RESUMO
The photocatalytic production of H2O2 has gained recognition as an economical and eco-friendly technology, but it suffers from limitations such as low production rates and difficulty in achieving high concentrations. This study was designed to overcome these limitations by preparing BiPO4 nanocrystals (BIP NCs) via high-temperature hydrolysis, and X-ray diffraction (XRD) and transmission electron microscopy (TEM) indicated that BIP NCs with particle sizes of 8.5 ± 3 nm were synthesized. In a photocatalytic performance test, only H2O and O2 were used to produce H2O2, resulting in an accumulation of H2O2 of up to 30.44 mM·g-1, as measured with the potassium titanium oxalate method; this value was 3.13 times greater than that of bulk BiPO4 (BIP-B). The resulting nanocrystals demonstrated superior electron-hole transport and separation efficiency compared to those of BIP-B, and H2O2 was formed in a one-step two-electron process. Furthermore, a film composed of a gas diffusion layer (GDL) and BIP NCs provided continuous accumulation of H2O2; a concentration of 7.23 mM was achieved after 96 h of reaction, and the stability of the film was confirmed by comparing scanning electron microscopy (SEM) images obtained before and after the reaction. Construction of a nanocrystalline structure to enhance the activities of photocatalysts and films and achieve continuous accumulation of H2O2 will provide insights into the photocatalytic production of highly concentrated H2O2.
RESUMO
BACKGROUND: Heme is an iron/porphyrin complex compound, widely used in the health care, food, and pharmaceutical industries. It is more advantageous and attractive to develop microbial cell factories to produce heme by fermentation, with lower production costs and environmentally more friendly procedures than those of the traditional extraction based on animal blood. In this study, Bacillus subtilis, a typical industrial model microorganism of food safety grade, was used for the first time as the host to synthesize heme. RESULTS: The heme biosynthetic pathway was engineered as four modules, the endogenous C5 pathway, the heterologous C4 pathway, the uroporphyrinogen (urogen) III synthesis pathway, and the downstream synthesis pathway. Knockout of hemX encoding the negative effector of the concentration of HemA, overexpression of hemA encoding glutamyl-tRNA reductase, and knockout of rocG encoding the major glutamate dehydrogenase in the C5 pathway, resulted in an increase of 427% in heme production. Introduction of the heterologous C4 pathway showed a negligible effect on heme biosynthesis. Overexpression of hemCDB, which encoded hydroxymethylbilane synthase, urogen III synthase, and porphobilinogen synthase participating in the urogen III synthesis pathway, increased heme production by 39%. Knockouts of uroporphyrinogen methyltransferase gene nasF and both heme monooxygenase genes hmoA and hmoB in the downstream synthesis pathway increased heme production by 52%. The engineered B. subtilis produced 248.26 ± 6.97 mg/L of total heme with 221.83 ± 4.71 mg/L of extracellular heme during the fed-batch fermentation in 10 L fermenter. CONCLUSIONS: Strengthening endogenous C5 pathway, urogen III synthesis pathway and downstream synthesis pathway promoted the biosynthesis of heme in B. subtilis. The engineered B. subtilis strain has great potential as a microbial cell factory for efficient industrial heme production.
