28 research outputs found
Applications of the Comprehensive Rural Village Development Project and Transportation Accessibility
ํ์๋
ผ๋ฌธ (๋ฐ์ฌ) -- ์์ธ๋ํ๊ต ๋ํ์ : ๋์
์๋ช
๊ณผํ๋ํ ๋๊ฒฝ์ ์ฌํํ๋ถ(์ง์ญ์ ๋ณด์ ๊ณต), 2021. 2. ์ด์ฑ์ฐ.The purpose of this research is to propose pragmatic policy evaluation models with scientific rigor and to empirically analyze the effectiveness of large-scale public policies implemented in rural areas of South Korea. The impacts of two policies, the Comprehensive Rural Village Development Project (CRVDP) and the improved transportation accessibility, on agricultural income are analyzed from the ex-post standpoint applying quantitative policy evaluation methods. This dissertation is composed of three empirical essays.
In the first essay, the impact of the CRVDP on agricultural income is analyzed using a quasi-experimental research design. The Heckman selection model was used to overcome selection bias, and the Blinder-Oaxaca decomposition method was employed to estimate the causal impact of the CRVDP. The results revealed that the project had a positive impact on raising farm households agricultural income. A higher probability of making agricultural income was found in the project implemented areas vis-ร -vis project not-implemented areas and in the period after the project implementation vis-ร -vis period before the project implementation.
The second essay attempts to analyze the impact of the CRVDP on agricultural income by sub-groups of rural population. The analysis was conducted by applying the propensity score matching and the double cohort model developed from the age-period-cohort framework. The results find that young farmers in their early-career stage experienced a significant increase in the probability of making a higher agricultural income with the implementation of the CRVDP. On the other hand, significant effect was not visible for the cohorts of middle-aged and elderly farmers at all experience levels.
The last essay explores the benefits of transportation infrastructural investments to the agricultural sector and rural areas. The paper examines the impact of changes in transportation accessibility over the course of 2005 to 2015 on agricultural income. The impact was analyzed from both micro- and macro-levels of farm households and rural autonomies utilizing the multilevel model and the spatial econometrics model. According to the results, a positive association was found in 2005, but the effect turned negative starting in 2010 which suggest that public investments in transportation accessibility had a meager or negative impact on agricultural income.
There has been a rising call to come up with evidence-based recommendations employing scientifically credible evaluation methods in the public sector. Under this context, this research presents pragmatic approaches to policy evaluation for using credible secondary data. In this research, practical yet rigorous policy impact evaluation methods were applied to Koreas rural sector where evaluation of policies using rigorous scientific methods remains relatively limited.๋ณธ ์ฐ๊ตฌ์ ๋ชฉ์ ์ ์ฒด๊ณ์ ์ด๊ณ ๊ณผํ์ ์ธ ์ ์ฑ
ํ๊ฐ๋ชจํ์ ๊ตฌ์ถํ๊ณ , ์ฐ๋ฆฌ๋๋ผ ๋์ด์ง์ญ์์ ์ํ๋ ๋๊ท๋ชจ ๊ณต๊ณต์ ์ฑ
์ ํจ๊ณผ์ฑ์ ์ค์ฆ์ ์ผ๋ก ๋ถ์ํ๋ ๋ฐ์ ์๋ค. ๋ณธ ์ฐ๊ตฌ์์๋ ๋์ ์ ํ๊ธฐ์ ์ํ๋ ๋ํ์ ๋์ด์ ์ฑ
์ธ ๋์ด๋ง์์ข
ํฉ๊ฐ๋ฐ์ฌ์
๊ณผ ๊ตํตSOC ํฌ์์ ๋ฐ๋ฅธ ๊ตํต์ ๊ทผ์ฑ ๊ฐ์ ์ด ๋์
์๋ ์ฆ์ง์ ๋ฏธ์น๋ ์ํฅ์ ์ ๋์ ํ๊ฐ๊ธฐ๋ฒ์ ํ์ฉํ์ฌ ์ฌํ์ ์ผ๋ก ๋ถ์ํ์๋ค. ๋ณธ ๋
ผ๋ฌธ์ 3๊ฐ์ ์ค์ฆ๋ถ์์ผ๋ก ๊ตฌ์ฑ๋๋ค.
์ฒซ ๋ฒ์งธ ์ค์ฆ๋ถ์์์๋ ๊ฐ์ฉ์๋ฃ๋ฅผ ๊ณ ๋ คํ์ฌ ํคํฌ๋ง์ ๋ณ๋ชจํ(Heckman Selection Model)์ ์ด์ฉํด ์ ํํธ์๋ฅผ ๋ณด์ ํ๊ณ , ํด์ฒด๊ธฐ๋ฒ(Decomposition Method)์ ํ์ฉํ์ฌ ๋์ด๋ง์์ข
ํฉ๊ฐ๋ฐ์ฌ์
์ํ์๋ฐ๋ฅธ ๋์
์๋ ์ฆ๋ํจ๊ณผ๋ฅผ ๋ถ์ํ์๋ค. ๋ถ์ ๊ฒฐ๊ณผ ์ฌ์
์ํ์ง์ญ์ ๋ฏธ์ํ์ง์ญ์ ๋นํด ๋์
์๋ ์ฆ๋ํจ๊ณผ๊ฐ ์กด์ฌํ๋ ๊ฒ์ผ๋ก ๋ํ๋ฌ์ผ๋ฉฐ, ์ฌ์
์ํ์ง์ญ์ ์ฌ์
์ํ ์ด์ ๋ณด๋ค ๋์
์๋์ด ์ฆ๊ฐํ์ฌ ์ ์ฑ
ํจ๊ณผ๊ฐ ์๋ ๊ฒ์ผ๋ก ํ๊ฐ๋์๋ค.
