13 1 Vol13 No1 1 2009 2 Life Science Research Feb 2009 1a 1b, 2, 1 416000 2 416000 3 410300 : (Cordyceps militaris), :, 6% 1% 25, ; 6% 1% 22, : ; ; ; ; ; : TQ92 : A : 1007-7847(2009)01-0065-06 Researches on Fermentation Engineering of Polysaccharide of Cordyceps militaris WANG Ju-feng 1 LI Hu-ming 2 3 1 College of Bio-resources and Environmental Sciences Jishou University Jishou 416000 Hunan China 2 Research Centre for natural Pharmacy Jishou University Jishou 416000 Hunan China 3 Hunan TGL Bio-engineering Co LTD Changsha National Bio-industrial Base Changsha 410300 Hunan China Abstract The influence of temperature the concentration of glucose and protein on the biomass and the polysaccharide yield of Cordyceps militaris in cultural conditions were investigated The results showed that these factors greatly infected the biomass the yield of the extrocellular polysaccharide intracellular polysaccharide and total polysaccharide In this experiment when cultural temperature was 25 and with 6% of glucose and 1% of protein in the media the biomass the yield of total polysaccharide and extrocellular polysaccharide reached the highest and the yield of polysaccharide of the mycelium intracellular polysaccharide reached the highest in the following conditions 22 6% of glucose and 1% of protein Key words temperature carbon source nitrogen source Cordyceps militaris fermentation Cordyceps militaris polysaccharide Cordyceps militaris Lex Fr Link Life Science Research 2009 13 1 065~070 Eumycota Ascomycotina Pyrenomycetes Sphaeriales Clavicipitaceae Cordyceps [1] Cordyceps sinensis 2008-03-04 : 2008-10-21 [2004]99 04JJ3057 1965- Tel 13739018786 E-mail wangchenyu888@126com
66 2009 2 d 22 25 28 [2,3] 135~140 r / min 5 d [4~10] 1 L 9 (3 4 ) Table 1 The election of factors and levels in L 9 (3 4 ) Factors Levels a: C/% b: N/% c: Temperature/ 1 2 5 22 2 4 3 25 3 6 1 28 132 5 d 10 000 r / min 10 min 2 60 133 5 d 10 000 r/min 10 min 3 95% 24 h 1 11 Cordyceps militaris 134 12 SIGMA3-18K 15 h 14 000 r / min SW-CJ-2FD 10 min 10 HPG-400H 15 h 3 THZ-82B 95% 65%~70% HVE-50 24 h AE100 100 ml - 135-13 131 Sevag 10% H 2 O 2 3 95% 24 h 100 ml - 250 ml 10 100 490 nm W/V L 3 4 9 1 2 10% 25 ± 1
1 : 67 2 3 K 1 K 2 K 3 k 1 k 2 k 3 2 Table 2 The results of data processing of the mycelium and polysaccharide of Cmilitaris Dry weight of mycelium Polysaccharides a: C/% b: N/% c: Temperature/ a: C/% b: N/% c: Temperature/ 38818 33375 37810 436702 441337 447337 33383 39007 47801 441476 493210 564591 40250 40069 26840 595480 539111 461730 12939 11125 12603 145567 147112 149112 11128 13002 15934 147159 164403 188197 13417 13356 8947 198493 179704 153910 R Comparison in factors Best project 6867 6694 c a b a 3 b 3 c 2 20961 158778 97774 a c b a 3 b 3 c 2 117254 K 1 1 K 2 2 K 3 3 k 1 k 2 k 3 K 1 K 2 K 3 R=max{K 1 K 2 K 3}-min{K 1 K 2 K 3} Notes K 1 is the sum total observed targets under 1st level K 2 is the sum total observed targets under 2nd level K 3 is the sum total observed targets under 3rd level k 1 k 2 k 3 is the mean value of K 1 K 2 K 3 respectively R=max{K 1 K 2 K 3}-min{K 1 K 2 K 3} K 1 K 2 K 3 k 1 k 2 k 3 R Table 3 Comparison in factors Best project 3 The results of data processing of the extracellular polysaccharide and intracellular polysaccharide of Cmilitaris a: C/% 295984 301490 407741 98661 100497 135914 111757 b: N/% 317925 337117 350173 105975 112372 116724 32248 c a b a 3 b 3 c 2 c: Temperature/ 260253 392161 352801 86751 130720 117600 131908 a: C/% 140718 139986 187739 46906 46662 62580 47753 b: N/% 123412 156093 188938 41137 52031 62979 65526 c b a a 3 b 3 c 1 c: Temperature/ 187084 172430 108929 62361 57477 36310 78155 K 1 1 K 2 2 K 3 3 k 1 k 2 k 3 K 1 K 2 K 3 R =max{k 1 K 2 K 3}-min{K 1 K 2 K 3} Notes K 1 is the sum total observed targets under 1st level K 2 is the sum total observed targets under 2nd level K 3 is the sum total observed targets under 3rd level k 1 k 2 k 3 is the mean value of K 1 K 2 K 3 respectively R =max{k 1 K 2 K 3}-min{K 1 K 2 K 3} 21 15571 g / L 9 2 R c 20961 a 6867 b 6694 c a b K a K 3 K b K 3 K c K 2 a 3 b 3 c 2 a 3 b 3 c 2 12~30 1 [11] 6% 1% 25 3~4 1
