetate utilizing bacteria were isolated from 120 kinds of soil sample. 2. Thirty one strains among 214 strains of glucose-acetate utilizing bacteria which were isolated, formed L-Glutamic acid of more than 10g/ℓ, and a best bacterial production strain of L-Glutamic acid was isolated from the sewage. 3. This strain was identified as Brevibacterium saccharolyticum sp by the standard method given in Manual Microbiological Methods. 4. The maximum production of L-Glutamic acid with isolated strains was resulted in 28g/ℓ when the experimental broth were made with 2% glucose, 2% sodium acetate, 1% ammonium acetate, and biotin 3∼4㎍/ℓ, pH 7.5. temp. 30℃.