Development of a two-step cultivation strategy for the production of vitamin B12 by Bacillus megaterium.
Development of a two-step cultivation strategy for the production of vitamin B12 by Bacillus megaterium.
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开发利用巨大芽孢杆菌生产维生素 B12 的两步培养策略。
DOI：
10.1186/s12934-014-0102-7
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发表时间：
2014-07-15
影响因子：
6.4
通讯作者：
Du G
中科院分区：
文献类型：
--
作者：
Mohammed Y;Lee B;Kang Z;Du G
维生素B12是一种迷人的分子，在许多生物体的代谢中起辅助作用，特别是影响DNA合成和调节，脂肪酸合成和能量产生。维生素B12的合成仅限于少数细菌和古细菌。因此，工业微生物发酵用于满足全世界对维生素B12的年度需求，并作为需要至少60个步骤的化学合成的替代方法，这是不经济的。巨大芽孢杆菌是维生素B12的生产者之一，是许多生物技术应用的理想宿主，也是工业生产多种酶的最佳工具之一。因此，建立了两步优化策略，以提高B生产维生素B12的产量。通过提供维生素B12生物合成的生产要求和合适的条件，使其转化为巨大的微生物。确定了B的最佳发酵条件。结果表明，与无参数对照（0.26 μg/L）相比，维生素B_（12）产量提高了759倍（204.46 μ g/L）。高效液相色谱-可变波长检测器-质谱联用技术已被用于维生素B12的形态鉴定和结果确认。我们开发了B的发酵工艺。通过提供合成维生素B12所需的补充物（CoCl 2、δ-氨基乙酰丙酸（ALA）和5，6-二甲基苯并咪唑（DMB））并将发酵过程分为三个阶段，此外，最佳的培养时间的三个发酵阶段进行了调查，并进行了减少数量的实验和评估的多个参数和它们的相互作用，通过使用统计实验设计和分析。所有这些策略已被证明成功地将维生素B12的产量提高到204.46 μg/l，并证明B. megaterium可能是工业生产维生素B12的良好候选者。
Vitamin B12 is a fascinating molecule which acts as a co-factor in the metabolism of many organisms, especially affecting DNA synthesis and regulation, fatty acid synthesis and energy production. The synthesis of vitamin B12 is limited to a few of bacteria and archaea. Therefore, industrial microbial fermentation is used to meet annual demands worldwide of vitamin B12 and as an alternative method to the chemical synthesis which requires at least 60 steps that is uneconomical. Bacillus megaterium is one of vitamin B12 producers and an ideal host for many biotechnology applications and being one of the best tools for the industrial production of several enzymes. Therefore, a two-step optimization strategy was established to produce high yield of vitamin B12 by B. megaterium through the provision of the production requirements and the suitable conditions for the biosynthesis of vitamin B12. We achieved the optimum conditions for the fermentation process of B. megaterium to produce high yield of vitamin B12 in a practical way based on statistical design and analysis which allowed vitamin B12 production to increase up to 759-fold (204.46 μg/l) as compared with control without parameters (0.26 μg/L). High performance liquid chromatography coupled to variable wavelength detector and mass spectrometry has been used to identify vitamin B12 forms and confirm the results. We developed the fermentation process of B. megaterium to enhance the production of vitamin B12 by providing the required supplements for the synthesis of vitamin B12 (CoCl2, δ-aminolevulinic acid (ALA) and 5,6-dimethylbenzimidazole (DMB)) and dividing the fermentation process into three stages. In addition, the optimum incubation times of the three fermentation stages were investigated and performed with reducing number of experimental and evaluated multiple parameters and their interactions by using statistical experimental design and analysis. All of these strategies has proven successful in enhancing the production of vitamin B12 up to 204.46 μg/l and demonstrated that B. megaterium could be a good candidate for the industrial production of vitamin B12.
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DOI：
10.1051/lait:2004035
发表时间：
2005-01-01
期刊：
LAIT
影响因子：
--
作者：
Murooka, Y;Piao, Y;Yamashita, M
通讯作者：
Yamashita, M
影响因子：
56.9
作者：
DRENNAN, CL;HUANG, S;LUDWIG, ML
通讯作者：
LUDWIG, ML
影响因子：
64.8
作者：
Taga, Michiko E.;Larsen, Nicholas A.;Walker, Graham C.
通讯作者：
Walker, Graham C.
影响因子：
3.9
作者：
Moore, Simon J.;Warren, Martin J.
通讯作者：
Warren, Martin J.
影响因子：
4.1
作者：
Raux, E;Lanois, A;Thermes, C
通讯作者：
Thermes, C