为提高发酵法生产腺苷的产量,解决现有发酵产腺苷周期短,葡萄糖转化成腺苷的效率较低等问题,以Bacillus subtilis XGL(8-AGr+ His-+xan-+SGr) 为供试菌株,进行膜透析发酵生产腺苷的研究。在5 L发酵罐中发酵生产腺苷,分别在36、60 h对发酵液进行2次膜偶联透析,使发酵周期延长至96 h,菌体OD600 nm提高到94.6,单批次产苷448.3 g,平均产苷44.2 g/L,糖苷转化率提高到25.1%。相比传统发酵工艺,膜偶联间歇透析发酵使得产苷周期延长36~40 h,菌体量增加29.6%,单罐产量增加298.4 g,糖苷转化率提高8.8%。通过膜透析使副产物乙偶姻的产量降低,更多的碳源流向腺苷合成途径。在发酵52 h时检测透析发酵中磷酸核糖焦磷酸(phosphoribosyl pyrophosphate,PRPP)转酰胺酶和腺苷琥珀酸合成酶酶活力,相比于正常发酵工艺均有所提高。得出通过膜偶联间歇透析发酵法生产腺苷能有效解除发酵液内产物的反馈抑制作用,降低副产物的质量浓度,延长发酵周期,有效提高发酵液的菌体量、腺苷产量和糖苷转化率。
In order to increase the yield of adenosine produced by fermentation and solve the problems of short adenosine production cycle and low efficiency of converting glucose into adenosine in the existing method, this paper uses Bacillus subtilis XGL (8-AGr+ His-+xan-+SGr) as the test strains to study the production of adenosine by membrane dialysis fermentation. Adenosine was fermented in a 5 L fermentor, and the fermentation broth was subjected to membrane-coupled dialysis twice at 36 h and 60 h. The fermentation cycle was extended to 96 h, with the OD600nm of the bacteria increased to 94.6, and produce 448.3 g adenosine in a single batch. The average adenosine production was 44.2 g/L. The rate of glucose conversion to adenosine increased to 25.1%. Compared with the traditional fermentation process, membrane-coupled batch dialysis fermentation prolonged the adenosine production cycle by 36-40 h, increased the biomass of bacteria by 29.6%, increased the yield of a single batch by 298.4 g, and increased the conversion rate of glucose to adenosine by 8.8%. Through membrane dialysis, the production of by-product acetoin was reduced, and more carbon sources flowed to adenosine synthesis pathway. The activity of amidophosphoribosyl transferase and adenylosuccinate synthase in the dialysis fermentation was detected at 52 h of fermentation compared with the normal fermentation process. Therefore, the production of adenosine by fermentation coupled with membrane dialysis can effectively relieve the feedback inhibition of the products in the fermentation broth, reduce the mass concentration of by-products, extend the fermentation cycle, and effectively increase the biomass of bacteria in the fermentation broth, adenosine production and adenosine conversion rate.
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