グリーンバイオ戦略フォーラム　

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2 i

3 1 C2 6 C ii

5 1.2 (JBA) JBA JBA 2007 JBA JKA - 2 -

6 2008 (NED) NED 3 JBA 2008 ( ) JBA JBA 2000 NED 2008 ( ) - 3 -

7 JBA - 4 -

8 - 5 -

9 C2 2.1 ( ) (RITE) CP15 30 (Copenhagen Accord) (Take note) CP ( ) (C2 ) (CH4 ) (N2 ) - 6 -

10 (HFCs) (PFCs) (SF6) 6 C2 ( ) C2 C2 C2 C2 UNFCCC (United Nations Framework Convention on Climate Change) UNFCCC ( ) C2 ( ) C2 C2 C2 C2 ( ) UNFCCC ( ) C2 ( ) ( )

11 C ( ) (Clean Development Mechanism) (Emissions Trading) (Joint Implementation) (Cool Earth 50) ( ) (2020 ) EU CP GDP C2 CP15 CP

12 ( ) CP CP UNFCCC - 9 -

13 JBA () (RITE) 3 12CP15 (Conference of the Parties) 30 (Copenhagen Accord ) (Take note) 1997CP % ()

14 () (C 2 2)( )(CH 4 4)( )(N 2 ) (HFCs)(PFCs) (SF6)6 C 2 ()C ( ) 2 C 2 C 2 C 2 ( UNFCCC(United Nations Framework Convention on Climate Change) UNFCCC( items/3800.php) (2) 7 C 2 UNFCCC () 1 C 2 C 2 6 (1) ( )(C 2 ()C 2 )C 2 () IPCC 8 (3) 2 C 2 C 2 C 2 XX

15 (4) 3 C 2 C 2 ) C 2 (1) 11-6% () ) () (39GtC 2 eq/yr), 4% EU, 13% 27%, 2% NZ, 2% 19906% EU19908% (EU, 27% = EU), 3%, 6%, 19%, 5%, 2%, 18% ) UNFCCC, IEA(International Energy Agency) 19900% (1990 = ) 19907% (2) 12 ) ,

16 () (CDM: Clean Development Mechanism) (ET: Emissions Trading) (JI: Joint Implementation) AB (1) 50Cool Earth 50(20075) 2050GHG (20081)(20086) 2050GHG 2050C26080% 2050C26080% % % (2) % %

17 (1990) 50% 6080% -EU202020%30% % %1990 (CP %) % % %20099 C 2 19 C 2 G8 G G % UNFCCC UNFCCC G % % G () I I EU REDD CDM CCS PEC LDCs ASIS REDDReducing Emissions from Deforestation and Degradation () LDCsLesast Developed Countries () ASISAlliance of Small Island States ()

18 CP % 25% 30% EU %30% 20%30% 14%25% % 3% 17% % 3% 22% 20005%25% +13%11% 11%30% %25% 20%25% +17%+25% 20054% +80% 4% * C 2 /GDP %45% C 2 /GDP2005 * 20%25% +366%+327% +105%+88% +373%+344% +142%+127% * C /2005 RITEGDPC 2 2 C2 (ppm) C2 (ppm () 2050 C2 (00%) C2eq.) () (00%) %) %) I II (GDP %) +5.5 (GDP %) (+)(-) III IV (-04) 0.5 (-12) + V VI IPCC6 IPCC IPCC 23 CP15 30( ) , MRV(// ) CP15 () % 2 () UNFCCC

19 2.2 C2 ( ) (RITE) C C C2 78, C2 6,000 C6 C5 C2 C3 C4 C2-16 -

20 C2 C2 C2 C2 C2 DE C2 1tC2 /t kgc 2 /kgresin LDPE LCI DENEDRITE LDPE tc2 /t 3,016 C2 2,100 2,

21 C2 C2 3,082 C2 2, ( ( ( C2 ( C

22 C ,300 4,900 C2 C C2 25 RITE

23 ) )

24 Lactobacillus paracasei LA1 LA1 L- 2) 60g/L LA1 40 ph 5.0 L- 9.77g/L ph 3) L- 120 / /

25 7.7 / 9.6 kl/ 15 / L- 4) ) Draths, K. M., Frost, J. W., J. Am. Chem. Soc., 113, 9361 (1991). 2) Nakasaki, K. et al., Environ. Sci. Technol., 33, 198 (1999). 3) Nakasaki, K., Adachi, T., Biotechnol. Bioeng., 82, 263 (2003). 4) (2000)

26 Lactobacillus paracasei subsp. paracasei LA1 CH3 ) ( -CH-C

27 - 24 -

28 Air filter 4. Jar fermentor 7. Peristaltic pump 2. Thermocouple 5. ph electrode 8. H2S4 (1N) 3. Temp. controller 6. ph controller 9. Na (5N)

