Vitamin B6 is one of the important water-soluble vitamins, which has been widely applied in biomedicine, feed, food ,and beauty industries. At present, oxazole method is usually used for the production of vitamin B6 in industry, which has safety risks and causes environmental pollution. In this study, we constructed a potential and environmental vitamin B6 cell factory using Escherichia coli as chassis cells. In order to enhance the fermentation yield of vitamin B6 in E. coli and reduce the biotoxicity of the intermediate metabolite 4-hydroxy-L-threonine (4HTP) to E. coli, the key enzymes were modified by constructing an integrated protein scaffold and combining with random mutation technology. The results showed that among four different sources of 4HTP dehydrogenase (PdxA), PdxA3 from Sinorhizobium meliloti had the highest activity, reaching a yield of 2.24 mg/L; random mutation targeting PdxA3 resulted in a 60% increase in vitamin B6 titer, with a yield of 3.92 mg/L; the protein scaffold optimized at a ratio of 1∶1∶2 resulted in a greatly improved, with an approximately 20-fold increase in yield, compared to the starting strain, and eventually reached 21.23 mg/L. It was found that the pathway enzyme with optimal source and optimal expression ratio could significantly increase titer. This study is based on protein scaffolding technology, which avoids excessive accumulation of toxic intermediates in the metabolic process and is important for the construction of E. coli vitamin B6 engineered strains.
WANG Tenghe
,
WANG Yanyan
,
LIU Linxia
,
QIAO Changsheng
,
ZHANG Dawei
. Synthetic scaffolds increased vitamin B6 biosynthesis in engineered Escherichia coli cells[J]. Food and Fermentation Industries, 2023
, 49(13)
: 17
-22
.
DOI: 10.13995/j.cnki.11-1802/ts.034781
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