(hyaluronic acid HA) -N- 10 1000 Dalton 200,000 3000~4000 ( ) 1
2.1 2.1.1 1943 Meyer Palmer 1 connective tissue polysaccharides mucopolysacchrides glycosaminoglycans" chondroitin sulfatedermatan sulfatekeratan sulfateheparan sulfate heparin [D-glucuronic acid1--3]n- [N-acetyl-D-glucosamine1--4] 2 2.1-1 3 爲 爲 爲 酶 2
ph 860 NaCl 0.11 M NaOH 37 50 5 産 酶 酶 酶 酶 酶 肽 酶 1937 産 4 5-7 Lancefield's A C 産 UDPUridin-5'-Diphosphate Lancefield's A C 産 S. Dysgalactiae, S. Equi, S. Equisimilis, S. Pyogenes S. Zooepidemics 2.1.2 10 5 ATP 2 NAD 1 acetyl-coa 8 D-glucuronic acid N-acetyl-D-glucosamine 3
Glucose Glucose-6-p Fructose-6-p Glucose-6-p UDP-Glucuronic AcidFructose-6-p UDP N-Acetyl Glucosamine 酶 2.1-2 2.1-2 HA HAS, Hyaluronate synthase; NOX, NADH oxidase; LDH, lactate dehydrogenase; PFL, pyruvate formate lyase; PDH, pyruvate dehydrogenase; ADH, alcohol dehydrogenase; AK, acetate kinase 4 HAS operon hasahasbhasc 酶 UDP-glucose dehydrogenaseudp-glucose pyrophosphorylase 酶 UDP-Glucuronic Acid UDP N-Acetyl Glucosamine UDP-glucose dehydrogenase UDP-Glucuronic Acid UDP-glucose pyrophosphorylase 9,10 A hasa hasb has operon hasahasb hasc 酶 酶 UDP-Glucuronic Acid UDP N-Acetyl 4
Glucosamine 11 2.1.3 12 NH COO - CH 爲 (1 µg/ml) 13 2 98 14 爲 15 16 (Extra Cellular Matrix) Fibroblast CD-44 receptor Aggregation Fibroblast CD-44 receptor 5
2.1.4 1937 産 Streptococcus zooepidemicus Streptococcus pyogenes Streptococcus dysgalactiaestreptococcal A groups Streptococcal C groups 17 6
X- NH 2 OHHCl 18 Streptococcus zooepidemicuc NUF-035 産 爲 2.11 10 6 Dalton 産 1.88 g/l 2.1.5 NaOH 37 50 % 19 酶, 酶 酶 酶 酶,, 肽 酶 ph 20 3 CH 3 COONapH 3 CH 3 COONa95 92 2 0.4 M NaCl 95 23 (> 95 %) (< 0.2 %) (10 6 ) 1 formaldehyde 0.02 dodecyl sulphate 16 formaldehydedodecyl sulphate 95 95 10000 rpm4 7
30 0.15 M NaCl 0.1 para-hydroxybenzoic acidmethyl ester 3 10000 rpm4 3 0.9 g 1 N HCl Toyo No.1 Toyo No.4 0.45 m 10 CPC cetylpyridinium chloride CPC COO NaCl 0.15 M NaCl 10 1 M NaCl 40 mesh activated of Florisil 4 g 200 ml 1M NaCl HA 1.5 95 300 ml 0.15M NaCl 0.45 m MWCO 6000-8000 4 4 0.01 M PBS buffer 37 2 RNase DNase10 l/ml 37 2 3 95 20 95 10000 rpm4 30 0.15M NaCl 10000 rpm 10CPC 37 15 4000 rpm 20 1M NaCl 40 10000 rpm 5 2 95 5 10000 rpm 5 1M NaCl 2 95 MWCO 6000-8000 4 4 21 95 10000 rpm4 8
30 0.15 M NaCl 10 CPC 37 15 4000 rpm 20 0.4 M NaCl 40 10000 rpm 5 2 95 97 22 2.1.6 爲 爲 爲 産 23 爲 14 爲 12 barrier 24 1990 Casado 1.5~2 10 5 0.4 9
hydrolipic barrier 25 14 12 26 93 55-100 Fibroblast CD-44 receptor 1960 1980 Miller 10