๋ ๋ฒ์งธ ์ค์ฆ๋ถ์์ ์ฑํฅ์ ์๋งค์นญ(Propensity Score Matching)๊ณผ APC(Age-Period-Cohort) ๋ชจํ์ ๊ธฐ์ดํ ์ด์ค์ฝํธํธ ๋ชจํ(Double Cohort Model)์ ์ ์ฉํ์ฌ ๋์
์ธ์ ์ฐ๋ น๊ณผ ์๋ ๊ฒฝ๋ ฅ์ ์ํด ๋ถ๋ฅ๋ ์ฝํธํธ๋ณ๋ก ๋์ด๋ง์์ข
ํฉ๊ฐ๋ฐ์ฌ์
์ ๋์
์๋ ์ฆ์ง ํจ๊ณผ๋ฅผ ๋ถ์ํ์๋ค. ๋ถ์ ๊ฒฐ๊ณผ ๋์ด๋ง์์ข
ํฉ๊ฐ๋ฐ์ฌ์
์ ๊ฒฝ๋ ฅ ์ด๊ธฐ๋จ๊ณ ์ฒญ๋
๋์
์ธ์ ๊ณ ์๋ ๊ฐ๋ฅ์ฑ์ ๋์ธ ๊ฒ์ผ๋ก ๋ํ๋ ๋ฐ๋ฉด, ์ค๋
๋ฐ ๋
ธ๋
๊ธฐ ๋์
์ธ ์ฝํธํธ์ ์์ด์๋ ๊ทธ ํจ๊ณผ๊ฐ ๋ฏธ๋ฏธํ๋ ๊ฒ์ผ๋ก ๋ถ์๋์๋ค.
์ธ ๋ฒ์งธ ์ค์ฆ๋ถ์์ ๋ค์ธต๋ชจํ(Multilevel Model)๊ณผ ๊ณต๊ฐ๊ณ๋๋ชจํ(Spatial Econometrics Model)์ ํ์ฉํ์ฌ ๊ตํตSOC ํฌ์์ ์ํ ๊ตํต์ ๊ทผ์ฑ ๊ฐ์ ์ด ๋์
์๋์ ๋ฏธ์น๋ ์ํฅ์ ๋ฏธ์์ ์์ค์ ๋๊ฐ๋จ์์ ๊ฑฐ์์ ์์ค์ ์ง์ญ๋จ์ ์ธก๋ฉด์์ ๋ถ์ํ์๋ค. ๊ฒฐ๊ณผ์ ๋ฐ๋ฅด๋ฉด 2005๋
์๋ ๊ตํต์ ๊ทผ์ฑ์ ๋ณํ๊ฐ ๋์
์๋์ ๊ธ์ ์ ์ธ ๋ฐฉํฅ์ผ๋ก ์์ฉํ ๊ฒ์ผ๋ก ๋ถ์๋์์ผ๋, 2010๋
์ดํ์๋ ๋ณ๋ค๋ฅธ ์ํฅ์ด ์๊ฑฐ๋ ๋ถ์ ์ ์ธ ๋ฐฉํฅ์ผ๋ก ์์ฉํ๋ ๊ฒ์ผ๋ก ๋ํ๋ฌ๋ค.
๋ณธ ์ฐ๊ตฌ๋ ๊ณต๊ณต์ ์ฑ
์ ์ค์ฅ๊ธฐ์ ์ฑ๊ณผ ๋ฐ ์ฌ์ ํฌ์
ํจ์จ์ฑ ํ๊ฐ์ ๋ํ ๊ฐ๊ด์ ์ด๊ณ ๊ณผํ์ ์ธ ๋ถ์์ด ์๊ตฌ๋๋ ์ํฉ์์ ์ด์ฉ ๊ฐ๋ฅํ ๊ฐ๊ด์ ํต๊ณ์๋ฃ๋ฅผ ๊ธฐ๋ฐ์ผ๋ก ์ฒด๊ณ์ ์ด๊ณ ์ ๋์ ์ธ ํ๊ฐ๋ชจํ ๋ฐฉ๋ฒ์ ์ ์ํ์๋ค๋๋ฐ ์ผ์ฐจ์ ์๋ฏธ๊ฐ ์๋ค. ๋ํ, ๊ธฐ์ดํต๊ณ ๋ฐ ์ ์ฑ์ ์งํ ์์ฃผ์ ์ ์ฑ
ํ๊ฐ๊ฐ ์ฃผ๋ฅผ ์ด๋ฃจ์๋ ๋์ด์ ์ฑ
๋ถ๋ฌธ์ ์ ๋์ ์ฌํํ๊ฐ๋ชจํ์ ์ ์ฉํ์ฌ ํ๊ฐ๊ฒฐ๊ณผ์ ํต๊ณ์ ํ๋น์ฑ์ ์ ๊ณ ํ์๋ค๋ ์ ์์ ์์๊ฐ ์๋ค.Chapter 1. Introduction 1
1.1. Recent Trends in Rural Korea 1
1.2. Towards Evidence-based Policymaking in the Rural Sector 6
1.3. Purpose and Scope of the Research 8
1.4. Structure of the Research 10
Chapter 2. Koreas Rural Development Policies 13
2.1. A Chronological Overview 13
2.2. The Comprehensive Rural Village Development Project 17
Chapter 3. The Impact of the Comprehensive Rural Village Development Project on Agricultural Income 21
3.1. Introduction 21
3.2. Methodological Challenges in Counterfactual Analysis 22
3.3. Methodology 24
3.4. Data and Variables 34
3.5. Empirical Results 37
3.6. Conclusion 54
Chapter 4. Decoupling the Impact of the Comprehensive Rural Village Development Project on Agricultural Income by Birth and Experience Cohorts 57
4.1. Introduction 57
4.2. Literature Review 58
4.3. Conceptual Framework 63
4.4. Theoretical Framework: The Double Cohort Model 67
4.5. Data and Variables 70
4.6. Methodology 73
4.7. Empirical Results 79
4.8. Conclusion 91
Chapter 5. Micro- and Macro-Level Investigations of the Impact of Transportation Infrastructure on Agricultural Income, 2005-2015 93
5.1. Introduction 93
5.2. Literature Review 95
5.3. Data and Variables 102
5.4. Methodology 107
5.5. Empirical Results 112
5.6. Conclusion 127
Chapter 6. Concluding Remarks 129
6.1. Summary of Findings and Policy Implications 129
6.2. Limitations of the Studies and Future Research 135
Bibliography 138
Appendix 154
Abstract in Korean 158Docto
Determination of goitrogenic metabolites in the serum of male Wistar Rat fed structurally different glucosinolates.
