甜菜碱对脱氮假单胞杆菌发酵产VB12影响.doc

甜菜碱对脱氮假单胞杆菌发酵产VB12影响[摘要] 通过研究不同甲基供体（甜菜碱和氯化胆碱）对脱氮假单胞杆菌发酵产VB12的影响，结果表明甜菜碱能显著促进VB12的生物合成；在此基础上考察了发酵培养基中甜菜碱浓度对VB12合成的影响，结果表明15 g/L的甜菜碱下VB12的合成量最高；进一步研究了甜菜碱对脱氮假单胞杆菌发酵过程的影响，结果表明发酵液中一定浓度的甜菜碱可以显著提高δ-氨基乙酰丙酸的合成量，从而促进VB12的合成。 [关键词] 甜菜碱；菌体生长；VB12合成；δ-氨基乙酰丙酸；脱氮假单胞杆菌 [中图分类号] R977.2[文献标识码]A [文章编号]1673-7210（2008）09（b）-012-04 Effect of betaine on the fermentation of vitamin B12 by Pseudomonas denitrificans WANG Zhen-guo1,CAO Yun-feng1,CHEN Xue-jun1,LI Yong-liang1,LI Kun-tai2, ZHANG Si-liang2* （1.Shijiazhuang Pharma.Group Hua Rong Pharmaceutical Co. Ltd., Shijiazhuang 050041, China；2. State Key Laboratory of Bioreactor Engineering，National Engineering Research Center for Biotechnology(Shanghai)，East China University of Science and Technology，Shanghai 200237，China） [Abstract] The effect of different methyl-group donors (betaine and choline chloride) on vitamin B12 production in shake flasks with Pseudomonas denitrificans was studied, and the results demonstrated that the addition of betaine to the fermentation medium could greatly stimulate biosynthesis of vitamin B12. Betaine concentration in fermentation medium was further investigated in shake flask, as a result, 15 g/L of betaine was the most beneficial for vitamin B12 synthesis. Moreover, the effect of betaine on the fermentation of P. denitrificans was studied, it was found that the δ-aminolevulinic acid (ALA) biosynthesis could be significantly improved when betaine was kept at certain concentration in the broth, thus the biosynthesis of vitamin B12 was greatly increased. [Key words] Betaine;Cell growth;Vitamin B12 biosynthesis;δ-aminolevulinic acid,;Pseudomonas denitrificans 维生素B12（VB12）通常用来表示钴胺素类化合物，它是一种重要的生物活性物质，可以用来治疗恶性贫血症，同时也是许多微生物和动物的生长因子[1]。自然界中，VB12的生物合成存在两种不同的途径，即好氧途径和厌氧途径，但是VB12的生物合成严格局限于一些微生物中[2]，目前用于商业化生产VB12的微生物主要有谢氏丙酸杆菌（Propionibacterium shermanii）、菲氏丙酸杆菌（P. freudenreichii）和脱氮假单胞杆菌（Pseudomonas denitrificans）等。P. denitrificans好氧合成VB12的途径已经详细阐明[3]。正是由于对VB12合成途径的了解，所以可以通过诱变和分子生物学等手段来改造菌株，已经使得P