27 vinyl sulfone Hylaform Viscoelastic Gel 2.2 2.2.1 fermentation fervere CO 2 11
DNA 苷 酶 DNA 26 2.2.2 20 30 (lag phase) 12
(log phase) (stationary phase) (decline phase) N hours 2.2-1 28 2.2.3 stirred-reactor kg / cm 2 13
0.3 Mpa brine d / D / circulatory revolve d / D 0.1~0.2 Mpa 14
ph air-lift reactor hold-up hold-up normal ameliorate packed formed liquid cycle membrane rolling-up 15
2.2.4 Batch culture Continuous culture Semi-continuous culture 28-30 16
ph lag phase log phase static phase ph turbidostat chemostat 17
ph medium 2.2.5 18
28-30 2.2-12.2-2 2.2-1 K Pa VVMcm 3 /h r/minrpm KW Pas /m 3 m 3 L DO K L a K L a K L a K L a l/h K L a 1/h /h /h /h /h 19
2.2-2 RNADNA mg/gdcw ATPADPAMP NADH O 2 CO 2 CO 2 RQ rh gdcw 1 /L u/mlg/l mg/gdcw mg/ml mg/100ml mol Fe 2 Mg 2 Ca 2 Na 1 NH 4 PO 4 3 2 SO 4 mg/gdcw go 2 /Lh go 2 g h ph mv 1DCWDry cell weight CO 2 20
ph ph ph ph ph ph ph ph ph ph 6.3~7.5 3~6 綫 7~8 ph ph H 2 SO 4 NaOH 21
22
23
4.1 4.1.1 (NUNR) (NIKONECLIPSE TE2000-U) -80 HPLC U-1500 UV-Vis Spectrophotometer HITACHI 5L (BIOTRON) SORVALL RC 5C PLUS HSIANG TAI CN-820 VortexSHIN KWANG 4 KANSIN KCC-1 (ASPIRATORA-3A TOKYO RIKAKIKAI) DENG YNG DOV-40 SV-3A DENG YNG AMICON 8400 HANIL MEGA 17R 24
4.1.2 4.1-1 Yeast extract Sigma Glucose Tryptone Sigma 95% Ethanol K 2 HPO 4 anhydrate MgSO 4 anhydrate NaCl Cabazole reagent Sigma D-glucuronic acid Sigma Borax Sigma Sigma MnCl 2 H 2 SO 4 Sigma Phenol Merck NaOH Merck RO 4.1.3 4.2 4.2.1 4.2.1.1 LB medium 25
Yeast Extract 10 g/l Tryptone5 g/l NaCl5 g/l Glucose30 g/l RO 1. 100 ml 2. 121 20 3. 3~5 15 4. 3~5 ml 5. 80 4 6. 7. 37 100 rpm 12~15 8. LB medium petri-dish 9. 37 15~17 4 4.2.1.2 yeast extract20 g/l NaCl1 g/l K 2 HPO 4 8 g/l MgSO 4 1 g/l Glucose30 g/l RO 1. 100 ml 121 20 2. 1~7 3. 37 100 rpm 15~17 OD 600nm 3~3.5 26
4.2.1.3 1. 3 L 2. 121 20 3. 100 ml 4. OD 600nm 4.2.2 1. 95 10000 rpm4 30 2. 37 24 3. 0.15 M NaCl 10000 rpm 4. 2 95 4000 rpm 30 5. 37 24 6. 7. 4 4 4.2.3 OD 4.2.3.1 OD OD 600nm OD 600nm OD 27
1. 15 ml 40 24 24 2. 5 L sample 10 ml 3. RO 10000 rpm 10 RO 10 ml 10000 rpm 10 4. 40 24 24 5. 4-1 4-1 OD OD OD 4.2.3.2 1. 2. 2 95 3. 4 4. - 31 1. g/100 ml 0.010.020.040.06 0.080.1 2. 0.5 ml 5% phenol 0.5 ml 2.5 ml 3. 10 4. 30 490nm 5. 4-2 28
OD OD600nm 4-1 OD 4-2 29
4.2.3.3 32-33 uronic acid Carbazole :, carbazole reagent,,,, 2.153 2.153, 4-3 1. : 1.1 1.2 1.3 Carbarzole 1.4 1.5 D-glucuronic acid 1.6 1.7 1.5 ml 1.8 UV 2. : 2.1 A:2.39 g 250 ml ( ) 2.2 B:0.125 g Carbazole 100 ml ( 4,12 ) 2.3 (g/ml):0.02 g D-glucuronic acid 20 ml 2.3.1 0,7.8125,15.625,31.25,62.5,125(g/ml) 2.3.2 : 1.5 ml, 2.4 3. : 3.1 18 1.5 ml, 3 1.5 ml, 3.2 1.5 ml 900l A, 4 3.3 150l 150l, 3.4 3.5 100 10 30
