R NSC 89 2311 B 9 6 89 8 1 9 7 31
has been noticed as functional food. This research is to investigate the influence of different environmental factors on the production of mycelium and polysaccharide in submerged culture of C. sinensis. The optimal medium condition for the mycelium and polysaccharide production is initial ph=6, glucose 5% and yeast extract 1%. The best carbon and nitrogen source are glucose and yeast extract, respectively. Adding NH 4 H 2 PO 4 is to inhibit the yield of polysaccharide. The optimal concentration of the MgSO 4 7H 2 O and olive oil to added is 5.6% and.12%. However, we study these 1 yeast extract.5 yeast extract + three factors for polysaccharide formation by.5% con steep liquor ph 5-7 RSM, we found the interaction of factors had NH 4 H 2 PO 4 significant on inhibition of polysaccharide formation..6.12 Keywords: Cordyceps sinensis, polysaccharide, Response Surface Methodology Abstract Dongchongxiachao is famous as crude drug in Chinese medicine expensively and rarely. The therapy and health claims include the benefits to lung and kidney, the enforcement of immunity and antitumor activity. The yield of Dongchongxiachao is limiting by the geographyc region and the condition of climate in the mainland of China leading to dear the price. The great potency of application in C. sinensis by fermentation 1~4 15~ 25 2~8 2~4(~6) 1
1 (1). (2). 1.84 8.4 25.32 (3). 28.9 4.1 2 (4). (5). 3 4 D- 5 6 Kiho 45 kda 62 28 1 SE-HPLC beta-1,3 7 (1). 118 PDB B A 9 1 11 23 1 rpm.81g/ml B ph 5~7 12 13 ph=6 C D 5 1 1 23 1rpm ph=6 Glucose 5 Yeast 1 extract 1 one-factor at (2). one time Guihua Guoshan 17 C. militaris 1951 Box Wilson 14 Surface A B Response Methodology RSM 15 16 data fitting technique (3). peptone corn steep liquor 2
yeast extract yeast extract A B yeast Y=66.18-33.6X 1-9.78X 2-1.4X 3 extract yeast extract.5 + corn steep liquor.5 yeast extract NH 4 H 2 PO 4 X 1 1 X 2 X 3 corn steep liquor yeast NH 4 H 2 PO 4 X 1 extract X 2 X 3 NH 4 H 2 PO 4 (4)..6.12 Acremonium persicinum NH 4 H 2 PO 4 -glucan (Glucan) (Olive oil) (Sunflower oil) 18 (.4kgm -3 ) 12~14 kgm -3 22~24 kgm -3 Olive oil 28 kgm -3 C/N ph beta-glucan 19 A ~ D Olive oil C/N ph A NH 4 H 2 PO 4.1 C/N 45 55 Olive oil.35.55ml B (NH 4 ) 2 HPO 4 ph 4. 5. Data not show C.1 D NH 4 H 2 PO 4 RSM (5). RSM 3
(1). 1991 1 5 42-44 (11). 1995 9 3 162-167 (12).Zhu., J.S., Halpern, G.M., and Jones, K. (1). 1995 1998a, The scientific rediscovery of the Chinese herbal medicine Cordyceps 12 77-79 sinensis part I. J. Altern. Complement. (2). 1995 Med. 4 3 289-33. (13).Zhu., J.S., Halpern, G.M., and Jones, K. 17 4-41 1998b, The scientific rediscovery of a (3). precious ancient Chinese herbal 1991 regimen Cordyceps sinensis part II. J. 14 11 Altern. Complement. Med. 4 4 35-37 429-457. (4). 1992 (14).Box, G.E.P., and Wilson, K.B. 1951, On the experimental attainment optimum conditions. J. Roy. Statist. Soc. B13 14 2 36-37 1-45. (5). Miyazaki, T., Oilawa,N., and Yamada, (15). 1992 H.1997, Studies on fungal 39 225-236 polysaccharides. XX. Galactomannan of Cordyceps sinensis. Chem. Pharm. Bull. (16). 