29 - 26 -

30 - 27 -

31 PCR-DGGE DNA lysozyme phenol ethanol RNase chloroform 10%SDS 16S rdna PCR polyacrylamide 6-12% denaturing gradient 0-100% Primer : 357F 517R Denature : 93-30s Annealing : 65,60,55-30s 30 cycles Extension : 72-60s TAcloning PCR DGGE PCRproducts E.coli DGGE PCR DNA Sequence PCRproducts

32 Rhizoctonia solani 2 Micrograph of GM

33 100 CTRL B A(=GM-21) SD-62 C Cultivation time (d) (B) a b c b a a Disease severit ty (%)

34 R. solani 2 2R. solani K1 3F. oxysporum f.sp.melonis F-Me-2 4F. oxysporum f.sp.redicis-lycopersici F-T

35 CP

36 (2-deoxy-scyllo -inosose=di) Bacillus circulans SANK DI DI BtrC DI DI

37 DI DI DI 95 DI DI DI DI DI 99 DI

38 DI Rh/Al % H 3 P 4 39% HI/Ac y. 59% C2 C2 C2 C2 C2 C2 DI 2-deoxy-scyllo-inosose DI HN

Bacillus circulansbutirosin B D-glucose 2- H P 3 H Glucose-6- phosphate p NAD + NADH H 2- P 3 H H DI H

Bacillus circulans DI Glucose Phosphotransferase System H 2- P 3 H D-glucose Glucose-6- phosphate 2- P

coli RNA polymerase H 2-deoxy-scyllo-inosose IPTG HexokineseBtrC DI D-glucose Hexokinase (ATP, Mg ++ )

39 Bacillus circulansbutirosin B D-glucose 2- H P 3 H Glucose-6- phosphate p NAD + NADH H 2- P 3 H H DI H BtrC) H 2-deoxy-scyllo-inosose H btra btrb btrc btrd btre H2N DI(btrC) NH 2 H2N NH butirosin B NH 2 Bacillus circulans DI Glucose Phosphotransferase System H 2- P 3 H D-glucose Glucose-6- phosphate 2- P 3 H NAD + NADH DI BtrC) H H T7 promoter T7 RNA polymerase btrc H lac Lac promoter o T7 gene 1 E. coli RNA polymerase H 2-deoxy-scyllo-inosose IPTG HexokineseBtrC DI D-glucose Hexokinase (ATP, Mg ++ ) H H BtrC (NAD +, Co ++ ) P 2-3 Glucose-6-phosphate 2-deoxy-scyllo-inosose High cost ne-pot : y. 78% DI DI DI DI

40 DI rich operator DI -ci Repressor DI 2YT M Trp promoter (h) Transcription ci Repressor No transcription operator DI Molasses-CSL Tryptophan M Trp promoter DI Transcription (h) No transcription ci Repressor DI synthase btrc gene btrc gene operator P L promoter operator P L promoter Negative element Positive element gapa promoter gada promoter TT btrc btrc TT (h) M pgap-btrc/pgad-btrc DI puc ori Ampicillin SDSPAGE CBB gapa (Glycelaldehyde-3-phosphate dehydrogenase A) gada (Glutamic acid decarboxylase A) P L promoter gada promoter btrc btrc Terminator Terminator 2YT Molasses-CSL P L -system P L -gada-system P L -system P L -gada-system DI DI DI glycogen biosynthesis glucose-1-p pgm glucose glucose-6-p pgi fructose-6-p glucose PTS zwf -6-6-P-gluconolactone Pentose phosphate pathway fructose-1, 6-P2 fdaa, fdab dihydroxyacetone-p glyceraldehyde-3-p tpia (5 rxns) pyruvate TCA cycle

41 WT (D660) Glucose-6-phosphate (mol/ g dry cell ) pgi pgm zwf 40 pgipgm ( (h) (h) pgizwf pgmzwf pgizwfpgm WT 1 1 pgi pgm zwf pgipgm lactose lacy lacz glucose glycogen biosynthesis (lactoseglucoseglk glucose-1-p pgm mannitol PTS mtld xylose glucose PTS, glk xyle glucose-6-p 6-P-gluconolactone xylf zwf xylg Pentose xylh pgi phosphate pathway xyla mannitol mannitol-1-p fructose-6-p xylulose-5-p xylulose xylb fructose PTS fructose fructose-1-p fruk fructose-1, 6-P2 fdaa, fdab dihydroxyacetone-p glyceraldehyde-3-p glpa, glpb, tia tpia (5 rxns) glpc, glpd pyruvate glycerol glycerol-3-p glpf glpk TCA cycle (D660) (D660) ( pgizwf pgmzwf pgizwfpgm (h) (h) WT pgi pgm zwf pgipgm pgizwf 1 (D660) (D660) pgmzwf pgizwfpgm (h) (h) (D660) (D660) (h) (h)