3.6 20 3.7 B 30l, 3.8 100 15 3.9 3.10 (VIS 530nm) (blank: 0 g/ml) (µg/ml) 4-3 31
4.2.3.4 GPC HPLCHigh Performance Liquid Chromatography SESize Exclusion IR TSK-GEL GMPWXL CARBOSep CORE GEL-87H3 100µl 0.45µm 60~70 0.5 ml/mincolumn 37 pump 25 psi dextran 4-2 4-4 4.2.3.5 25 0.847 1. 25 2. 25 25 3. 25 32
4-2 Dextran Log M.W. M.W (RT)min 3.698 5000 52.77 4 10,000 51.04 4.30 20,000 49.33 4.698 50,000 46.95 5 100,000 44.43 5.30 200,000 42.37 5.602 400,000 40.03 5.903 800,000 38.03 6.149 1,414,000 37.13 4-4GPC 33
5.1 5.1.1 5-7,18 16,18 Yeast extract 20 g/lk 2 HPO 4 8 g/lnacl 1 g/l MgSO 4 1 g/lglucose 60 g/l 37 100 rpm 5.1-1 100 rpm OD 600nm 3.643.58 100 rpm 100 rpm 5.1.2 K 2 HPO 4 K 2 HPO 4 K 2 HPO 4 0 g/l8 g/l16 g/l 5.1-2 K 2 HPO 4 K 2 HPO 4 0 g/l8 g/l16 g/l OD 600nm 1.653.644.95 K 2 HPO 4 K 2 HPO 4 8 g/l 16 g/l K 2 HPO 4 16g/L K 2 HPO 4 8 g/l 34
( 5.1-1 100 rpm OD600nm (hours) 5.1-2K 2 HPO 4 0 g/l8 g/l16 g/l 35
5.1.3 MgSO 4 MgSO 4 MgSO 4 0 g/l1 g/l2 g/l 5.1-3 MgSO 4 0 g/l1 g/l2 g/l OD 600nm 3.413.643.58 MgSO 4 1 g/l MgSO 4 1 g/l 5.1.4 NaCl NaCl 0 g/l1 g/l2 g/lnacl 5.1-4 NaCl 0 g/l1 g/l2 g/l OD 600nm 3.63.643.8 NaCl 36
OD600nm ( 5.1-3MgSO 4 0 g/l1 g/l2 g/l NaCl ( 5.1-4NaCl 0 g/l1 g/l2 g/l 37
5.1.5 MnCl 2 MnCl 2 0 g/l1 g/lmn DNA MnCl 2 5.1-5 MnCl 2 MnCl 2 0 g/l1 g/l OD 600nm 3.642.95 1 g/l MnCl 2 MnCl 2 MnCl 2 5.1.6 yeast extract bacto-tryptone yeast extract 20 g/lyeast extract 10 g/l+tryptone 10 g/ltryptone 20 g/l 5.1-6 yeast extract yeast extract 20 g/lyeast extract 10 g/l + tryptone 10 g/ltryptone 20 g/l OD 600nm 3.64 3.332.84 yeast extract tryptone yeast extract yeast extract yeast extract 38
( 5.1-5Mn 0 g/l1 g/l ( 5.1-6 yeast extract 20g/L yeast extract 10g/L+tryptone 10g/L tryptone 20g/L 39
5.1.7 60 g/l 60 30 g/l 5.1-7 OD 600nm 60 g/l30 g/l 3.644.165 30 g/l 30 g/l ( 5.1-7 30 g/l60 g/l 40
5L 16 25 40 yeast extract 20 g/lmgso 4 1 g/l K 2 HPO 4 8 g/lnacl 1 /Lglucose 60 g/l 5 L 3 L 25 33 37 40 OD 200 rpmod 1.5~2 250 rpmod 8~9 350 rpmod 13 400 rpm OD 0.5 vvmod 8~9 1 vvmph 7~7.2 12 6 OD 600nm 3.2~3.5 100 ml 5.2.1 5.2.1.1 25 5.2-1 25 30 28 OD 600nm 14.42 25 10~13 26 25 25 5.2.1.2 5.2-2 OD 600nm 33 5~6 12 25 33 25 41
( 5.2-1 25 ( 5.2-2 33 42
5.2.1.3 37 5.2-3 16 15 OD 600nm 18.68 37 6 12 6 1.032 18.68 33 25 33 5.2.1.4 40 5.2-4 40 13 12 OD 600nm 17.56 40 5 9 5 0.772 17.56 40 37 33 43
( 5.2-3 37 ( 5.2-4 40 44
5.2.2 Cabazole assay 4.2.2 1~5 5.2.2.1 25 5.2-5 7.18 g/l5.2-1 10 OD 5.2-1 10 4 g/l 25 3 g/l 5.2.2.2 33 5.2-6 17 5.2-2 10 33 25 7.32 g/l 45