ํ์๋
ผ๋ฌธ (์์ฌ)-- ์์ธ๋ํ๊ต ๋ํ์ : ์ํ์์ํ๊ณผ, 2014. 8. ๊ถํ์ .๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ (glucosinolate)๋ ์ญ์ํ๊ณผ ์๋ฌผ์ ๋ค๋ ํจ์ ๋์ด ์๋ ์ด์ฐจ๋์ฌ์ฐ๋ฌผ๋ก ๊ฐ์์ ์ ๋ถ์ ์ ์ธ ์ํฅ์ ๋ฏธ์น๋ ๊ฒ์ผ๋ก ์๋ ค์ ธ ์๋ค. ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ์ ์ฒด๋ด ํก์์ ๊ดํ ์ฐ๊ตฌ๋ฅผ ์ดํด๋ณด๋ฉด ๋๋ถ๋ถ์ด organic isothiocyanates๋ฅผ ๊ฒฝ๊ตฌํฌ์ฌ ํ๊ฑฐ๋, ์๋ฌผ์ฒด ๋ด myrosinase์ ํจ๊ป ๋ค์ํ ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ๊ฐ ํฌํจ๋ ์๋ฌผ์ฒด๋ฅผ ํฌ์ฌํ์๋ค. ๋ฐ๋ผ์ ๊ฐ๋ณ์ ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ์ ํก์ ๋ฐ ๋์ฌ๋ฅผ ์์ธกํ๋๋ฐ ์ด๋ ค์์ด ์๋ค. ๋ณธ ์ฐ๊ตฌ์์๋ ๋ซ๋ (rat)์ ์์ํ๊ฒ ๋ถ๋ฆฌํ 4์ข
์ ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ (sinigrin, progoitrin, glucoerucin, glucotropaeolin)๋ฅผ ๊ฒฝ๊ตฌํฌ์ฌ ํ์์ผ๋ฉฐ, ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ๊ฐ ๋ฐ๊ฐ์์ ์ํฅ์ ๋ฏธ์น๋ ๋ถํด์ฐ๋ฌผ๋ก ๋์ฌ๋๋์ง ํ์ธํ๊ธฐ ์ํด thiocyanate ion, cyanide ion, organic isothiocyanates, organic nitriles, organic thiocyanates์ ๋๋๋ฅผ ํ์ฒญ์์ ์ธก์ ํ์๋ค. Thiocyanate ion์ ๋น์๋ฒ์ผ๋ก ์ธก์ ํ์์ผ๋ฉฐ, cyanide ion์ CI-GC-MS๋ฅผ ์ฌ์ฉํ์ฌ ์ธก์ ํ์๋ค. Organic isothiocyanates์ ๊ทธ ๋์ฌ์ฒด๋ cyclocondensation assay๋ก ์ธก์ ํ์๋ค. Organic nitriles์ organic thiocyanates๋ EI-GC-MS๋ฅผ ์ฌ์ฉํ์ฌ ์ธก์ ํ์๋ค. Progoitrin๊ตฐ์ ์ ์ธํ ๋ชจ๋ ๊ตฐ์์ ๋์ฌ์ฐ๋ฌผ ์ค thiocyanate ion์ ๋๋๊ฐ ๊ฐ์ฅ ๋๊ฒ ์ธก์ ๋์๋ค. Progoitrin๊ตฐ์์๋ organic isothiocyanates (goitrin)๊ฐ ๊ฐ์ฅ ๋์ ๋๋๋ก ๊ฒ์ถ๋์๋ค. Glucoerucin๊ตฐ์์๋ ๊ฐ์์ ์ข
๋ฐ์์ ์ํฅ์ ๋ฏธ์น๋ ๋ฌผ์ง์ ์์ฑ์ด ์๋์ ์ผ๋ก ์ ์ ํธ์ด์๋ค. ๋ณธ ์ฐ๊ตฌ์์ ์ธก์ ํ ํฌ์ฌ 5์๊ฐ ๋ด ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ๋ ๋๋ถ๋ถ ์ต์ข
๋ถํด์ฐ๋ฌผ์ธ thiocyanate ion์ผ๋ก ๋์ฌ๋๋ ๊ฒ์ด ํ์ธ๋์์ผ๋ฉฐ, ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ ๊ณ์ฌ์ฌ ๊ตฌ์กฐ์ ๋ฐ๋ผ thiocyanate ion ์์ฑ๋์ด ๋ค์ํ ๊ฒ์ผ๋ก ๊ด์ฐฐ๋์๋ค. ๊ณ์ฌ์ฌ ๊ตฌ์กฐ๊ฐ ์์ ํ ํ์ ์์ด์จ์ ํ์ฑํ ์ ์๋ ๊ฒฝ์ฐ์ ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ์ ๋ถํด๊ฐ ์ด์ง๋์ด ๊ฐ์์ ์ข
๋ฐ์์ ์ํฅ์ ๋ฏธ์น๋ thiocyanate ion์ ์์ฑ ๋ํ ์ฆ๊ฐํ๋ ๊ฒ์ผ๋ก ๋ณด์ธ๋ค. ๋ฟ๋ง ์๋๋ผ ์๋ฌผ์ฒด ๋ด myrosinase์ ์์ฉ ์์ด ๊ฐ์์ ์ข
๋ฐ์์ ์ํฅ์ ๋ฏธ์น๋ ๋์ฌ์ฒด๊ฐ ํ์ฑ๋ ์ ์์ผ๋ฏ๋ก, ์กฐ๋ฆฌ ๊ณผ์ ์์ ๋ฐ์ํ๋ ์๋ฌผ์ฒด ๋ด myrosinase ๋ถํ์ฑํ๋ ์ญ์ํ๊ณผ ์๋ฌผ์ ํฌํจ๋ ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ์ ๋ฐ์์์ ํจ๊ณผ์ ์ํฅ์ ์ฃผ์ง ์์ ๊ฒ์ผ๋ก ์ฌ๋ฃ๋๋ค. ํนํ sinigrin๊ณผ progoitrin์ ๊ฒฝ์ฐ ๊ตญ๋ด ์ญ์ทจ๋์ด ๋์ ํธ์ธ๋ฐ ๊ฐ๊ณต์กฐ๊ฑด์ ๊ฑฐ์น ํ ์ญ์ทจ ์ ์ฌ์ ํ ๊ฐ์์ ์ข
