4 6 8 10 12 14 16 18 20 22 24
32 34 36 38 40 42 44 46 48 2008    2 2007 2007   2009 ---
pET
BL21( DE3) Takahashi [S. Takahashi et al. Bi osci. Bi otechnol.  Bi ochem  70 ( 12) ,  2913-2918 ( 2006 ) Na-Ile-Hs-Pro-Phe-Hs-Leu*Val -Ile-Thr- H s-Lys- (Dp) - D Arg-D Arg-N ூ  (*, [S. Takahashi et al. ) Biosci. Biotechnol. Biochem 71(10), 2610-2613 (2007)] Sep-Pak Vac C18 35cc (Ml l i pore) ... ... SA-402 NaO 1% MSG Q 5% 2008　 2 . . . 2007 - 0 05 Ω 2 7. 9× 4 3× 10cm 12 5 37. 5g 5 100 - 0 OBMa 50g 100g 10 0 OBNPa
: : : ( 2)
2007 2003   2007 --------------- --- 1)
2%
2%
2008 - 2
(1) 2003 2007 2007 1) 2) 3) 4) 5) 1) 2) 3) 4) 5) 6) 7) 2008    2 support of the st 2007 2006　2007 1) ( 10nh) ( ） (Ω ( 20g) ) (ACP) (
2 E coli 0   Q 1% E coli (3 O× 10 ⁄ m) 1000 (10nh) ( 20g) E coli E coli E coli W ( E   col i ) ( 50g 0. 25% ( ) ) (Q 035g) ( 100g) 3 5% (A E 0. 25% 15% 2 460g 4: 1 500g 3 5kg 5 3 (F)
2 1)公Q QZ% E coli Q 5% （
Q 1% E coli W ( 
 
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2 12 )
F 14.51 5% F F

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3 ... such a similar and services and services. 2007 2006　2007 (1) 2006 1) ) 2) 15% a) b) lkg 2kg 1kg 60 3 5 3 (2) 2007 1) ) ( Ω Ω ( 20g) ( 10nh) (
2 E coli 0  E coli 1000 E coli         ( 100μ l) ( 20g E coli (3 0× 10 ⁄ m) ( 10nh) ( ) (Q 007g) E coli E coli ) ( 50g) ( ) ( Q Q5g) ( 100g) 50nh ) 3.5% lkg ( 100g) ( E  coli 3) ( 1) - 2) 0 25% ( ) 2 (A E 460g 0. 25% (F) 4: 1 5 3 500g 3 5kg 2

(1) 2006
1) BO 1. 5 2) (ユ 207 1) Q 5% Q QZ%
Q 1% 2) 2006 22 F ( 12  )  F F F A B C D E F 2 10 ) F 02 2 20 25 10 1% B C D F 2 2 14.51 5% F --- --- 1 3 5g 10nh 3 5 2nh,  ME1. 2nh,  SDSO 4g 1M 1OnL SDS- PAGE 8nh 5 105 3 1 80 1 80 2 pH6 8   1nL   BPB1ng, 2 80 Q 1
2 DTT --- --- 1 SDS pH 8   1n1L   BPB2ng, 4nL 1OnL SDS- PAGEと
2 1 80 5 1M 1OnL pH6 8   1n1.   BPB1ng, 90 2 2nh,  ME1. 2nh,  SDSO 4g SDE- PAGE SS

昭和
昭和20 30.0 2 3 5 4 DTT 2- 1c ... .2007 --- 2  ... 2008    2 (2) 2007 2003　2007 support and services and services are allowed to the services of the services of the services. (RSA) RSA RSA Strept onyces B cereus   Aerononus 16s Strept onyces PCR S. cerevi si ae C Streptonyces  RSA Strept onyces RSA Aerononus Q 5% 1% 0 5%  CaO 21OnM RSA 27C 120rpm 35℃ 90% 30C 70% pH6 0   40°C ) ( RSA RSA RSA Strept onyces Aerononus ) ( Strept onyces RSA RSA ( ) Bi asci . Bi ot echnol . Bi ochem, 69(6) , 1073- 1081(2005) 2005- 143440) ( ( 2005- 208480) Aeronous ( ) ( ) 2008    2 ( 2) 2007 2003　2007 (RSA) RSA RSA Streptonyces B cereus Aeronanus 16s Strept onyces POR S. cerevi si ae C Streptonyces  RSA Streptonyces 1% Aerononus RSA 0.5% CaO 210nM 27℃ 120rpm Q 5% RSA 35℃ 90% 30C 70% pH6 0 40C Bi asci . Bi ot echnol . Bi ochem, 69(6) , 1073-1081(2005) 2005- 143440) ( 2005- 208480) ( RSA （ ） RSA Strept onyces RSA Aeronous RSA RSA RSA RSA RSA ... ---
11 2007
19 ACK139 EFA) 1) 18 2) 3) 3- A 3-B 1) 18 18 2) ACK139 3) 3-A 3-B 19 30 12  29 2 13
20 2007/6/27 2007/8/10 2007 2007/11/14 2007 2007/9/20 --- (4) JA 11 . . . A 13 18 8 124 135 B ACK139 ACK139 AL- 1 C D A ACK139 B ACK139 ACK139 C D 6 HLC 30 2 13 16 41 12 29 ACK139 11 51, 698-702( 2004) 2005 2005 53, 388-393 H17. 7. 15 - 57 - H16 4 23 52 17 53 H17.8 30 H17. 11. 10 H17. 11. 18 H17. 12 1 F 53 H18 8 29 H19 6 27 H9 8 10 2007 H9. 11. 14 2004 2007 2008 --- 2 --- 2007 2003 2007 --- --- Paeni baci l l us  sp   B38 ( Paeni dase, ) D Q 5% Q 3%NaO  pH 7. O 30 1% 1% Suc- [ D Asp] - MCA 380 nm 460 nm [S. Takahashi et al., J. Biochem 139(2), 197-202 (2006)] 50 nMTris-HI, pH 8 Q, Q Q2% Tween 20, Q Q2% NaNg Paeni baci l l us  sp. B38 DA SDS- PAGE PMF PCR S1016
