BackgroundGinsenoside Rg3(S) and compound K (C-K) are pharmacologically active components of ginseng that promote human health and improve quality of life. The aim of this study was to produce Rg3(S) and C-K from ginseng extract using recombinant Lactococcus lactis.MethodsL. lactis subsp. cremoris NZ9000 (L. lactis NZ9000), which harbors β-glucosidase genes (BglPm and BglBX10) from Paenibacillus mucilaginosus and Flavobacterium johnsoniae, respectively, was reacted with ginseng extract (protopanaxadiol-type ginsenoside mixture).ResultsCrude enzyme activity of BglBX10 values comprised 0.001 unit/mL and 0.003 unit/mL in uninduced and induced preparations, respectively. When whole cells of L. lactis harboring pNZBglBX10 were treated with ginseng extract, after permeabilization of cells by xylene, Rb1 and Rd were converted into Rg3(S) with a conversion yield of 61%. C-K was also produced by sequential reactions of the permeabilized cells harboring each pNZBgl and pNZBglBX10, resulting in a 70% maximum conversion yield.ConclusionThis study demonstrates that the lactic acid bacteria having specific β-glucosidase activity can be used to enhance the health benefits of Panax ginseng in either fermented foods or bioconversion processes.
This study aimed to produce a pharmacologically active minor ginsenoside F2 from the major ginsenosides Rb1 and Rd by using a recombinant Lactococcus lactis strain expressing a heterologous β-glucosidase gene. The nucleotide sequence of the gene (BglPm) was derived from Paenibacillus mucilaginosus and synthesized after codon optimization, and the two genes (unoptimized and optimized) were expressed in L. lactis NZ9000. Codon optimization resulted in reduction of unfavorable codons by 50% and a considerable increase in the expression levels (total activities) of β-glucosidases (0.002 unit/mL, unoptimized; 0.022 unit/mL, optimized). The molecular weight of the enzyme was 52 kDa, and the purified forms of the enzymes could successfully convert Rb1 and Rd into F2. The permeabilized L. lactis expressing BglPm resulted in a high conversion yield (74%) of F2 from the ginseng extract. Utilization of this microbial cell to produce F2 may provide an alternative method to increase the health benefits of Panax ginseng.
The release of silver (Ag) nanoparticles (NPs) into sewage streams has heightened concerns about potential adverse impacts on wastewater treatment processes. Here, we show that the rate constants of both biological nitrification and organic oxidation decreased exponentially with an increase in the Ag NP concentration, but nitrification was more severely inhibited than the organic oxidation even at low Ag NP concentrations (<1 mg Ag L(-1)) in batch experiments. The long-term exposure effects of Ag NPs on activated sludge bacteria were evaluated in sequencing batch reactors (SBRs) fed with two different substrates favoring heterotrophic and autotrophic bacteria. From a continuous operation for 50 days, it was found that heterotrophic bacteria in the organic removal process have higher tolerance to Ag NPs than do nitrifying bacteria. The effects of Ag NPs on the microbial community in both SBRs were analyzed using 16S ribosomal ribonucleic acid (rRNA) gene sequences obtained from pyrosequencing. The results showed that the level of microbial susceptibility is different for each type of microorganism and that the microbial diversity decreased dramatically after continuous exposure to Ag NPs for 50 days, resulting in a decrease of wastewater treatment efficiency.
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