๋ฐ์ ๋ฌผ์ง์ ์์ฑ ๊ฐ๋ฅ์ฑ์ด ์๋นํ๋ฏ๋ก, ์ด๋ค์ ์ํ ๋ฐ๊ฐ์์ ์ํฅ์ ์ค์ด๊ธฐ ์ํด ๊ฐ๊ณต ๋ฐ ์กฐ๋ฆฌ๋ฒ ๊ฐ์ ์ ๋ํ ์ฐ๊ตฌ๊ฐ ํ์ํ ๊ฒ์ด๋ค.Glucosinolates (GLSs) are abundant in cruciferous vegetables and reported to have anti thyroidal effects. Four GLSs (sinigrin, progoitrin, glucoerucin, and glucotropaeolin) were administered orally to rats, and the breakdown products of GLSs (GLS-BPs: thiocyanate ions, cyanide ions, organic isothiocyanates, organic nitriles, and organic thiocyanates) were measured in serum. Thiocyanate ions were measured by colorimetric method, and cyanide ions were measured with CI-GC-MS. Organic isothiocyanates and their metabolites were measured with the cyclocondensation assay. Organic nitriles and organic thiocyanates were measured with EI-GC-MS. In all treatment groups except for progoitrin, thiocyanate ions were the highest among the five GLS-BPs. In the progoitrin treated group, a high concentration of organic isothiocyanates (goitrin) was detected. In the glucoerucin treated group, a relatively low amount of goitrogenic substances was observed. The metabolism to thiocyanate ions happened within five hours of the administration, and the distribution of GLSs varied with the side chain. GLSs with side chains that can form stable carbocation seemed to facilitate the degradation reaction and produce a large amount of goitrogenic thiocyanate ions. Because goitrogenic metabolites can be formed without myrosinase, the inactivation of myrosinase during cooking would have no effect on the anti-nutritional effect of GLSs in cruciferous vegetables.๊ตญ๋ฌธ์ด๋ก
๋ชฉ์ฐจ
List of tables
List of figures
โ
. ์๋ก
โ
ก. ๋ฌธํ๊ณ ์ฐฐ
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ก.1. ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ์ ํน์ฑ
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ก.2. ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ ๋ถํด์ฐ๋ฌผ์ ์์ฑ
โ
ก.3. ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ์ ์ฒด๋ด ๋ถํด ๋ฐ ํก์
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ก.4. ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ์ ์ฒด๋ด ํจ๊ณผ
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ข. ์ฌ๋ฃ ๋ฐ ๋ฐฉ๋ฒ
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ข.1. ์คํ ์ฌ๋ฃ
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ข.1.1. ์๋ฃ
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ข.1.2. ์์ฝ ๋ฐ ๊ธฐ๊ธฐ
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ข.2. ์คํ ๋ฐฉ๋ฒ
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ข.2.1. ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ์ ์ถ์ถ ๋ฐ ๋ถ๋ฆฌ
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ข.2.2. ๋๋ฌผ์คํ
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ข.2.3. ํ์ฒญ ๋ด ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ ๋์ฌ์ฐ๋ฌผ ๋ถ์
โ