( ( 5.2-525 ( ( 5.2-633 46
5.2.2.3 37 5.2-7 16 5.2-3 8.03 g/l 33 25 5.2.2.4 40 5.2-8 13 25 25 33 7.9 g/l 37 47
( ( 5.2-737 ( ( 5.2-840 48
5.2.3 5.2.3.1 25 25 10~13 5~6 37 OD 600nm 18.68 25 OD 600nm 14.42 33 OD 600nm 14.8640 OD 600nm 17.56 37 40 25 33 37 40 5.2.3.1 5.2-6~8 5.2-2~4 25 7.18 g/l 33 7.32 g/l37 8.03 g/l40 7.9 g/l 37 40 49
5.3 0 g/l30 g/l60 g/l 4.2.2 1~7 5.3.1 0 g/l glucose 37 5.3-1 6 2 4 OD 2.9 5.3-23 2.58 cps OD 5.3-3 5.3-4 3 1.11 g/l 5.3-5 OD 5.3-6 5.3-7 3 GPC 22~27 1.54 10 8 ~2.9 10 7 Dalton 24.46 6.8 10 7 Dalton 50
( 5.3-1 cps ( 5.3-2 51
5.3-3 OD ( 5.3-4 52
5.3-5 ( 5.3-6 53
20 15 52.49 Intensity (mv) 10 5 24.46 0 0 5 10 15 20 25 30 35 40 45 50 55 60 Retention Time (min) 5.3-7 3 54
5.3.2 30 g/l 30 glucose 37 5.3-8 10 7 OD 15.42 5.1-7 ph 60 g/l 5.2-3 6 4 5.3-9 9 196 cps OD 5.3-10 4 7 OD 5.3-11 9 6.61 g/l 5.3-12 5.3-13 8 0 30 g/l 5.3-14 9 GPC 20~30 2.99 10 8 ~1.08 10 7 Dalton 28.13 2.01 10 7 Dalton 55
(hours) 5.3-8 30 g/l cps (hours) 5.3-9 30 g/l 56
5.3-10 30 g/l OD ( ( 5.3-11 30 g/l 57
5.3-12 30 g/l (hours) 5.3-13 30 g/l 58
20 15 28.13 Intensity (mv) 10 5 41.63 0 0 5 10 15 20 25 30 35 40 45 50 55 60 Retention Time (min) 5.3-1430 g/l 9 59
5.3.3 60 g/l 60 glucose 37 5.3-15 17 11 OD 19 37 5.2-3 6 5.3-16 13 708 cps OD 5.3-17 6~11 OD 10~18 5.3-18 13 11.09 g/l 5.3-19 5.3-20 17 OD 5.3-1511 13 5.3-21 13 GPC 20~35 2.99 10 8 ~2.05 10 6 Dalton 29.59 1.23 10 7 Dalton 60
(hours) 5.3-15 60 g/l (hours) 5.3-16 60 g/l 61
5.3-17 60 g/l OD ( ( 5.3-18 60 g/l 62
(g/l) 5.3-19 60 g/l ( 5.3-20 60 g/l 63
40 35 29.87 29.92 29.60 30 25 Intensity (mv) 20 15 10 5 43.39 0 0 5 10 15 20 25 30 35 40 45 50 55 Retention Time (min) 5.3-2160 g/l 13 64
5.3.4 0 g/l30 g/l60 g/l 5.3-1 0 g/l 5.3-8 30 g/l 5.3-15 60 g/l 2 4 6 5.3-1 2.9 6 5.3-8 5.3-8 30 g/l 7 OD 600nm 15.42 7 5.3-15 60 g/l 13 OD 600nm 19 11 5.3-13 30 g/l 8 5.3-15 OD 600nm 19 5.3-22 60 g/l 17 60 g/l 5.3-4 0 g/l5.3-11 30 g/l 5.3-18 60 g/l 1.11 g/l6.61 g/l 11.09 g/l 60g/L 5.3-7 0 g/l5.3-14 30 g/l 5.3-21 60 g/l GPC 1.54 10 8 ~2.9 10 7 Dalton2.14 10 8 ~1.08 10 7 Dalton 2.99 10 8 ~2.05 10 6 Dalton 6.8 10 7 Dalton2.01 10 7 Dalton 1.23 10 7 Dalton 5.3-21 65
60 g/l 66
5.4 5.3 5 L 3 L 5.4.1 60 g/l glucose60 g/l glucose 400 ml 3 ml/min1.17 10 3 kgglucose/h 37 OD 600nm 8.08 8 5.4-1 15 12 14 OD 16.74 5.3-15 60 g/l 5~6 5.4-2 406 cps OD 5.4-3 OD 9~10 5.4-4 13 10.41 g/l 5.3-18 1 g/l 5.4-5 8 g/l 8 g/l 5.4-6 5.4-7 13 GPC 20~34.5 2.99 10 8 ~2.42 10 6 Dalton 29.44 1.3 10 7 Dalton 67
( 5.4-1 60 g/l glucose3 ml/min (hours) 5.4-2 60 g/l glucose3 ml/min 68