1992 25 12 3324-3328. (6). Kiho, T., Hui, J.I., Yamane, A., and 39 3-18 Ukai, S. 1993, Polysaccharides in fungi. (17).Guiha, Y., and Guoshan, W. 1988, XXXII. Hypoglycemic activity and Apreliminary report on growth habit of Chemical properties of the Cordyceps militaris. Edible Fungi. 4 polysaccharide from the culture 11-111. mycelium of Cordyceps sinensis. Biol. Pharm. Bull. 16 12 1291-1293. (18).Stasinopoulos, S. J., and R. J. Seviour. 199. Stimulation of exopolysaccharide (7). 1993 - production in the fungus Acremonium SE-HPLC persicinum with fatty acids. Biotech. Bioeng., 36 778-782. 1-3 beta 1 2 477-499 (8). Kiho, T., Yamane, A., Hui, J.I., Usui, S., and Ukai, S. 1996, Polysaccharides in fungi. XXXVI. Hypoglycemic activity of the polysaccharide CS-3 from the culture mycelium of Cordyceps sinensis and its effects on glucose metabolism in mouse liver. Biol. Pharm. Bull. 19 2 U., and Wagner, F. 1991. A high affinity 294-296. Zn 2+ uptake system controls growth and (9). biosynthsis of an extracellular,branched 1995-1,3-B-1,6-glucan in Sclerotium 97 rolfsii ATCC 155. xp. Mycol., 15, 6 3 178-179 181-192. 4 (19).Reeslev, M., and Jensen, B. 1995. Influence of Zn 2+ an Fe 3+ on polysaccharide production and mycelium/yeast dimorphism of Aureobasidium pullulans in batch cultivations. Appl. Microbiol. Biotechnol., 42 91-915. ().Pilz, F., Auling, G., Stephan, D., Rau,
.82 A. B.. (g/ml).78.76.74 (mg/ml) (mg/ml).72 4 6 8 1 12 14 (day) 3 4 5 6 7 ph 1 C. 1 D. 1 1 1 (mg/ml) (mg/ml) (mg/ml) 1 (mg/ml) 2 4 6 8 1 1 2 3 Glucose ( % ) Yeast extract ( % ) A PDB 23 1 rpm B ph 3 4 5 6 7 C 1 3 5 7 1 D.5 1 1.5 2 3 5
A. (mg/ ml) 15 5 (mg/ml) Glucose 5% Sucrose % Glucose 2.5% Sucrose 2.5% Glucose % Sucrose 5% B. (mg/ml) (mg/ml) Glucose 5% Maltose % Glucose 2.5% Maltose 2.5% Glucose % Maltose 5% A B 6
1 A. (mg/ml) (mg/ml) yeast extract 1% peptone % yeast extract.5% peptone.5% yeast extract % peptone 1% B. (mg/ml) (mg/ml) yeast extract 1% corn steep liquor % yeast extract.5% corn steep liquor.5% yeast extract % corn steep liquor 1% yeast extract peptone A yeast extract corn steep liquor B 7
A. 1 1 1 B. 1 1 (mg/ml) 15 5 (mg/ml) (mg/ml) (mg/ml)..1.3.5 1...1.3.5 Olive oil (%) NH 4 H 2 PO 4 (%) (mg/ml) 1 C. 1 1 (mg/ml) (mg/ml) 1 D. 1 1 (mg/ml)..1.3.5..1.3.5 (NH 4 ) 2 HPO 4 (%) MgSO 4. 7H2 O (%) A Olive oil B NH 4 H 2 PO 4 C (NH 4 ) 2 HPO 4 D MgSO 4 7H 2 O 8
RSM NH 4 H 2 PO 4 MgSO 4 7H 2 O Oliver oil Polysaccharide No. (g/ ml) (g/ ml) (ml/ ml) (mg/ ml) 1.1.1.2 93.14 2.5.1.2 65.34 3.1.5.2 85.42 4.5.5.2.88 5.1.1 1. 91.86 6.5.1 1. 33.93 7.1.5 1. 61.48 8.5.5 1. 57.36 9 (C).3.3.6 7.49 1 (C).3.3.6 75.38 No. 9 (C) 1(C) 9
NH 4 H 2 PO 4 MgSO 4 7H 2 O Oliver oil Polysaccharide No. (g/ ml) (g/ ml) (ml/ ml) (mg/ ml) 1.3.3.6 62.25 2.18.26.52 66.37 3.6.22.44 68.43 4.18.36 75.64 5.14.28 75.64 6.1..79 7.6.12 17.56 8.2.4 99.87 9 79.76 1
MgSO 4 7H 2 O Oliver oil Polysaccharide No. (g/ ml) (ml/ ml) (mg/ ml) 1.4.7 99.32 2.8.7 15.76 3.4.17 92.63 4.8.17 112. 5.2.12 85.59 6.1.22 86.19 7.6.2 113.23 8.6.2 17.3 9.6.12 121.98 1.6.12 123.1 11.6.12 117. 12.6.12 1.77 13.6.12 119.28 11