ข.2.3.1. Thiocyanate ion ์ ๋ ๋ถ์
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ข.2.3.2. Cyanide ion ์ ๋ ๋ถ์
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ข.2.3.3. Organic isothiocyanates ์ ๋ ๋ถ์
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ข.2.3.4. Organic nitriles, organic thiocyanates ์ ๋ ๋ถ์
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ข.2.4. ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ ํฌ์ฌ์ ๋ฐ๋ฅธ ๊ฐ์์ ํธ๋ฅด๋ชฌ ๋ณํ
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ข.2.5. ํ์ก ์ํํ์ ๊ฒ์ฌ
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ข.2.6. ํต๊ณ๋ถ์
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ฃ. ๊ฒฐ๊ณผ ๋ฐ ๊ณ ์ฐฐ
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ฃ.1. ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ์ ์ถ์ถ ๋ฐ ๋ถ๋ฆฌ
โ
ฃ.2. ํ์ฒญ ๋ด ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ ๋์ฌ์ฐ๋ฌผ ๋ถ์
โ
ฃ.3. ๊ธ๋ฃจ์ฝ์๋๋ ์ดํธ ํฌ์ฌ์ ๋ฐ๋ฅธ ๊ฐ์์ ํธ๋ฅด๋ชฌ ๋ณํ
โ
ฃ.4. ์ฅ๊ธฐ ์ค๋ ์ธก์ ๋ฐ ํ์ก ์ํํ์ ๊ฒ์ฌ
โ
ค. ์์ฝ ๋ฐ ๊ฒฐ๋ก
โ
ฅ. ์ฐธ๊ณ ๋ฌธํ
AbstractMaste
์ฌ์กฐํฉ ํจ๋ชจ๋ฅผ ์ด์ฉํ ํฌ๋๋น๊ณผ ๊ฐ๋ฝํ ์ค๋ก๋ถํฐ 2,3-butanediol ์์ฐ
ํ์๋
ผ๋ฌธ (์์ฌ)-- ์์ธ๋ํ๊ต ๋ํ์ : ํ๋๊ณผ์ ๋ฐ์ด์ค์์ง๋์ด๋ง์ ๊ณต, 2016. 2. ์์งํธ.2,3-Butanediol (2,3-BD)๋ ์ฐ์
์ ํ์ฉ๋๊ฐ ๋์ ํ๋ซํผ ํํ ์์ฌ์ด๋ค. 2,3-BD๋ ํํ์ ์์ฐ์ด ๊ฐ๋ฅํ์ง๋ง ์ ๊ฐ ๋ณ๋๋ฌธ์ ์ ์ง๊ตฌ์จ๋ํ ๋ฌธ์ ๋ก ์ธํด ์ต๊ทผ ๋ฐ์ด์ค ๊ธฐ์ ์ด ๊ฐ๋ฐ๋จ์ ๋ฐ๋ผ ์๋ฌผ๊ณตํ์ ์ผ๋ก ์์ฐ์ด ์ฃผ๋ชฉ ๋ฐ๊ณ ์๋ค. 2,3-BD์ ์๋ฌผ๊ณตํ์ ์์ฐ์๋ ์ฃผ๋ก ๋ฐํ
๋ฆฌ์๊ฐ ์ฌ์ฉ๋๋๋ฐ ๊ณ ์์จ๋ก 2,3-BD๋ฅผ ์์ฐํ ์ ์์ง๋ง ์ด๋ค ๋๋ถ๋ถ์ด ๋ณ์์ฑ ๋ฐํ
๋ฆฌ์๋ก ๋ถ๋ฅ๋๊ธฐ ๋๋ฌธ์ ์์ ๊ณผ ์ฐ์
ํ ์ธก๋ฉด์์ ๋๋ ์์ฐ ๊ณต์ ๊ตฌ์ถ์ด ์ด๋ ต๋ค. ๊ทธ ๋์์ผ๋ก GRAS (Generally Recognized As Safety) ๋ฏธ์๋ฌผ๋ก์ ์์ ํ๋ค๊ณ ์๋ ค์ง Saccharomyces cerevisiae๋ฅผ ์ด์ฉํ 2,3-BD ์์ฐ์ ์ฃผ๋ชฉํ ํ์๊ฐ ์๋ค. ํ์ง๋ง S. cerevisiae๋ ์์ฐ ์ํ์์๋ 2,3-BD๋ฅผ ๊ฑฐ์ ์์ฐํ์ง ๋ชปํ๊ณ ์ํ์ฌ์ ์ฃผ๋ก ์์ฐํ๊ธฐ์ ๊ณ ํจ์จ์ 2,3-BD๋ฅผ ์์ฐํ๊ธฐ ์ํด์๋ ๋์ฌ๊ณตํ์ ๋ฐฉ๋ฒ์ ์ด์ฉํ ์ฌ์กฐํฉ S. cerevisiae์ ๊ตฌ์ถ์ด ์๊ตฌ๋๋ค.
๋ํ, ๊ฒฝ์ ์ ์ธก๋ฉด์ผ๋ก ๋ณผ ๋ ํด์ ๋ฐ์ด์ค๋งค์ค๋ก๋ถํฐ 2,3-BD๋ฅผ ์์ฐํ๋ ๊ฒ์ด 2,3-BD์ ์ง์ ๊ฐ๋ฅํ ์์
ํ๋ฅผ ์ํด ํด๊ฒฐํด์ผ ํ ์ฃผ์ ์ฐ๊ตฌ ๊ณผ์ ์ด๋ค. ํ์ง๋ง S. cerevisiae๋ ํด์ ๋ฐ์ด์ค๋งค์ค์ ๋ค๋ ์กด์ฌํ๋ ํฌ๋๋น๊ณผ ๊ฐ๋ฝํ ์ค์ค๋ฅผ ๋์์ ์๋ชจํ์ง ๋ชปํ๊ณ , ํฌ๋๋น์ด ๋ชจ๋ ์๋ชจ๋ ํ์์ผ ๊ฐ๋ฝํ ์ค์ค๊ฐ ์ด์ฉ๋๋ ํฌ๋๋น์ ์ํ ์ ์ฌ ์ต์ (catabolite repression)์ ๋จ์ ์ ๊ฐ์ง๋ค.
์ ํ์ฐ๊ตฌ์์ ์ํด S. cerevisiae์ ์ฃผ์ ๋์ฌ ์ฐ๋ฌผ์ธ ์ํ์ฌ ์์ฑ์ ์ต์ ํ๊ณ ์ pyruvate decarboxylase ํ์ฑ์ด ์์ ํ ์ ํด๋ ํจ๋ชจ (SOS2)์, SOS2 ๊ท ์ฃผ๋ฅผ ํฌ๋๋น ๋ฐฐ์ง์์ ์์ฅ์ด ๊ฐ๋ฅํ๋๋ก ํ๊ธฐ ์ํด ์ด๋ณผ๋ฃจ์
์ํจ ํจ๋ชจ (SOS4)๋ฅผ ์ ์ํ ๋ฐ ์๋ค. ์ด๋ณผ๋ฃจ์
์ ๊ณผ์ ์์ SOS4 ๊ท ์ฃผ๋ S. cerevisiae ๋ด ํฌ๋๋น signal regulator์ ์ญํ ์ ํ๋ MTH1 ์ ์ ์์ ์๋ฏธ๋
ธ์ฐ ์ผ๊ธฐ์์ด์ ๋์ฐ๋ณ์ด๊ฐ ๋ฐ์ํ์๋ค.