5.4-3 60 g/l glucose3 ml/min OD ( ( 5.4-4 60 g/l glucose3 ml/min 69
5.4-560 g/l glucose3 ml/min (hours) 5.4-6 60 g/l glucose3 ml/min 70
12 29.44 10 Intensity (mv) 8 6 4 2 45.80 48.61 0 0 5 10 15 20 25 30 35 40 45 50 55 60 Retention Time (min) 5.4-760 g/l glucose3 ml/min13 71
5.4.2 60 g/l glucose60 g/l glucose 400 ml 6 ml/min2.34 10 3 kgglucose/h 37 OD600nm 7.6 8 1 5.4-8 13 11 OD 17.86 5.4-1 3 ml/min 5.4-9 12 445 cpsod 5.4-10 9~11 5.4-11 12 10.70g/L5.3-18 5.4-4 60 g/l3 ml/min 0.5 g/l 5.4-12 8 g/l 5.4-12 910 3.65 g/l 6.43 g/l 3 100 cps 1012 6.43 g/l 10.27 g/l4 g/l 300 cps 5.4-13 3 ml/min 5.4-14 12 GPC 25~38 5.6 10 7 ~7.5 10 5 Dalton 29.41 1.31 10 6 Dalton 72
(hours) 5.4-8 60 g/l glucose6 ml/min (hours) 5.4-9 60 g/l glucose6 ml/min 73
5.4-10 60 g/l glucose6 ml/min OD (hours) 5.4-11 60 g/l glucose6 ml/min 74
cps g/l 5.4-12 60 g/l glucose6 ml/min (hours) 5.4-13 60 g/l glucose6 ml/min 75
50 40 30.71 30 Intensity (mv) 20 10 42.23 0 0 5 10 15 20 25 30 35 40 45 50 55 60 Retention Time (min) 5.4-14 60 g/l glucose6 ml/min 12 76
5.4.3 60 g/l glucose 60 g/l glucose 200 ml 20 ml/min7.8 10 3 kgglucose/h 37 OD 600nm 7.06 9 5.4-15 22 13 OD OD 21 20.06 5.4-1 3 ml/min 5.4-8 6 ml/min OD 600nm 5~6 8~13 13~21 5.4-16 776 cps 12 11 13 OD 161715 OD 5.4-17 5.4-18 20 12.52 g/l 5.3-18 60 g/l 5.4-4 60 g/l 3 ml/min 5.4-11 60 g/l 6 ml/min 1.5~2 g/l 5.4-19 10~15 5.4-20 5.4-21 20 GPC 25~35 5.6 10 7 ~2.05 10 6 Dalton 31.30 0.7 10 7 Dalton 77
(hours) 5.4-15 60 g/l20 ml/min cps (hours) 5.4-16 60 g/l20 ml/min 78
5.4-17 60 g/l20 ml/min OD g/l (hours) 5.4-18 60 g/l20 ml/min 79
5.4-19 60 g/l20 ml/min (hours) 5.4-20 60 g/l20 ml/min 80
160 Intensity (mv) 140 120 100 80 60 40 31.88 31.03 20 0 38.89 41.99 0 5 10 15 20 25 30 35 40 45 50 55 Retention Time (min) 5.4-21 60 g/l20 ml/min 20 81
5.4.4 30 g/l glucose 30 g/l glucose 400 ml 3 ml/mins1.17 10 3 kgglucose/h 37 OD 600nm 6.73 6 5.4-22 12 8 9 OD 17.64 6 8 5.4-1 60 g/l3 ml/min 1 5.4-23 9 461 cpsod 5.4-24 12~18 5.4-25 10 12.34 g/l5.4-4 60 g/l3 ml/min 2 g/l 5.4-26 5.4-27 6~8 5.4-28 9 GPC 21~34 2.14 10 8 ~2.8 10 6 Dalton 30.05 1 10 7 Dalton 82
( 5.4-22 30 g/l glucose 30 g/l glucose3 ml/min cps ( 5.4-23 30 g/l glucose 30 g/l glucose3 ml/min 83
5.4-24 30 g/l glucose 30 g/l glucose3 ml/min OD ( ( 5.4-25 30 g/l glucose 30 g/l glucose3 ml/min 84
5.4-26 30 g/l glucose 30 g/l glucose3 ml/min (hours) 5.4-27 30 g/l glucose 30 g/l glucose3 ml/min 85
20 30.05 15 Intensity (mv) 10 5 45.47 0 0 5 10 15 20 25 30 35 40 45 50 55 60 Retention Time (min) 5.4-28 30 g/l glucose 30 g/l glucose3 ml/min 9 86
