Use of fulvic acid-like compounds from pulp-derived black liquor for enhancing the selenium content of peanut buds.

BACKGROUND: Cleaner production involving the extraction of useful material from the black liquor by-product of straw pulp would be environmentally beneficial and would permit increased wastewater usage. RESULTS: The fulvic-acid-like components of pulp black liquor (PFA) with molecular weights below 10 kDa were isolated. The chemical and physiological characteristics of PFAs were investigated. Selenite can enhance the selenium nutrition level of crops, but excessive selenite may be toxic to plant growth. In order to explore how to increase selenite tolerance and selenium accumulation in peanut, the effects of PFA on selenium-associated properties in peanut seedlings were examined by growing seedlings with sodium selenite (0, 5, 15, and 25 mg·L- 1 Na2SeO3, 15 mg·L- 1 Na2SeO3 solution containing 60 mg-C/L PFA, and 25 mg·L- 1 Na2SeO3 containing 60 mg-C/L PFA). CONCLUSION: The results showed that with 15 mg·L- 1 Na2SeO3, PFA significantly increased both the total and hypocotyl fresh weight of the seedlings but reduced the fresh weight of the root. PFA also effectively promoted the conversion of Se from inorganic to organic compounds in the root and hypocotyl, increased the soluble total sugar and soluble protein contents of the hypocotyl, and thus improved the edible quality and food safety of the selenium-enriched peanut buds. The results suggest that PFA can be used as an innovative bio-based substance for selenium-enriched sprout vegetable production.

Enhancement of Production of D-Glucosamine in Escherichia coli by Blocking Three Pathways Involved in the Consumption of GlcN and GlcNAc.

D-Glucosamine is a commonly used dietary supplement that promotes cartilage health in humans. Metabolic flux analysis showed that D-glucosamine production could be increased by blocking three pathways involved in the consumption of glucosamine-6-phosphate and acetylglucosamine-6-phosphate. By homologous single-exchange, two key genes (nanE and murQ) of Escherichia coli BL21 were knocked out, respectively. The D-glucosamine yields of the engineered strains E. coli BL21ΔmurQ and E. coli BL21ΔnanE represented increases by factors of 2.14 and 1.79, respectively. Meanwhile, for bifunctional gene glmU, we only knocked out its glucosamine-1-phosphate acetyltransferase domain by 3D structural analysis to keep the engineered strain E. coli BL21glmU-Δgpa survival, which resulted in an increase in the production of D-glucosamine by a factor of 2.16. Moreover, for further increasing D-glucosamine production, two genes encoding rate-limiting enzymes, named glmS and gna1, were coexpressed by an RBS sequence in those engineered strains. The total concentrations of D-glucosamine in E. coli BL21 glmU-Δgpa', E. coli BL21ΔmurQ', and E. coli BL21ΔnanE' were 2.65 g/L, 1.73 g/L, and 1.38 g/L, which represented increases by factors of 8.83, 5.76, and 3.3, respectively.

Development of mazF-based markerless genome editing system and metabolic pathway engineering in Candida tropicalis for producing long-chain dicarboxylic acids.

Candida tropicalis can grow with alkanes or plant oils as the sole carbon source, and its industrial application thus has great potential. However, the choice of a suitable genetic operating system can effectively increase the speed of metabolic engineering. MazF functions as an mRNA interferase that preferentially cleaves single-stranded mRNAs at ACA sequences to inhibit protein synthesis, leading to cell growth arrest. Here, we constructed a suicide plasmid named pPICPJ-mazF that uses the mazF gene of Escherichia coli as a counterselectable marker for the markerless editing of C. tropicalis genes to increase the rate of conversion of oils into long-chain dicarboxylic acids. To reduce the ß-oxidation of fatty acids, the carnitine acetyltransferase gene (CART) was deleted using the gene editing system, and the yield of long-chain acids from the strain was increased to 8.27 g/L. By two homologous single exchanges, the promoters of both the cytochrome P450 gene and the NADPH-cytochrome P450 reductase gene were subsequently replaced by the constitutively expressed promoter pGAP, and the production of long-chain dicarboxylic acids by the generated strain (C. tropicalis PJPP1702) reached 11.39 g/L. The results of fed-batch fermentation showed that the yield of long-chain acids from the strain was further increased to 32.84 g/L, which was 11.4 times higher than that from the original strain. The results also showed that the pPICPJ-mazF-based markerless editing system may be more suited for completing the genetic editing of C. tropicalis.