๋ณธ ์ฐ๊ตฌ์์๋ ํด์ ๋ฐ์ด์ค๋งค์ค์ ๋ค๋ ์กด์ฌํ๋ ํฌ๋๋น๊ณผ ๊ฐ๋ฝํ ์ค์ค๋ฅผ ํจ์จ์ ์ผ๋ก ์ด์ฉํ๋๋ก ํ์ฌ, 2,3-BD์ ์์ฐ์ ์์ด ๋์ ์์จ๊ณผ ์์ฐ์ฑ์ ์ป๋ ๊ฒ์ด ์ต์ข
๋ชฉํ์ด๋ค. ๋จผ์ , pyruvate decarboxylase ํ์ฑ์ด ์ ํด๋๊ณ ์ด๋ณผ๋ฃจ์
๋์ด MTH1 ์ ์ ์์ ๋์ฐ๋ณ์ด๊ฐ ์กด์ฌํ๋ ๊ท ์ฃผ (SOS4)์ 2,3-BD ์ํฉ์ฑ ๊ฒฝ๋ก๋ฅผ ๋์
ํ ๊ท ์ฃผ (BD4)์ ํ๋ถ์ ๋ฐฐ์์ ํตํด ํฌ๋๋น๊ณผ ๊ฐ๋ฝํ ์ค์ค์ ๋์ ์๋ชจ๋ฅผ ํ์ธํ์๋ค. ๋ฐ๋ผ์ ์ด ๊ฒฐ๊ณผ๊ฐ MTH1 ์ ์ ์์ ๋์ฐ๋ณ์ด์ ์ํ ํจ๊ณผ์ธ์ง๋ฅผ ํ์ธํ๊ณ ์ ์ด๋ณผ๋ฃจ์
์ด ์ผ์ด๋์ง ์์ ๊ท ์ฃผ (SOS2)์ ์ธ์์ ์ผ๋ก MTH1 ์ ์ ์์ ๋์ฐ๋ณ์ด๋ฅผ ๊ฐํ์ฌ ๊ตฌ์ถํ ๊ท ์ฃผ (SOS2_Mth1)๋ฅผ ๋ณธ ์ฐ๊ตฌ์ ์ฌ์ฉํ์๋ค. ์ด ๋ ๊ท ์ฃผ์ 2,3-BD ์ํฉ์ฑ ๊ฒฝ๋ก๋ฅผ ๋์
ํ ๊ท ์ฃผ๋ฅผ ๊ตฌ์ถํ์๊ณ , ํ๋ถ์ ๋ฐฐ์ ๊ฒฐ๊ณผ MTH1์ ๋์ฐ๋ณ์ด๊ฐ ์กด์ฌํ๋ ๊ท ์ฃผ (BD2M)๊ฐ ํฌ๋๋น๊ณผ ๊ฐ๋ฝํ ์ค์ค๋ฅผ ๋์์ ์๋ชจํจ์ผ๋ก์จ MTH1 ์์ผ๋ํ์
๊ท ์ฃผ (BD2)์ ๋นํ์ฌ 2,3-BD์ ์์ฐ์ฑ์ด ์ฝ 41% ์ฆ๊ฐํ ์์น๋ฅผ ๋ณด์๋ค. ์ด ๊ท ์ฃผ์ ์ ํํ ๋ฐํจ๋ฅ๋ ฅ์ ๋ณด๊ธฐ ์ํ์ฌ fed-batch fermentation์ ์ํํ์๊ณ , ๋ง์ฐฌ๊ฐ์ง๋ก ๋ ๋น์ ๋์ ์๋ชจํ์ฌ ์ฝ 75.5 g/L์ 2,3-BD๋ฅผ ์์ฐํ์์ผ๋ ๋ฐํจ ํ๋ฐ๋ถ๋ก ๊ฐ์๋ก ๊ฐ๋ฝํ ์ค์ค์ ์๋ชจ ์๋๊ฐ ๋๋ ค์ง๋ ์์์ ํ์ธํ ์ ์์๋ค.
๋ ๋ฒ์งธ๋ก, ๊ฐ๋ฝํ ์ค์ค ์๋ชจ ์๋๋ฅผ ์ฆ์ง์ํค๊ธฐ ์ํด ๊ฐ๋ฝํ ์ค์ค ๋์ฌ๊ฒฝ๋ก ๋ด์ ์กด์ฌํ๋ 5๊ฐ์ ์ ์ ์๋ฅผ ๊ฐ๊ฐ ๊ณผ๋ฐํ ์์ผ ๊ท ์ฃผ๋ฅผ ๊ตฌ์ถํ์๋ค. ์ด๋ค์ ๊ฐ์ง๊ณ ํ๋ถ์ ๋ฐฐ์์ ์งํํ์๊ณ , phosphoglucomutase๋ฅผ ์ํธํํ๋ PGM2 ์ ์ ์๋ฅผ ๊ณผ๋ฐํํ ๊ท ์ฃผ (BD2M_PGM2)์์๋ง ์์ผ๋ํ์
๊ท ์ฃผ (BD2M)์ ๋นํ์ฌ ์ฝ 13% ์ฆ๊ฐํ ๊ฐ๋ฝํ ์ค์ค ์๋ชจ ์๋๋ฅผ ๋ํ๋ด์๋ค. ์ด ๊ท ์ฃผ์ ์ ํํ ๋ฐํจ๋ฅ๋ ฅ์ ๋ณด๊ธฐ ์ํ์ฌ fed-batch fermentation์ ์ํํ์๊ณ , ์์ผ๋ํ์
๊ท ์ฃผ์ ๋นํด ์ฝ 42% ์ฆ๊ฐํ ๊ฐ๋ฝํ ์ค์ค ์๋ชจ ์๋๋ฅผ ํตํด ์ฝ 39% ์ฆ๊ฐํ 2,3-BD์ ์์ฐ์ฑ์ ํ์ธํ ์ ์์๋ค.
์ธ ๋ฒ์งธ๋ก, pyruvate decarboxylase ๊ฒฐ์ฌ ํจ๋ชจ์ ๋จ์ ์ธ ํ์์์ ์์กด์ ์ธ ์์ฅ ์ฒด๊ณ๋ฅผ ๊ทน๋ณตํ๊ณ ์ ์ ํ์ฐ๊ตฌ์์ ์ํด ํ์ธ๋ Candida tropicalis ์ ๋์ pdc1 ์ ์ ์๋ฅผ ๋์
ํ๊ณ , ๋ณดํจ์ ๋ถ๊ท ํ์ผ๋ก ์ธํด ๋ถ์ฐ๋ฌผ๋ก ๊ธ๋ฆฌ์ธ๋กค์ด ๋ค๋ ์ถ์ ๋๋ ๋ฌธ์ ์ ์ ๋ณด์ํ๊ณ ์ ์ ํ์ฐ๊ตฌ์ ์ํด ํ์ธ๋ Lactococcus lactis ์ ๋์ noxE ์ ์ ์๋ฅผ ๋์
ํ ๊ท ์ฃผ๋ฅผ ๊ตฌ์ถํ์๋ค. ์ด์ ๋ฐฐ์์ ์กฐ๊ฑด๊ณผ๋ ๋ฌ๋ฆฌ ์ด ๊ท ์ฃผ (BD2M_PGM2_Ctnox)์ ๋ฐฐ์์์๋ ์ธ๋ถ ํ์์์ผ๋ก ์ํ์ฌ์ ๊ณต๊ธํ์ง ์๊ณ ์งํํ์๋ค. ํ๋ถ์ ๋ฐฐ์ ๊ฒฐ๊ณผ, ์์ผ๋ํ์
๊ท ์ฃผ (BD2M_PGM2)์ ๋นํ์ฌ ์ฝ 19% ๊ฐ์ํ ๊ธ๋ฆฌ์ธ๋กค์ ์์จ์ ํ์ธํ ์ ์์์ผ๋ฉฐ, ํ์์์ ๋น์์กด์ ์ธ ์์ฅ์ด ๊ฐ๋ฅํด์ง์ ํ์ธํ์๋ค. ์ด ๊ท ์ฃผ์ ์ ํํ ๋ฐํจ๋ฅ๋ ฅ์ ๋ณด๊ธฐ ์ํ์ฌ fed-batch fermentation์ ์ํํ์๊ณ , ์ต์ข
์ ์ผ๋ก ์ฝ 115.7 g/L์ 2,3-BD๋ฅผ ์์ฐํ์์ผ๋ฉฐ ๋ถ์ฐ๋ฌผ์ธ ๊ธ๋ฆฌ์ธ๋กค์ ์์จ์ด 0.08 gglycerol/gsugars๋ก ๋งค์ฐ ๋ฎ์ ์์น๋ฅผ ๋ณด์๋ค. ๋ณธ ๋ฐํจ์์์ 2,3-BD ์์ฐ ์์จ์ 0.47 g2,3-BD/gsugars, 2,3-BD ์์ฐ์ฑ์ 1.75 g/L/h๋ฅผ ๋ํ๋ด์๋ค.