5.4.5 30 g/l glucose 30 g/l glucose 200 ml 20 ml/min7.8 10 3 kgglucose/h 37 OD 600nm 9.33 6 5.4-29 13 9 10 OD 18.55.4-15 60 g/l 20 ml/min 4 5.4-30 10 495 cps OD 5.4-31 OD 12~18 5.4-32 12 10.42 g/l 3 ml/min12.34 g/l 5.4-33 5.4-34 30 g/l 5.4-35 12 GPC 20~34.5 2.99 10 8 ~2.4 10 6 Dalton 29.56 1.2 10 7 Dalton 87
5.4-29 30 g/l glucose 30 g/l glucose20 ml/min cps ( 5.4-30 g/l glucose 30 g/l glucose20 ml/min 88
5.4-31 30 g/l glucose 30 g/l glucose20 ml/min OD (hours) 5.4-32 30 g/l glucose 30 g/l glucose20 ml/min 89
5.4-33 30 g/l glucose 30 g/l glucose20 ml/min (hours) 5.4-34 30 g/l glucose 30 g/l glucose20 ml/min 90
20 15 29.17 29.56 Intensity (mv) 10 5 48.60 0 0 5 10 15 20 25 30 35 40 45 50 55 Retention Time (min) 5.4-35 30 g/l glucose30 g/l glucose20 ml/min 12 91
5.4.6 5.4.1 60 g/l 60 g/l 3 ml/min 5.4.2 60 g/l 60 g/l 6 ml/min 5.4.3 60 g/l 60 g/l 20 ml/min 5.4.4 30 g/l 30 g/l 3 ml/min 5.4.5 30 g/l 30 g/l 20 ml/min 5.3 30 g/l 4 60 g/l 5~6 5.4.1 12 OD 16.745.4.2 11 OD 17.865.4.3 13 OD 20.06 5.4.4 8 OD 17.645.4.5 9 OD 18.5 60 g/l 5.4.4 5.4.5 8~9 120 g/l 5.4.15.4.2 5.4.3 11~13 60 g/l 60 g/l 5.4.4 30 g/l 30 g/l 3 ml/min 5.4-22 4 9 OD 600nm 17.64 10 12.34 g/l 5.4-45.4-115.4-185.4-25 5.4-32 5.4-4 13 10.41 g/l 5.4-11 12 10.70 g/l 5.4-18 20 12.52 g/l 5.4-25 9 12.34 g/l 5.4-32 12 10.42 g/l 5.4.3 60 g/l 60 g/l 20 ml/min 92
20 5.4.4 30 g/l 30 g/l 3 ml/min 0.18 g/l 5.4.4 5.4.4 5.4.3 60 g/l~120 g/l 5.4-75.4-145.4-215.4-285.4-35 5.4-7 2.99 10 8 ~2.42 10 6 Dalton 5.4-14 5.6 10 7 ~7.5 10 5 Dalton 5.4-21 5.6 10 7 ~2.05 10 6 Dalton 5.4-28 2.99 10 8 ~2.8 10 6 Dalton 5.4-35 2.99 10 8 ~2.4 10 6 Dalton 1.3 10 7 Dalton1.31 10 6 Dalton0.7 10 7 Dalton1 10 7 Dalton1.2 10 7 Dalton 5.4.1 5.3 93
5.5 K 2 HPO 4 5.1.2 K 2 HPO 4 K 2 HPO 4 OD 600nm K 2 HPO 4 yeast extract 20 g/lk 2 HPO 4 8 g/lmgso 4 1 g/lnacl 1 g/lglucose 60 g/l 5 L 3 L ph 7~7.2 OD 200 rpmod 1.5~2 250 rpmod 8~9 350 rpmod 13 400 rpm OD 0.5 vvmod 8~9 1 vvm 5.5.1 16 g/l K 2 HPO 4 37 5.5-1 17 OD 14.8 11 K 2 HPO 4 5.3-15 K 2 HPO 4 8 g/l 3 K 2 HPO 4 5.5-2 51 cps 10 OD 5.5-3 7~12 5.5-4 13 10.23 g/l 5.5-5 5.5-6 5.5-7 13 GPC 22~33.5 1.54 10 8 ~3.3 10 6 Dalton29.93 1.1 10 7 Dalton 94
( 5.5-1 K 2 HPO 4 16 g/l ( 5.5-2 K 2 HPO 4 16 g/l 95
5.5-3 K 2 HPO 4 16 g/l OD ( ( 5.5-4 K 2 HPO 4 16 g/l 96
5.5-5 K 2 HPO 4 16 g/l (hours) 5.5-6 K 2 HPO 4 16 g/l 97
50 40 29.93 30 Intensity (mv) 20 10 0 0 5 10 15 20 25 30 35 40 45 50 55 Retention Time (min) 5.5-7 K 2 HPO 4 16 g/l 13 98
