1. 毕业设计（论文）的内容和要求

内容： 1、查阅文献，熟悉维生素k2的相关研究 2、熟悉实验室基本操，作安全并规范操作实验室仪器设备 3、在发酵初期添加不同种类、浓度的维生素k2前体，找出最适前体及添加量。

4、选出最优前体物质及添加量进行不同时间的考察（发酵初期0h，对数生长期12h，稳定期24h，发酵末期48h），选出最优添加时间。

5、在最优条件下，经过物理方法处理（升高温度提高膜的流动性，超声）和化学方法处理（表面活性剂）来促进前体的吸收，最终提高mk-7产量。

2. 参考文献

[1] Shearer MJ, Bolton 2 Smith C. The UK food data 2 base for vitamin K and why we need it[J]. Food Chemistry, 2000, 68(2): 213-218. [2] Kazuko H, Kohji N , Yoshihisa S, et al . Nuclear vitamin K2 binding protein in human osteoblasts [J]. Biochem Pharmacol ,1999,58 (10) : 1631-1638. [3] Chiang C C, Huang T N, Lin Y W, et al. Enhancement of 4-Acetylantroquinonol B Production by Supplementation of Its Precursor during Submerged Fermentation of Antrodia cinnamomea[J]. Journal of agricultural and food chemistry, 2013, 61(38): 9160-9165. [4] Harsch M J, Benkwitz F, Frost A, et al. New precursor of 3-mercaptohexan-1-ol in grape juice: Thiol-forming potential and kinetics during early stages of must fermentation[J]. Journal of agricultural and food chemistry, 2013, 61(15): 3703-3713. [5] Yoshiki T, Hisataka T. Excretion of menaquinone-4 by a mutant of Flavobacterium sp. 238-7 in a Detergent- supplemented culture [J]. Agric Biol Chem, 1988, 52(2): 449-454. [6] Yoshiki T, Hisataka T. Extracellular production of menaquinone-4 by a mutant of Flavobacterium sp.238-7 with a detergent-supplemented culture [J]. J Ferment Bioeng, 1989, 67(2): 102-106. [7] Hisataka T, Takashi S, Chiranun D, et al. Menaquinone-4 production by a sulfonamide-resistant mutant of Flavobacterium sp. 238-7 [J]. Agric Biol Chem, 1989, 53(11): 3017-3023. [8] Hisataka T, Shunbun K, Yoshiki T. Enzymatic alteration in the shikimate pathway during derivation of menaquinone-4 producing mutants of Flavobacterium sp. 238-7 [J]. Agric Biol Chem, 1991, 55(3):769-773. [9] Aydin Berenjian, Raja Mahanama, Andrea Talbot, Ray Biffin, Hubert Regtop, Peter Valtchev, John Kavanagh, and Fariba Dehghani, 'Efficient Media for High Menaquinone-7 Production: Response Surface Methodology Approach', New Biotechnology, 28 (2011), 665-72. [10] Puri A, Iqubal M, Zafar R, et al. Influence of physical, chemical and inducer treatments on menaquinone-7 biosynthesis by Bacillus subtilis MTCC 2756[J]. Songklanakarin Journal of Science Technology, 2015, 37(3). [11] Bentley R, Meganathan R. Biosynthesis of vitamin K (menaquinone) in bacteria[J]. Microbiological reviews, 1982, 46(3): 241. [12]聂洪勇,黄伟坤, 唐英章等.维生素及其分析方法[M].上海:上海科学技术文献出版社 ,1987.114-116. [13]许静,姚振乐,杨小娇等.原生质体电融合选育维生素K2高产菌株[J]. 食品工业, 2012,2:80-83. [14]李拖平, 郭梅, 王娜等. 枯草杆菌发酵生产VK2的技术工艺条件初探[J]. 食品科学, 2008, 29(11): 379-382. [15]刘国庆,赵肖,胡卫华等.一种利用纳豆芽孢杆菌生产维生素 K2的方法. 公开号: CN102808005A.

3. 毕业设计（论文）进程安排

起讫日期 设计（论文）各阶段工作内容 备 注 12.30-01.30 查阅文献，完成开题报告 01.30-03.01 完成英文翻译 03.01-05.30 完成实验工作 06.01-06.10 完成毕业论文