PCR 322
D
(Paeni dase) 3 D 19 9 14 Satoru N rasava,  Motomitsu Kitaoka,  H deyuki  Kobayashi,  and Saori  Takahashi,  O oning of novel D aspart yl endopeptidase, Paenidase, fromprokaryote. International Meeting of the International Proteolysis Society 2007 19 10 23 Patras, Greece, Satoru N rasava,  Mitomitsu Kitaoka,  H deyuki  Kobayashi,  and Saori  Takahashi,  Prinary structure of novel D aspart yl endopeptidase, paenidase, fromprokaryote. 80 19 12 13 30 D (Paeni dase) 2008 27 20 3 2008    2 2003 2007 --- 1% 1% Q 5% 30 0. 3%Na0 1. 5% pH 7. O 30 24 70 30 30 D Suc- [D Asp] - pNA (DAEP) DAEP Paeni baci l l us sp   B38 DAEP SDS- PACE PMF PCR DAEP 4, 100 1, 100 16s rDA Paeni baci l l us  sp   B38 ( Paeni dase,   Paeni baci l l us  sp.   D Aspart yl   Endopept i dase) N9 F70 S2262 S2502 S1016 S1017 PCR 322
2006- 271275
Y. Li u, S. Takahashi, H Ogasavara, H G Seo, M Kavagoe, F. H rasava, N Gou, Y. Leno, T. Kaneda, and T. Sugi yana, Protect i on of hepat ocyt es fromapopt osi s by a novel subst ance fromactinonycetes culture nedium Bioned Res. 26(1), 9-14, 2005 S. Takahashi, H Qgasavara, K Hvatashi, K Hbri, K Hata, T. Tachibana, Y. Itoh, and T. Sugi yana, Paeni dase, a novel D aspart yl endopept i dase fromPaeni baci l l us sp. B38 Purification and substrate specificity. J. Biochem 139(2), 197-202, 2006 Saori Takahashi, Hronobu Qgasavara, Keishi Hata, Kazuyuki H vatashi, Kazuyuki Hbri, Tadanori   Tachi bana, Yoshifuni Itoh, and Toshihiro Sugiyana, Bacteri al   D aspart yl 21 endopept i dase. 10 17 D D 17 D – Paeni dase 6
2 D ( Paeni dase) D 3 19 14 9 Satoru N rasava,  Mt om t su Ki t aoka,  H deyuki  Kobayashi,  and Saori  Takahashi,  O oni ng of novel D aspart yl endopept i dase, Paeni dase, fromprokaryote. International Meting of the International Proteolysis Society 2007 Patras, Greece, 19 10 23 Satoru N rasava,  Mt om t su Ki t aoka,  H deyuki  Kobayashi,  and Saori  Takahashi,  Prinary structure of novel D aspart yl endopepti dase, paeni dase, fromprokaryote. 30 80 19 12 13 D ( Paeni dase) 2008 20 27 3 2008    2 2007 --- 2003   2007 --- 1. 8
Q 5
1. 8 1. 8 65: 25: 4 TLC 1448 30 40
804 1. 7 / 45nh
Ω 07 5 H    3 24 Cer TLC YPD HB 5 H 14( 2) , 52- 60( 2007) 436, 19-21( 2007) -------- 2008     2 ... 2007 2003 2007 --- --- H cerevi si ae Saccharonyces 2008 2  (5) 2007 2007 2009 . . . (RSA) RSA B cereus Aeronous Paeni baci I I us 1) ( Aerononus Paeni baci l l us But yl -    DEAE-    OMToyopearl Toyopearl - HM5( S) Sonagyi - Nel son RSA Aeronous ( 2) 90% 30°C 70% Paeni baci l l us pH 6 0 40C 35℃ ( 1) Aerononus RSA Paeni baci I I us N Bi asci . Bi ot echnol . Bi ochem, 69(6) , 1073- 1081(2005) 2005- 143440) ( 2005- 208480) (
Paeni baci l l us ... 2008    2 ( 1 ) . . . 2007 2009 20-100μ m Saccharonyces  cerevi seae Pichia stipitis 3 1. 80% cerevi seae 59( g/L) 1 1 S. 230L 3 2 (2 3 3 P.stipitis 3 4 2 Pichia stipitis Pignal 3 5 P. stipitis S. cereviseae 2 3 6 8( wv% 3
3 7.