68 2009 6 1 R 111757 32248 c a b K c K 2 a 3 b 3 c 2 1 25 28 [15] 25 2 [12] [13] [14] [16] 25 6% 1% 25 mycclium / g L -1 17 15 13 11 9 7 20 22 24 26 28 30 Temperature / 1 Fig1 The relationship of temperature and the mycelium yield of Cmilitaris 22 4 3 R 131908 polysaccharide / g L -1 22 20 18 16 14 12 10 08 06 04 02 0 Total polysaccharide 0 2 4 6 8 C / % 3 Fig3 The relationship of nitrogen source and the 2 Fig2 The relationship of carbon source and the extracellular polysaccharide the intracellular polysaccharide and the total polysaccharide content of Cmilitaris K K a K 3 K b K 3 C/N [17] 3 6% 1% 1584 7 g / L 25 polysaccharide / g L -1 20 18 16 14 12 10 08 06 04 02 0 Total polysaccharide 0 1 2 3 4 5 6 N / % extracellular polysaccharide, the intracellular polysaccharide, and the total polysaccharide content of Cmilitaris
1 : 69 polysaccharide / g L -1 20 16 12 08 04 Total polysaccharide 0 20 22 24 26 28 30 Temperature / 4 Fig4 The relationship of temperature and the extracellular polysaccharide, the intracellular polysaccharide, and the total polysaccharide content of Cmilitaris 23 1% 25 3 R c 78155 b 65526 a 47753 3 c b a K c K 1 31 187084 b K 3 188938 a K 3 187739 6% a 3 b 3 c 1 2 3 4 22 6% 1% 22 a 3 b 3 c 2 9 227503 6% 1% 25 2 [15] 3 4 25 2 a 3 b 3 c 2 6% [18] 5 d 1% 24 2 a R 158778 c b 32 R 117254 97774 a c b 3 K a 3 b 3 c 2 25
70 2009 3 231-233 [7] action of 22 [8] 33 Fungus Journal 1993 15 6 41-42 [9] WAGNAR H Progress in Medicinal and Economic Plant[M] London Academic Press 2002 20-22 [10] [J] QIU De-kai SHEN Min The treatment study on Cordyceps polysaccharide lipin body to chronic second grade hepatitis[j] China Digestion Magazine 1991 11 2 69-71 [11] [M] 5 d 5 d LI Hao WU Bai-chang LI 7 d Chun-nan The Manual Cultivation and the Depth Development of Cordyceps[M] Beijing Science and Technology Litera- 8 d ture Publishing House 2003 37-43 [12] [J] SHANG De-jing WANG Xiao-min The study on Cordyceps in in-depth ferments[j] Agriculture Biological Technology Journal 1998 15 6 48-49 [13] Y3 [J] JIANG Xiao-lu GE Bei-lei The study on 5 d in-depth fermentation condition of Cordyceps militaris Y3 [J] Bio-engineering Progresses 1997 5 6 31-35 [14] (References): ification 1998 12 1 35-40 [1] [M] SHAO Li- [15] ping Fungus Taxonomy[M] Beijing Chinese Forestry Publishing House 1984 109 [2] [J] TANG Xiao-yun Progresses of Cordyceps militaris[j] Basic Unit Chinese Native Medicine Magazine 2002 16 4 50-53 [3] [J] LI Nan GONG Chang- progresses of the Cordyceps[J] Chinese Medicine 1999 30 [J] ZHOU Shi-wen XU Chuan-fu Immunity pharmacological polysaccharide[j] China Biochemistry Medicine Magazine 1994 2 15 143-147 [J] CHEN Ming Anti-tumor progress of fungus polysaccharide[j] Edible [J] LI Xin XU Lei The study on the optimization of mycelium culture medium and fermentation condition of Cordyceps militaris[j] Biological Technology Not- [J] LI Zongjun WEN Qing-ying The study on asexual of Cordyceps militaris The mycelium culture in fermentation and the extract of the extracellular polysaccharide[j] Hunan Agricultural College Journal 1998 24 5 375-379 [16] [J] LI Xin XU Lei The optimization of fermentation process hong ZHANG Hong The manual cultural technique and the in the polysaccharide of Cordyceps militaris[j] Chemical Metallurgy healthy product development of Cordyceps militaris [J] 1998 19 3 254-259 Changchun Normal School Journal 2001 20 1 36-37 [17] [4] [J] [J] ZHAO Ming-wen WU XU Zhou-shan ZHOU Xiao-yan The progress of Cordyceps polysaccharide[j] Industry Microorganism 2000 30 1 56-57 Yan-na LI Yu-xiang et al The study on optimization condition of Cordyceps militaris extracellular polysaccharide in fermentation[j] China Edible Fungus 2000 19 4 30-32 [5] - [J] [18] YUAN Jian-guo The research and development of the function factor - Cordyceps polysaccharide[j] Wuxi Light Industry University Journal 1999 18 5 102-105 [J] WEN Lu XIA Ming SONG Huw ei et al The metabolism of cordycepin and adenosine of Cordyceps militaris in fermentation[j] Microbiology Notification) 2005, [6] 32 3 91-94 [J] GUO Hai-ping YANG Zhi-min The pharmacological