Journal
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 106, Issue 17, Pages 5479-5493Publisher
SPRINGER
DOI: 10.1007/s00253-022-12110-x
Keywords
Carboxyspermidine dehydrogenase; Carboxyspermidine decarboxylase; NADPH self-sufficient; Spermidine; E; coli whole-cell catalytic system
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Funding
- National Key Scientific Instrument and Equipment Development Project of China [2013YQ17052504]
- Program for Changjiang Scholars and Innovative Research Team in the University of Ministry of Education of China [IRT_15R55]
- seventh group of Hundred-Talent Program of Shanxi Province
- Key Project of Research and Development Plan of Shaanxi [2017ZDCXL-SF-01-02-01]
- Natural Sciences Foundation of Jiangsu Province [BK20210471]
- Natural Sciences Foundation of Guangdong Province [2021A1515110263]
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A novel efficient whole-cell biocatalytic method for the synthesis of spermidine was designed and constructed. This method utilizes co-expression of key enzymes and optimized conditions to achieve high yield and productivity of spermidine.
Spermidine is an important polyamine that can be used for the synthesis of various bioactive compounds in the food and pharmaceutical fields. In this study, a novel efficient whole-cell biocatalytic method with an NADPH self-sufficient cycle for spermidine biosynthesis was designed and constructed by co-expressing homoserine dehydrogenase (HSD), carboxyspermidine dehydrogenase (CASDH), and carboxyspermidine decarboxylase (CASDC). First, the enzyme-substrate coupled cofactor regeneration system from co-expression of NADP(+)-dependent ScHSD and NADPH-dependent AfCASDH exactly provides an efficient method for cofactor cycling. Second, we identified and characterized a putative CASDC with high decarboxylase activity from Butyrivibrio crossotus DSM 2876; it showed an optimum temperature of 35 degrees C and an optimum pH of 7.0, which make it better suited for the designed synthetic route. Subsequently, the protein expression level of each enzyme was optimized through the variation of the gene copy number, and a whole-cell catalyst with high catalytic efficiency was constructed successfully. Finally, a yield of 28.6 mM of spermidine was produced in a 1-L scale of E. coli whole-cell catalytic system with a 95.3% molar conversion rate after optimization of temperature, the ratio of catalyst-to-substrate, and the amount of NADP(+), and a productivity of 0.17 g center dot L-1 center dot h(-1) was achieved. In summary, this novel pathway of constructing a whole-cell catalytic system from L-homoserine and putrescine could provide a green alternative method for the efficient synthesis of spermidine.
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