Vol. 40 No Journal of Jiangxi Normal University Natural Science Nov ITS-rDNA. Acremonium sp. AChEI

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1 40 6 Vol 40 No Journal of Jiangxi Normal UniversityNatural Science Nov SF * ITS-rDNA AChEI SF4 Acremonium sp Acremonium sp SF4 SF4 AChEI r min d AChEI g L % AChEI R A DOI /j cnki issn ITS-rDNA SF4 Acetylcholinesterase Inhibiters AChEI Alzhe- AChEI imer s disease AD AChEI AChEI AChEI AD AChEI 1 1 AChEI SF4 2-6 AChEI ATCI DTNB 7-8 AChE Sigma HupA Sabouraud's dextrose broth SDB 12 Cazpek broth CB 13 Potato dextrose broth PDB Huperzia serrata AChEI SF BAB

2 r min d ITS-rDNA CTAB DNA 15 PCR 7 AChE ITS-rDNA 3 7Neighbour-joining r min d ml ml PDB 3 d AChE r min d 500 ml 120 ml d % AChE AChE 3 95% 2 d mg ml PBS ph 7 7 SF4 PDA 1 mg ml d 20 μg ml ~ 35 mm AChE AChE G L Ellman I I = OD - OD 1 100% OD % PDB SDB CB r min - 1 SF4 10 d Hyphomycetes Moniliales Moniliaceae Amerosporoideae of Moniliaceae Botrytideae AChE 2 SDB Acremonium SF4 PDA 2 SF4

3 6 SF SF4ITS-rDNA sp P42E5 99% 3 ITS-rDNA ITS1 ITS4 ITS-rDNA SF4 549 bprdna GU985205Blast Acremonium sp Acremonium Acremonium sp SF4 3 SF4 ITS-rDNA 2 3 SF4 AChE AChEI 4 SF4 PDB AChE PDB SBD 11 23% AChE CB SBD 46 94% SF4 AChEI AChE PDB SF4 AChE SBD SF4 AChE 1 SF4 SBD PDB CB / g L ± ± ± AChE /% ± ± ± SBD SF4 AChE AChE % AChE AChE 28 SF4 AChEI SF4 25 AChE AChE AChEI

4 % SBD 2 SF4 / / g L ± ± ± AChE /% ± ± ± SBD r min r 7-8 min SF4 AChEI 80 r min - 1 AChEI 3 AChEI AChE SF4 150 r min SF4 SF4 AChEI SBD / r min / g L ± ± ± AChE /% ± ± ± 1 26 ITS-rDNA SF Acremonium sp Acremonium sp SF d SF g L d 4 1Blusztajn J K Berse B The cholinergic neuronal phenotype in Alzheimer s disease J Metabolic Brain Dis- 3 AChE ease d 2Zhao Qin Tang Xican Effects of huperzine A on acetylcholinesterase isoforms in vitrocomparison with 4 SF4 tacrine /d / g L ± ± ± AChE /% ± ± ± J PDB r J min d ± g L - 1 AChE ± r min d ± 0 017g L - 1 AChE ± 3 01% SF4 AChEI 8% r min d ± 0 017g L - 1 AChE ± 3 01% donepezil rivastigmine and physostigmine J European Journal of Pharmacology / Ma Xiaoqiang Gang D R The Lycopodium alkaloids J Natural Product Reports Ma Xiaoqiang Tan Changheng Zhu Dayuan et al Hu-

5 6 SF4 573 perzine A from Huperzia speciesan ethnopharmacolgical 2007 review J Journal of Ethnopharmacology M Wang Ya Zeng Qinggui Zhang Zhibin et al Isolation and 14 M 1979 characterization of endophytic Huperzine A-producing fun- 15Guo L D Hyde K D Liew E C Y Identification of endophytic fungi from Livistona chinensis based on morphology gi from Huperzia serrata J Journal of Industrial Microbiology & Biotechnology and rdna sequences J New Phytologist Yan Riming Zhang Zhibin Wang Ya et al Efficient strategy for maintaining and enhancing the huperzine A pro- 16Ellman G L Courtney K D Andre V A new and rapidcolo duction of Shiraia sp Slf14 through inducer elicitation rimetric determination of acetylcholinesterase activity J J Journal of Industrial Microbiology & Biotechnology Biochemical Pharmocology g5 9Stierle A Strobel G Stierle D Taxol and taxane production J by Taxomyces andreanae endophytic fungus of Pacific Yew J Science M 10Stone J K Bacon C WWhite J F An overview of endophytic microbesendophytism defined M Bacon C W 19Rivera-Chaávez J Figueroa M Gonzaález MDC et al α White JFJr Microbial Endophytes New YorkMarcel Dekker 2000 with Hintonia latiflora J Journal of Natural Products 11Wang Ya Gao Boliang Li Xixi et al Phylogenetic diversi Glucosidase inhibitors from a Xylaria feejeensis associated ty of culturable endophytic fungi in Dongxiang wild rice 20Liu Ling Gao Hao Chen Xulin et al Brasilamides A-D Oryza rufipogon Griffdetection of polyketide synthase sesquiterpenoids from the plant endophytic fungus Paraconiothyrium brasiliense J European Journal of Organic gene and their antagonistic activity analysis J Fungal Biology Chemistry M The Identification and Optimization of Fermentation Conditions of Acetylcholinesterase Inhibiters-Producing Endophytic Fungi Strain SF4 LI Xixi 1 2 YAN Riming 2 ZHANG Zhibin 2 YANG Huilin 2 ZHU Du 2 3 WANG Ya 3 4* 1 Science and Technology College Jiangxi University of Traditional Chinese Medicine Nanchang Jiangxi China 2 Key Laboratory of Protection and Utilization of Subtropic Plant Resources of Jiangxi Province Jiangxi Normal University Nanchang Jiangxi China 3 Key Lab of Bioprocess Engineering of Jiangxi Province College of Life Sciences Jiangxi Science and Technology Normal University Nanchang Jiangxi China 4 Key Laboratory for Research on Active Ingredients in Natural Medicine of Jiangxi Province Yichun University Yichun Jiangxi China AbstractThe acetylcholinesterase inhibitersachei-producing endophytic fungi strain SF4 was identified as Acremonium sp SF4 based on its morphological characteristics and ITS-rDNA sequence analysis The effects of fermentation medium temperature rotation speed and cultural time on acetylcholinesterase AChEinhibitory activity and AChEI metabolites amount were analyzed It was indicated that the appropriate fermentation condition wassabouraud dextrose brothsdbmedium r min - 1 for 14 d Under the optimized conditions of fermentation the amount of AChEI metabolites and AChE inhibitory activitycytotoxic activity is g L - 1 and 60 27% respectively higher than initial conditions These results can lay the foundation for the further separation and purification of the AChEI metabolites Key wordsendophytic fungiacetylcholinesterase inhibitersacetylcholinesteraseidentificationfermentation