์ด๋ฅผ ํตํ์ฌ ์ฌ์กฐํฉ S. cerevisiae์ธ BD2M_PGM2_Ctnox ๊ท ์ฃผ๋ ํด์ ๋ฐ์ด์ค๋งค์ค๋ก๋ถํฐ 2,3-BD๋ฅผ ํจ์จ์ ์ผ๋ก ์์ฐํ ์ ์๋ ๊ท ์ฃผ์์ ์ฆ๋ช
ํ์๋ค.
์ฃผ์์ด : 2,3-Butanediol, pyruvate decarboxylase, ๊ฐ๋ฝํ ์ค์ค, ํด์ ๋ฐ์ด์ค๋งค์ค, ํผํฉ๋น ๋ฐํจ, mth1, phosphoglucomutase, ํฌ๋๋น์ ์ํ ์ ์ฌ์ต์ , ๋ณดํจ์ ๋ถ๊ท ํ2,3-Butanediol (2,3-BD) is a platform chemical with extensive industrial applications since it can be converted into other valuable chemicals by dehydration, dehydrogenation, ketalization and esterification. Especially, 1,3-butadiene, a dehydration product of 2,3-BD, is a main substance used for producing synthetic rubber. Most microbial fermentations for 2,3-BD production have been focused on pathogenic bacteria, which makes large-scale fermentations difficult in terms of safety and industrialization. As an alternative, 2,3-BD production by a GRAS (Generally Regarded As Safe) microorganism Saccharomyces cerevisiae would be suitable.
Also, production of 2,3-BD from marine biomass is one of the key issues for economic viability of bio-based chemicals. Among various sugars, glucose and galactose are of special interest because they are abundant in marine biomass. One of the most important concerns in mixed sugar fermentations is the catabolite repression that represses the enzyme expression to utilize sugars other than glucose.
Pyruvate decarboxylase(Pdc)-deficient S. cerevisiae (SOS2) was constructed to eliminate ethanol production. Then, the evolved SOS2 strain (SOS4) was obtained by serial cultivation in excess glucose medium for cell growth on glucose medium. Among the chromosomal mutations of the SOS4 strain, a single nucleotide polymorphism was found on the MTH1 gene which functions as a glucose signal regulator.
Efficient 2,3-BD production using engineered strains capable of effective fermentation from glucose and galactose is the objective in this study.
First, the 2,3-BD-producing Pdc-deficient BD4 strain with the MTH1 mutation was tested for a performance of 2,3-BD production in a batch fermentation, resulting in simultaneous utilization of glucose and galactose. Therefore, in order to evaluate if the MTH1 mutation affects the fermentation performances in a mixture of glucose and galactose, the evolved pdc-deficient strain (SOS2) and engineered strain by a point mutation on the MTH1 gene (SOS2_Mth1) were used. The SOS2 and SOS2_Mth1 strains containing the alsS gene encoding ฮฑ-acetolactate synthase and the alsD gene encoding ฮฑ-acetolactate decarboxylase both from Bacillus subtilis and overexpression of the endogenous BDH1 gene coding for 2,3-BD dehydrogenase were constructed (BD2 and BD2M) and tested for a performance of 2,3-BD production in batch fermentation under oxygen-limited conditions. The BD2M strain which is the MTH1 mutant was able to co-ferment both sugars, resulting in increased 2,3-BD productivity by 41% compared to the control strain, BD2 strain. To evaluate the fermentation aspects of the BD2M strain in a bioreactor, fed-batch fermentation was carried out to obtain 75.5 g/L of 2,3-BD by simultaneous consumption of glucose and galactose. However, in the rest of the fermentation, galactose consumption rate was substantially reduced compared with glucose consumption rate.
Second, to increase galactose uptake rate in 2,3-BD production, the GAL10, GAL1, GAL7, PGM1 and PGM2 genes involved in the Leloir pathway were overexpressed into the BD2M strain. These five consutructed strains were tested for a performance of galactose consumption in batch fermentation under oxygen-limited conditions. Only the strain for overexpression of the PGM2 gene, the BD2M_PGM2 strain obtained 13% increased galactose uptake rate. Also, the BD2M_PGM2 strain resulted in 42% increased galactose uptake rate and 39% improved 2,3-BD productivity compared to the control strain, BD2M strain in a fed-batch fermentation.