5.5.2 8 g/l K 2 HPO 4 8 g/l K 2 HPO 4 6 ml/min 100 ml 17 2.16 10 3 kgglucose/h 37 OD 5.38 5 17 5.5-8 10 OD 8 16.62 5.5.3 5.3.15 8 g/l K 2 HPO 4 K 2 HPO 4 5.5-1 16 g/l K 2 HPO 4 5.3-15 8 g/l K 2 HPO 4 K 2 HPO 4 5.5-1 3~4 K 2 HPO 4 5.3-15 5.5-9 328 cps 8 g/l K 2 HPO 4 5.3.16 OD 5.5-10 OD 5.5-11 9 12.55 g/l 5.5-12 5.5-13 5.5-14 9 GPC 20~31.5 2.99 10 8 ~6.5 10 6 Dalton 27.09 2.8 10 7 Dalton 99
(hours) 5.5-8 8 g/l K 2 HPO 4 (hours) 5.5-9 8 g/l K 2 HPO 4 100
5.5-10 8 g/l K 2 HPO 4 OD (hours) 5.5-11 8 g/l K 2 HPO 4 101
5.5-12 8 g/l K 2 HPO 4 (hours) 5.5-138 g/l K 2 HPO 4 102
50 40 30 Intensity (mv) 20 10 27.09 41.28 0 0 5 10 15 20 25 30 35 40 45 50 55 60 Retention Time (min) 5.5-14 8 g/l K 2 HPO 4 9 103
5.5.3 K 2 HPO 4 8 g/l K 2 HPO 4 16 g/l 8 g/l 5 L 5.3.3 K 2 HPO 4 8 g/l 5.3-15 8 g/l K 2 HPO 4 K 2 HPO 4 5.5-1 16 g/l K 2 HPO 4 5.5-8 8 g/l K 2 HPO 4 8 g/l K 2 HPO 4 6 3 3~4 K 2 HPO 4 5.5-8 5.3-15 11 11 7 13 12 8 1914.8 16.62 5 L K 2 HPO 4 K 2 HPO 4 K 2 HPO 4 5.3-18 8 g/l K 2 HPO 4 K 2 HPO 4 5.5-4 16 g/l K 2 HPO 4 5.5-11 8 g/l K 2 HPO 4 8 g/l K 2 HPO 4 11.09 g/l10.23 g/l 12.55 g/l K 2 HPO 4 K 2 HPO 4 5.3-21 8 g/l K 2 HPO 4 K 2 HPO 4 5.5-7 16 g/l K 2 HPO 4 5.5-14 8 g/l K 2 HPO 4 8 g/l K 2 HPO 4 2.99 10 8 ~2.05 10 6 Dalton1.54 10 8 ~3.3 10 6 Dalton 2.99 10 8 ~6.5 10 6 Dalton 1.23 10 7 Dalton1.1 10 7 Dalton 2.8 10 7 Dalton 5.3-21 8 g/l K 2 HPO 4 K 2 HPO 4 5.5-14 8 g/l K 2 HPO 4 8 g/l K 2 HPO 4 104
K 2 HPO 4 105
5.6 5.1.1 yeast extract 20 g/lk 2 HPO 4 8 g/lmgso 4 1 g/l NaCl 1 g/lglucose 60 g/l 5 L 3 L ph 7~7.2 OD 200 rpmod 1.5~2 250 rpmod 8~9 350 rpmod 13 400 rpm OD 0.5 vvmod 8~9 1 vvm 5.6.1 37 5.6-1 14 11 OD 10.7 5 5.3-15 19 5.6-2 36 cps OD 5.6-3 5.1-2 16 g/l K 2 HPO 4 51 cps 5.6-4 13 6.52 g/l 5.6-5 5.6-6 5.6-7 13 GPC 23~35 1.1 10 8 ~2 10 6 Dalton 31.61 6.3 10 6 Dalton 106
( 5.6-1 (hours) 5.6-2 107
5.6-3 OD (hours) 5.6-4 108
5.6-5 (hours) 5.6-6 109
100 80 60 Intensity (mv) 40 31.61 20 45.04 0 0 5 10 15 20 25 30 35 40 45 50 55 Retention Time (min) 5.6-7 13 110
5.6.2 37 OD 9.05 6 5.6-8 10 4~5 13 OD 11 16.1 OD 19 5.6-9 10 489 cps OD 5.6-10 5.6-11 9 14.93 g/l 5.3-18 60 g/l 2~3 g/l 5.6-12 5.6-13 5.6-14 9 GPC 25~37 5.6 10 7 ~1 10 6 Dalton 31.35 6.9 10 6 Dalton 111
( 5.6-8 (hours) 5.6-9 112
5.6-10 OD (hours) 5.6-11 113
5.6-12 (hours) 5.6-13 114
100 80 29.57 31.35 60 Intensity (mv) 40 20 0 0 5 10 15 20 25 30 35 40 45 50 55 Retention Time (min) 5.6-14 9 115
5.6.3 5.3.3 5.6.1 5.6.2 5.3-15 5.6-1 5.6-8 5.3-15 60 g/l 13 19 5.6-1 11 10.7 5.6-8 11 16.1 11 10 10 5.6-1 5.6-8 6 5.3-15 5.3-15 5 5.3.3 60 g/l 708 cps 5.3-18 60 g/l 5.6-4 5.6-11 11.09 g/l6.52 g/l14.93 g/l 4 g/l 1/2 5.3-21 5.6-7 5.6-14 2.99 10 8 ~2.05 10 6 Dalton1.1 10 8 ~2 10 6 Dalton5.6 10 7 ~1 10 6 Dalton 1.23 10 7 Dalton6.3 10 6 Dalton6.9 10 6 Dalton 5.6-6 5.6-13 116