whole crop Fl anmul i na vel ut i pes FV2 4 P.stipitis 2007- 185456 Si mil t aneousl y 1. 13 h European congress of Biotechnology(Barcelona) sacchari f i cat i on and bi oet hanol  product i on f rompowder  of  Japanese  cedar 2
2008 2  2007 1g 30nh ( AR) NADPH AR ( pig skin) 485nm ( A ast ri di umhi st al yt i cum 515nm DPA qui none 492nm L- DOPA N P 585nm 530nm DPPH 62 AR 100μ g/nh Ω 1μ g/nh 70% 10μ g/nh AR 1μ g/nh 70% AR AR 10 AR AR
19
cDVA PCR pET32a EIHRNI BL21(DE3) 4M 25g 300m
115 30 Sep-Pak Vac 35cc 10,000xg, 30 ( Na-Ile-Arg-Pro-Phe-Hs-Leu-Val - I l e- Thr- H s- Lys- DhpNe
Saori  Takahashi ,  Kazuyuki  Hri ,  Misanori  Kunagai ,  and Saburou Valkabayashi ,  Hınan reni n i nhi bi t ory act i vi t y  i n  l egunes. J. Bi ol. Macronal. 7(3), 49-54 (2007). Saori  Takahashi,  Kazuyuki  Hbri,  Masanori  Kunagai,  and Saburou Vakabayashi,  Renin i nhi bi t or  i n l egunes.  Int ernati onal  Conference on Food Fact ors  f or  Heal t h Pronot i on 2007 23 19 ( I COFF 2007) 10 2008
3 19 2007 19
19 2 5 6 5 8 4 20 20 20　2 5-8 2008 19 17 19 GANOTM “ l [ 1. , Lupane 9- 14 (2006)
9 ,42,60-61  2007 - Hata K., Mukaiyama T., Tsujimura N., Sato Y., Kosaka Y., Sakamoto K., Hori K., Differentiation-Inducing Activities of Lupane Triterpenes on a Mouse Melanoma Cell Line, Cytotechnology, 52, 151- 158 (2006) Sasaki H., Kawarasaki S., Sugawara M, Hata K., Effects of Lupeol on Visceral Fat Weight and Serum Lipoprotein Profile in High- Fat Diet Fed Mice. J. Clin. Biochem Nutr., in press. Hata K.,  Ogawa S.,  Makino M,  Mukaiyama T.,  Hori K.,  Iida T.,  Fujimoto Y.,  Lupane Triterpenes with a Carbonyl Group at C- 20 Induce Cancer Cell Apoptosis. J. Nat. Med , in press 2 2 NTS 2006 Mukaiyana T., Tsujimira N., Ohtaka S., Kosaka Y., Hata K., Hori K., Sakamoto K., Anti- melanogenic  Activity of Ergosterol  Peroxide from Ganoderma  Lucidum on a Mouse  Melanoma Cell Line, The Proceedings of the JAACT Kyoto, in press Hata K., Mukaiyama T., Tsujimura N., Sato Y., Kosaka Y., Sakamoto K., HoriK., Differentiation-Inducing Activities of Lupane Triterpenes from Lactuca Indica on a Mouse Melanoma Cell Line, The Proceedings of the JAACT Kyoto, in press 3
19 JST 19 18 19 ( 7 Hata K., Mukaiyama T., Tsujimira N., Sato Y., Kosaka Y., Sakamoto K., Hori K., Differentiation-Inducing Activities of Lupane Triterpenes on a Mouse Melanoma Cell Line, Cytotechnology, 52, 151- 158 (2006) Hata K.,  Ogawa S.,  Makino M,  Mukaiyama T.,  Hori K.,  Iida T.,  Fujimoto Y.,  Lupane Triterpenes with a Carbonyl Group at C- 20 Induce Cancer Cell Apoptosis. J. Nat. Med, in press