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【文献标题】Rational design of S-adenosylmethionine decarboxylase SpeD and spermidine synthase SpeE for green synthesis of spermidine

【作者】Ziyue Zhao 1, Dian Zou 1, AnYing Ji et.al

【作者单位】Huazhong Agricultural University

【文献中引用产品】

微生物S腺苷甲硫氨酸脱羧酶(SAMDC)ELISA试剂盒    活性

微生物亚精胺合成酶(SPDS)ELISA试剂盒     活性

【关键词】S-adenosylmethionine decarboxylase ，Spermidine synthase，Bacillus，amyloliquefaciens，Spermidine，Rational design，Metabolic engineering

【DOI】doi.org/10.1016/j.ijbiomac.2025.144680

【影响因子(IF)】8.4

【出版期刊】《International Journal of Biological Macromolecules》

【产品原文引用】

After fermentation for 48 h, the bacterial suspension was centrifuged at 8000 g for 10 min, resuspended in PBS. The cell suspension was then subjected to ultrasonication, followed by centrifugation at 8000 g for 10 min. The supernatant was collected as the crude enzyme solution. The SpeD enzyme activity was detected using the S-adenosylmethionine decarboxylase (SAMDC) ELISA kit (Shanghai Huding Biotechnology Co., Ltd). The specific steps were as follows: Standards and samples were dispensed into the wells at the bottom of the enzyme plate. The plate was

subsequently sealed with a film and incubated at a temperature of 37 ◦C for 30 min. Following this, the plate was washed five times with a specified washing solution. Except for the blank wells, 50 μL of an enzyme-labeled reagent was introduced into each well. The plate was then resealed and further incubated at 37 ◦C for another 30 min. This washing step was repeated another five times afterwards. Next, 50 μL of

both chromogenic agents A and B were added to each well and mixed gently. The plate was incubated in the dark at 37 ◦C for 10 min to allow for color development. The reaction was terminated by adding 50 μL of stop solution (turning the blue color to yellow), and the OD450 was measured. The SpeE enzyme activity was detected using the spermidine synthase (SPDS) ELISA kit (Shanghai Huding Biotechnology Co., Ltd). The specific operational procedures were identical to those employed for

the SpeD enzyme. Each group was tested in triplicate.

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