117
5.7 100 L 5.7.1 yeast extract 20 g/lk 2 HPO 4 8 g/lmgso 4 1 g/lnacl 1 g/lglucose 60 g/l glucose 60 g/l 0.3~0.4 kg/cm 3 OD 200 rpmod 1.5~2 250 rpmod 8~9 350 rpmod 13 400 rpm OD 0.5 vvmod 8~9 1 vvm 37 ph 7~7.2 60 L 3 LOD 12.34 37 60 L 3 LOD 12.34 5.7-1 11 8 OD 19.72 7 2 100 L 5 L 60 g/lod 19 5.7-2663 cps OD 5.7-3 100 L 100 L OD 5 L 5.7-4 9 12.42 g/l 5 L 1 g/l 5.7-5 8 g/l 8 g/l 5.7-6 5.7-7 9 GPC 23~37 1.1 10 8 ~1.05 10 6 Dalton 32.04 5.4 10 6 Dalton 118
(hours) 5.7-1100 L (hours) 5.7-2100 L 119
5.7-3100 L OD (hours) 5.7-4100 L 120
5.7-5100 L (hours) 5.7-6100 L 121
(mv) Intensity 100 80 60 40 20 0 0 5 10 15 20 25 30 35 40 45 50 55 32.04 Retention Time (min) 5.7-7100 L 9 44.28 122
100 L 5 L 5.3-155 L 60 g/l 5.7-1100 L 100 L 5 L 100 L OD 5 L 100 mlod 3.4100 L 3 LOD 12.34 5 L 30 100 L 20 100 L 100 L 100 L 5 L 5.3-155 L 5.7-1100 L 1919.72 5 L 100 L 17 11 11 7 100 L 12.42 g/l 5.3-185 L 11.09 g/l 100 L 5 L 5 L 5.3-215 L 5.7-7100 L 2.99 10 8 ~2.05 10 6 Dalton1.1 10 8 ~1.05 10 6 Dalton 1.23 10 7 Dalton5.4 10 6 Dalton 5 L 100 L 100 L 123
1. yeast extract 20 g/lnacl 1 g/lk 2 HPO 4 8 g/lmgso 4 1 g/lglucose 60 g/l 2. 25 37 25 26 OD 14.4237 12 OD 18.68 37 8.03 g/l 37 3. 4. 30 g/l 60 g/l 0.1 g/l 120 g/l 60 g/l 60 g/l~120 g/l 5. 120 g/l60 g/l 20 ml/min7.8 10 3 kg/h OD 600nm 20.06 6. 6.52 g/l 14.93 g/l 7. K 2 HPO 4 124
GPC 8 g/l K 2 HPO 4 8 g/l K 2 HPO 4 2.8 10 7 Dalton 8. 100 L 5 L 100 L 12.42 g/l 5 L 11.09 g/l 1 g/l GPC 100 L 5.4 10 6 Dalton 5 L 1.23 10 7 Dalton 100 L 5 L 5 L 9. 6-1 HA hours hoursod 600nm g/l Dalton Dalton 25 13 26 14.42 7.18 33 5~6 12 14.86 7.32 37 5~6 12 18.68 8.03 40 5~6 9 17.56 7.9 0 g/l 2 3~4 2.6 1.1 1.54 10 8 ~2.9 10 7 6.8 10 7 30 g/l 4~5 7 15.42 6.62 2.14 10 8 ~1.08 10 7 2.01 10 7 60 g/l 6 11 19 11.09 2.99 10 8 ~2.05 10 6 1.23 10 7 60 g/l 60 6 12 16.74 10.41 2.99 10 8 ~2.42 10 6 1.3 10 7 g/l3 ml/min 60 g/l 60 6 11 17.86 10.7 5.6 10 7 ~7.5 10 5 1.31 10 6 g/l6 ml/min 60 g/l 60 6 13 20.06 12.52 5.6 10 7 ~2.05 10 6 7 10 6 g/l20 ml/min 30 g/l 60 4 8 17.64 12.34 2.14 10 8 ~2.8 10 6 1 10 7 g/l3 ml/min 30 g/l 60 4 9 18.5 10.42 7.9 10 7 ~2.4 10 6 1.2 10 7 g/l20 ml/min 4 10 10.7 6.52 1.1 10 8 ~2 10 6 6.3 10 6 4 10 16.1 14.93 5.6 10 7 ~1 10 6 6.9 10 6 K 2 HPO 4 16 g/l 3 11 14.8 10.23 1.54 10 8 ~3.3 10 6 1.1 10 7 K 2 HPO 4 8 4 7 16.62 12.55 2.99 10 8 ~6.5 10 6 2.8 10 7 g/l 8 g/l 100 L 1~2 7 19.72 12.42 1.1 10 8 ~1.05 10 6 5.4 10 6 125
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