42 Ớᐲ (D D660) 㙃㑆 (h) 䉫䊦䊦䉮䊷䉴Ớᐲ ᐲ (%) 㙃㑆 (h) 㙃㑆 (h) 30 䉫䊦䉮䊷䉴-6-䊥䊮 20 䉫䊦䉮䊷䉴Ớᐲ 㙃㑆䋨h䋩 24 DI glucose pgi zwf pgm pgm zwf pgi zwf pgi pgm zwf pgm pgi WT 䋳䋮㙃ᶧਛ䈻䈱DI䈱 DI ቯᕈ䈫䈋ᄢ 㙃 ઙ䈱ᦨㆡൻ DI ቯᕈ䈫䈋ᄢ 㙃 ઙ䈱ᦨㆡൻ 0 40 㙃㑆 (h) 䊙䊮䊆䊃䊷䊦Ớᐲ 20 ኋਥ pgi zwf Vector plex-btrc 2㬍YT䇮5% 2㬍YT 5% Glucose ph glu ucoseớ Ớᐲ(g/L L) 䊙䊮䊆䊃䊷䊦䊙䊦Ớᐲ (%) 3.5 Glucose-6-ph G hosphate Ớᐲ (Pmol/ g dry cell ) ᦛ DIỚᐲ(g/L) D pgi 㪫㪚㪘 zwf ㊁ pgiන უ ㊁䊐䊦䉪䊃䊷䉴䋶䋭䊥䊮㙃㑆㙃㑆 (h) (h) 㙃㑆 (h) Phosphoglucose isomerase 㑆䋨h䋩 30 㑆䋨h䋩 ph ph3(4 ) ph3(30 ) ph3(60 ) ph5(4 ) ph5(30 ) ph5(60 ) p ( ) Phosphoglucomutase 䉫䊦䉮䊷䉴 pgi, zwf, pgm ㊀ უ pgiන უ pgi, zwf ㊀ უ 䋶䋭䊥䊮 pgm WT pgi pgi/ zwf pgi/ zwf/ pgm pgi WT 䊕䊮䊃䊷䉴䊥䊮㑆䋨h䋩 DI䈲 ᕈ ઙ䇮ૐ ઙ䈪 ቯᕈ䈏㜞䈇䇯 20 ph ᄙ 㘃 วᚑ Glucose-6-phosphate dehydrogenase 䉫䊦䉮䊷䉴䋱䋭䊥䊮 ᄢ DI䈱 ᐲ䈍䉋䈶pH䈮ኻ䈜䉎 ቯᕈ D-mannitol D-glucose 50 6 Glucose50PTS DI合成酵素 DI 䉫䊦䉮䊷䉴䈎䉌DI䈻䈱ᄌ ല 䈱 DIỚᐲ (g/l) DIỚᐲ (g/l) DIỚᐲ (g/l) 100 DI (䋦䋦) D I (䋦) DIỚᐲ (g/l) ( 䉫䊦䉮䊷䉴䇮䊙䊮䊆䊃䊷䊦 ሽਅ䈮䈍䈔䉎㊁䈫䉫䊦䉮䊷䉴-6-䊥䊮㑐ㅪ უ 䈱セ DI (䋦)

43 DI DI(g/L) 0 ph h 30 ph ph ph DI(g/L) pgizwf Vector pgap-btrc/pgad-btrc 2YT5% Glucose0.5% Mannitol h 25pH:6.0 DI DI 45g/L /, DI (g/l) DIDI ph3 DI H + CH3C ml/min (h) 100 g DI 90 g 100% DIDI DI

44 DIDI 1 H-NMR Profile DI 13 C-NMR Profile

45 ppm 1 3kg

46 9,000 / , PLA (PBS) PLA

47 PDC Pseudomonas sp. SG4502 BDF PHA R

49 neo-symbiotic microorganism, NSM NSM

50 ,000kl ha 250 ha 40 ha

51 1ha t 7 8 t t 20 t ha 10 ha 20 t ha 200 t 1,

52 kl 21 kl kl kl ( ) HP

53 16 40 C (18 ) C ;2005)

55 1 D L 2 L- L L- L- 3 PHA PHA ICI 1980 Ralstonia eutropha PHB R. eutropha

56 PHA 1,3-1,3-PD 1. 1,3-PD Klebsiella pneumoniae 1,3-2. TCA 3. Clostridium IPA 10 1,

57 p- - - DI 1. p- p- 2. DI -6- DI 2-6- DI

58 EU BT GHG

59 GHG

62 - 59 -

63 ASPEX

64 TEL 03(5541)2731

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Microsoft Word - 本文.doc 中國大陸節能減碳策略之探討財團法人中技社民國 98 年 12 月中國大陸節能減碳策略之探討計畫主持人 : 顧洋教授委託機關 : 財團法人中技社受託機關 : 國立台灣科技大學助理 : 吳佳盈劉祐誠本報告內容係研究單位之觀點, 不代表