Finally, to decrease the accumulation of glycerol and to solve the C2-dependent growth in 2,3-BD-producing Pdc-deficient S. cerevisiae, the NADH oxidase (noxE) gene from Lactococcus lactis and the pyruvate decarboxylase 1 (pdc1) gene from Candida tropicalis were expressed in the BD2M_PGM2 strain. To test for a performance of 2,3-BD production, batch and fed-batch fermentation was carried out. Unlike other conditions of fermentations in this study, this cultivation was conducted without addition of ethanol due to expression of the pdc1 gene. The resulting strain (BD2M_PGM2_Ctnox) obtained 19% reduced yield of glycerol and was able to grow without addition of ethanol in a batch fermentation. Also, in a fed-batch fermentation, the resulting strain produced 116 g/L of 2,3-BD from glucose and galactose with a low glycerol yield (0.08 gglyerol/gsugars) with 0.47 g2,3-BD/gsugars of yield and 1.75 g/L/h of productivity in a fed-batch fermentation. These results suggested that the BD2M_PGM2_Ctnox strain is suitable for producing 2,3-BD from marine biomass for industrial applications
Keywords : 2,3-butanediol, pyruvate decarboxylase (Pdc), galactose, marine biomass, mixed sugar fermentation, mth1, phosphoglucomutase (Pgm2), catabolite repression, redox balanceI. INTRODUCTION 1
1. 2,3-Butanediol 1
2. Marine biomass as feedstock 4
3. Galactose metabolism in S. cerevisiae in glucose and galactose mixed fermentation 6
4. 2,3-Butanediol production using bacteria 10
5. 2,3-Butanediol production using S. cerevisiae 14
6. 2,3-Butanediol production using pyruvate decarboxylase deficient S. cerevisiae 17
7. Research objectives 24
II. MATERIALS AND METHODS 25
1. Reagents 25
2. Strains and plasmids 26
2.1. Strains 26
2.2. Plasmids 29
3. DNA manipulation and transformation 35
3.1. Enzymes 35
3.2. Polymerase chain reaction (PCR) 35
3.3. Preparation of plasmid DNA and bacteria genomic DNA 35
3.4. Transformation of E. coli 36
3.5. Isolation of DNA fragments and DNA sequencing 37
3.6.Yeast transformation 37
4. Media and culture conditions 38
4.1. Media 38
4.2. Batch fermentations 38
4.3. Fed-batch fermentations 39
5. Analysis 41
5.1 Dry cell weight 41
5.2 Metabolite detection 41
III. RESULTS AND DISCUSSIONS 42
1. Construction of the 2,3-BD producing Pdc-deficient S. cerevisiae strain by simultaneous consumption of glucose and galactose 42
1.1. Confirmation of the effect of mutant MTH1 on co-consumption of glucose and galactose 42
1.2. Influence of glucose and galactose ratio on 2,3-BD fermentation by the BD2M strains 48
1.3. Fed-batch fermentation of the BD2M strain in glucose and galactose mixed sugars 51
2. Overexpression of galactose metabolic enzymes to increase galactose uptake rate in 2,3-BD poroduction 54
3. Expression of L. lactis noxE and C. tropicalis pdc1 in the Pdc-deficient S. cerevisiae 60
IV. CONCLUSIONS 67
V. REFERENCES 69
๊ตญ ๋ฌธ ์ด ๋ก 80Maste
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ผ๋ฌธ(์์ฌ)--์์ธ๋ํ๊ต ํ๊ฒฝ๋ํ์ :ํ๊ฒฝ๊ณํํ๊ณผ ๋์ ๋ฐ ์ง์ญ๊ณํ์ ๊ณต,2001.Maste
Ag/C3N4 ๊ด์ด๋งค๋ฅผ ์ด์ฉํ ๋ฉํธ๋ฉ๋ฅด์บ ํ์ ์ ํ์ ์ ๊ฑฐ ๋ฐ ๋ฉ์ปค๋์ฆ ๊ท๋ช
MasterIn general, air pollutants such as volatile organic compounds (VOCs) are not degraded by the CN based catalysts generating O2- radicals, but only by WO3 based catalysts generating OH radicals. In this study, we found that CN based catalysts can selectively degrade methyl mercaptan, which is a typical odorous compound, under visible light. Modification of carbon nitride with silver nanoparticles significantly enhanced the adsorption capacity for CH3SH using interaction between Ag and sulfur atoms. Additionally, deposited Ag nanoparticles as co-catalyst increased the photoconversion efficiency and production of reactive radical species. On the basis of the control test of oxygen and water in the carrier gas and EPR analysis, it was revealed that the responsible reactive species for CH3SH decomposition was O2- radical. Mass balance of
removed sulfur atoms and FT-IR analysis showed that Ag/CN degraded methyl mercaptan to gaseous dimethyl disulfide (CH3SSCH3) and sulfate deposited on the surface of catalyst. Thus, Ag/CN is proposed as an effective and appropriate photocatalyst to remove CH3SH under visible light
A discussion for directions of a Korean Textbook for Listening skills based on the spoken grammar
Correlations in Appearance, Job-seeking Stress and Job-seeking Anxiety in Undergraduates according to Gender Differences
Purpose: The purpose of this study was to evaluate the differences between the level of satisfaction and concern regarding appearance according to gender, the importance of appearance in job interviews, and the influence appearance has on job-seeking stress and anxiety under the ever-increasing pressure on students to secure employment so that appropriate psychological interventions can be provided to undergraduates. Methods: The survey was conducted on 170 third and fourth year undergraduates in Gyeonggi province from August 31st to September 6th, 2012 to figure out the correlations and differences in variables of appearance satisfaction and concern, and the importance of appearance in job interviews and job-seeking stress and anxiety according to the differences in gender. Data were analyzed by descriptive statistics, independent t-test, two-way ANOVA, and Pearsons correlation analysis. Results: 1. There was difference between the genders for appearance satisfaction and concern. The level of satisfaction was higher in male students while concerns regarding appearance were higher in female students. There was no gender difference regarding the importance of appearance in job interviews, job-seeking stress, and job-seeking anxiety. 2. For the concerns in appearance, there was a significant difference between the genders in appearance management, and gender and the importance of appearance in job interviews showed significant interactive effects. Stress and anxiety from job-seeking did not show significant differences between the genders, but there were common significant interactive effects between gender and grades, gender and employment, and gender and the importance of appearance in job interviews. 3. Both male and female students showed higher job-seeking stress as their job-seeking anxiety was higher. Job-seeking anxiety was higher in male students who had higher concern in appearance, and in female students who placed more emphasis on the importance of appearance in job interviews. Conclusion: As female students perceived appearance in job interviews as important more than male students, their job-seeking anxiety was higher, while male students who showed higher concern in appearance also showed higher level of anxiety in job-seeking. As job-seeking anxiety was higher, job-seeking stress was also higher according to the entire participants, based on this result, methods to decrease job-seeking anxiety should be suggested